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National Bureau of Fish Genetic Resources(NBFGR), set up at Allahabad in 1983

under the aegis of Indian Council of AgricultureResearch, has evolved into a premier institutionto undertake research on diverse issues relatedto cataloguing, characterization andconservation of fish biodiversity. The Bureau’spermanent infrastructure was developed atCanal Ring Road, Telibagh, Lucknow, U.P. in1999 comprising an administrative block,laboratories, farm and residential complexcovering an area of 52 acres. The Bureau hasdeveloped state of the art facilities and expertisein several research areas including,development of fish databases, geneticcharacterization, genomics and proteomics,fish germplasm and habitat inventory, risksanalysis of exotic species, diagnostics for OIEnotified pathogens, aquatic microbes and otherareas of germplasm conservation with specialfocus on threatened, prioritized and exotic fishspecies. During the year under report, theresearch activities were conducted through 24Institutional, 17 externally - funded and 3contract research projects.

The existing database on finfish diversityof India was updated by adding informationabout 39 additional fish species reported fromIndian waters. The database now containsinformation on 2358 indigenous finfishesreported from Indian waters belonging to 868genera under 225 families of 39 orders. Achecklist of 169 catfishes belonging to 55 generaof 15 subfamilies under 14 families, reportedfrom Indian waters was prepared. A databaseon fish genetic diversity named as ‘FishKaryobank’ was developed that containsinformation on karyomorphology, molecularmarkers and nucleotide sequences of selectedfish species.

Under the programme on molecular

EXECUTIVE SUMMARY

characterization of wild populations of selectedcultivable fish species, partial Cyto b fragment(307 bp) was sequenced in Labeo rohita, Catlacatla, Cirrhinus mrigala and Clarias batrachus formitochondrial gene in 146, 200, 80 and 159individual samples, respectively, collected from9, 12, 8 and 11 rivers, respectively, to determinethe genetic variability. In Channa maruliussamples from 15 localities, 842 bp of ATPase 8and ATPase 6 mitochondrial gene wasamplified and analyzed to determine geneticvariation in wild population. Completemitochondrial DNA of Pangasius pangasius andC. batrachus was sequenced. Studies on bio-prospecting of genes and alleles for abiotic stresstolerance: Hypoxia tolerance in C. batrachusgave encouraging results. Species-specificsignatures of 11 finfish and shellfish speciesenlisted in Schedule-I of the Indian Wildlife(Protection) Act, 1972 were generated, to aidin their conservation and curb the illegal tradeof these species.

Work on development of microsatellitemarkers for genetic variability studies in Penaeus(Fenneropenaeus) indicus was continued. Eightyone polymorphic microsatellites in F. indicuswere developed through cross-amplificationfrom other penaeids. The polymorphicmicrosatellite DNA markers developed in thisstudy will be useful for commercial shrimpbreeding and selection programmes and geneticstudies of both wild and cultured stocks of F.indicus. Species-specific DNA profiles of eightHAB (7 Dinoflagellates: Ceratium furca,Neoceratium fusus , Dinophysis caudata, D.acuminata, D. fortii, Prorocentrum lima andNoctiluca scintillans; and 1 Haptophyta:Phaeocystis globosa) were developed that will behelpful in accurate and timely identification ofthese species.

Taxonomic ambiguity was resolved in

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exotic Pacu (Piaractus brachypomus) andPiranha (Pygocentrus nattereri) caught fromPeriyar and Chalakkudy Rivers near Manjali,Ernakulam, Kerala. The work on DNAbarcoding of Indian fish species was continued.A total of 102 sequences for samples offreshwater and marine finfishes were submittedto NCBI GenBank.

A new work on protein profiling in Indianmajor carps was initiated. Conventional andmolecular cytogenetic characterization of ninemarine fish species namely, Scarus ghobban,Sargocentron diadema, Neonippon summara,Lutjanus vitta, Abudefduf saxatilis, Plectorhinchusnigrus, Siganus canaliculatus, Liza parsia andSillago sihama was done. Based upon thesequences of ITS 2 regions, the phylogenetic treewas constructed that revealed Chaetodon chollareand Plectorhinchus nigrus were more closelyrelated, followed by, Liza parsia, Pomacentrusaquilus and Siganus canaliculatus.

Investigations of cadmium inducedalteration in DNA integrity and lipidperoxidation response in Labeo rohita,estimation of bioaccumulation of selected heavymetals and assessment of genotoxicity inducedby tannery effluents in fishes collected fromRiver Ganga, gave significant results. Theresults showed presence of heavy metals,especially chromium, which was above thepermissible limits of FAO/WHO (1984). Therewas significantly higher DNA damage andmicronuclei induction in fishes collected fromtannery effluent discharge site of River Ganga,as compared to other sites. Besides, nuclearabnormalities like blebbed, notched, bi-nucleated and lobed nuclei were also observedin the erythrocytes of fish species.

The work on fish germplasm exploration,assessment, cataloguing and conservation inNorth Eastern Region was continued incollaborative mode. Four new species namely,Glyptothorax ater, G. pantherinus, Batasiomizoramens and Hara koladynensis were

discovered from the N-E region. Studies on fishbiodiversity and aquatic environment of Kenand Betwa rivers were continued with a viewto generate baseline information prior tointerlinking of the two rivers. The studyrevealed that fish diversity of both the riversconsisted of a total of 67 fish species belongingto 49 genera under 21 families. River Betwashowed higher richness of 63 species belong to45 genera under 20 families, as compared toKen which showed 57 species representing 42genera under 20 families. The relativeabundance of exotic species in Ken and Betwarivers ranged from 11.7% to 45.6 % in catchesand was recorded highest in River Betwa.

A new work was initiated for raisingmonoclonal antibodies to serumimmunoglobulins of C. catla and evaluated theirapplicability for use as marker of humoralimmunity. Similarly, a new work ondevelopment of in vitro cellular model forassessment of innate immunity parameters ofLabeo rohita, was initiated. Large - scale fishmortality due to Aphanomyces invadans infectionwas documented and analysed in small scalefish farms of Uttar Pradesh. Work on isolationand characterization of Flavobacterium speciesfrom fish and aquatic environment anddevelopment of cell lines from prioritizedspecies was continued. Work oncryopreservation Puntius sarana subnasutusindicated that the extender NBFGR 9B with 10%DMSO can be used for large-scale cryostorageof the milt of this species.

A new diagnostic product, in the form ofthree monoclonal antibodies (MAbs) againstserum immunoglobulins of Channa striatus wasdeveloped for monitoring of fish health. Thenew diagnostic product has potentialapplication in immunological studies fordetection and isolation of Ig+ cells to studyontogeny, distribution and functions of reactivelymphocyte populations. This product can beeffectively utilized to elucidate the effect of

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environmental factors such as pH, salinity,temperature, dissolved oxygen, pollutants etc.on humoral immune response in fish. Theseapplications of the new product can be usefulimmunological tool in monitoring health statusof cultured C. striatus and have commercialvalue.

A beautiful public aquarium wasestablished at the Institute with a purpose tospread awareness about rich fish diversity ofthe country, which was inaugurated byDr. S. Ayyappan, Secretary, DARE andDirector General ICAR on 19 November,2010. The Xth Agricultural ScienceCongress was successfully held at NBFGRduring 10-12 February, 2011 in which over700 delegates from all over the countryparticipated.

The Institute celebrated its 28th FoundationDay on 12 December, 2010. A Farmers -Entrepreneurs -Scientists Interaction Meet wasorganized on this occasion to promote adoptionof new technologies in freshwater aquacultureand the dignitaries presented Annual InstituteAwards for the year 2009-10 to the staffmembers of the Institute for their outstandingcontributions and also to the selectedprogressive farmers/entrepreneurs of thecountry. International Biodiversity Day wascelebrated on 22 May, 2010. A number oftechnical workshops/trainings including, an‘Expert Consultation on Marine Biotechnologyand Biodiversity Conservation’, at InternationalCentre, Goa during 21-22 April, 2010 and asensitization training programme on‘Bioinformatics Tools and Resources in FisheriesResearch’ during 24 –29 January, 2011, aNational Seminar on “Biodiversity, Ecosystemsand Climate Change: The Challenge Ahead”during 4-6 December, 2010 and a Partners Meet

on “Application of Bioinformatics in FisheriesDomain” under Component I of the NationalAgricultural Innovation Project entitled“Establishment of National AgriculturalBioinformatics Grid in ICAR” on 29 January,2011 were organized at NBFGR. Besides, aseries of short-term training programmes on‘Aquaculture Technologies and ProductivityEnhancement’ were organized in which a totalof 208 progressive fish farmers from differentdistricts of U.P. and Bihar were trained. A totalof 260 lakhs of quality spawn of Indian majorcarps was produced under the ICAR Mega SeedProject and sold to the fish farmers. The Institutealso participated in several exhibitions/aqua-fairs in different parts of the country.

The Research Advisory Committeemeeting of NBFGR was organized under thechairmanship of Prof. T.J. Pandian, FormerNational Professor, ICAR on 15 March, 2011.The mid-term meeting of the Institute ResearchCouncil was conducted under thechairmanship of Dr. J.K. Jena, Director on 1November, 2010. The Institute observed a HindiPakhwada and organized Hindi Day andInternational Women’s Day programmes.National festivals - Independence Day andRepublic Day were celebrated with greatenthusiasm in the Institute.

The Institute published 47 research papersin peer-reviewed journals, contributed 6chapters to books/proceedings and 6 populararticles in national magazines. Besides, 2training manuals and 11 extension bulletinswere published by the scientists of the Institute.The NBFGR library added 339 documentscomprising 139 books, 149 serials and 51 annualreports. Against a target of Rs.13 lakhs, theBureau generated revenue of Rs. 13.18 lakhs,during the year under report.

4

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5

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INTRODUCTION

Brief HistoryIndia possesses rich fish germplasm

resources with large number of endemic species.However, many of the species are reported tobe declining in their areas of naturaldistribution. It was in this context that theGovernment of India approved establishmentof the National Bureau of Fish GeneticResources at the end of Sixth Five Year Plan toprovide scientific input for conservation of fishgermplasm resources of the country.Sanctioned in December, 1983 under the IndianCouncil of Agricultural Research, NBFGRstarted functioning as an independent instituteinitially at Allahabad, which later shifted toLucknow during 1994. With possession of asprawling 52 acres campus, the Bureau,however, occupied its magnificently builtadministrative and laboratory facilities onlyduring 1999. Since then several newinfrastructure facilities including hatchery, wet

laboratories, public aquarium, guest house, staffquarters and above all, required experimentaltanks and ponds have been created, satisfyingthe need of research and other amenities. TheNBFGR has a fully equipped ARIS cell whichprovides round the clock Internet facilitythrough V-SAT and a broadband connectionvia a LAN system to all scientists and sectionat their work place. The Bureau has a welldeveloped library with more than 5700 bookscatering the need of scientists and students. TheBureau, over the years, has created excellentinfrastructure, state of the art facilities andexpertise in several research areas including,development of fish databases, geneticcharacterization, gene banks, fish germplasmand habitat inventory, risks analysis of exoticspecies, diagnostics for OIE notified pathogens,aquatic microbes and other areas of germplasmconservation with special focus on threatened,prioritized and exotic fish species.

MANDATE Collection, classification and cataloguing of fish genetic resources of the country.

Maintenance and preservation of fish genetic material for conservation of endangeredfish species.

Evaluation and valuation of indigenous and exotic fish species.

VISIONAssessment and conservation of fish genetic resources for intellectual property

protection, sustainable utilization and posterity.

MISSIONCollection, cataloguing and documentation of fish genetic resources using operational

strategies of partnership and cutting-edge technologies.

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Bio

tech

nolo

gy

8

STAFF POSITIONThe overall staff position as on 31st March, 2011 is given below:

Financial StatementAllocation of funds and expenditure incurred during the year 2010-2011.

(Rs. in lakhs)

S. N. Category of posts Post created Staff in position

Post vacant (out of created posts)

1. Research Management (Director)

01 01 --

2. Scientific 40 29 11

3. Technical 36 35 01

4. Administrative 20 20 -

5. Supporting 20 19 01

Total 117 104 13

Budget Allocation Expenditure

Plan 490.00 489.99

Non Plan 773.00 772.05

Total

Revenue generation Target Achieved

13.00 13.18

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REASEARCH ACHIEVEMENTS

5.1 CATALOGUING OF FISH GENETICRESOURCES OF INDIA

The collection and cataloguing ofinformation on fish genetic resources of Indiais an important mandate of NBFGR. During theperiod under report, the Bureau continued itsefforts towards collection and cataloguing ofthe fish genetic resources of India. The existingdatabase on finfish diversity of India wasupdated by adding information about 39additional fish species reported from Indianwaters. The details of these species are given inTable 1. The database now contains informationon 2358 indigenous finfishes belonging to 868

genera under 225 families and 39 orders (Table2). Besides, more information on morphology,habitat preference, economic value,distribution, etc., was also added on the speciesalready included in the database. The databaseincludes information on several fields including,scientific name, synonyms, common name, localname, fin-formula, Indian and globaldistribution, morphology, coloration, habitat,fishery information, economic importance,commercial utilization, references, taxonomy,digital images and diagrams, habitat, feedingand breeding behavior, migratory behaviour,genetic information and the conservation statusof individual fish species.

Table 1. Additional fish species added in the Database

S.No. Species Author Family Distribution

1 Amblyceps laticeps McClelland, 1842 Amblycipitidae India: Assam, Khasi hills

2 Amblyceps tenuispinis Blyth, 1860 Amblycipitidae India: Ghazipur, Uttar Pradesh.

3 Channa bleheri Vierke, 1991 Channidae India, South East Asia (Oriental region)

4 Cyprinus nicholsi Mysers Cyprinidae North East India

5 Exostoma vinciguerrae Regan, 1905 Sisoridae India

6 Gagata itchkeea Sykes, 1839 Sisoridae India: Peninsular India. Cauvery, Krishna and Narmada rivers. Not found in Myannmar

7 Gagata youssoufi Ataur Rahman, 1976 Sisoridae India: Ganga at Patna. Tezpur in Assam. Bangladesh: Brahamaputra near Mymensingh and Meghna river near Chandpur

8 Garra compressus Kosygin & Vishwanath, 1998

Cyprinidae Manipur (Chindwin basin)

9 Garra gravelyi Annandale, 1919 Cyprinidae Southern Shan States in Myanmar, Ganga and Brahmaputra basin

10 Gerres phaiya Iwatsuki & Heemstra, 2001

Gerreidae Southwest coast of India, Bengal Bay and Andaman Sea.

11 Glyptothorax botius Hamilton, 1822 Sisoridae Ganges River drainage

12 Glyptothorax sufii Asghar Bashir & Mirza, 1975

Sisoridae India, Pakistan: Kanganpur, Punjab and Sind

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S. No. Species Author Family Distribution

13 Gogangra viridescens Hamilton, 1822 Sisoridae India: Assam, Bihar and North Bengal. Bangaladesh

14 Gogangra laevis Ng, 2005 Sisoridae Drainage of lower Brahmaputra in Eastern India

15 Gudusia variegate Day Clupeidae India: UP, Gorakhpur

16 Hemibagrus maydelli Rossel, 1964 Bagridae India: Bhima falls, wadgaon, Maharashtra state; Krishna river system, Tungabhadra river, Nagarjunasgar, Andhra Pradesh

17 Hemibagrus menoda Hamilton, 1822 Bagridae India: Brahmaputra, Ganga, Godavari and Mahanadi river system. Bangladesh. Mainly confined to peninsular India. All records from Myanmar are referable to H. peguensis

18 Hemibagrus microphthalmus

Day, 1877 Bagridae India: Manipur. Myanmar: along Ayeyarwaddy velly. Thialand: Salween river

19 Hemibagrus punctatus Jerdon, 1849 Bagridae India: Mainly Cauvery river system in south India up to Bhavani River in Tamil Nadu. Also in rivers in Mysore, Western Ghats and adjoining hill ranges

20 Myersglanis jayarami Vishwanath & Kosygin, 1999

Bagridae India: Manipur, Laniye river of Chindwin Basin

21 Nangra assamensis Sen & Biswas, 1994 Glyptosterninae Brahmaputra river near Jorhat, Assam

22 Nangra carcharhinoides Roberts & Ferraris, 1998

Sisoridae India: Ganga river at Patna, Bihar. (Specimens procured from fish market; precise location unknown)

23 Nemacheilus denisoni mukambbikaensis

Menon Sisoridae Kollur, Karnataka

24 Neotropius mitchelli Gunther, 1864 Balitoridae India: Western Ghats in Kerala

25 Neotropius atherinoides Bloch, 1794 Schilbeidae Throughout India (expect West Coast) Bangladesh. Pakistan

26 Osteobrama cotio cumna Day Schilbeidae North East India

27 Pareuchiloglanis kamengensis

Jayaram, 1966 Cyprinidae India: Kameng and Norgum river, Kameng divison, Arunachal Pradesh

28 Psenopsis intermedia Piontrovskiy, 1987 Glyptosterninae Western Indian Ocean: Saya de Malha Bank and Kenya

29 Psilorhynchus gracilis Rainboth, 1983 Centrolophidae Tributaries of Barak in Manipur, Bangladesh: tributaries of Brahmaputra (All Brahmaputra basin)

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S.No. Species Author Family Distribution

30 Pterocryptis afghana Gunther, 1864 Psilorhynchidae India: Darjeeling Himalaya and Khasi hills in Assam. Locality Afghanistan

31 Pterocryptis gangelica Peters, 1861 Siluridae India: Darjeeling Himalaya. Manipur. Bangladesh

32 Puntius ater Linthoingambi & Vishwanath 2007

Cyprinidae Manipur valley (Chindwin basin)

33 Puntius khugae Linthoingambi & Vishwanath 2007

Cyprinidae Manipur, Khuga river (Chindwin basin) at Churachandpur district

34 Rita macracanthus Ng, 2004 Bagridae Indus River drainage in Afghanisan, Pakistan, and northeastern India

35 Schismatogobius deraniyagalei

Pethiyagoda & Kottelat, 1989

Gobiidae Western Ghats, India

36 Silurus torrentis Kobayakawa Siluridae Arunachal Pradesh, India

37 Sisor rheophilus Ng, 2003 Sisoridae India: Middle and upper parts of Ganga in Bihar, U.P. May be found in Bangladesh too.

38 Sisor torosus Ng, 2003 Sisoridae India: middle parts of the Ganga in Bihar and Yamuna river at Delhi

39 Sperata aorella Blyth, 1858 Bagridae India and Bangladesh, Ganga river Delta extending upto Bihar

Table 2. Fish diversity of India fishes belonging to 115 genera under 34 families

and 13 orders. Besides, the Institute alsodeveloped an album which containsphotographs of 144 indigenous freshwateraquarium fishes. This album will be useful tovarious stakeholders associated withornamental fish trade, research and policymaking.

Taxonomic Information System:The work on development of a Taxonomic

Information System was continued. Digitalidentification protocols based on the keymorphological attributes for seven species ofthree families viz., Cyprinidae, Clariidae andPangasidae were attempted and tested usingthe software developed. The protocols werefound to be valid. Distinguishing features basedon morphometric, meristic and other diagnosticcharacters like body proportions to standardlength and head length, fin rays count; lateral

Category of Fishes

Ecosystem No. of Fish Species

Freshwater 877 Brackishwater 113 Marine water 1368

Native fishes

Total 2358 Exotic fishes 291 Grand Total 2649

A checklist of 169 catfishes belonging to

55 genera of 15 subfamilies under 14 families,reported from Indian waters was prepared,after checking the synonyms of each fish speciesfrom all available sources. The checklist offreshwater fishes reported from North EasternIndia was revised. It now contains 316 fishspecies belonging to 124 genera under 37families and 13 orders. Similarly, the checklistof freshwater fishes reported from WesternGhats was also revised which now includes 312

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line scales with types, scutes count, number ofgill-rakers, etc. were selected as key charactersof the selected groups. The data on selectedcharacters for 29 fish species were collectedfrom the published literature, books andwebsites and added in the databases. Theupdated taxonomic information system fortotal 214 fish species was tested, validated anduploaded on the administrative local domainof NBFGR, which is running successfully. It wasalso able to identify total 87 fish species basedupon their few important trait information inthe field/laboratory.

Updated checklist of macro fauna andmacro flora of Gulf of Mannar MarineBiosphere Reserve

Updated checklists of macro fauna andmacro flora of Gulf of Mannar MarineBiosphere Reserve (3144 species) were prepared(Table 3) which will help in sustainableutilization and management of resources of theregion. The accuracy of the species names wasascertained with the help of the experts ofCMFRI and ZSI.

Table 3. List of Macro fauna and flora of Gulf of Mannar Marine Biosphere

Macro fauna 1. Class Mammalia (11 species) 1.1) Dolphins (4 species) 1.3) Whales (5 species) 1.2) Dugong (1 species) 1.4) Sea Otter (1 species) 2. Class Reptilia (25 species) 2.1) Sea Snakes (20 species) 2.2) Marine Turtles (5 species) 3. Class Pisces (585 species) 3.1) Elasmobranchs (55 species)

I. Phylum Chordata (621 species)

3.2) Teleosts (530 species) 1.Sub phylum: Cephalochordata (6 species) 2. Sub phylum: Urochordata (58 species)

II. Phylum: Protochordata (65 species)

3. Sub phylum: Hemichordata (1 species) III. Phylum: Echinodermata (263 species) IV. Phylum: Chaetognatha (Arrow worms) (17 species)

Class: Crustacea (638 species) 1. Copepoda (239 species) 2) Amphipoda (52 species) 3) Ostracoda (57 species) 4) Isopoda (18 species) 5) Decapoda (41 species) 6) Leptostraca (Cirripedia) (1 species) 7) Schizopoda (26 species) 8) Anomura (35 species)

V. Phylum: Arthropoda (638 species)

9) Brachyura (169 species)

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‘Fish Karyobank’ databaseA database on fish genetic diversity named

as ‘Fish Karyobank’ was developed under theaegis of National Agricultural BioinformaticsGrid (NAIP), ICAR. The database presentlycontains information on karyomorphology,molecular markers and nucleotide sequences ofselected fish species. The database will beenriched with the genetic information on otheraquatic organisms such as crustaceans,molluscs, echinoderms, etc. The Scientists,research workers and students will have freeaccess to the information. The information onfish chromosomes and molecular geneticaspects will serve as a useful reference forresearchers working on molecular genetics,phylogenetics, cyto-taxonomy, markerdevelopment, chromosomal manipulation,physical gene mapping, fish diversityassessment and conservation. The database willhave the facility to upload relevant informationat the users end.

Barcode of Indian Fishes (BIF) DatabaseThe BIF system is a unique resource for

molecular taxonomy of Indian fishes based onthe sequence diversity in small segment of

mitochondrial DNA i.e. cytochrome c oxidasesubunit I. The BIF was improved to a wellaccumulated and multipurpose advancedversion with incorporation of additionalfeatures, including browser applications, aswell as, larger numbers of dataset records andthe ‘administrator part’. Currently, BIF has 1706barcode records pertaining to 342 speciesincluding, fishes, molluscan and crustaceans.Besides above, physical information wasextracted from FishBase (with reference no.)and incorporated in the corresponding records.BIF database consists of powerful browserapplications i.e taxonomy browser, map viewer,species identification and statistics. The‘taxonomy browser’ provides accessibility toview the taxon/subtaxon information for allspecies, and permits the user to download therecords relevant to selective taxon/subtaxon.The ‘map viewer’ application estimates thegenetic divergence and ancestral relationshipamong the species and genus. ‘Speciesidentification’ browser identifies the fish speciesbased on the existing information in BIF withthe help of Blastn program and intelligentqueries. ‘Statistics’ application reveals thecurrent status of database associated withnumber of COI sequences, families and its

Macro fauna 1) Polyplacophora (11 species) 2) Opisthobranchia (60 species) 3) Pulmonata (5 species) 4) Gastropoda/Prosobranchia (450 species) 5) Bivalvia/Pelecypoda (213 species)

VI. Phylum: Mollusca (769 species)

6) Cephalopoda (30 species) VII. Phylum: Annelids (69 species) Polychaetes (69 species)

1) Corals ( 254 species) 2) Gorgonids (15 species)

VIII. Phylum: Coelenterata/Cnidaria (285 species)

3) Other coelenterates (Sea anemones) (16 species) IX. Phylum: Porifera/Sponges (271 species) Macro flora XI. Sea weeds (133 species) XII. Sea grasses (13 species)

Total Species (Macro fauna and flora) of Gulf of Mannar Marine Biosphere : 3144

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species and also representthe statistics for habitatand IUCN Red List Statusfor all existing species inBIF databse. The‘administrator part’ in theBIF system has been madefully automated (based ona simple mouse clicking) to:a) analyze the publiclysubmitted data in BIF andapprove/discard thesubmission; b) upload newdatasets for databasepopulation; and c)manipulate, updating ofrecords and managing the database in thebackend.

5.2 GENETIC CHARACTERIZATIONDocumentation of genetic variation among

vast finfish and shellfish resources of thecountry is of vital importance for evolvingconservation and aquaculture strategies forlong-term sustainability of the resources.Realizing this, NBFGR has been carrying outstudies on population genetics of prioritizedspecies so as to facilitate the conservation andmanagement of different natural stocks of thesespecies.

Molecular characterization of wildpopulations of selected cultivable fish species

Labeo rohita

Partial Cytochrome b MitochondrialGene

Mitochondrial gene partial Cyto b fragment(307 bp) was sequenced in 146 individualsamples collected from nine different rivers todetermine the genetic variability. A total of 278positions were found to be conserved with 32haplotypes and 17 parsimony informative sites.The average frequencies of four nucleotides for

all the samples of Labeo rohita were: A: 28.00%;T: 29.20%; C: 28.30% and G: 14.50%.Nucleotide sequences of cytochrome b wereA+T rich (57.20%) with transition totransversion ratio was 4.154 (Fig.1).Transitional substitutions were detected morecommonly than transversional ones. Thenucleotide diversity (ð) for all the ninepopulations was found to be 0.004 andhaplotype diversity (Hd) was 0.709. Analysisof Molecular Variance (AMOVA) of ninepopulations indicated that out of total variation,only 16.12% was contributed due to variationamong populations and 83.88% wascontributed due to within population variation.The FST value was found to be significant 0.161(P< 0.05).

Catla catla

Partial cytochrome b mitochondrial geneIn C. catla partial Cyto b fragment (307 bp)

was sequenced, for mitochondrial gene 200individual samples, collected from 12 differentrivers, to determine the genetic variability. Atotal of 271 positions were found to beconserved with 36 haplotypes and 13parsimony informative sites (Fig. 2). Theaverage frequencies of four nucleotides for all

Fig. 1. Haplotype network obtained from different populations (Cyto. b) analysis ofL. rohita

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Fig. 2. Haplotype network obtained from different populations (Cyto b) analysis ofC. catla

the samples of C. catla were: A: 28.10%; T:30.7%; C: 27.10% and G: 14.10% Nucleotidesequences of cytochrome b were A+T rich(58.17%) with transition to transversion ratio2.96. Transitional substitutions were detectedmore commonly than transversional ones. Theoverall nucleotide diversity (ð) for the all the 12populations was found to be 0.002 andhaplotype diversity (Hd) was 0.474. Thehaplotype diversity varied from 0.284 inGhaghra to 0.710 in Son, while nucleotidediversity varied from 0.001 in Tons, Satluj,Brahmaputra and Ghaghra to 0.004 inChambal and Gomti. AMOVA of 12populations indicated that out of total variation,only 10.72% was contributed variation amongpopulations whereas, 89.28% was contributeddue to variation within populations. The FSTvalue was found to be significant (0.107).

Microsatellite DNA analysis in C. catlaThe mean number of alleles per locus were

8.123 (Bhagirathi), 7.00 (Satluj), 7.63(Mahanadi) and 5.13 (Bhagirthi). The expectedheterozygosity (He) values ranged from 0.7269(Bhagirathi) to 0.8081 (Mahanadi), whereas,observed heterozygosity (Ho) varied from

0.6125 (Bhagirathi) to0.7284 (Mahanadi). Noevidence of linkagedisequilibrium wasdetected at any locuspair comparisons in anypopulation.

Private alleles in C.catla

Private alleles werefound at loci Lro37(Mahanadi), Lro43(Satluj), Lro25(Mahanadi, Satluj),Lr35 (Bhagirathi andSatluj), Lr45(Brahmaputra) and

Lro33 (Brahmaputra).

Cirrhinus mrigalaPartial cytochrome b mitochondrialDNA gene

Partial region of Cytocrome b fragment(307 bps) was successfully amplified andsequenced for mitochondrial DNA in C. mrigla.Eighty individuals belonging to eight differentpopulations of C. mrigla including Satluj (15individuals), Brahamputra (15 individuals),Ghagra (3 individuals), Gomti, (20 individuals),Sharda, (3 individuals), Ganga, (8 individuals),Betwa (8 individuals) and Bhagirathi (9individuals) were amplified and sequenced.DNA sequences were aligned using ClustalWand then analysed for sequence composition.Molecular diversity indices, geneticdifferentiation and FST (Population specific andpopulation pair-wise) values were calculatedusing Arlequin 3.11. Population specific FST inSatluj was 0.72715, Brahmaputra 0.72015,Ghagra 0.74494, Gomti 0.70920, Sharda0.74494, Ganga 0.69318, Betwa 0.72860 andBhagirathi 0.68898. Total FST was found to be0.71514. A total of 17 haplotypes were observed

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Fig. 4. Haplotype network between 11 populations of Clarias batrachus

Fig. 3. Haplotype network obtained from 8 different populations(Cyto b) analysis of C. mrigala

in Cytochrome b sequence analysis (Fig. 3).Percentage of variation among population was71.51, whereas, it was 28.49 within population.

Microsatellie DNA analysis inC. mrigala

Fifteen microsatellite loci thatwere found polymorphic weresequenced and submitted to NCBI. Totest the suitability of these loci forpopulation studies, 24 individuals(from two locations each from Satlujand Brahmaputra) were genotypedmanually for all the fifteen loci. Themean of expected heterozygosity was0.6262 (Satluj) and 0.6289(Brahmaputra), while, observedheterozygosity for Satluj was 0.3771and for Brahmaputra 0.4291.Significant allelic homogeneity wasobserved at seven loci Lr35*, Lr36*,Lro26*, Lro34*, Lro40*, Lro41* andLro44* (P < 0.05).

Clarias batrachus

Mitochondrial DNA: Cytochrome bGene

Cytochrome b region was successfullyamplified in populations of Varanasi,Sandeela, Guwahati (Paltan Bazaar), Tista,Udaipur, Yamuna Nagar Lakxer, BegulDam, Kolkata, Jaunpur, Lucknow andMalegaon. Sequencing was completed for159 individuals. For mitochondrial gene,partial Cyto b fragment (307 bp) wassequenced in 159 individual samplescollected from 11 different rivers todetermine the genetic variability. A totalof 285 positions were found to beconserved with 21 haplotypes and 19parsimony informative sites (Fig. 4). Theaverage frequencies of four nucleotides forall the samples of Clarias batrachus were:A: 26.80%; T: 30.40%; C: 26.50% and G:16.30%. Nucleotide sequences of

cytochrome b were A+T rich (57.20%) withtransition to transversion ratio 3.514.Transitional substitutions were detected morecommonly than transversional ones. The

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haplotype diversity (Hd) for the all the 11population ranged from 0.179-0.712 (Varanasi- Jaunpur) and nucleotide diversity (ð) rangedfrom 0.0007-0.003 (Varanasi- Jaunpur). Thehaplotype h01 was the most common andobserved in Lucknow, Jaunpur, Varanasi andSandeela. AMOVA of 11 populations indicatedthat out of total variation, 78.39% wascontributed variation among populations and21.61% was contributed variation to withinpopulations. The FST value was found to besignificant 0.78393 (P< 0.05) and populationspecific FST ranged from 0.7419-0.80.

Macrobrachium rosenbergiiSampling of M. rosenbergii from Kerala (41

samples from Kumarakom, Kottayam andKanakkankadavu, Chalakkudy River); Gujarat(68 samples) and Maharashtra (32 samples)continued during the period and Trusslandmarks were identified and measurementstaken. Pleopods collected from the samples afterrecording the morphometric and Trussmeasurements, total weight, sex and gonadalcondition of the specimens were preserved in95% ethanol. Additional 17 single-locusmicrosatellite loci were successfully amplifiedwith 4-14 alleles in each locus. Occurrence ofrepeats in the cross-amplified polymorphicmicrosatellite loci were confirmed bysequencing the PCR products in forward andreverse directions in the sequencing facility.None of the loci showed significant linkagedisequilibrium for all pairs of loci (P>0.05).Fifteen microsatellite loci are now being usedfor population genetic analysis of M. rosenbergii(M. dacqueti) from Kerala, Karnataka,Maharashtra and Gujarat.

Genetic variability in wild Channamarulius population based on ATPase 6/8 region of mtDNA

In the samples of C. marulius from 15localities, 842 bp of ATPase 8 and ATPase 6

mitochondrial gene was amplified andanalyzed to determine genetic variation in wildpopulation. ATPase 8 region was detected from1-168 bp of the sequence and ATPase 6 regionspanned from 159-842 bp. An overlappingregion of 10 bp was also seen from 159-168 bp.The two regions were analysed together forvariation in this study. For the averagefrequencies of four nucleotides for all the 131samples across 15 populations of C. marulius,transition to transversion ratio was 3.417.ATPase 6&8 sequence data generated 35haplotypes. Samples from river Teesta, Sone,Chambal, Sindh, Gomti and Tons exhibitedpopulation specific haplotypes. Most commonhaplotype h01 was found across 10 populationsnamely Mahanadi, Teesta, Yamuna,Brahmaputra, Ghaghra, Sharda, Godavari,Sindh, Gomti and Tons. The next most commonhaplotype was h02 which was present insamples from Ganga and Sindh. ExceptYamuna, Ghaghra and Godavari, allpopulations exhibited population specifichaplotypes. Total 47 variable sites wereidentified and 23 of them were parsimonyinformative. Mean no. of polymorphic loci wasestimated to be 4.53 and mean nucleotidediversity (pi) was 0.00358, haplotype diversity(Hd) was found to be 0.889 and variance of Hdwas 0.00037. Mean diversity for entirepopulation (d) was 0.0036. Mean interpopulation diversity was 2.147. Coefficient ofdifferentiation for entire samples was 5.992.AMOVA within 15 populations of C. maruliusrevealed that out of total variation, nosignificant variation (-5.28%) was presentamong the groups, 63.78% variation wasobserved among populations within groups i.e.total 63.78% of variation was seen among thepopulations. Variation within the populationwas 41.49%. Population structuring wasrevealed by FST value of 0.58 (Fig.5).

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Mitochondrial genome of two Indiancatfish species

Complete mitochondrial DNA of Pangasiuspangasius (Family: Pangasidae) and Clariasbatrachus (Family: Claridae) was sequenced. InP. pangasius and C. batrachus, wholemitochondrial DNA was of 16476 and 16571bp, respectively. The base compositions in boththe species were as follows: In P. pangasiusT=25.1%, C= 28.7%, A= 30.5% and G=15.7%,whereas in C. batrachus T=25.0%, C= 27.4%,A= 32.3% and G= 15.4. The mean geneticdistance, based on whole mitochondrialsequence, between the two catfish species wasfound to be 0.1755.

Bio-prospecting of genes and alleles forabiotic stress tolerance: Hypoxiatolerance in Indian catfish, Clariasbatrachus

Oxygen is of fundamental importance formany terrestrial, as well as, aquatic livingorganisms. Among teleost, catfishes showhypoxia tolerance as evident from theirphysiological adaptations, which includerespiratory, as well as, airbreathingadaptations. The Indian species of air-breathing

catfish Clarias batrachus lives infreshwater pools which usually have lowO2 and high CO2 content. The fish is afacultative airbreather at normoxia andshows a negative correlation between air-breathing frequency and dissolvedoxygen. In this study, physiologicalparameters for hypoxia in C. batrachuswere defined and molecular responses tohypoxia tolerance were explored.

Critical oxygen tensionCritical oxygen tension (pCrit) was

calculated based on CO2 and DO values.The resultant graph of CO2 v/s DO showsa steady decline in metabolism of fish with

time. R2 value indicated linear regression inmetabolism of fish with decrease in dissolvedoxygen concentration and there was no pointof sharp decrease in metabolic rate with varyingDO. Calculated pCrit for C. batrachus was found0.98 mg/l at 25°C.

Physiological parameters for hypoxiaHematological and physiological/

biochemical parameters were determined atpCrit for 0,1,2,3,6 and 12 hrs in liver, muscle,gill and serum tissues. Hemoglobin andhematocrit had significantly increased at 12hrfollowing hypoxia (Fig. 6). Serum glucose levelhad increased significantly but tends to

Fig .5. Neighbour – Joining tree of population pair-wise FST valuesbased on ATPase 6/8 mitochondrial gene in Channa marulius

Fig. 6. Change in Hemoglobin and Hematocrit at pCritwith different Hypoxic times

* Significant increase in Hb and Hct at 6 and 12 hr ofhypoxia

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Fig. 7. KEGG orthology analysis of EST differentiallyexpressed under hypoxia

maintain at its control level after 12 hr ofhypoxia. Serum lactate concentration hadchanged following 0 and 6 hr of hypoxic stress.There was no significant change reported intotal protein content. Lactate dehydrogenseactivity had changed significantly following 0and 6hr of hypoxia in liver and gill. Superoxidedismutase activity significantly increased in gillat the onset of hypoxia while it was decreasedin liver and serum at all experimental hours.Catalase activity significantly increased in gilland serum following 3 and 6hrs of hypoxia,respectively. No significant change was reportedfor total glutathione level.

Identification of differentiallyexpressed genes under hypoxia

Forward and reverse subtracted librarieswere constructed for six tissues (brain, heart,liver, muscle, spleen and head kidney) undersix different time intervals (0, 1, 2, 3, 6 and 12hrs)of hypoxic stress at pCrit. A total of 2020colonies were randomly selected and sequencedfrom forward and reverse subtracted librariesof all six tissues. A total of 1801 EST (ExpressedSequence Tags) sequences were obtainedsuccessfully and analysed. Clubbing all ESTsequences from all tissue, clustering andassembly of EST’s revealed 1036 unique EST’sconsisting of 192 contigs and 844 singletons. Atotal of 536 ESTs were functionally annotated,while 468 were assigned as purely novel genes(no or non-significant match) among all the six

tissues. KEGG orthology analysis of all EST’srevealed that 191 genes were differentiallyexpressed (up- and down-regulated collectively)in all the tissues examined which affects majormetabolic pathways and processes (Fig.7). Atotal of 377 functionally annotated genes werenot assigned to any major standard pathwaysand considered to be putative novel for hypoxiatolerance in catfish. For validation of differentialexpression and identification of novel genes forhypoxia tolerance, 13 genes were selected forthe normalization analysis, out of which 2genes were selected as to be used ashousekeeping genes. On the other hand, 57genes from all tissues (up and down-regulated),including 24 purely novel ones, were selectedfor validation of differential gene expressionanalysis.

Genetic characterization ofcommercially important fishes from theWestern Ghats and marine ecosystem

PomfretsThe work on genetic characterization of

pomfrets was continued during the periodunder report. Samples of silver pomfret (Pampusargentius) were collected from Maharashtra(Mumbai - 32 nos), Kerala (Munambam - 41nos) and Tamil Nadu (Tuticorin - 32 nos) coastsand DNA was extracted. Fourteen of the fifteenpolymorphic microsatellite markers developedthrough cross-amplification during theprevious year (2009-10) were used forpopulation genetics. All primer pairs amplifiedonly a single locus with 2-6 alleles in each locus.The allele size ranged from 104-338 base pairsand PCR reactions were carried out employingthe microsatellite primers. The amplifiedproducts were electrophoretically analyzedthrough 10% non-denaturing polyacrylamide(19:1 Acrylamide and bisacrylamide) gel andvisualized through silver staining. The alleleswere designated according to PCR product size

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S. No. Species COI 16S rRNA Schedule-I. Part 2 (A) - Fishes

1 Rhincodon typus (5)* (646 bp) (546 bp) 2 Rhynchobatus djiddensis (2) (640 bp) (520 bp) 3 Epinephelus lanceolatus (1) (650 bp) (560 bp) 4 Hippocampus kuda (4) (648 bp) (580 bp) 5 H. trimaculatus (4) (650 bp) (580 bp) 6 Fistularia petimba (2) (635 bp) (585 bp)

Schedule-I. Part 4 (C) - Echinodermata: Sea cucumbers (All Holothurians) 7 Holothuria scabra (3) (640 bp) (510 bp) 8 H. atra (2) (640 bp) (640 bp) 9 Thelenota ananas (1) (640 bp) (530 bp)

Schedule-I. Part 4 (B) -Molluscs 10 Tridacna maxima (2) (490 bp) (465 bp) 11 T. squamosa (2) (520 bp) (459 bp)

Table 4. Generation of species-specific molecular signatures of endangered aquatic species[under Indian Wildlife (Protection) Act, 1972]

*Figure in parenthesis indicates no of samples sequenced

relative to molecular marker (pBR322DNA/MspI digest). None of the loci showedsignificant linkage disequilibrium (P>0.05). Itwas therefore assumed that allelic variation atmicrosatellite loci could be consideredindependent. The estimated null allelefrequency was not significant (P<0.05) at all in14 tested loci using different algorithms inMICRO-CHECKER, indicating the absence ofnull alleles. In initial data analysis, the observedheterozygosity values were in the range of 0.047- 0.239. The probability test provided theevidence that the observed allele frequencies insix loci significantly deviated (P<0.05) from thatexpected under Hardy-Weinberg equilibrium inthree populations.

Amplification of ATPase 8/6 region insilver pomfret

The complete ATPsynthase 6 and 8 geneswere sequenced from 4 samples each from threedifferent geographic locations. The genes werePCR amplified using primers ATP8.2L8331 (5’–AAA GCR TYR GCC TTT TAA GC-3’) andCOIII.2H9236 (5’ – GTT AGT GGT CAK GGGCTT GGR TC - 3’). The cleaned up products

were bidirectionally sequenced in thesequencing facility in AB 3730 XL capillarysequencer (Applied Biosystems) which yieldedfinal edited sequence of 842 bp and 5haplotypes. More samples will be sequencedfrom different populations and data analyzedfor stock structure analysis.

Generation of species-specific molecularsignatures of endangered aquatic speciesand other marine species

Wildlife protected speciesSpecies-specific signatures of 11 finfish and

shellfish species enlisted in Schedule-I of theIndian Wildlife (Protection) Act, 1972 weregenerated, to aid in their conservation and curbthe illegal trade of these species (Table 4).

Molecular signatures (16SrRNA and COI)of marine finfish and shellfishes, including ninespecies of economically important crabs suchas, Scylla serrata, S. tranquebarica, Portunuspelagicus, P. sanguinolenticus, Charybdis cruciata,C. lucifera, C. ranulate, C. natator, Podophthalmusvigil; and 35 elasmobranchs of Indian waters

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including, 10 deep sea sharks, were alsogenerated in collaboration with CMFRI, Kochiand CIBA, Chennai. In addition to 16S rRNAand COI genes, for the whale shark (Rhincodontypus), species-specific partial sequenceinformation of cytB (589bp) and RAG2 (935bp)were also generated.

Resolving taxonomic ambiguities anddeveloping species-specific molecularsignatures in large barbs

Taxonomic ambiguity exists among thecultivable large barbs of Peninsular India.Therefore, to resolve the taxonomic disputes,the following species (Table 5) were collectedfrom the type localities & other areas with aview to generate morphological andmolecular signatures of these species, jointlywith the ZSI-SRS. The specimens wereaccurately identified; measurements taken andstudy is in progress to develop molecularsignatures (16SrRNA, COI, cytB, RAG2/ITS2regions) of these species.

Microsatellite markers for geneticvariability studies in Penaeus(Fenneropenaeus) indicus

Work on development of microsatellitemarkers for genetic variability studies in Indianwhite shrimp, Penaeus (Fenneropenaeus) indicuswas continued. Eighty one polymorphicmicrosatellites in F. indicus were developedthrough cross-amplification from otherpenaeids. The putative microsatellites developedwere cloned, sequenced and confirmed tocontain repeats. The suitability of the identifiedpolymorphic loci for stock identification wasevaluated on a heterogenous collection of 23 F.indicus samples from India and Oman. Thenumber of alleles per locus ranged from 5 to 14and the observed heterozygosities from 0.237to 0.889 (Table 6). Following the sequentialBonferroni adjustment, the probability test didnot detect any significant deviation in allelefrequencies from that expected under (P<0.001)Hardy–Weinberg equilibrium. None of the locishowed significant linkage disequilibrium

*Figure in parenthesis indicates no of samples collected

Table 5. Details of collection of cultivable large barbs of Peninsular India for developingspecies-specific molecular signatures

S. No Species of Peninsular barbs Location and river basin 1 Barbodes carnaticus (Jerdon) (10) Srirangapatanam-Cauvery River and

Chalakkudy River

2 Puntius jerdoni (Day) (6) Dakshina Kannada – Nethravathi River

3 P. dobsoni (Day) (4) Shimoga, and Chalakkudy River

4 P. pulchellus (Day) (2) Dakshina Kannada – Nethravathi River and Tungabhadra (Krishna River)

5 Gonoproktopterus curmuca (Hamilton) (5) Tungabhadra (Krishna River)

6 G. kolus (Sykes) (3) Deccan – Tungabhadra (Krishna River)

7 G. (=Hypselobarbus) kurali Menon and Rema Devi (5)

Kumaradhara, Nethravathi River and Chalakkudy River

8 G. thomassi (Day) (4) Chalakkudy River

9 G. mysorensis (Valenciennes) (2) Srirangapatanam-Cauvery River

10 G. periyarensis (Raj) (1) Periyar Lake – Periyar River

22

Gujarat Manglore Kochi Tuticorin Andaman Persian Gulf Red Sea Gujrat --- 0.075* 0.072* 0.064* 0.125* 0.008 0.010 Mangalore 0.061* --- 0.000 0.005 0.097* 0.155* 0.172* Kochi 0.088* 0.002 --- 0.000 0.091* 0.111* 0.147* Tuticorine 0.079* 0.011 0.003 --- 0.120* 0.173* 0.141* Andaman 0.292* 0.119* 0.158* 0.185* --- 0.212* 0.208* Percian 0.007 0.149* 0.231* 0.211* 0.239* --- 0.007 Gulf Red Sea 0.012 0.288* 0.225* 0.198* 0.260* 0.002

Table 7. Pair-wise FST (above diagonal) and RST (below diagonal) among seven populations ofF. indicus with 15 microsatellite loci (* p<0.005)

(P>0.05) and occurrence of nullalleles (P< 0.05). The polymorphicmicrosatellite DNA markersdeveloped in this study will beuseful for commercial shrimpbreeding and selectionprogrammes and genetic studies ofboth wild and cultured stocks ofF. indicus.

The suitability of 15 of the 81identified polymorphic loci forstock identification was evaluated

on a heterogenous collection of 23 F. indicussamples from Kochi. Individual shrimpgenotypes for each locus were determined. Thedata was analysed using software Genetix 4.02to obtain allele frequencies, mean number ofalleles per locus, expected (He) and observed(Ho) heterozygosity values. Tests for conformityto Hardy–Weinberg expectations wereperformed by the Markov chain method withparameters dememorization = 1000, batches =100 and iteration = 100 (Genepop Version 3.3d,probability test, Raymond and Rousset, 1998)(Table 7). The data was also analysed forgenotype linkage disequilibrium between pairsof loci in a population using the GENEPOP Ver.3.3d programme, based upon the nullhypothesis that genotypes at one locus areindependent of genotypes at other locus. Thegenotypes of the loci deviating from Hardy-Weinberg Equilibrium (HWE) were tested forexpected frequency of null alleles using MICRO-

Table 6. Summary of microsatellite markersdeveloped in Fenneropenaeus indicusParameter No./unit Total no. of markers screened for cross-priming

392

No. of loci cross-amplified in Fenneropenaeus indicus

81

Percentage of cross-amplification 20.7% No. of alleles 5-14 Average no. of alleles 11.75 Molecular weight/Allele size range 142-392 bp Type of repeat Mononucleotide repeat Di-nucleotide repeat Tri-nucleotide repeat Tetranucleotide repeat Pentanucleotide repeat Hexanucleotide repeat

2 46 19 12 1 1

Type of repeat Simple/perfect Compound Complex

43 28 10

Fig. 8. Three distinct genetic stocks of in Fenneropenaeus indicus

23

CHECKER (http://www.microchecker.hull.ac.uk/). The number of alleles per locusranged from 5 to 14 and the observedheterozygosities from 0.237 to 0.889. Followingthe sequential Bonferroni adjustment, theprobability test did not detect any significantdeviation in allele frequencies from thatexpected under (P<0.001) Hardy–Weinbergequilibrium. None of the loci showed significantlinkage disequilibrium for all pairs of loci(P>0.05). It was, therefore, assumed that allelicvariation at microsatellite loci could beconsidered independent. The estimated nullallele frequency was not significant (P<0.05) atall 15 tested loci using different algorithms inMICRO-CHECKER, indicating the absence ofnull alleles. The microsatellite DNA markersdeveloped in this study were used for geneticdiversity analysis of stocks of F. indicus fromIndian seas, Persian Gulf and Red Sea. Initialdata analysis indicated occurrence of 3 distinctgenetic stocks: 1) Dakshina Kannada, KeralaCoasts and Gulf of Mannar; 2) AndamanIslands; 3) Gujarat Coast, Persian Gulf and RedSea, thus, indicating the need for stock-specificmanagement strategies for the naturalpopulations of F. indicus (Fig. 8). The data willalso be of immense use in future selectivebreeding programme of F. indicus.

Genetic characterization of commonHAB species from Indian waters

Species-specific DNA profiles of eight HABspecies (7 Dinoflagellates: Ceratium furca,Neoceratium fusus , Dinophysis caudata, D.acuminata, D. fortii, Prorocentrum lima andNoctiluca scintillans; and 1 Haptophyta:Phaeocystis globosa) were developed that will behelpful in accurate and timely identification ofthese species. DNA extraction protocol for allthe above species was cetyltrimethylammoniumbromide (CTAB) and chloroform/isoamylalcohol method. The extracts were stored at -200 C until used for PCR. The genes were PCR

amplified using primers from related species.The cleaned up products were bidirectionallysequenced in the sequencing facility in AB 3730XL capillary sequencer (Applied Biosystems).The summary of results is given in Table 8.

Table 8. HAB species and details of theirspecies-specific DNA signatures

Genetic divergence * Cf & Nf = 3.29%, # 5.18%; @ Dc & Da =2.7%; Dc & Df = 3.47%; Da & Df = 2.29%; $ Da & Df = 3.67%

Identification of HAB samplesThe samples provided by the Centre for

Marine Living Resources and Ecology (CMLRE),Kochi (three samples – one from Andamanwaters and the other two from North-WestArabian Sea) were found to be Noctilucascintillans, based on partial sequenceinformation of two genes. Two cyanobacterialsamples from South-East and North-Westcoasts were found to be Trichodesmiumerythraeum, based on partial sequenceinformation of three genes and not the toxicspecies, T. thiebautii.

Taxonomic ambiguity resolved in exoticCharaciform fish Pacu and Piranha

Taxonomic ambiguity prevails in the exoticfishes, red-bellied Piranha and red-bellied Pacuas both look very much alike in coloration andmorphology in younger stages. Exotic fishcaught from Periyar and Chalakkudy Rivers

Species-specific genes/ sequences and edited length

HAB Species 18SrRNA (~610bp)

COI (~580bp)

Ceratium furca * # Neoceratium fusus * #

Dinophysis caudata @ No amplification

D. acuminata @ $ D. fortii @ $ Prorocentrum lima Noctiluca scintillans Phaeocystis globosa No amplification

24

near Manjali, Ernakulam, Kerala were earlierreported as red-bellied Piranha by manyagencies. These specimens were examined indetail by NBFGR. None of the sampled fish hadcharacteristic sharp triangular shaped teeth ofPiranha, but showed molar-like square-shapedteeth. Gut content of the fishes were alsoexamined and it included mostly leaves andnuts of some plants, semi-digested fish flesh,scales and juvenile bivalve shells (Villoritacyprinoides) indicating that the fish is anomnivore. These observations gave anindication that the sampled fish were not thepredatory Piranha. To confirm the correcttaxonomic position of the fish and to resolveambiguity, partial sequence analysis ofmitochondrial 16S rRNA (621bp) and COI(655bp) genes were carried out. The sequenceanalysis revealed that the species is red-belliedPacu (Piaractus brachypomus) and not the red-bellied Piranha (Pygocentrus nattereri) asreported earlier. The genetic divergencebetween Piaractus brachypomus and Pygocentrusnattereri was found to be 3.8 - 4.0% with COIand 1.7- 2.0% with 16S rRNA (Fig. 9).

DNA barcoding of Indian fishesThe work on DNA barcoding of Indian fish

species was continued. A total 169 tissuesamples of 70 marine fish species were collectedfrom Goa and Mangalore landing centers for

DNA barcoding as given below:

Fishes collected from Goa1. Congresox talabonoides2. Gerres filamentosus3. G. abreviatus4. Lactarius lactarius5. L. splendens6. Liza dussumieri7. L. macrolepis8. L. parsia9. L. waigiensis10. Mugil cephalus11. Nematalosa nasus12. Nemepterus japonicus13. Pomadasys maculatus14. Psettodes erumei15. Sardinella longiceps16. Scatophagus argus17. Secuter insidiator18. Selaroides leptolepis19. Siganus oramin20. Sillago sihama21. Sphyraena obtusata22. T. jarbua23. Therapon theraps24. Thryssa hamiltoni25. Trichurus trichurus

Fig. 9. Phylogentic relationship (NJ Tree) among pacu and piranha species based on sequence information of 16SrRNAgene (621bp)

25

Fishes collected from Mangalore1. Ablennes hians2. Aluterus monoceros3. Anadontostoma chacunda4. Apogon quadrifasciatus5. Arius maculates6. Caranx kalla7. C. malabaricus8. C. sexfasciatus9. Cephalopholis spp.10. Chirocentrus nudus11. Congersox talabon12. Crossorhombus valderostratus13. Cynoglossus aral14. Decapterus russelli15. Drepane punctata16. Dussumieria acuta17. Eleutheronema tetradactylum18. Eppinephelus diacanthus19. Escualosa thoracata20. Fistularia petimba21. Goniolosa manmina22. Hemiramphus unifasciatus23. Rastrelligar kanagurta24. Rhina ancylostoma25. Rhynchobatus djiddensis26. Sardinella longiceps27. Saurida tumbil28. S. undosquamis29. Scomberoides tol30. Scomberomorus guttatus31. Pleuronectus orientalis32. Scomberomorus lineolatus33. Siganus oramin34. Sphyraena langsar35. Tenualosa toli36. Therapon theraps37. Trachinocephalus myops38. Trachinotus blochi39. Upenus sulphurius,

40. Valamugil buchanani41. Xenentodon cancilla42. Pterois russelli43. P. zebra44. Rachicentron canadum45. Filimanus heptadactylus46. Diodon hystrix47. Priacanthus hamrur48. Nemipterus japonicas49. Pampus argentius50. Platax pinnatus,51. Lactarius lactarias52. Liza waigiensis53. Lagocephalus inermis54. Lutjanus semicinctus55. L. argentimaculatus56. L. fulviflama57. L. rivulatus58. Megalaspis cordyla59. Megalops cyprinoides60. Mene maculate61. Mobula diabolus

In addition to the above, 72 tissue samplesof 18 freshwater fish species were collected fromDevlond, Bihad Nadi Sirmor, Chhoti Pul,Bichhia, Ramnagar, Bakia Dam near Rewa(M.P.). These fishes were: Ompok pabda, O.bimaculatus, Labeo rohita, Heteropneusteus fossilis,Puntius sarana, Cirrhinus reba (Labeo ariza),Oreochromis mossambicus, Labeo bata,Eutropiichthys vacha, Clupisoma garua, Channapunctatus, Nandus nandus, Labeo boggut, Garagotyla, Xenentodon cancila, Mystus tengara,Notopterus notopterus and Ailia cheeh.

DNA isolation, PCR amplification andsequencing were completed for all the samples.A total of 102 sequences for samples offreshwater and marine finfishes were submittedto NCBI GenBank (Accession numbersHQ219094 - HQ219175; HQ219184 -HQ219203).

26

Fig.10. Agrose gels showing amplified products by: (A) OPAS decamer primers, DAX1 and DMRTA2 gene from male (M)and female (F) Clarias batrachus genomic DNA

A DAX1 and DMRTA2

Genetic stock structure elucidation inTenualosa ilisha and Channa striatus

Molecular markers are powerful tools todetect genetic uniqueness of individuals,populations or species. It is now possible to scorenuclear and mitochondrial genetic markers andidentify variants and strains. A study wasundertaken with the aim to develop geneticmarkers to determine genetic stock structure ofcommercially important species, Tenualosa ilishaand Channa striatus using microsatellite,mtDNA and molecular cytogenetic markers. Sixsamples each of T. ilisha and C. striatus werecollected and DNA was isolated from thetissues. In both the species, the cytochrome bregion of mtDNA was amplified afterstandardization and submitted for sequencing.For cross species amplification of microsatellitesin T. ilisha and C. striatus, primers were searchedfrom closer species in literature. A total of 70microsatellite primers were finalized to be usedin two species.

Population genetics and molecularcharacterization of selected species ofNorth-Eastern region

One hundred and ten fish species werecollected and molecular identification and

phylogenetic studies using mtDNA markers(COI, 16S rRNA and cytochrome b) werecontinued for Channa spp, Mystus spp andDanio spp. Additionally, in order to characterizethe population traits of Ompok bimaculatus, atotal of 97 individuals were collected fromGomoti and Muhuri rivers of Tripura, out ofwhich 77 tissue samples were processed forDNA isolation and PCR conditions werestandardized for mitochondrial DNA regions.A total of 70 primers were finalized for crossspecies amplification of microsatellite loci.Standardization of polymorphic microsatelliteloci based on cross species amplification wasdone for N. hexagonolepis and O. belangeri. Theprimers were screened for the respectivepopulations.

Development of sex specific markers inClarias batrachus

For the identification of sex specificmarkers in C. batrachus, 40 decamer primerswere used, out of which, 4 primers gaveamplification in both male and female. Oneprimer was found useful to give sex specificamplification. Seven primers for sex relatedgenes were designed for amplification in C.batrachus, and two genes namely, DAX1 andDMRTA2 gave amplification (Fig. 10).

27

Phylogenetic analysis in catfish speciesThe catfish are an important group of fishes

having widespread geographic distribution.Out of 28,000 fish species found worldwide,there are around 2,867 species of catfishbelonging to 446 genera under 35 families. 202species belonging to 52 genera under 13 familiesare reported from India. There are existencesof higher phyloigenetic relationship amongthem, but systematic uncertainty occurs atseveral taxonomic levels in catfish species. Thepresent study was aimed to construct thephylogeny among the group based on DNAsequence data from three nuclear gene regionsnamely, ITS 2, S7 ribosomal protein intron 1(S71) and ribosomal protein L18 (RPL18) ofselected catfishes.

Sequence comparison of the ITS region iswidely used in taxonomy and molecularphylogeny due to the high copy number ofrRNA genes easy to amplify even from smallquantities of DNA, and of high degree ofvariation even between closely related species.The ribosomal protein (r-protein) genes areconsidered housekeeping genes, with a stronglycoordinated constitutive expression. Thesegenes have become widely used as markers forphylogenetic studies and comparativegenomics, but they have not been available infish. Similarly, S7 r-protein gene is adiscontinuous protein-coding gene consists of7 exons and 6 introns in series spanning about6 kb in human, although the introns lengthvaries considerably. The 1 st intron is anappropriate genetic marker for phylogeneticreconstruction of the taxa. Introns appear toharbor a much greater degree of genetic

polymorphism within and between species thanexons because of lower evolution pressure.

The ITS 2 sequences of Glyptothorax specieswere analyzed using Kimura-2 parameter (K2P)in MEGA (version 4.0.2) software. Theneighbor-joining tree was generated, whichgrouped G. brevipinnis, G. dakpathari and G.garhwali in one cluster, while G. granulus andG. ngapang in other, showing comparativeintimacy (Fig. 11). The pairwise distance amongthese species was computed with averagedistance of 0.037, ranged from minimumdistance of 0.020 (between G. brevipinnis andG. dakpathari) to maximum of 0.063 (betweenG. granulus and G. garhwali). The sequenceanalysis of other species is under progress.Ribosomal protein L18 (RPL18) gene region wasamplified using suitable primers and sequencedin Mystus cavasius, M. aor, Sperata seenghala,Clarias batrachus, Glyptothorax and Ompok spp.The S7 intron 1 gene region was amplified inM. cavasius, M. aor, S. seenghala, C. batrachusand Wallago attu using suitable primers.

Protein profiling in Indian major carpsProteomics research has enormous

applications in human and animal research.However, proteomic studies in fisheries scienceare quite scanty, especially in India. Massspectral-based proteomic technologies areideally suited for the development of proteinbiomarkers in the absence of any priorknowledge of quantitative changes in proteinlevels. Keeping in view of its emergingimportance in fisheries, new initiative has beentaken to commence proteomics research atNBFGR, Lucknow in collaboration with

Biophysics Department,AIIMS, New Delhi. The gilltissues from normal Labeorohita and specimensexposed to cadmiumchloride were analyzed forprotein profiling using 2-Dgel electrophoresis. The gelstained with silver clearlyFig.11. Neighbor joining tree showing comparative relatedness among five

Glyptothorax spp.

Glyptothorax brevipinnis Glyptothorax dakpathari

Glyptothorax garhwali Glyptothorax granulus

Glyptothorax ngapang

81

100

0.005

28

revealed distinct protein spots in the treatedspecimens. The protein spots were analyzedusing ESI Mass Spectrometry. Further studiesare being carried out to make protein profile ofIndian major carps. Sample preparations forproteomic analysis of gill, liver, heart, kidneyand muscle tissues of L. rohita, Catla catla andCirrhinus mrigala were made, followed by 2-Delectrophoresis of protein samples of thesetissues. The protein spots are being analyzedusing ESI Mass Spectrometry.

Cytogenetic studies in freshwater andmarine species

Conventional cytogenetic characterizationof marine fishes

Live specimens of marine fish species viz.,Scarus ghobban, Sargocentron diadema, Neonipponsummara, Lutjanus vitta, Abudefduf saxatilis,Plectorhinchus nigrus, Siganus canaliculatus, Lizaparsia and Sillago sihama were collected fromVizhinjam, Kerala. The specimens were injuvenile stage and the sex was unidentifiableby visual examination. Metaphasecomplements were obtained from anteriorkidney after intramuscular administration of0.05% colchicine using conventional hypotonictreatment – fixation (methanol: acetic acid) –flame drying technique. The chromosome slideswere stained with 6% Giemsa in Phosphatebuffer. The chromosomes slides were stainedfor conventional karyotyping and chromosomebanding (Fig.12). The karyotypes wereprepared from the cells exhibiting the completechromosome complement and possessingcharacteristic morphology. Homologouschromosomes were paired based on theirmorphology and centromeric position andarranged in decreasing order of their size usingLieca CW4000 Karyo software. The diploidchromosome number in Scarus ghobban was(2n=48) and the karyotype formula was10m+4sm+12st+22t, whereas, the diploidchromosome number in other marine speciesnamely, Sargocentron diadema, Neonipponsummara, Lutjanus vitta, Abudefduf saxatilis,

Plectorhinchus nigrus, Siganus canaliculatus, Lizaparsia and Sillago sihama were 48 withtelocentric chromosomes. The CMA3 and Ag-NOR staining revealed presence of one pairsignals in N. summara, S. diadema, L. vita, S.ghobban and on two pairs in S. canaliculatusand Z. carnutus (Fig. 13).

Molecular cytogenetic characterization ofmarine fishes

Genomic DNA was isolated from ethanolpreserved blood cells of the collected marinespecies. Several rDNA primers were used foramplification of different regions of ribosomalgenes, viz. 18S, 28S and ITS 2. The PCRamplified products of various rDNA weresequenced. The length of various regions ofrDNA is given in Table 9.

Table 9. Length of various regions of rDNAin selected marine fishes

S. N.

Description Species Size (bp)

1. 18S rDNA, partial Abudefduf sexfasciatus 910 2. 18S rDNA, partial Chaetodon collare 1393 3. 18S rDNA, partial Liza parsia 1887 4. 18S rDNA, partial Plectorhinchus nigrus 1404 5. 18S rDNA, partial Scarus ghobban 1411 6. 18S rDNA, partial Signus canaliculatus 970 7. 18S rDNA, partial Sillgo sihama 418 8. ITS 2 Plectorhinchus nigrus 658 9. ITS 2 Pomacentnus aquilus 473 10. ITS 2 Liza Parsia 612 11. ITS 2 Signus canaliculatus 696 12. ITS 2 Plectorinchus

flavomaculatus 692

13. ITS 2 Scarus ghobban 559 14. ITS 2 Chaetodon chollare 632 Based upon the sequences of ITS 2 regions,

the phylogenetic tree was constructed usingMEGA (version 5.05) software that revealedChaetodon chollare and Plectorhinchus nigruswere more closely related followed by Lizaparsia, Siganus canaliculatus and Pomacentrusaquilus (Fig. 14).

29

Fig. 12. Giemsa stained karyotypes of selected marine fishes

30

Fig. 13. CMA3 stained metaphase spreads of selected marine fishes

Fig. 14. Phylogenetic relationship among five marine fishes based on ITS-2 region

Plectorhinchus nigrus

Chaetodon chollare

Liza Persia

Signus canaliculatus

Pomacentnus aquilus

9954

0.00.10.20.30.4

31

Fig. 15. Lipid peroxidation in gills of L. rohita

Fig.16. DNA damage in gills of L. rohita

Genotoxicity studies

Investigation of cadmium inducedalteration in DNA integrity and lipidperoxidation response in Indian majorcarp, Labeo rohita

Cadmium (Cd) is a pervasive toxicant thathas been recognized as one of the most toxicheavy metals and also hazardous to aquaticecosystem. The dispersion of Cd in theenvironment has increased over the pastdecades due to its widespread industrial use.This metal has many cellular impacts; it triggersinduction of some proto-oncogene, and inhibitsDNA repair system and several enzymesinvolved in oxidative stress responses. Keepingthis in view, a study was undertaken, followingthe OECD guideline No. 203 in the static testconditions, to investigate the effect of Cd onlipid peroxidation and genotoxicity in Indian

major carp, L. rohita. The healthy andlaboratory acclimatized specimens wereexposed in vivo to three test concentrations ofcadmium chloride, viz., sublethalconcentrations I (SL-I; 1/4th of LC50 =33.55 ml),II (SL-II; 1/2nd of LC50 = 67.10 ml) and III (SL-III; 3/4th of LC50= 100.65 mg).

Exposure of L. rohita to test chemical for96 h caused a significant (P<0.05) induction ofLPO in gills (Fig. 15). Maximum amount ofTBARS formed at highest concentration after96 h exposure. The LPO level in gills was164.69% at the highest concentration andexposure time, when compared to control. Theextent of tissue LPO in the treated groupsincreased in a dose- and time-dependentmanner.

The single strand DNA breaks representedin the form of % tail DNA increasedsignificantly (P<0.05) with increasing chemical

32

Fish/ tissue Chromium Cadmium Lead Copper L. calbasu (muscle) 0.8870* 0.1497 0.9642 1.0980 L calbasu (gill) 0.2391* <0.05 0.7126 0.3741 P. sophore (muscle) 0.4411* <0.05 0.7671 0.7040 P. sophore (gill) 1.2268* 0.2169 1.4483 0.9896 M. vittatus (muscle) 0.9617* 0.1046 0.0378 0.3603 M. vittatus (gill) 0.3628* 0.0599 0.7823 0.2330

Table 10. Heavy metal concentrations (µg/g) in tissues (fresh weight) of fishes collected frompolluted site of river Ganga

*Values represent higher concentration of heavy metals as set up by FAO / WHO

Fig. 17. Comet cells showing DNA damage in erythrocytes of: a. C. punctatus; b. P. sophore; c. S. bacaila; and d. M.vittatus collected from polluted site

concentrations and exposure durations (Fig.16).The highest DNA damage observed at highestconcentration and exposure duration. Thelowest DNA damage was observed in gills at24 h and there was gradual non linear increasein the DNA damage with progression of theexperiment.

Estimation of bioaccumulation ofselected heavy metals in fishes collectedfrom polluted sites of Ganga River

Analysis of heavy metal concentrations,especially chromium, cadmium, lead andcopper in fish tissue and water samples wereperformed using AAS. Muscle and gill tissuesof L. calbasu, P. sophore and M. vittatus, weighing2 g each, were taken for bioaccumulationstudies. The results of bioaccumulation studiesshowed the presence of aforesaid heavy metals,especially chromium (Table 10), which wasabove the permissible limits of FAO/WHO(1984).

Assessment of genotoxicity induced bytannery effluents in fishes collectedfrom river Ganga

The specimens of Channa punctatus,Salmostoma bacaila, Puntius sophore andMystus vittatus were collected from threedifferent sites i.e., upstream (Site A),tannery effluent discharge site (Site B) anddownstream (Site C) of the river Ganga, nearKanpur, for assessment of genotoxic andclastogenic effects of polluted river water inblood cells, using comet assay and micronucleus(MN) test. There was significantly (P<0.01)higher DNA damage and micronuclei inductionin fishes collected from tannery effluentdischarge site B, as compared to sites A and C(Figures 17-19). Other than micronuclei,nuclear abnormalities like blebbed, notched, bi-nucleated and lobed nuclei were also observedin the erythrocytes of above mentioned fishspecies (Fig. 20).

33

Fig. 18. Percentage of tail DNA (±S.E.) in erythrocytes ofC. punctatus, S. bacaila, P. sophore and M. vittatus collectedfrom three sites of river Ganga

Fig. 19. Micronuclei frequencies (%) in erythrocytes offish specimens collected from different sites of river Ganga

Fig. 20. Micronuclei test showing micronuclei and other nuclear abnormalities (a-f) in the erythrocytes of Salmostomabacaila collected from Site B

34

Fig. 21. Effect of potassium dichromate on superoxide dismutase enzyme activity (units/min/mg protein) in liver andkidney of C. carpio fingerlings

The results reaffirmed that water from theeffluent discharge site was more genotoxic dueto the presence of chromium containing tanneryeffluent which resulted in higher rate of DNAdamage and micronuclei formations, as well as,other nuclear abnormalities. The study providesinformation on pollution status of the riverGanga due to disposal of tannery effluents inthe river and also acute toxicity status of theriver.

Assessment of chromium inducedoxidative stress in liver and kidney cellsof Cyprinus carpio fingerlings

Fish, being an important inhabitant ofaquatic ecosystem, is exposed to multi-pollutionstates and, therefore, considered as modelorganism for eco-toxicological studies of aquaticpollutants, including toxicity of metals. Amongthe heavy metals, hexavalent chromium, Cr(VI), is predominant in the environment andcan induce oxidative stress in aquaticorganisms. In this study, oxidative stressresponse was investigated through antioxidant

enzymes activities induced in liver and kidneytissues of C. carpio fingerlings, following 96 h invivo exposure at three test concentrations ofpotassium dichromate (70.75, 141.50 and212.25 mg) along with control. The tissuesamplings were done at 24, 48, 72 and 96 hpost-exposure.

The results indicated that the exposedspecimens experienced oxidative stress ascharacterized by significant variation in theantioxidant enzyme activities as compared tocontrol. The exposures lead to the productionof reactive oxygen species (ROS) andconsequent modulation in antioxidant enzymesactivities, such as superoxide dismutase (SOD).In the subject tissues of treated specimens, theactivity of SOD increases with progression ofthe experiment (Fig.21). The findings of thisstudy will be helpful in organ-specific riskassessment of Cr (VI) induced oxidative stressin fishes.

Values in treatment groups weresignificantly higher (P<0.05) from thecontrol.

Superoxide dismutase in Liver

Exposure in hours Exposure in hours

Superoxide dismutase in Kidney

35

5.3 IN SITU CONSERVATION

Fish germplasm exploration,assessment, cataloguing andconservation in North-Eastern Region

The North-Eastern region of India,comprising the states of Assam, Meghalaya,Manipur, Arunachal Pradesh, Mizoram,Tripura, Nagaland and Sikkim possesses aunique potential of fishery resources and isconsidered as one of the hot spots of freshwaterbiodiversity in the world. In view of this, thework in the north-eastern region was continuedin collaborative mode, involving localcollaborators from the region. The priority wasgiven for persuing research programmes onbiodiversity exploration, germplasm collection,maintenance and preservation of fish geneticmaterial through gene banking. The details ofthe work programme, along with theachievements for the period underreport, are given below:

Fish germplasm explorationand evaluation

Fish germplasm explorationand collection was carried out in therivers of Kaladan drainage (28species) covering Koladyne River,Mat River, tributary of theKoladyne; Ganga-Brahmaputradrainage (24 species) coveringKarnaphuli River; Brahmaputradrainage (17 species) coveringBrahmutra River, Noa-DehingRiver, Dikrong River and Chindwindrainage (15 species) in the valleyand hill districts of Manipur(Fig.22). Four new species namely,Glyptothorax ater, G. pantherinus,Batasio mizoramens and Harakoladynensis were discovered duringthe study period (Fig. 23).

Extensive field surveys were conducted inthe unexplored areas of East and West Bank ofthe Daying Ering Memorial Wildlife Sanctuary(a protected area) in East Siang district ofArunachal Pradesh and identified 10 feederrivers and streams. A total of 35 fish specieswere recorded from this protected area so far,of which 10 species were reported for the firsttime in the river Siang. In River Brahmaputra,a total of 117 species were collected from 10sampling locations and certain physiochemicalparameters were estimated. Fish diversityexploration of Jinari River resulted in 75 species,whereas, in Simsang River, a total of 53 specieswere recorded. Out of the total species recordedfrom Simsang River, 20 species were identifiedas inhabitants of high altitude streams andrivers, along with, some endemic species (e.g.Chaudhuria khajurial, Macrobrachium assamensepeninsulare, etc). The exploration andassessment were also conducted in River Tista

Fig. 22. Map of North-Eastern India showing collection sites

36

of Sikkim covering 7 different altitudinal zoneswhich resulted in a total of 20 fish species, outof which, some species of genus Glyptothorax,Garra, Nemacheilus and Barilus are stillunidentified.

Phylogenetic studies of the catfish superfamily Sisoridae

Phylogenic characterization was carriedout for the genus Amblyceps and Glyptothoraxbased on the morphology and anatomy. The

results were depicted using cladogram whichformed an independent separate group for A.mangois while, A. arunachalensis and A.tuberculatum formed a common clade. Acommon clade was also shared by A. apangi andA. torrentis which, revealed a closer relation tothat of A. arunachalensis and A. tuberculatum(Fig. 24). In Glyptothorax, a distinct group wascreated by G. cavia and G. chindwinica while theremaining species bifurcated into botius andtelchitta group.

Fig. 23. New species identified from the North-eastern region of India

Fig. 24. Cladogram of Amblyceps of North-Eastern India

37

Osteological inventory of Glyptothoraxspp.

Osteological studies were conducted forthe eight species of Glyptothorax based on themorphological characters of the premaxilla,dentary, Weberian lamina, infraorbital series,vomer and frontal bones. In G. botius, G.granulus, G. manipurensis, G. ngapang, G. striatusand G. ventrolineatus, the premaxilla consistsonly of proximal and distal tooth plates, theanterior portion of the dentary was slender andits dorsal surface beared villiform teeth. Thelateral extension of posterior portion ofweberian lamina terminates at the level of the

lateral margin of its anterior portion, and thefrontal had a shallow orbital notch (Fig. 25). InG. cavia and G. chindwinica, the premaxillaconsists of proximal, distal and posteriorelements on the roof of the oral cavity; theanterior portion of the dentary beared posteriorextension of dentary tooth-plate; the lateralextension of the posterior portion of Weberianlamina extended almost to the distal tip of thefifth parapophysis; there were nine or teninfraorbital bones with a longer and broaderbody of the lacrimal; greatly enlarged vomerinehead; and frontal beared a deep orbital notch(Fig. 26).

Fig.25. Weberian lamina: A - ventral view of G.chindwinica; B- dorsal, C - ventral of G. ngapang; D -ventral view of G. botius

Fig. 26. Lateral view of intraorbital series: A - G.chindwinica; B - G. ngapang

38

Habitat inventory of Murrel speciesIntensive field studies were conducted to

explore the natural habitat pattern of murrelsin Assam and their various micro and macro-habitat features were identified (Fig. 27). Fieldstudies were also conducted in some places ofoccurrence of Channa aurantimaculata and C.stewartii. They were recorded in associationwith different habitat zones in the water bodiesof Assam, viz., tributaries, wetlands, ponds,ditches, swamps, phragmite grassland,inundated depression of silted river beds,inundated wetland shoreline and high landdepression near inundation zones. A suitablehabitat was identified for the four endemic andthreatened species of murrels(C. aurantimaculata, C. bleheri, C. barca andC. stewartii) in Chandubi tectonic wetland

located in a protected reserved forest under theDepartment of Forest, Govt. of Assam. Thewetland habitat of murrel was consisting ofmyriads of submerged and underground treestumps, which proved to be the bestenvironment for the hole dwelling murrels. Thecauses of population decline of these fourmurrel species in the studied zones wereexamined during the study. The study revealedthat several factors like removal of Nal chaparior phragmites grassland cover by the nearbyvillagers for cultivation of crop, capturing of thefishes from inside the holes (micro-habitat) bythe skilled fishers, EUS (Epizootic UlcerativeDisease Syndrome) and shrinkage of waterbodies, etc may be responsible for decline ofpopulation of these important endemic species(Fig. 28).

Fig. 27. Field studies on natural habitat pattern of endemic snakeheads in Assam

39

Studies on fish biodiversity and aquaticenvironment of Ken and Betwa rivers: Anassessment prior to river inter-linking

In order to fulfill the increasing waterdemand and to mitigate flood and drought inIndia, the country’s first river interlinkingproject on Ken-Betwa Rivers has been plannedamong the states of Uttar Pradesh and Madhya.The Ken–Betwa interlink (KBI) involves buildinga dam at Daudhan 2.5 km upstream of existingGangau weir on River Ken and diverting thewater to Betwa through a link canal of 231.45km. River Ken, from its origin in the Jabalpurdistrict of Madhya Pradesh, covers a total lengthof 427 km while, River Betwa originates in theRaisen district of Madhya Pradesh and coverstotal length of 590 km. The KBI area is spreadin the Panna and Chhatarpur district ofMadhya Pradesh. Prior to interlinking of boththe rivers, the studies were continued to collectthe baseline information on the status ofdiversity, species composition, abundance,richness, distribution, identification of potentialbreeding grounds, along with, aquaticmicrohabitat characteristics. The data werecollected from 5 sites each in rivers Ken (K1 -K5 ) and Betwa (B1 - B5) to synthesize the levelof species and expected habitat change/lossafter interlinking and remedial measures.

Environmental characterization and fishhabitat use pattern

Habitat sampling was conducted at 50 sub-sites of both the rivers and data were analyzedfor 26 fish species in Betwa and 24 fish speciesin Ken. Out of total 17 environmental variables,fifteen environmental variables in Betwa andeleven environmental variables in Ken wererecorded as significant contributors in shapingthe environmental structure of both the rivers.As revealed by the canonical correspondenceanalysis (CCA), the most importantenvironmental variables for the fish assemblagein both the rivers were water flow, depth andsubstrate (Fig. 29 a & b). For example, thepresence and abundance of the featherback(endangered), which was collected in goodabundance i.e. >20% in both the rivers wasstrongly associated with sand substrate, deepwaters and presence of rangeland land use. Itwas found that in River Betwa, the majority ofspecies including Catla catla, Chitala chitala,Walllago attu, Bagarius bagarius, Ompokbimaculatus and Eutropiicthys vacha were mostlyassociated with average of parameters for thesite sampled, whereas, in Ken River, thesespecies were associated with specific habitattypes. For example, major carp, C. catla wasassociated with the sites contained deep waters

Fig. 28. EUS in snakeheads

40

Fig.29. Canonical correspondence analysis showing correlation between species composition and environmental variablesin: (a) Betwa River and (b) Ken River

41

with slow water current and higher percentageof sandy substrate, whereas, the large sizedmigratory species like C. chitala, W. attu, B.bagarius were associated with the sitescontained high conductive waters and higherconcentrations of dissolve oxygen. The habitatpreferences of these fishes in River Ken towardsspecific habitats supports our assumptions ofthe present intact ecological set up in River Kenthan in River Betwa which, at present, is facingvarious natural and anthropogenic threats tothe habitat and fish diversity.

Fish germplasm exploration andassessment

The exploration of fish diversity of both therivers consisted of a total of 67 fish speciesbelonging to 49 genera under 21 families. RiverBetwa showed higher richness of 63 speciesbelonging to 45 genera under 20 families, ascompared to Ken which showed 57 speciesrepresenting 42 genera under 20 families. TheShannon-Weiner diversity index indicated adistinct relationship with overall speciesrichness for both the rivers. In Betwa, themaximum diversity index was observed inupper stretch, as compared to lower stretch. Itranged from 1.89 to 3.51, whereas, in Ken, themiddle stretch showed maximum diversityranging between 2.73 to 3.18. The speciesrichness index ranged between 1.12 to 4.31 inBetwa, whereas, it ranged between 3.22 to 4.44in River Ken. The highest species richness wasrecorded from the upstream in Betwa, whereas,from midstream in River Ken. In both the rivers,the Community Dominance Index (CDI)showed differences between the sampling sitesand recorded highest at B3 of river Betwa andK2 of river Ken. The relative abundance of thespecies across all sampled sites revealed thatmost of the fish species are relatively rare, while,a few species dominated an area in terms oftheir numerical abundances. A total of 43species had an abundance of more than 100individuals out of the total 15,008 individuals

in River Betwa, whereas in Ken, it was foundin 26 species out of total 12,374 individuals.Analysis of biological indices were continuedfor the parameters like length-weightrelationship, age and growth, sex ratio, gonado-somatic index, fecundity, and length at firstmaturity of the prioritized fishes, from both therivers. GIS (Geographic information system)maps were prepared based on diversity indexand species richness for different sampling sitesof both the rivers, which showed higherdiversity and species richness in the upstreamof Betwa and middle stream of Ken. The otolithstructure of freshwater mugil R. corsula wereanalysed using SEM (Scanning electronmicroscope) and significant linear relationshipswere recorded with TL, FL, and SL of the fish(Fig. 30).

Fig. 30 Scanning electron microscope (SEM) structure ofotolith of R. corsula

Among exotic fishes, Oreochromismossambicus (Peters), Cyprinus carpio (Linnaeus)and Clarias gariepinus (Burchell) were recordedfrom various sampling sites. From the total catchamong all the exotics, the relative abundanceof the exotic species in Ken and Betwa riversranged from 11.7% to 45.6 % and was highestin River Betwa. The highest distributional rangewas recorded in C. carpio which was distributedin the downstream section of River Betwa andmidstream to downstream regions of Ken River.C. gariepinus was restricted to upstream (B2) ofBetwa River.

42

New morphotypeAn analysis was conducted to compare the

population structure of Labeo boggut, animportant food fish from varied eco-habitatconditions of both rivers (Fig. 31). Univariateanalysis of variance (ANOVA) revealedsignificant differences with varying degreesbetween the two population samples for 8, out

of 12, standardized morphometricmeasurements (only IOL, AFL, DFL and CFLwere not significantly different, P>0.05). Inorder to determine which morphometricvariables most effectively differentiate the twopopulations, the contributions of variables toprincipal components were examined. The firstthree principal components were extractedfrom 12 morphometric characteristics. Out ofthese, pre- dorsal length was found to be themost important character, which discriminatedboth populations. On the second principalcomponent (PC2), the inter orbital length,pectoral fin and caudal fin lengths being highlycorrelated. Inter orbital length was found to bethe PC2 while, anal fin length was the PC3

responsible to distinguish these two populations(Fig.32).

Length-weight relationshipLength-weight relationship (LWR) of 1683

individuals, covering 22 fish species of 16genera under 9 families, were studied fromboth the rivers, Betwa and Ken. All the datawere log transformed and least squares linearregressions performed on the transformed databy Graph pad Prism 5 with W as the dependentvariable, following the well known length–weight relationship log W = log a+b log L.Maximum value of ‘b’ was found in L. dyochilus(3.97), whereas, it was minimum in S. aor (2.31).Most of the studied species showed positiveallometric growth (b=3.00), except, M. Cavasius(2.91 to 2.94), S. aor (2.31 to 2.82), R. corsula(2.94) and M. armatus (2.40), which showednegative allometric growth in both the rivers(b=3.00). All relations were highly significant(P<0.001), with value of r>0.93.

Stock assessment of selected speciesThe ELEFAN-I program estimated

asymptotic length (L8) and growth co-efficient(K) of the von Bertalanffy Growth Formula(VBGF) was maximum for W. attu (1.5),followed by L. fimbriatus (1.4) and R. corsula(0.99). The calculated growth performanceindex (') was maximum for W. attu (3.52),followed by L. calbasu (3.45), R. corsla (3.22) andL. gonius (3.14). Minimum ' value was

Fig. 32. Contribution of morphometric variables to the canonical functions

Fig. 31 Labeo boggut collected from River Betwa

43

observed in L. fimbriatus (2.56), whereas, for L.boggut it was 22.58 cm and 0.53/ year for RiverBetwa and 23.63 cm and 0.610/ year for RiverKen. The calculated growth performance index(') of L. boggut in River Ken (2.53) was higherthan Betwa (2.43). The total mortality coefficient(Z), natural mortality (N) and fishing mortality(F) of 12 selected fish species were estimated byusing the length converted catch curve. Thetotal mortality per year was higher for T. tor(2.8/ year); whereas, it was lower for E. vacha(1.41/year). Natural mortality wasconsiderably higher for L. fimbriatus (2.12), whileit was notably lower for S. aor (0.90) and E.vacha (0.90). The exploitation level (E) of T. tor(0.61), P. sarana (0.61), L. calbasu (0.59), E. vacha(0.52) and L. dyochilus (0.51) was higher thanthe optimum level of exploitation (E= 0.50). Thisis a further indication that most of the riverfishery were above the optimal exploitation andif this scenario continues it would come underintense fishing pressure. Recruitment patternof all studied species was continued throughoutall months during the study period, with twomajor peaks in T. tor (June and August), C.mrigala (June and September), L. calbasu(February and August) L. dyochilus (May andOctober), L. gonius (April and July), W. attu(June and August), and E. vacha (April andOctober) (Fig. 33).

Prioritization of sites for biodiversitymanagement

Based on extensive explorations in the tworivers planned for interlinking, the criteria ofhabitat conservation and restoration of therespective sampling sites were developed. Thepriority for biodiversity management throughhabitat conservation should be given to thosesites having relatively higher percentage ofthreatened species. The higher number ofthreatened species was found in B2 and B4 ofRiver Betwa and K3, K4 and K5 of River Ken,hence, these may be considered for habitatconservation. The remaining sites like B5, B3,and B1 in Betwa, and K2 and K4 in Ken withrelatively lower percentage of threatenedspecies must be considered for biodiversitymanagement through restoration, whichinclude maintenance and habitat improvementof those areas. Based on the study, a generalprotocol for the restoration and conservationof threatened fish diversity in the post-interlinking phase was outlined for both therivers.

Ontogeny of Ompok bimaculatus larvaldigestive system

O. bimaculatus has been declared as theState Fish of Tripura and is categorized as anendangered species according to the IUCNcriteria. At present, high mortality rate of larvaeduring larval rearing is the most seriousbottleneck for commercial production of thisspecies. The information on ontogenicdevelopment of its digestive system may providebasis to understand the nutritional requirementsof the larvae and develop appropriate diets forlarviculture. Therefore, studies on ontogenesisof digestive system of O. bimaculatus duringlarval development were continued. Accordingto the morphological and histologicalcharacteristics of the digestive organs, thedevelopment and differentiation of the digestivesystem in O. bimaculatus could be divided intothree different phases: (1) an intense period ofcell proliferation and morphogenesis of the

Fig 33 Recruitment pattern of ten selected species fromRiver Ken and Betwa

44

digestive organs; (2) a period in which digestivestructures differentiated and achieved theirdefinitive morphological and histologicalorganization, and finally, (3) a stagecharacterized by the growth in size and volumeof the digestive organs. According to enzymeanalysis data, a progressive shift in activity fromalkaline to acid proteases was observed aftertwo weeks of larval development, reflecting thatalkaline proteases were not longer then the maindigestive enzymes involved in protein digestionafter the development of gastric glands andonset of acidic digestion.

Neuroendocrine regulation ofreproduction in threatened fishes(a) Neuroendocrine regulation ofovarian maturation in the seabass, LatescalcariferHypothalamus

Production of viable eggs is essential forsurvival and propagation of any species. Therehas been growing interest in teleost reproductionduring the past because of a number ofcommercial, as well as, conservation reasons.Hypothalamus in the vertebrate braincomprises groups of neurosecretory cells thatmediate the endocrine responses of the animalleading to adjustments to the environmentalchanges through modulating secretion ofvarious releasing (-RH), as well as, inhibitinghormones (-IH). Hypothalamus also containsreceptors specifically sensitive to the hormonewhich, in turn, regulates its activity throughfeedback mechanism. There are increasingevidences that in fishes also, hypophysialfunctions are modulated by the hypothalamicneurohormones but the regulatory mechanismsare not yet clearly understood. Seabass (Latescalcarifer) has been identified as one of thecommercially important aquaculture species inIndia. Though success has been achieved ininduced breeding and mass-scale seedproduction, neuroendocrine mechanism relatedto the reproduction of the species has not yetbeen clearly defined. Therefore, a study was

initiated to record the changes occurring inneurosecretory cells of hypothalamus andgonadotrophs of the threatened fishes forinducing early ovarian maturation forenhanced seed production.

Female L. calacrifer (total length 855-1050mm; total weight 8.5-11.0 kg) were segregatedand divided into 6 groups based on their ovarianmaturation. 5-6 µm thick sections were cut andstained in (i) lead hematoxylin (PbH)-periodicacid Schiff’s reagent (PAS)-orange G (OG), (ii)Herlant tetrachrome, (iii) Mallory’s triple, (iv)alcian blue (AB)-PAS-orange G, (v) aldehydefuchsin (AF) and (vi) chrome-alum-hematoxylin-phloxine (CAHP). Hypothalamo-neurosecretory complex of the Asian seabasscomprised nucleus preopticus (NPO), nucleuslateralis tuberis (NLT) and their axonal tracts.NPO was a paired structure situated on eitherside of the third ventricle dorsal to the opticchiasma. The horizontal limb of NPO comprisedsparsely distributed neurons whereas theneurosecretory cells were closely packed in thevertical aspect of pre-optic area. Based on sizeof the neurosecretory cells, NPO was furtherdivided into dorsal pars magnocellularis (PMC)formed of larger neuronal cells and a ventralpars parvocellularis (PPC) comprising smallercells. NPO was highly vascularized structureand its neurosecretory cells were positive toaldehyde fuchsin (AF) chrome-alum-hematoxylin-phloxine (CAHP) and acidfuchsin (Figs. 34-35). Neurons of PMC and PPCwere generally bipolar and contributed beadedaxons to form left and right neurohypophysialmain tracts. The neurosecretory cells of NPOwere laden with the secretory material in thematured specimens while they exhibitedexcessive vacuolization during spawning phaseof the ovarian cycle. NLT cells, distributed inthe ventral floor of brain adjacent to thepituitary stalk, were negative to AF but stainedreadily with acid fuchsin and CHAP. Theseneurosecretory cells were variously shaped andtheir size ranged from very small to large withpolymorphic nuclei (Fig. 36). The neurons wereusually bipolar but a few uni- or multipolar cells

45

were also seen. Based on the distribution andsize, NLT was divided into pars anterior, parsposterior and pars inferior. This structure washighly vascular and several neurons were seenin close association with the blood vessels.Neuro-hypophysial tract entered the pituitarygland through the infundibulum. Theneurosecretory cells of NLT exhibitedheightened activity during pre-spawning phaseof the reproductive cycle. Few acid fuchsinpositive globule-like structures were also seenin the NLT of maturing female seabass (Fig. 37).

GonadotrophsIn Lates calcarifer, the gonadotrophs were

localized in the proximal pars distalis (PPD)

and exhibited cyclical changes in activity inrelation to ovarian maturation. The PPDconsisted of two types of cells - acidophilsstainable with erythrosine and orange G as wellas cynophils stainable with PAS, alcian blueand aniline blue. The former predominatedduring resting phase of reproductive cycle whileactivity of the latter cells fluctuated widely withthe maturity stage. During maturing phase,granulation of the gonadotrophs was initiated(Fig. 38) while these were partiallydegranulated during pre-spawning stage (Fig.39). During spawning period, there wasdegranulation as most of the cynophil cellsdepicted excessive vacuolization due to releaseof the gonadotripin in blood circulation whilein preparatory and immature phases, the

Fig. 34. NPO of immature Lates calcarifer Fig. 35. NPO of maturing seabass showing vascularization

Fig. 36. NLT of maturing seabass depicting accmulationof granules

Fig. 37. NLT of maturing seabass exhibiting acidfuchsin+ve globules

46

gonadotrophs exhibited inactivity or low profileof activity with scanty accumulation ofsecretory granules (Fig. 40). Though somevesicular structures with acid fushsin+vecolloid-like material in their lumina wereencountered in the pituitary gland of L.calcarifer, its physiological/ phylogeneticsignificance is not yet known.

(b) Neuroendocrine regulation ofovarian maturation in golden mahseer,Tor putitora

Attempt was made to record the changesoccurring in hypothalamus, pituitary andovary of the golden mahseer in relation to

ovarian maturation. We could collect the femalespecimens in stage 2 and 3 of ovariandevelopment. 5-6 µm thick paraffin sections ofbrain and pituitary were stained in (i) leadhematoxylin (PbH)-periodic acid Schiff’sreagent (PAS)-orange G (OG), (ii) Herlanttetrachrome, (iii) Mallory’s triple, (iv) alcian blue(AB)-PAS-orange G, (v) aldehyde fuchsin (AF)and (vi) chrome-alum-hematoxylin-phloxine(CAHP) while ovarian sections were stained inhematoxylin-eosin, bromophenol blue andtoluidine blue (Fig. 41).

Though the basic cytoarchitectural patternof the hypothalamus of Tor putitora wascomparable to those of cyprinid teleosts but it

Fig. 38. PPD maturing seabass exhibiting granule-ladencynophil cells

Fig. 39. PPD matured seabass showing partiallydegranulated cynophils

Fig. 40. PPD of seabass during spawning with degranulatedcynophils

47

differed considerably from the hypothalamusof the L. Celcarifer. The pituitary gland of thegolden mahseer was divisible intoadenohypophysia and neurohypophysis. Theformer was further divisible into roastral parsdistalis (RPD), proximal pars distalis (PPD) andpars intermedia (PI) arranged one after the otherin antero-posterior axix. In PPD, somatotrophs(STH), prolactin. lasctotrophs (LTH) andgonadotrophs (GtH) cells were localized (Fig.42). Histological studies of the ovaries revealedoocytes in stage 2 and 3 of development.Further studies are in progress.

Role and potential of fisherfolkorganizations and NGOs in co-management of aquatic resources

The study on the role, performance andpotential of different types of fisherfolkorganizations and NGOs in the managementof aquatic resources, was continued. During theyear under report, current status of fisherfolkorganizations and NGOs working in fisheriesmanagement in Andhra Pradesh wasdocumented. Relevant policy and programmedocuments from different offices of the statefisheries agencies from Andhra Pradesh statewere collected and studied. The state regulatoryframework for fisheries management in theAndhra Pradesh was reviewed. Fisherfolk

organizations working in the form of fishingcooperative societies in Andhra Pradesh werereviewed and their functions were documented.Discussions were held with concerned officialsfor selection of sites for detailed case studies. Threesites (Vijayawada, Rajahmundary and Kakinada)were selected for detailed micro-level case studiesin Andhra Pradesh. Visits were made to theselected sites and primary information wascollected from the identified sites.

Formulation of cost effective feeds forClarias batrachus during seed rearing

Experiments were carried out to evaluatedifferent feed supplements during rearing ofClarias batrachus in order to minimize the costtowards supplementary feeding. The fry raisedfrom induced breeding was initiallyacclimatized with provision of moist feedcontaining goat intestine, wheat flour, soybeanmeal and vitamin & mineral mix mixed in aratio of 45: 15: 5: 1 w/w. Five different feedswere formulated and growth study was carriedout for 12 weeks rearing period with fry of C.batrachus in 15 plastic pools of 300 l capacity(Fig.43). The results recorded in terms of SGR,FCR and survival at the end of the 12th weekdemonstrated better efficiency of feed F- 4,followed by F-3 and F-5 (Table 11).

Fig. 42. PPD of immature Tor putitoraFig. 41. Examining ovarian

48

Morphological and biologicalcharacterization

Age and growth analysisThe scales from 509 Labeo rohita, 453

Cirrhinus mrigala, 192 Catla catla and 32 L.fimbriatus were extracted and preserved for ageand growth analysis. The detailed result of ageand growth analysis in 76 L. rohita, 84 C.mrigala and 46 C. catla is depicted in figures 6,7 and 8.

L. rohitaSpecific rate of linear growth showed

downward trend with increase in the age groupin all the studied rivers in case of L. rohita. Thestudy revealed the composition of age classes1+ to 8+ in case of rivers viz. Betwa and Ken(Fig. 44A). The value of specific rate of weight

increase was also observed in the decliningtrend from all the rivers (Fig. 44B).

C. mrigalaAnalysis of specific rate of linear growth

showed downward trend with increase in theage group in all the studied rivers in case of C.mrigala, except rivers Son and Tons where thevalue increased slightly. The age classes fromfive populations were composed of 1+ to 7+

(Fig.45A). The value of specific rate of weightincrease was also observed in the decliningtrend from all the rivers, except rivers Son andTons which went parallel to the value specificrate of linear growth (Fig.45B).

C. catlaSpecific rate of linear growth showed

downward trend with increase in the age groupin all the stocks of C. catla. Maximum age classes(8+) were recorded from River ChambalFig.6A). The value of specific rate of weightincrease was also observed in the decliningtrend from all the rivers (Fig. 46B).

Clarias batrachusThe attempts were made to study the age

and growth from otolith in C. batrachus fromthree populations (Fig. 47). The study indicatedthe age classes upto 2+ years.

Table 11. The growth performance of the grow-out of Clarias batrachus

Same alphabet in superscript in a column represents no significant difference in weight gain.* = p< 0.01 ; **= p< 0.05. The results are of triplicate sets of feeding trial.

Feed Initial weight

(g)

Final weight (g)

4th week

Final weight (g)

8th week

Final weight (g) 12th week

Specific growth rate

(SGR) after 12 weeks

Survival (%) FCR

F-1 2.2+ 0.1 4.3+ 0.3a 10.2+ 0.1a 14.6+ 0.2a 14.76+0.9a 76 + 1.5a 3.23+0.27c

F-2 2.4+ 0.4 4.1+ 0.3a 10.1+ 0.1a 14.7+ 0.4a 14.64+1.0a 68 + 3.4b 3.11+0.17c

F-3 2.1+ 0.3 5.3+ 0.1 b* 11.4+ 0.2b 16.9+ 0.3b** 17.62+0.6b** 75 + 1.2a 2.65+0.14b**

F-4 2.5+ 0.6 5.4+ 0.2b* 12.6+ 0.3b* 18.9+ 0.3c* 19.52+0.7c* 79 + 2.3c 2.24+0.12a*

F-5 2.7+ 0.9 4.8+ 0.4c** 10.3+ 0.2a 17.2+ 0.2b** 17.26+0.4b** 64 + 1.5b 2.02+0.07a*

Fig. 43. Growth study of Clarias batrachus fry in pools

49

Fig. 44A-B. Different growth parameters of L. rohita from rivers of Ganga basin

A B

Fig. 45A-B. Different growth parameters of C. mrigala from rivers of Ganga basin.

Fig. 46A-B. Different growth parameters of Catla catla

A B

A B

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Fig 47. Scanning electron microscopic structure of Otolithof C. batrachus

Explorations of fish germplasmresources

Exploratory surveys on germplasmexploration were conducted in the three districtsnamely, Chamba (Himachal Pradesh), Udaipur(Rajasthan) and Adilabad (Andhra Pradesh)(Figures 48-50). In district Chamba, altogether15 sites were explored covering an altitude ofapproximately 6000 ft to 1800 ft in the mainchannel of River Ravi alongwith its tributariesand 2 reservoirs. In total, 79 fish samples of 6species belonging to genus Schizothorax,Crossocheilus, Aspidoparia, Oncorhynchus, Salmo

and Cyprinus were collected across all thesampling sites. Among physio-chemicalparameters, the water temperature rangedbetween 7.5°C in the upper stream (Holi) to21.01°C in the downstream (Ranjit Sagar Dam)of Ravi. A low dissolved oxygen (D.O.) (7 ppm)was recorded in Chaned stream (a tributary ofRavi), while high quantity of D.O. (8.24 ppm)was recorded in the main stream of the RiverRavi near Kariyaan. Although, the pH did notvary much (8.16- 9.06); the conductivity (79-350µS/Cm) and TDS significantly varied (39-180 ppm) across all the aquatic bodies of theregion. In district Udaipur, a total of 171samples of 36 species were collected in JaismandLake, Som River, TD River (a tributary of Som),Nadeshwar Lake, Bari River, Fatehsagar Lakeand Banas River. Indian major carps; minorcarps i.e., Labeo dyocheilus, L. gonius, L. boggut,L. calbasu. etc., and catfishes including, Sperataaor, Ompok bimaculatus, Wallago attu etc. wererecorded among the dominant species of thedistrict. The major physio-chemical parametersof the district includes: pH (8.18-9.36), D.O.(6.5–5.1mg/L), TDS (206-501 ppm) andConductivity (442-1001 µS/cm). In district

Adilabad, 5 rivers/reservoir/ wetland i.e.,Penganga, Satnala,Godavari, Sriram SagarDam and Kondapurwetland area were explored.Over 250 samples of 46species including Tor spp,Labeo spp, Ompok spp andChanna spp and werecollected across the studyarea. Significant variationswere recorded among majorwater quality parametersincluding pH (8.2 - 9.7), D.O.(5.9–19.3mg/L), TDS (221-261ppm) and conductivity(442-525 µS/cm).

Fig.48. Fish germplasm exploration survey in Chamba

51

Fig.50. Fish germplasm of aquatic resources in Adilabad

Fig.49. Fish germplasm exploration survey in District Udaipur

52

5.4 EX SITU CONSERVATION

Cryopreservation of milt of Puntiussarana subnasutus

Puntius sarana subnasutus is a medium sizedcarp, having culture potential in the peninsularstates of India (Fig. 51). Its juveniles haveornamental value too. It is distributedthroughout the Western Ghats. The species hasgood demand as food fish among local people.Due to over-exaltation and habitat alternation,Puntius sarana subnasutus is declining in manyof its habitats. Hence, this fish was selected forcryopreservation of milt as a part of ex-situconservation programme.

Spawners were collected from theMuringoor wetlands (Fig. 52), nearIrinjalakkuda (Trichur District, Kerala) andwere brought to the laboratory in oxygen filledbags and kept in cement tanks foracclimatization prior to the experiment. Maleswere separated and kept in an aquarium. Themature male fishes were given Ovapriminjection @0.2 ml/kg body weight and milt wascollected from individual fish after 12 hours ofthe injection. The pooled milt showing morethan 75% motility was used forcryopreservation. The milt quality was checkedusing tap water as an activator and the durationmotility of sperm ranged 80-90 seconds.Spermatocrit of the milt ranged 32-34% andsperm density 7.7-8.2 x 108 SPZ. Six extendercompositions used earlier for carps were testedand finally four extenders (code named asNBFGR 3B, NBFGR 6B, NBFGR 7B and NBFGR9B) were selected for milt cryopreservation.

Dimethyl sulphoxide (DMSO) 10% was usedas cryoprotectant in all the extenders. Afterstorage for 24 hours in liquid nitrogen, thestraws were taken, thawed at 370C and motilitywas checked. Motility score was less in all theextenders compared to fresh milt and motilitytime was maximum 30 seconds.

The fertility trial was carried out usingthawed milt. Four mature female fishes weregiven Ovaprim injection @0.4 mg/kg bodyweight. After 12 hours of injection, the eggswere collected by stripping. About 200 ml(approximately 200 eggs) of eggs were takenand fertilized with thawed milt from a singlestraw. Fresh milt was used as control.Immediately after pouring the milt, water wassprinkled over the eggs for activation. Aftertwo minutes, the egg mass was washed withfreshwater and kept for hatching, eggs wereslightly sticky after fertilization. Fertilizationrate was calculated after eight hours andhatching rate was calculated 30 hours afterfertilization. The experiment was carried outin triplicate and the summary of results is givenbelow.

The percentage of fertilization andhatching was used for comparing the efficacyof different extenders. Fertility rate in theextender NBFGR 9B showed values close tocontrol (49.7±0.2%), followed by NBFGR 7B(48.1±0.7%), NBFGR 6B (45.9±0.9%) andNBFGR 3B (35.2±1.5%) (Table 12). In hatchingalso, the extender NBFGR 9B was close toFig. 51. Puntius sarana subnasutus

Fig.52. Collection site of Puntius sarana subnasutus

53

control (42.3±1.5%), followed by NBFGR 7B(41.3±0.9%), NBFGR 6B (27.9±1.0%) andNBFGR 3B (23.2±2.1%). It is observed that theextender NBFGR 9B with 10% DMSO can beused for mass cryostorage of the milt of Puntiussarana subnasutus.

Live gene banking (LGB) and captivepropagation of selected species

The facility of LGB developed incollaboration with the Department of Fisheries,Govt. of Assam was strengthened and facilityof a new deep tube was made functional. Livesamples of Chitala chitala, Ompok pabo, Anabastestudineus and Puntius sarana were collectedfrom tributaries of Brahmaputra namely, riversPichala, Subansiri, Sesha and Bogapara andBhelaimara wetlands in the Lakhimpur districtand stocked for brood stock development. Incontinuation of previous work, successfulcaptive breeding of C. chitala and O. pabo wasachieved, along with rearing of fry andfingerlings. A total of approx. 60,000 fry/fingerlings of C. chitala and O. pabo wereproduced. Additionally, breeding trial ofChanna aurantimaculata in the earthen pondwith Ovaprim @3 ml/kg bw in female and 2ml/kg bw in male was successful. Spawningwas noticed in small clusters of spawnsnumbering 500-800 nos/cluster during June,2010 in the live gene bank facility of theDepartment of Zoology, Gauhati University,Guwahati. During the field studies for theMurrels in Assam, a total of 56 nos C.aurantimaculata and 18 nos C. stewartii werecollected and stocked in gene bank maintained

at Gauhati University, Guwahati.

Under another programme with TripuraUniversity, a total of 97 individuals of Ompokbimaculatus were collected from Gomoti andMuhuri Rivers of Tripura. The spawningactivity of the species was noticed at a lengthranging from 230-353 mm.

Tissue repository collectionsTissue repository is an ongoing programme

of NBFGR and the fish tissues can be used forvarious purposes in future as a source of geneticmaterial. Tissue repository will be useful forreferring the related species in case of anyambiguity. During the period under report,more than 200 tissue samples were collectedfrom 75 marine specimens from the west coastof India and more than 170 tissue samples werecollected from 65 freshwater species from theWestern Ghats (Table 13). The tissue samplescollected in 95% ethanol are maintained in thetissue repository.

Fish Seed ProductionThe seed production of Indian major carps

was continued under the ICAR Mega SeedProject. A total of 260 lakh spawn wasproduced. Emphasis was given on productionof seed from riverine brood stock that wasraised at the fish farm by stocking fingerlingscollected from riverine sources. The seed wassold in the form of spawn, fry and fingerlingsto the farmers of different districts of U.P.Fig. 53. Revenue of Rs.2.53 lakh was generatedfrom seed sale.

Table. 12. Fertility trial with cryopreserved milt of Puntius sarana subnasutus (mean±SD)Extender Total eggs Fertilization % Hatching % % as that of control

3B 318 ± 5.3 35.2 ±1.5 23.2±2.1 49±4.5 6B 305 ±8.5 45.9 ±0.9 27.9±1.0 59±2.5 7B 312 ±5.6 48.1±0.7 41.3±0.9 87±2.0 9B 314 ±8.5 49.7 ±0.2 42.3±1.5 89±2.6

Control 390 ±7.5 50.8 ±0.5 47.5±1.8 100

54

S. No. Place River Basin/ coast No. of species

No. of Accessions made

Freshwater 1. Athirampally, Kerala Chalakkudy River 12 30 2. Pune, Maharashtra Mula-Mutta & Indrayni, Krishna River basin 10 22 3. Kolhapur, Maharashtra Panchganga, Krishna River basin 8 20 4. Hosanur, Karnataka Thungabhadra, Krishna River basin 10 25 5. Thirthahalli, Karnataka Bhadra, Krishna River basin 12 35 6. Mysore, Karnataka Cauvery River basin 13 38 Marine 7. Mangalore, Karnataka West coast 41 122 8. Trivandrum, Kerala West coast 20 68 9. Tuticorin, Tamilnadu East coast 14 27

Table 13. Details of surveys and tissue repository collections in 2010 –11

Fig. 53. Carp seed ready for sale under ICAR mega seedproject

Fig. 54. Experimental seed production of L. dyocheilus atNBFGR

NBFGR Live Gene Bank, LucknowThe stock of sixteen prioritized fish species

was maintained at the Live Fish Gene Bank ofthe institute. These include, Catla catla, Labeorohita, L. dyocheilus, L. calbasu, L. bata, Cirrhinusmrigala, Puntius sarana, Clarias batrachus ,Heteropneustes fossilis, Channa marulius, C.striatus, Tor putitora, Sperata seenghala, Mystusvittatus, Chitala chitala and Notopterusnotopterus. Successful experimental breedingand larval rearing of Labeo dyocheilus , C.marulius, C. striatus and Ompok bimaculatus wascarried out (Fig. 54). A good number of brood

stocks of these species are available for furtherbreeding in the next season.

Freshwater prawn farmingFarming practice of M. rosenbergii was

continued. The seed (PL-10) was procuredfrom the Rohtak center of CIFE and stocked ina pond prepared @50,000/ha in monoculture.This practice continued specially fordemonstration to aqua-farmers of the regionduring short-term training programmes.Average weight of 35 g could be achieved atthe end of November, 2010 with 70%survival.

55

5.5 EXOTICS AND QUARANTINE

Experimental infection and detection ofAphanomyces invadans in common carp

Common carp (Cyprinus carpio) isconsidered to be one of the resistant fish speciesto Epizootic Ulcerative Syndrome (EUS). Innatural EUS outbreaks in India, there havebeen several reports of common carp gettingaffected by EUS, but the diagnostic featuresconforming to the OIE diagnostic manual havenot been demonstrated in their reports. Inexperimental infection studies with commoncarp, it was observed in NBFGR laboratory thatabout 20% of the normal common carp(unpublished data) also develop infection anddie with typical EUS pathology. It may be thatsome of the common carps may be in immuno-compromised condition and, as a result, theinjected zoospores might be taking upper hand,thus, getting germinated and the germinatedhyphae getting proliferated and finally resultingextensive pathology/mortality.

To test this hypothesis, twenty numbers ofjuveniles of common carp were artificiallyimmuno-compromised by injection with aknown immunosuppressant i.e. Triamcinoloneacetonide @100 µg/g of fish and afterwardsexperimentally infected with A. invadanszoospores. Twenty fish injected with only

autoclaved pond water served as control. Afterexperimental infection, two fish from eachgroup were sampled regularly at 1, 3, 6, 9, 12,15 days of post injection. DNA was extractedfrom each sample using a DNeasy Tissue Kit(Qiagen) and was amplified using A. invadansspecies - specific primers. In experimentalgroup all the fishes were positive for A. invadansand with progression of infection there wasincrease in intensity of bands (Fig. 55A). Thisindicated that as the infection progressed, thehyphal numbers in the infected tissue were alsoincreased. As a result, more amount of A.invadans DNA were present in the tissuesamples. Histology of the tissue samples of themoribund fishes (Fig. 55B) indicated extensivenecrosis of the muscle fibers due to thepenetrating fungal hyphae (Fig. 55C). In viewof the present findings, it will be interesting toexamine the susceptibility of common carpduring natural outbreaks of EUS because thepresent study provides evidence that commoncarp in immuno-compromised condition aresusceptible to EUS.

Large-scale fish mortality in UttarPradesh due to EUS

Consequent to the report of heavymortality due to diseases twenty fish farms indifferent parts of Uttar Pradesh were surveyed.

Fig. 55 (A). Detection of Aphanomyces invadans in experimentally infected common carp.(Lane1- 100 bp marker (fermentas); Lane 2, one day of post injection (dpi) with A. invadans zoospores; Lane 3, three dpi;Lane 4, six dpi; Lane 5, nine dpi; Lane 6, twelve dpi; Lane 7, fifteen dpi (moribund common carp)Fig. 55 (B). Moribund common carps with gross visible lesionsFig. 55 (C). Histology of lesion area of moribund common carp showing necrosis of muscle fibers (arrowheads) due topenetration of fungal hyphae (arrows)(Grocott-H&E, x400)

56

Most of them were small-scale farms in thewetland regions of the state. During survey,large-scale mortality ranging from 20 to 50%of the total stock was observed in almost all thefish farms. (Fig. 56) Even though the grossvisible lesions of the fishes were typical of EUSlesions, for confirmatory diagnosis, ulceratedfishes from all the farms were collected andbrought to the laboratory for further study.

Total 97 samples representing 12 species(Catla, rohu, mrigal, common carp, bighead,Puntius, snakehead, Wallago, Mastacembelus,Colisa, Mystus and Glossogobius) were collected.Through Grocott’s Methenamine Silver stainingfungal hyphae were observed in 95.8% of thesamples (N=93) (Fig. 57). Through use of PCRassay, A. invadans was detected in all speciesexcept common carp. A. invadans was

Fig. 56. Large-scale mortality of fishes in EUS affected farms of Uttar Pradesh

57

Fig 57. Histology of lesion area showing necrosis of muscletissue (arrow heads) due to invasion of fungal hyphae(arrows) (Grocott-H&E,400X)

successfully isolated from the ulcerated mrigaland Puntius. The growth characteristics and themorphology of the isolates was similar to thatdescribed for already reported A. invadans. Formolecular analysis of the isolates, DNA wasextracted from the isolates using DNeasy tissuekit (Qiagen) and PCR amplified (amplicon size-564bp within internal transcribed space regionITS1 and ITS2). The PCR product wassequenced and resulting sequences wereassembled using the alignment program andcompared with other known sequences of A.invadans. The sequence analysis revealed 100%homology with the A. invadans sequences inGeneBank (accession numbers EU422990,FM999229, DQ403202, FM999231 andAF396684). This result was consistent withearlier findings that all the isolates of A.invadans in different parts of world are part ofone fungal strain that has been colonised in all

the affected countries in matter of decades andresulted in spread of EUS. This is the first reportof isolation of A. invadans in India.

In this study, histological preparations,fungal isolations and molecularcharacterization were used to identify A.invadans and provide evidence about large-scalefish mortality in cultured farms due to A.invadans infection. Preliminary survey oneconomic losses due to EUS outbreaks indicatedthat farmers lost approximately Rs. 60,000 to1, 80,000/ per farm (Table 14). This observationis against the general perception that EUSincidence has declined and no longer a problemin aquaculture farms in the country.

Production of monoclonal antibodies (MAb)to serum immunoglobulins of Channastriatus and Labeo rohita and theirapplication in immunoassays

Flow cytometry: Quantification of Ig bearingcells in blood, spleen, kidney and thymus

Mononuclear cells (MNCs) from blood,spleen, kidney and thymus of C. striatus wereindividually analyzed for FSC (Forward scatter)and SSC (side scatter) patterns representing sizeand granularity of cells, respectively.Lymphocytes were presumed to be representedby medium sized cells with less granularity, inall analyzed lymphoid tissues. Themononuclear cell population in all analyzedtissues had relatively homogenous FSC (200–400) properties [Fig. 58(A), (C), (E) & (G)]. The

Name of site/farms

Water area (Ha)

Total number of fish stocked

Total (apprx.)

production

Approx rate/kg

(Rs)

Total income (approx.)

Total fish lost due to EUS

(approx.)

Total economic loss (approx.) (Rs)

Dhandel 17 10 lakhs fry 10,000 kg 60 6 lakhs 3000 kg 1.8 lakhs Mitauli 5.5 30,000 fingerlings 2,000 kg same 1.2 lakhs 1000 kg 0.6 lakh Raipur 6.2 10 lakhs fry 10,000 kg same 6 lakhs 2000 kg 1.2 lakhs

Table 14: Estimation of direct economic losses in a few EUS affected farms

Note: Information has been based on the discussions with the farmers and visual estimates

58

Fig. 58. Quantification of Ig+ mononuclear cells in blood and lymphoid organs of Channa striatus by flow cytometry[FSC/SSC dot plot of mononuclear cells of blood, spleen, kidney and thymus, respectively, showing gated lymphocytes(A, C, E, G), Fluorescence histogram of gated mononuclear cells in blood (B), spleen (D), kidney (F) and thymus (H)without MAb (black line) and with C9 MAb (grey line)]

distribution of putative B-lymphocytes in bloodand lymphoid tissues of adult C. striatus wasmeasured by incubating the cells with C9 MAband subsequently, staining with anti-mouse IgGFITC conjugate. Representative flow cytometryhistograms of negative control and C9 MAbstained lymphocytes in blood, spleen, kidneyand thymus are shown in Fig. 1(B), (D), (F) and(H), respectively. The percentage of Ig-positivecells in blood, spleen, kidney and thymus wasfound to be 18.2%, 27.7%, 10.3% and 0.5% ofgated lymphocytes, respectively.

Indirect Immunoperoxidase test:Demonstration of Ig+ cells in blood andlymphoid organs

In the spleen sections, Ig+ cells wereobserved in white and red pulp of spleen. Thesecells were scattered predominantly as singlecells and sometimes as cluster of a few cells.The concentration of Ig+ cells was more inmantle layer of white pulp. In sections of headkidney, the Ig+ cells were scattered mainly in

lymphoid tissue and occasionally in thehematopoietic tissue of head kidney. In trunkkidney, the Ig+ cells were detected mainly inperitubular area near the basement membrane.The Ig+ cells occurred singly or in small clustersof 2-3 cells. The reaction was occasionallyobserved in glomeruli and blood vessels. Overall,less number of Ig+ cells was observed in kidneysections, then in spleen sections. No reactivityof C9 MAb was demonstrable in thymussections. In blood smears, MAb reacted withround cells that had large nucleus. No reactivityof MAb was observed with erythrocytes andgranulocytes. In control slides, Ig+ cells werenot observed in spleen, kidney and thymussections and blood smears.

Detection of anti-Edwardsiella tardaantibodies

C9 MAb was used as detector antibodiesin the indirect ELISA for measuring thehumoral antibody response in C. striatus. Anelevated and measurable immune response to

59

E. tarda was observed in the sera of immunizedfish as compared to 0-day serum. An almostlinear decline in specific antibody level wasobserved with an increase in serum dilution.

Development of monoclonal antibody-based marker for monitoring humoralimmune response in Catla catla

Catla catla is a commercially importantspecies for freshwater aquaculture. In thepresent study, C. catla is being targeted forraising monoclonal antibodies to its serumimmunoglobulins and evaluating theirapplicability for use as marker of humoralimmunity. C. catla samples were immunizedwith 100 µg of bovine serum albumin (BSA)emulsified with Freund’s complete adjuvant byintra-peritoneal route and boosted repeatedlywith BSA emulsified with Freund’s incompleteadjuvant at 2 weeks intervals. Blood wascollected from hyper-immunized fish fromcaudal vein and serum was separated. Indirecthaemagglutination (IHA) test was used toassess the humoral immune response inimmunized fishes. The post-immunization IHAtitre in the test group ranged from 1:16 to 1:128,whereas, the titre was <1:2 in pre-immunization serum and control group. Anti-BSA antibodies were purified from serum byBSA-CL Agarose column. A single peak wasobserved on elution with glycine buffer. Theeluted fractions were concentrated withCentriplus YM - 100 filter. The concentrated Igfraction showed an IHA titre of 1:128. Purityof purified and concentrated immunoglobulinswas checked by reducing and non-reducingSDS-PAGE. In reducing SDS-PAGE, two bandscorresponding to heavy (HC) and light chain(LC) were observed. In non-reducing gradientSDS-PAGE, one high molecular weight bandof 842 kDa was observed which was assumedto be tetrameric form of immunoglobulin. Thepurified immunoglobulin was used forimmunization of BALB/C mice. Two micewere immunized with 100 µg of purified Catla

Ig emulsified with Freund’s complete adjuvantby intra-peritoneal route and were boosted 2times with Catla Ig emulsified with Freund’sincomplete adjuvant at 14 days intervals.

Development of in vitro cellular modelfor assessment of innate immunityparameters of Labeo rohitaIsolation and culture of macrophages

Healthy L. rohita (250–300 g) wereobtained from NBFGR and local fish farm andwere maintained in FRP tanks foracclimatization. Peritoneal macrophages wereharvested from the peritoneal cavity of theeuthanized fish by injecting and withdrawingsterile PBS (Phosphate Sulfer Saline). Thisprocedure was repeated several times and theharvested cell suspensions of different fisheswere pooled in centrifuge tubes on ice.Mononuclear cells (containing the macrophagesand lymphocytes) were separated from othercells by density gradient centrifugation. Theisolated mononuclear cells were washed withPBS by centrifugation and suspended in tissueculture medium at a concentration of 105cell/ml. The isolated cells were seeded in the tissueculture flasks for attachment of macrophages.The non-adherent cells were removed bythoroughly washing the flasks with PBS after24 hours. The remaining cells were allowed togrow at 300C till confluency. Confluentmonolayer was sub-cultured by trypsinizationto obtain passage 1. At 5, 10 and 15 sub-cultures(Fig. 59), the macrophages were cryopreservedat –1960C. Growth conditions for culture ofperitoneal macrophages were also studied. Atotal of 16 attempts were made for isolation andculture of peritoneal macrophage, of whichonly one attempt could result in confluency ofthe cells. Six attempts were also made to isolateand culture macrophages from head kidney ofL. rohita, of which only one attempt could resultin confluency of the cells. These cells are beingsub-cultured at regular intervals.

60

Functional characterization of culturedmacrophage

The phagocytic activity of the culturedperitoneal/kidney macrophages was testedwith Saccharomyces cerevisiae. Macrophages(5×105 cells) were cultured for 24 h in tissueculture medium. Next day, the cultures wereinoculated with heat killed yeast suspensionand incubated for 90 minutes in a humidchamber at 300C. After the incubation period,the cells were washed twice in PBS, followedby fixing with 100% methanol and stainingwith Giemsa staining solution. The resultsindicate that the cultured peritonealmacrophages possessed phagocytic activity(Fig. 60).

Isolation and characterization ofFlavobacterium species from fish andaquatic environment

Isolation of Flavobacterium columnareA total of 32 fish samples showing disease

symptoms were collected from different fishfarms and processed for bacterial isolation. Allthe tested fish had gross lesions resemblingcolumnaris disease. Skin swabs were streakedaseptically or dilutions of homogenized tissuewere plated on to cytophaga agar platescontaining 5 ìg/ml neomycin sulfate and 200units/ml of polymixin B. The plates wereincubated at 250C for 3-4 days to obtain visiblebacterial growth. Yellow to green colonies withentire or rhizoid edges were selected forpurification. The purified isolates were initiallysubjected to Gram’s staining, catalase, oxidase,motility and O-F tests, and later to Griffinmethod of screening for identification of F.columnare.

F. columnare was isolated from one of thediseased C. catla sample and presumptiveidentification was carried out on the basis offive characteristics, as described by Griffin,(1992). The colonies on Cytophaga agar werepale yellow, spreading with rhizoid margins.The biochemical results indicate that the isolatewas flexirubin positive, Congo red positive,

Fig. 60. Demonstration of phagocytic activity of peritoneal macrophages using yeast cells(Black arrow heads showing engulfed yeast cells; white arrow heads are the nucleus of the macrophages)

Fig. 59. Culture of peritoneal macrophages of L. rohita at15 passages

61

resistant to neomycin resistant, gelatinasepositive and chondroitinase positive.

DNA isolation from F. columnare was doneby the method of Marmur et al. (1961).Amplication of 16s rRNA was done byuniversal primers to obtain approx 1500bpamplicon. The PCR product was eluted fromthe gel and cloned into cloning vector pTZ 57RTby TA cloning method. The cloned plasmid wassequenced using M13 primers to obtain approx1500bp nucleotide sequence. The sequencechromatogram was analysed by Bioeditsoftware and the resulting consensus sequences(~1500 bp) were compared with those availablein RDP 10 database by use of the SEQ MATCHprogramme to determine 16S rRNA genesequence similarities with its nearest neighbors.The results revealed identity of the isolate wasapprox 98% with F. columnare strain EK 28(AB016515) and LP8 (Genbank Accession No.AB015480).

Isolation of protease producing bacteriafrom aquatic environment

A total of 56 bacterial isolates collectedfrom different fish farms were screened forprotease production on skimmed milk agarplates. Out of these, nine isolates showedprotease activity at 40C, 200C and 370C. Theprotease activity of these isolates was quantifiedand expressed in units. These isolates weresubjected to molecular identification by 16srDNA sequencing. The results showed that theisolates belonged to genus Bacillus, Aeromonasand Pseudomonas. Out of the three, Bacillusgroup showed maximum activity (14 units),followed by Aeromonas (8 units) at 370C.

Development of cell linesCell cultures were initiated from several

tissues of the ornamental barb Puntius denisoniiincluding heart, swim bladder and caudal fintissues. The cells migrated from the differenttissue fragments and grew well and formed a

monolayer during the first month. Out of threeselected tissues/organs, one cell line could beestablished from caudal fin (Table 15).Emergence of growing cells from swim bladderand heart occurred 25 and 22 days after theexplantation of minced tissue fragments,respectively. The swim bladder and heart cellsconsisted of epithelial-like and long fibroblastic-like cells, respectively (Fig.61E and F.).However, these swim bladder and heart cellsshowed poor survival and died following sub-cultivation. The heart and swim bladder cellscould not be maintained after 12th and 11th

passage, respectively. However, the cells fromthe caudal fin grew continuously. In the initialpassages, fin cells were composed of aheterogeneous mixture of fibroblastic-like andepithelial-like cells (Fig. 61D). After 15 sub-cultures, fin cells were predominantlyfibroblastic-like cells (Fig. 61G, H and I). Thedifferent morphological characteristics of thecell lines developed from the different tissuesof the P. denisonii are given in Table 16. Inprimary culture, fin cells adhered well andachieved confluence in 6 days at 28°C. Cellswere sub-cultured in L-15 medium with 20%FBS (Fetal Bovine Serum) every 4-5 days for theinitial 10 passages. For the first 15 passages,50% culture medium was replaced with freshmedium at 4 day intervals. After 15 th

subcultures, cells were sub-cultured at a ratioof 1:3 at 3-4 days interval, and FBS was reducedto 15% in the L-15 culture medium. The fin cellshas been sub-cultured more than 52 times sinceinitiation and are designated as red-line torpedofin (RTF) cell line.

Optimal growth temperature for RTF cellsranged 26–30°C. However, maximum growthwas obtained at 28°C (Fig. 62A). RTF cells werealso able to spread and grow well at 20°C,although it took 48 h for the cells to becomewell attached (Fig. 62D). When incubated at30°C, the cells proliferated fast during the first48 h, but the growth and proliferation sloweddramatically, and the cells appeared to agerapidly (Fig. 62E). Although cells remained

62

viable during the test period when incubatedat 37°C, cell growth was minimal andindividual cells looked abnormal (Fig. 62F). Thegrowth rate of RTF cells increased as the FBSproportion increased from 2 to 20% at 28°C.Cells exhibited poor growth at 5%concentrations of FBS, relatively good growthat 10% but maximum growth occurred withthe concentrations of 15 and 20% FBS.

The details of the formation of themonolayer of RTF cells at varyingconcentrations of FBS, NBCS and equalmixture of FBS and NBCS (Newborn Calfserum) are given in Table 17. RTF cells in earlypassage changed shape depending on the kindof sera with which they were treated. Thenumber of cells increased at a steady rate ineach medium during the culture period. Thecells in FBS-supplemented medium weretypically fibroblastic-like, and sharply outlined(Fig. 62A). The morphology of the cells in NBCSand FBS supplemented medium were same asin the earlier case with slight change in theirhealth status (Fig. 62B). In the presence of NBCSalone, the cells were epithelial-like and lessclearly outlined, extending dendritically (Fig.62C). Plating efficiency for RTF cell line wasdetermined at seeding concentrations of 200,500 and 1000 cells. Moderately low platingefficiencies were observed with RTF cells (10.5(±0.5), 13.07 (±2.53) and 14.37 (±5.66) %),respectively (Fig. 63).

The RTF cells were cryopreserved at 10th,20th and 30th passages. The cells were recoveredafter six months from storage and grew toconfluency within 5 days. The average viabilityof the cells after cryopreservation was estimatedto be 80% with the same morphology. Thediploid karyotype of RTF cells is shown in Fig.64A and consists of 25 pairs of telocentric(2n=50) chromosomes. The results ofchromosome counts of 100 metaphase platesfrom RTF cells at passage 25 revealed that inthe 32% of the cells, the chromosome numbervaried from 38 to 54 (Fig. 64B) and about 68%

of the cells had a diploid chromosome numberof 2n=50.

The RTF cells were resistant to the marineviral nervous necrosis (VNN) virus.  Nocytopathic effect (CPE) was observed in the cellsup to 2 weeks of observation and even after 10blind passages. The ECPs from Vibrio choleraeMTCC 3904 and Aeromonas hydrophila (data notshown as the strain was not procured from anyreferral culture collection centers) proved to becytotoxic for all RTF cell lines. Cytotoxic effectscould be observed within 10 h after inoculation.The morphological changes the detected inthese cell lines were rounding, detaching andfinally monolayer destruction (Fig. 62G, H andI). To verify the origin of RTF cell line, DNAwas isolated from the RTF cells at their 25th

passage. Amplification and sequencing of the16S rRNA and COI genes from the RTF cell linesand P. denisonii muscle tissue revealed 562bpand 642bp edited sequences (Fig. 65) in allsamples. Subsequent comparative analysis ofthe identified sequences revealed a 100% matchfor RTF and P. denisonii muscle tissue, as well

P. denisonii Details (Number of explants/cell line) Heart Fin Swim

bladder Number of explant 20 40 15 Contamination of the explant 3 28 8 No radiation of the explant 10 4 3 Radiation of the explant 7 8 4 Formation of monolayer 2 5 2 Contamination of the monolayer

0 2 0

Number of cultures capable of subculturing

2* 3** 1***

Number of cultures still being cultured

0 1# 0

Table 15. Details of cell line developmentfrom the different tissues of P. denisonii

*One of the two cell lines could not be sub-cultured after12th passage and the other after 10th passage.**Two cell lines could not be sub-cultured after 10thpassage.***Cell line could not be sub-cultured after 11th passage.#Current passage level is 52.

63

Fig.61.Photomicrographs of Puntius denisonii cells derived from different tissues[A: Fin explant showing radiation of cells (100X); B: Heart explant showing radiation of cells (200X); C: Swim bladderexplants showing radiation of cells (200X); D: Heterogeneous populations of both epithelial like and fibroblastic likecells of fin cells (100X); E: Monolayer of heart cells at 8th passage (200X); F: Monolayer of swim bladder cells at 10th passage(200X); G: Monolayer of RTF cells at 5th passage (100X); H: Monolayer of RTF cells at 20th passage (100X), and I: Monolayerof revived RTF cells 6 months after cryopreservation (100X)]

as, a 99% match to known P. denisoniimitochondrial DNA sequences in the GenBankat NCBI. These sequences have been submittedto GenBank (COI accession number-GU566029).

Table 16. Morphological characteristics of celllines established from P. denisonii

Tissue source Initial

growth Morphology

Caudal fin ++++ Fibroblastic -like Swim bladder +++ Epithelial - like Heart ++ Long fibroblastic – like

S. No.

Animal sera

Concentrat-ion

(10%)

Number of cells seeded

Mono layer

forma-tion

(days)

Health Status of the cells

Number of cells after 10

days (X 105 cells/ ml)

1 FBS 10

1 X 105/ml

8

+++++ 6.4

2 NBCS 10

1 X 105/ml

12

++

3.2

3 Equal mixture of FBS and NBCS

10

1 X 105/ml

10

+++

4.6

++++ Rapid growth and formed over 75% cell monolayer.+++ Extensive outgrowth and formed large-size cellcolonies.++ Outgrowth with small-size colonies surrounding tissueexplants.

Table 17. Effect of Fetal Bovine Serum andNewborn calf serum on growth of RTF cellsat 30th passage

64

Fig.62. RTF cellular morphology at different animal serum, growing temperatures and cytotoxic effects of ECP of Vibriocholerae MTCC 3904[A: RTF cells at 28 °C and 10% FBS (100X); B: RTF cells at 28 °C and 5% NBCS and 5% FBS (100X); C: RTF cells at 28 °C and10% NBCS (100X); D: RTF cells at 20 °C (100X); E: RTF cells at 30 °C (100X); F: RTF cells at 37 °C (100X); G: RTF cells at 1 daypost inoculation of Vibrio cholerae MTCC 3904 ECP; H: RTF cells at 2 day post inoculation, and I: RTF cells at 3 day postinoculation]

A)

B)

Fig.63. Growth response of the RTF cell line to (a) selected temperature and (b) selected foetal bovine serum (FBS)concentrations

65

Fig.65: A) PCR amplification of ~ 600-bp and ~ 700-bp sequences of the P. denisonii genome using universal oligonucleotideprimers of the 16S rRNA and CO1 genes; respectively (Lane 1 & 2: RTF cells; lane 3&4: P. denisonii tissue); with CO1primer (lane 5 & 6: RTF cells; lane 7& 8: P. denisonii tissue); M: marker (100 bp DNA ladder) B) Nucleotide sequencesof the 562 bp and 642 bp fragments amplified using oligonucleotide primers of the 16S rRNAand CO1 genes of P.denisonii

Fig.64. Chromosome analysis of RTF cells:(A) Karyotype of RTF cells (passage 25), and (B) Metaphase chromosome numbers of RTF cells at passage 25

Maintenance of other cell linesCell lines developed earlier from other

species viz., Epinephelus merra, Pristolepisfasciata, Puntius fasciatus and Cyprinus carpio koiwere also maintained during the year.Currently the passage levels of the cell lines ofabove species were 35, 84, 21 and 60,respectively.

New species attempted for cell cultureCell cultures were initiated from 40

explants of Puntius chalakkudiensis including 12heart and 28 caudal fin tissues and 74 explantsof caudal fin of Tetraodon travancoricus (Table18). No cell lines could be established from thesespecies and further attempts are being made todevelop cell lines during 2011-2012.

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Monitoring of the imported exotics fishconsignments

The inspection of the imported YY (GMT)Oreochromis niloticus fry for their health statuswas carried out by the NBFGR scientists in thequarantine facility at Rajiv Gandhi Centre forAquaculture, MPEDA, Vijayawada centre. Sixrandom samples collected from the quarantinefacility were screened for Koi Herpes virusdisease, an OIE listed disease and foundnegative. The species status of the imported frywas confirmed through DNA barcoding (655bp COI) and partial sequence information of16S rRNA (552 bp).

Exploration of protozoan and monogeneanparasites among carps and catfishes

Periodic sampling of selected carp andcatfish species was done from the followingplaces in U.P.: Raju Hatchery, Haidergarh;Anas Hatchery, Faizabad; R.K. Fish Farm,Prakash Hatchery, Deva Farm, Barabanki;Pankaj Matsya Beej Utpadan, Lucknow; HarunFish Farm, Barabanki; Ramkumar Fish Farm,Unnao; Rahat Fish Farm, Salon, Raebareli;Aasiwan, Unnao; Butler Palace pond,Mohanlalganj, Barabirwa, Dubagga, Dallyganjand Mawaiya markets of Lucknow.

Screening of fishes and isolation of

Tetraodon travancoricus

Puntius chalakkudiensis

Parameters

Fin Heart Fin Number of explant 74 12 28 Contamination of the explant

65 7 18

Radiation of the explant

7 2 4

No radiation of the explant

2 3 6

Formation of monolayer

0 1 2

Contamination of the monolayer

0 0 1

Continuation of subculture

0 0 1

Passage level 0 0 5

parasites was done under stereo zoommicroscope with the help of needles, fine brushand micro-pippettes. The live parasites wereobserved under microscope and fixed inMalmberg’s soln., 5% formaline, methanol andglycerol, and stained with various dyes(temporary, permanent slides). Photographs oflive and fixed parasites were taken andmeasurements of parasites images and analysiswas done through a software NIS-E-BR, Nikon.Identification of the parasites was done usingon the basis of their morphological andmorphometric features with the help oftaxonomic keys by Satyu Yamaguti and Z.Kabata (1985).

Prevalence of parasitesA total of 343 fishes were screened; out of

which, 125 were found infested/infected withmonogenean and protozoan parasites. Theprevalence of parasites was 36.44% (Table 19).The intensity of monogeneans was highest(about 200) in Carassius auratus (Fig. 66):

The following parasites were identifiedfrom the selected fishes (Fig. 67):

Protozoans: Trichodina, Ichthophthiriusmultifilis, Tripartiella, Pliestophora,Tetrahymena, Chilodonella, Piscinoodinium,Ichthyobodo necator, Hexamita,Trypanosoma, Apiosoma, Thelohanellus,Epistylis, Hennegua, Myxidium, Myxobolusclarii and other species of Myxobolus. Monogeneans: Gyrodactylus elegans,

Table 18 Details of cell cultures from Tetradontravancoricus and Puntius chalakkudiensis

S. No. Name of Fish Prevalence (%) 1 Labeo rohita 30.86 2 Cirrhinus mrigala 17.64 3 Catla catla 27.27 4 Cyprinus carpio 23.07 5 Ctenopharyngodon idella 18.91 6 Hypophthalmichthys molitrix 15.38 7 Carassius auratus 46.29 8 Puntius ticto 42.42 9 Clarius batrachus 38.46 10 C. gariepinus 16.66 11 Heteropneustes fossilis 8.33 12 Mystus sp. 21.05

Table 19. Species-wise prevalence ofparasites

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Fig. 67. A few of the parasites isolated from the selected fishes

Dactylogyrus intermedius, D. extensus, D.anchoratus, Paradactylogyrus vastator, andGyrodactylus sp.; Digeneans: Diplostomum spathaceum,Allocreadium, Orientocraedium. Nematodes: Procamallanus, Camallanusand Capillari. Cestode: Lytocestus sp. and other species(to be identified). Acanthocephalan: Acanthosentis sp. Copepods: Argulus, Ergasilus andLernaea and some worms, cysts, eggs andlarvae of parasites.Fig. 66. High intensity monogeneans in a small piece of

gill scrape of Carassius auratus

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Effect of aromataze inhibitors on gonadal protein and lipid

0

2

4

6

8

10

12

C T-1 T-2 L-1 L-2

Gon

adal

pro

tein

(mg/

g)

0

0.5

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2

2.5

Gon

adal

lipi

d (u

g/g)

Protein mg/g Lipid µg/g

Production of monosex/sterile commoncarp, Cyprinus carpio, using aromataseinhibitors

Since regulation of the aromatase enzymein fish profoundly influences the sexualdevelopment, the role of Aromatase inhibitors(AIs) on gonadal development and sexdifferentiation in C. carpio was studied.Fingerlings of Cyprinus carpio were exposed todifferent doses (100 mg/kg and 200 mg/kgfeed) of tamoxifen (TA) and letrozole (LET) for60 days, maintaining them in glass aquaria of125 l capacity. The survival, specific growth rate(SGR%), length-weight and gonado-somaticindex (GSI) were observed after the treatmentof TA and LET and the changes were comparedwith that of control and also between treatmentgroups so as to evaluate the effect of differentaromataze inhibitors on the growth, survivaland gonadal development.

Different biochemical changes such ashaemoglobin (g%), total erythrocytes count(REC), total leucocytes counts (TLC), serumprotein, serum cholesterol, serum triglyceride,gonadal protein and gonadal lipid wereestimated after the treatment of aromataseinhibitors. The changes in the haemoglobin,REC and haematocrit (%) were lower to control,while, TLC was higher, however, thedifferences were not significant (Fig. 68). Thegonadal as well as serum protein and lipid wereexamined in normal, as well as, tamoxifen (T1& T2) and letrozole (L1 & L2) treated fishes.There was a decline in gonadal protein and lipidlevels as compared to the control group (Fig.69). The plasma cholesterol levels were alsofound to be depressed after AIs treatments.However, this effect was more pronounced inthe high dose of TA and LET (T2 & L2). Theserum triglycerides were slightly elevated in thelow dose of TA (T1) and LET (L1), however, itdeclined in high dose of TA (T2) and LET (L2)(Fig. 70).

Fig. 68 Haematological changes in C. carpio afteraromatase inhibitors treatment

Fig. 69: Effect of aromataze inhibitors on gonadal proteinand lipid in C. carpio

Fig. 70. Plasma cholesterol and triglycerides of C. carpioafter TA and LET treatments

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Histological changes associated with thegonadal development i.e., the testicular andovarian structure, were observed. The testiculardevelopment in about 38% fishes was foundmore advanced (spermatocytes) in high doseof TA or LET than those in the control(spermatogonium). There was reduced ovariangrowth and increased atresia suggesting thatbiosynthesis of estradiol is essential formaintenance and continued development ofoocytes during ovarian development (Fig.71).In case of LET treatment, however, thechanges were more pronounced than incase of TA treatment. In case of low dose ofLET, spermatogenic germ cells wereextensively proliferating and previtellogenicoocytes were found to undergo atresia in the

intersexual gonads indicating that the sexchange was processing with the treatment ofAIs.

The activity of the aromataze inhibitors(TA and LET) on the sexual development wasdetermined by assessing the estrogen levels, bothin gonadal tissue, as well as in plasma of fishes.Gonadal as well as plasma 17ß-oestradiol in theTA and LET treated fish was estimated byELISA method using 17ß-oestradiol rabbitpolyclonal antibody (Fig.72). The change in 17ß-oestradiol level was observed during and afterthe treatment of aromatize inhibitors for 60days. The changes in the gonadal 17ß-oestradiol levels were correlated with that ofhistological changes in different treatmentgroups.

Fig.5. Microphotographs of C. carpio showing various developmental stages. A- Undifferentiated gonad, B. Appearance ofthe testicular structures. C. Ovarian cells and D. Initiation of vitellogenesis

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17 beta estradiol level after treatment of aromatase inhibitors

0

50

100

150

200

250

C TA-1 TA-2 LA-1 LA-2

pg/m

l

Ecological impact assessment of Clariasgariepinus

The introduction and transfer of alien fishspecies among continents, regions and nationshas often had significant impacts on therecipient aquatic ecosystem. There has beenmuch concern resulting from intensive or semi-intensive aquaculture practices of Africancatfish, Clarias gariepinus in India which hadled to environmental problems in many aquaticecosystems. In view of this, a study wasundertaken to study the impacts of this alienfish species in Uttar Pradesh.Data on the availability of C.gariepinus was generatedthrough a benchmark surveyof fish markets and also fromgrow-out farms in Gorakhpur,Varanasi, Allahabad, Kanpur,Mau and Lucknow divisions ofUttar Pradesh (Fig.73). Apartfrom its culture in differentareas, data on its catches fromrivers such as Yamuna, Gomti,Varuna and Ganga and alsofrom lakes and canals was alsogenerated. Occasionally this

fish was caught from rivers Yamuna, Gangaand Gomti. A total of 37 surveys wereconducted in different places to record theavailability of C. gariepinus. The contribution ofthis fish in different fish markets was observedwhich ranged from 10% -40% in differentmarkets of U.P.

A total of 45 fish ponds culturing C.gariepinus were surveyed in different districtsof U.P. The production data were recorded andthe economics of culture was worked out. Therewas large variability in the productivity of C.

Fig. 72. Plasma and gonadal 17-â estradiol level after AIs treatments to C. carpio

Fig. 73. Total landing of C. gariepinus in fish markets of different cities

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gariepinus which ranged from fewkg to 30 t/ha or more.

The size of C. gariepinusmonoculture farms ranged from0.02 ha to 3 ha. Data on the physico-chemical parameters such astemperature, pH, dissolved oxygen,free carbon dioxide, turbidity andtotal alkalinity were recorded andfound existence of very poorhusbandry and hygiene conditions.The fish was found to tolerate hardphysico-chemical conditions andhigh ammonia levels. Food andfeeding of the fish in the culture farms werestudied and the generated data for differentfeed types was analysed with respect to thegrowth and overall production of the fish(Fig. 74).

Biological information such as level ofcannibalism, survival, growth, maturity andcompatibility with other local fish species were

Fig. 74. Production of African catfish with different types of diets

recorded from the grow-out farms and alsofrom laboratory experimental conditions,keeping the fish in aquaria and plastic pools.Data on the length and weight of C. gariepinuswere collected from various grow out farms todetermine the condition/factors and fitness ofthe fish in different physico-chemical conditions.

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‘Ganga Aquarium’ Inaugurated byDr. S. Ayyappan

Named after the River Ganga, a beautifulaquarium has been established in picturesquelush green surroundings of NBFGR campus ina finely architectured circular buildingspreading over an area of 450 m2. It is one ofthe largest and most beautiful aquarium in thecountry and displaying recreational andeducational value of fishes with respect toconservation and commercialization. It wasinaugurated by Dr. S. Ayyappan, Secretary,Department of Agricultural Research andEducation, Ministry of Agriculture, Govt. ofIndia and Director General, Indian Council ofAgricultural Research (ICAR) on 19 November,

2010. Dr. B. Meenakumari, Deputy DirectorGeneral (Fisheries), ICAR, New Delhi; Dr. H.P. Singh, Deputy Director General(Horticulture), ICAR, New Delhi; Dr. S.A.H.Abidi, Former Member, Agriculture Scientists’Recruitment Board, New Delhi; Dr. W. S. Lakra,Director, Central Institute of FisheriesEducation, Mumbai and several otherdignitaries also graced the programme.

In his address Dr. Ayyappan opined thatthe new aquarium is a unique and one of thebest aquariums in the country. The DirectorGeneral appreciated the efforts of NBFGR andopined that this new venture will help inincreasing public awareness regarding fishdiversity and educating the children and

IMPORTANT EVENTS AND MEETINGS

Glimpses of the inauguration of Ganga aquarium by Dr. S. Ayyappan, Secretary, DARE and DG, ICAR

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general public. Dr. J.K. Jena, Director, NBFGRinformed that the aquarium has been housedwith well decorated forty-two large aquariacontaining over 100 fish species belonging tothe continents of Africa, Asia and North &South America. He further stated that effortshave been made to display fishes of ornamental,sport, food and medicinal importance, so thatit can provide useful information about fishesfor general awareness of the public. One of themain attractions of Ganga Aquarium is anornately carved central fountain with a poolthat harbours brightly coloured fishes and acentral plinth graced by four marble carved fishmermaids.

The aquarium displays a wide variety offishes ranging from deadly piranhas to mostbeautiful feng shui (flower horn) and largepacu/catla/mahseer to tiny tetras. Amongstimportant exotic ornamentals, goldfishes(bubble-eye, celestial, comet, blackmoor,shubunkin, red oranda), koi carp, angels,alligator gar, sting ray, tetras, ghostfish, firemouth, malawi cichlids, malayan angels,gorami’s including albino variant, silver shark,tiger barbs, oscars, discus, albino severum, scats,tiger sharks, live bearers, senegal, silver dollar,red-tail shark, red-tail catfish,sucker mouth catfish, etc glitterthe well decorated aquaria. TheIndian ornamentals arerepresented by denisoni barb(Kerala Queen), flamingo barb,loaches, eels, etc. The major foodfish fauna of indigenous varietiesis represented by Indian majorcarps, catfishes and snakeheads,whereas, that of exotics by Chinesecarps and catfishes. Mahseers andfeatherbacks have been displayedas important sport fishes of thecountry. Marine life has beendisplayed in four aquaria thathouse quoran fish, butterfly fish,damsels and lionfish, the reef

fauna comprising of yellow tail, chaetodonts,wrasse, clownfishes, sea anemones and starfish.

NBFGR organised X th AgriculturalScience Congress at Lucknow

The Xth Agricultural Science Congresswas held at National Bureau of Fish GeneticResources, Lucknow during 10-12 February,2011 under the aegis of National Academy ofAgricultural Sciences (NASS), New Delhi,which was organized by NBFGR, Lucknow, incollaboration with IISR, Lucknow and CISH,Lucknow. The theme of the Congress was Soil,Plant and Animal Health for Enhanced andSustained Agricultural Productivity. TheCongress was inaugurated by His ExcellencyShri B.L. Joshi, Governor of Uttar Pradesh andShri Harish Rawat, Hon’ble Minister of Statefor Agriculture, Government of India presidedover the inaugural function. Other dignitariespresent on the dais were: Padma Vibushan Dr.R.S. Paroda and Dr. Mangala Rai, FormerDirector Generals of ICAR, New Delhi; Dr. R.B.Singh, President, NASS, New Delhi; Dr. AnwarAlam, Secretary, NASS, New Delhi; Dr. W.S.Lakra, Director, Central Institute of Fisheries,Mumbai and Dr. J.K. Jena, Director, NBFGR,

Shri B.L. Joshi, His Excellency, Governor of UP; Shri Harish Rawat, Hon'bleMinister of State for Agriculture; Dr. R.S. Paroda, Former DG, ICAR andDr. R.B. Singh, President, NAAS during inauguration ceremony of the Congress

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Lucknow. A galaxy of over 700 delegates fromdifferent parts of the country, including topagricultural experts of the country, Directorsof national and international organizations,policy makers, administrators, scientists,industry personnel, teachers and students fromIndian universities, progressive farmers andentrepreneurs participated in the Congress.

In his presidential address Shri HarishRawat complimented agricultural scientists,policy makers and farmers of the country forself-sufficiency in food grain production andadoption of new technologies. He urged thatin order to achieve evergreen revolution andcalled for ensuring judicious conservation ofsoil, water and biodiversity resources,upgradation of genetic resources and optimalutilization of natural resources through

improvement in management practices. Heurged the scientific community to fill existinggap in transfer of technology and create newmodels of management for diversifiedentrepreneurship in agriculture and alliedenterprises.

Inaugurating the Congress His ExcellencyShri B.L. Joshi, Governor of Uttar Pradeshopined that the Congress is a very importantevent as it has brought together the best brainsof the country in the field of agriculturalsciences. He expressed that Congress shouldcome out with strategies and solutions to theemerging new challenges to the agriculturesector in the country. He urged the scientificcommunity to prepare a comprehensivenational bio-security plan to maintainrobustness of the system. He also emphasized

Shri Harish Rawat, Hon'ble Minister of State forAgriculture, Govt. of India addressing the Congress

The dignitaries releasing the Souvenir of the Congress

Shri B.L. Joshi, His Excellency, Governor of U.P.,delivering the Inaugural Address

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Shri B.L. Joshi, His Excellency, Governor of UP and Shri Harish Rawat, Hon'ble Minister of State for Agriculture;inaugurating the 'Agro Vision 2011'(Left) and a view of the exhibition (Right)

A view of the audience A view of the Congress venue

that informal education of farmers and ruralyouths is essential to promote agri-basedenterprises and enhance agriculturalproductivity.

The Souvenir of X Agricultural ScienceCongress on “Soil, Plant and Animal Health forEnhanced and Sustained AgriculturalProductivity” was released by His ExcellencyShri B.L. Joshi, Governor of Uttar Pradesh. Thebook of Abstracts of X Agricultural ScienceCongress on “Soil, Plant and Animal Health forEnhanced and Sustained AgriculturalProductivity was released by Shri HarishRawat, Hon’ble Minister of State forAgriculture, Government of India. NAASawards were also presented on this occasionby His Excellency Shri B.L. Joshi.

The Congress included a plenary session,seven technical sessions and two evening

lectures by eminent personalities from differentfields of agricultural sciences. In the plenarysession chaired by Dr. R.B. Singh, President,NASS, on ‘Eco-technologies for LivelihoodSecurity’, Dr. Mangala Rai delivered keynoteaddress on “Efficiency mediated sustainableenhancement measures”. Other keynotespeakers during the plenary were: Dr. ThomasA. Lumpkin, Director General, CIMMYT,Mexico on ‘Global food security: Challengesand opportunities for agro-eco technologies andSouth Asia’; Dr. Robert Zeigler, DirectorGeneral, IRRI, Philippines on ‘Global Foodsecurity in the fate of climate change’ and Dr.B. Hanumaiah, Vice Chancellor, B. R.Ambedkar University, Lucknow on ‘Oil, plantand animal health for enhanced and sustainedagricultural productivity’. In the eveninglectures, Dr. R.S. Paroda spoke on ‘Concernsof food security, soil health and climate

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Technical sessions in progress

change’, whereas, Dr. K.L. Chadha addressedon ‘Way forward and policy actions towardshorticulture led transformation of agrarianeconomy in the eastern Indo-gangetic plains’.

Technical presentations were made underseven sessions focusing on: Status of farmhealth and assessment of losses, Biosecuritywith special reference to emerging exoticdiseases and pests, Diagnostics and healthmanagement, Risk assessment andmanagement, Climate change and its impacton farm health, Unleashing the agriculturalpotential of eastern India and an Interactivesession with farmers, bankers, industry andother stakeholders. In the interactive sessionover 60 progressive farmers from several statesincluding, U.P., M.P., Jharkhand, Orissa,Assam, Rajasthan, Gujrat and Uttarakhandparticipated and shared their innovations,experiences, expectations and problems withthe scientific fraternity. A grand exhibition wasalso organized for the benefit of the delegatesin which 45 exhibitors including, ICARInstitutes and industries displayed theirtechnologies and products.

Three Poster sessions were also organizedduring the Congress in which 167 posters werepresented, of which, 10 posters were presentedwith the Best Poster Award by Dr. R.B. Singh,President, NAAS during Valedictory function.Besides, an Elocution Competition was also heldon 11 February, 2011 on the theme ‘Soil – Plant– Animal Health: Safety and Security’ in whichthe winner students from different zones of the

country participated in the final contest. DuringValedictory function Dr. R.B. Singh, President,NAAS complemented the organizers, speciallyNBFGR, for successfully hosting the Congressand making excellent arrangements for such ahuge academic gathering. He profuselythanked Dr. W.S. Lakra, Convener and Dr. J.K.Jena, Co-convener of the Congress forsuccessful organization of the Congress. Dr. J.K.Jena, Director, NBFGR and Co-convener of theCongress expressed his gratitude to thePresident and Executive Council, NAAS forhaving faith in NBFGR’s capability to host theCongress. Dr. Jena complimented theorganizing committee and whole NBFGR stafffor their immense untiring efforts, despite allodds, to make the Congress a mega success.

NBFGR Celebrates 28th Foundation DayNBFGR celebrated its 28th Foundation Day

on 12 December, 2010. A Farmers -Entrepreneurs -Scientists Interaction Meet wasorganized on this occasion to promote adoptionof new technologies in freshwater aquaculture.On this occasion, Dr. S. N. Dwivedi, FormerAdditional Secretary, Govt. of India was theChief Guest whereas, Dr. Dilip Kumar, FormerDirector, CIFE, Mumbai and Dr. B.N. Singh,Former ADG (Fisheries), ICAR, New Delhi werethe Guests of Honour. Dr. P. Das, FormerDirector, NBFGR presided over the function. Inhis welcome address, Dr. J.K. Jena, Director,NBFGR apprised about the salientachievements of the Institute. The Chief Guest,

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while expressing satisfaction over the researchand infrastructure development at NBFGR,opined that the Bureau has emerged as a Centerof Excellence in research related to conservationand management of fish genetic resources.

Dr. Dilip Kumar lauded the initiativestaken by NBFGR in registering the fishgermplasm of the country and development ofthe repositories of genomic resources. He alsostressed upon the need to build researchcapacities in the newer areas of fish genomicsresearch and biosecurity. Dr. P. Dasemphasized upon to formulate adequateconservation and management strategies fordeclining natural aquaticgenetic resources of thecountry. He further suggestedbringing wild natural geneticresources under culturefisheries to diversify andimprove the livelihood of thefarmers and economicresources of the country.Technical lectures weredelivered by NBFGR scientistsfollowed by discussion withthe participating farmers/entrepreneurs. On theoccasion, the dignitariespresented Annual Institute

Awards for the year 2009-10 to thestaff members of the Institute fortheir outstanding contributions andalso to the selected progressivefarmers/entrepreneurs of thecountry. The dignitaries also releasedNBFGR publications.

Mid-term Institute ResearchCouncil Meeting

The mid-term IRC meeting ofthe Institute was conducted underthe chairmanship of Dr. J.K. Jena,Director on 1 November, 2010. Theprogress of all ongoing projects was

reviewed by the IRC and important suggestionsemerged from the discussion. Detailedpresentations on all the projects were made bythe respective Principal Investigators/Co-PI.

Independence Day CelebratedA flag-hoisting ceremony was observed on

the Independence Day. Dr. W.S. Lakra,Director hoisted the National Flag in thepresence of other staff members of the Bureau.On this occasion, a cultural programme wasalso organized for the NBFGR staff and theirfamilies.

An entrepreneur speaking during Farmers-Entrepreneurs-ScientistsInteraction Meet on the occasion of NBFGR Foundation Day

A view of the Republic Day celebration

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Republic Day CelebratedA flag hoisting ceremony was observed on

the Republic Day. Dr. J.K Jena, Director hoistedthe National Flag in the presence of other staffmembers of the Bureau. On this occasion, acultural programme was also organized for theNBFGR staff and their families.

Research Advisory Committee (RAC)Meeting

The RAC meeting of NBFGR wasorganized on 15 March, 2011 under thechairmanship of Prof. T.J. Pandian, FormerNational Professor, ICAR, Madurai. Dr. A.D.Diwan, Former Assistant Director General(Marine Fishery), ICAR, New Delhi; Dr. Y.Basavaraju, Professor, College of Fisheries,Mangalore and Dr. Madan Mohan, AssistantDirector General (Marine Fishery), ICAR, NewDelhi participated as expert members of theRAC. In his introductory remarks, the RACChairman, Prof. Pandian appreciated NBFGR’sresearch achievements. He called upon thescientists to take up programmes in emergingfrontier areas of research. He urged thescientists to publish research papers ininternational journals having high impactfactor.

Dr. J.K. Jena, Director, NBFGR welcomedthe Chairman and members of the newlyformed RAC and apprised the Committee onthe salient achievements made during 2010-11.

Based on the recommendations of previousRAC meeting held on 18 March, 2010, an actiontaken report was presented by Dr. K.K. Lal,Head, PME Cell, NBFGR. In the continuedsession, the Head of the Divisions presentedsalient achievements made under differentidentified research themes during the year. TheRAC reviewed progress of all the ongoingresearch projects of the Institute and providedsignificant inputs to improve the researchprogrammes.

International Biodiversity DayCelebrated

The Institute celebrated the InternationalBiodiversity Day on 22 May, 2010. On thisoccasion, an educational programme to createawareness about biodiversity and issues relatedto its conservation was organized for the benefitof school children. A total of 80 students alongwith their teachers from different reputedschools of Lucknow participated in theprogramme. Dr. Mangla Rai, the FormerDirector General, ICAR and President NAAS;Dr. C.D. Mayee, Chairman, ASRB, New Delhiand Dr. A.N. Mukhopadhyaya, Former Vice-Chancellor, Assam Agriculture Universityaddressed the Participants as Guest whereas,invited technical lectures were presented byProf. R.K. Sinha, Professor, Zoology, PatnaUniversity and Prof. B.C. Choudhary, Head,Endangered Wildlife Division, Wildlife Instituteof Dehradun. The dignitaries also released

Institute publications on theoccasion.

‘Fish Karyobank’ DatabaseReleased

NBFGR has developed adatabase on fish chromosomaldiversity named as ‘FishKaryobank’ under the aegis ofNational AgriculturalBioinformatics Grid (NAIP),ICAR. Dr. P. Das, FormerDirector, NBFGR released theA view of the RAC meeting chaired by Prof. T.J. Pandian

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database in public domain on the 28thFoundation Day of the Institute on 12December, 2010. The database containsinformation on 160 species of 41 families under10 orders. The information includes fishclassification, locality of fish collection,chromosome number, karyotype formula,authors, reference, photographs of fish,metaphase spreads and/or karyotypes ofselected species. The database will be enrichedwith the fish cytogenetic information foundacross the globe including, information of otheraquatic organisms such as crustaceans,molluscs, echinoderms, etc. Scientists, researchworkers and students will have free access tothe information. The information on fishchromosomes and allied molecular cytogeneticaspects will serve as a useful reference forresearchers working in conventional/molecularcytogenetics, karyo-evolutionary studies,cytotaxonomy, marker development,chromosomal manipulation, physical genemapping and fish diversity conservation. Thedatabase will also have the facility to uploadthe relevant information at the users end.

Hindi Day and Hindi Pakhwadaobserved

A function was organized on 14, Sept.2010 to celebrate the Hindi Day. Chief GuestDr. W.S. Lakra, Director CIFE, Mumbaiinaugurated the Hindi Day programme bylighting the lamp. The Institute also observed aHindi Pakhwada during 15-29 September, 2010during which seven Hindi competitions wereorganized among the staff of the Institute topromote the use of Hindi in official work. Allthe winners were given prizes by Dr. H.Ravishankar, Director, Central Institute ofSubtropical Horticulture, Rehmankheda,Lucknow, the Chief Guest of the Valedictoryfunction. Mrs. Mamta Chakraborti, PersonalAssistant won the prize for the Best HindiCompetitor – 2010. The Hindi programmeswere coordinated by Dr. L.K. Tyagi, HindiOfficer and Shri Akhilesh Mishra, TechnicalOfficer.

Chief Guest Dr. W.S. Lakra, Director CIFE, Mumbaichairing the Hindi Day programme

Dr. H. Ravishankar, Director, CISH, Lucknow and ChiefGuest of the Valedictory function giving prize for BestHindi Competitor–2010 to Mrs. Mamta Chakraborti

International Women’s Day celebratedThe Institute celebrated the International

Women’s Day. Dr Nishi Pandey was the ChiefGuest of the function organized on thisoccasion. The programme was coordinated byDr. R. Abidi, Principal Scientist and Head,Women Cell, NBFGR.

A view of Women's Day programme

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AWARDS AND RECOGNITIONS

NBFGR Annual Report 2009-2010 wasAwarded “Best Annual Report of theYear” by the ICAR in the small Institutescategory.

Dr. J.K. Jena, Director was conferred withDr. S.Z. Qasim Medal-2011 of BiovedResearch Society for his contributions inthe filed of aquaculture, during 13th IndianAgricultural Scientists’ and Farmers’Congress on 19 February, 2011 at BiovedResearch Society, Allahabad.

Dr. K.K. Lal, Head of the Division wasconferred with the B.G. Gupte Gold Medalof Bioved Research Society during 13th

Indian Agricultural Scientists’ andFarmers’ Congress on 19 February, 2011at Bioved Research Society, Allahabad.

Dr. Peyush Punia, Head of the Divisionwas conferred with the Fellowship Awardof Bioved Research Society during 13th

Indian Agricultural Scientists’ andFarmers’ Congress on 19 February, 2011at Bioved Research Society, Allahabad.

Dr. A.K. Pandey, Principal Scientist

received Prof. M.C. Dash Gold Medal ofthe Zoological Society of India, Bodh-Gayaduring 21st All India Congress of Zoologyand National Seminar on BiodiversityConservation with Special Reference toFisheries and its Management for Food,Livelihood and Environmental Securityorganized during 21-23 December, 2010at Central Inland Fisheries ResearchInstitute, Barrackpore.

Dr. A.K. Pandey, Principal Scientist wasconferred with Dr. R.S. Paroda Gold Medalof Bioved Research Society during 13th

Indian Agricultural Scientists’ andFarmers’ Congress on 19 February,2011 at Bioved Research Society,Allahabad.

Dr. A.K. Singh, Principal Scientist wasawarded “Honorary Fellow” of the Societyof Life Sciences in March, 2011.

Shri Karan Veer Singh, Scientist (SS), wasawarded “Pioneer in GenomicsEducation” class of 2010 at OcimumBiosolutions and Gene Logic, Hyderabad.

Dr. R.S. Patiyal,Technical Officer was conferredwith the Fellow of The Academyof Environmental Biology for hiswork on Live Gene Bank duringNational Consultation onBiodiversity of High AltitudeResource Conservation andUtilization at DCFR, Bhimtal on29 September, 2010. Dr. A. Gopalakrishnan,Principal Scientist was nominatedas the Member, Committee forPreparation of Master Plan ofKerala University of Fisheries andOcean Studies (KUFOS), Kochi,Kerala.

Shri Sharad Pawar, Hon'ble Union Minister of Agriculture and Food ProcessingIndustries alongwith Shri Harish Rawat, Hon'ble Minister of State forAgriculture, presenting the "Best Annual Report 2009-10 Award" for ICARSmall Institutes to Dr. W.S. Lakra and Dr. J.K. Jena, Former Director andDirector, NBFGR, respectively

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Best Division/Section/Centre/Cell of the Institute ARTU, Chinhat Best Scientist of the Institute Dr. Vindhya Mohindra, Principal Scientist Best Young Scientist of the Institute Dr. M. Goswami, Sr. Scientist Best Scientist/ Staff in Institution Building Dr. P.P. Srivastava, Sr. Scientist Award for Technology Transfer/ Commercialization at Institute

Dr. Lalit Kumar Tyagi, Scientist (SS)

Best Extension Worker of the Institute Shri Amit Singh Bisht, Technical Officer (T-5) Best Technical Staff of the Institute Shri Ravi Kumar, Technical Officer ( T-5)

Shri Rajesh Kumar, Lab. Asst. Best Administrative Staff of the Institute Shri Swapan Debnath, Sr. Clerk

Shri P.K. Awasthi, Sr. Clerk Best Supporting Staff of the Institute Shri Indrajeet Yadav, SSG-IV Best Hindi Worker of the Institute Smt. Mamta Chakraborty, PA Best Research Student/ SRF/JRF/ RA of the Institute

Shri Gokhlesh Kumar, SRF

Award for the School Going Children of the Employee of the Institute

Master Viraj Sahai, S/o Dr. P.P. Srivastava

The Annual Institute Awards for the Year 2009-10, were presented to the following staff indifferent award categories:

Best Centre of the Institute Award for 2009-10 to ARTU,Chinhat being received by Dr. P.K. Varshney, Incharge

Dr. Vindhya Mohindra, Principal Scientist receivingAward for the Best Scientist of the Institute 2009-10

Shri Ravi Kumar, Technical Officer receiving Award forBest Technical Staff of the Institute 2009-10

Shri Amit Singh Bisht, Technical Officer receiving Awardfor Best Extension Worker of the Institute 2009-10

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Expert Consultation on MarineBiotechnology and BiodiversityConservation

NBFGR, Lucknow in collaboration withNational Institute of Oceanography (NIO), Goaand Aquatic Biodiversity Conservation Society(ABCS), Lucknow organized an ‘ExpertConsultation on Marine Biotechnology and

Biodiversity Conservation’, at InternationalCentre, Goa during 21-22 April, 2010. The mainobjective of this consultation was to deliberateon the status of research, set priorities for futureand develop strategies and action plan involvingall stakeholders in the marine sector. TheConference had keynote address, invitedpresentations and panel discussion on thefollowing indicative themes: (i) MarineBiodiversity: Issues related to loss, conservationand utilization; (ii) Genetics of marineorganisms; (iii) Marine molecular biology andbiotechnology; (iv) Genetic resources for coastalaquaculture and mariculture development; (v)Marine ornamental fish species: resourcesutilization, conservation and diversification; (vi)marine protected areas, sanctuaries and

biodiversity parks; (vii) Bio-security and exoticspecies in coastal aquaculture; (viii) DNABarcoding of marine life and its role in fish tradeand conservation of endangered fish species;(ix) Ecosystem based marine fisheriesmanagement; (x) Climate change and its impacton marine and coastal fisheries; and (xi) Marinefisheries policy and regulations forsustainability of resources. The consultation

was attended by more than 50eminent professionals, policymakers, scientists and teachers.

International Consultationon DNA Barcoding

An International Consulta-tion on DNA Barcoding wasorganized by the Central Instituteof Fisheries Education, Mumbai,in collaboration with NBFGR,Lucknow and AquaticBiodiversity ConservationSociety, Lucknow at NASC, NewDelhi during 06-07 November,

2010. Over 60 researchers, policy makers andother professionals participated in theconsultation.

Partners meet on Application ofBioinformatics in Fisheries Domain

A Partners meet on “Application ofBioinformatics in Fisheries Domain” wasconducted under Component I of the NationalAgricultural Innovation Project entitled“Establishment of National AgriculturalBioinformatics Grid in ICAR (NABG)” on 29January, 2011 to discuss the issues in fishbioinformatics and to develop a road map forresearch in fisheries domain .The meeting wasattended by Dr. J.K. Jena, Director, NBFGRLucknow, Dr. U.N. Dwivedi, Pro-Vice

WORKSHOPS/SYMPOSIA/TRAININGS ORGANIZED

Inaugural session in progress

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A view of the Partners Meet on Application of Bioinformatics inFisheries Domain

Chancellor, University of Lucknow, Dr. AnilRai, CPI-NABG IASRI New Delhi, Dr. N.S.Nagpure, CCPI NABG, Prof I. J. Singh, Dean,Dept. of Fisheries Biology, GBPUAT, Pantnagar,Dr. Sukanta Banik, Bioinfomatics Co-ordinator,Tripura University and several other eminentscientists from all over the country. The various

recommendations emerged from thedeliberations were: preparation of inter-institutional research project proposals,installation of high end computing facilities atvarious ICAR centres and proper disseminationof knowledge on bioinformatics tools andresources.

National Seminar on “Biodiversity,Ecosystems and Climate Change”

A National Seminar on “Biodiversity,Ecosystems and Climate Change: TheChallenge Ahead” was organised by NationalInstitute of Ecology (NIE), New Delhi at

National Bureau of Fish GeneticResources, Lucknow during 4-6December, 2010. Dr. J.K. Jena,Director, NBFGR welcomed theparticipants. The dignitariesoffered the floral tributes toProfessor Ram Deo Misra, theFounding Father of IndianEcology. Professor J.S. Singh ofBanaras Hindu University wasawarded the 2010 Prof. R.D.Misra Birth Centenary Lecture.The inaugural function waschaired by Prof. R.S. Tripathi. Dr.P.V. Dehadrai, President, of NIE,New Delhi, Drs. Brij Gopal andP.S. Pathak, Vice Presidents, NIE,

New Delhi also graced the occaission. ProfessorJ.S. Singh delivered the birth centenary lectureentitled “Ecology in India: Retrospect andProspects”. Dr. K.K. Vass delivered the keynoteaddress on Diversity of fish genetic resourcesin India. The programme also included invitedpresentations by experts and students’

Glimpses of National Seminar on Biodiversity ecosystems and climate change

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presentations. The valedictory session waschaired by Dr. P.V. Dehadrai, President NIEand Dr. K.K. Vaas, Dr. P.S. Pathak, Dr. BrijGopal and Dr. J.K. Jena participated as panelist.The participants discussed the outcome of theseminar and consolidated the finalrecommendations.

Sensitization training programme onBioinformatics Tools and Resources inFisheries Research

A sensitization training programme on‘Bioinformatics Tools and Resources in FisheriesResearch’ was organized during January 24 –29, 2011 at NBFGR, Lucknow. The objective ofthe training was to disseminate the knowledgeabout the application of bioinformaticsresources on biological data and to promoteinter-disciplinary research groups with focus onfish computational research. The training wasinaugurated by Dr. C. S. Nautiyal, Director,NBRI, Lucknow. A total of 21 participantsincluding scientists/faculty members fromresearch institutes and universities participatedin the training programme. The programme

constituted of brainstorming lectures onfunctional genomics, proteomics,metabolomics, designing of biological softwares,computational skills in bioinformatics, in silicomicroarray data analysis, in silico taxonomy,protein 3D structure prediction, tools forprotein and gene annotation, primer designingand phylogenetic analysis in reference tofisheries research. Hands-on training on aboveaspects was also arranged for the participantsduring afternoon sessions. Dr. Bangali Baboo,National Director, National AgriculturalInnovation Project (NAIP), New Delhi was theChief Guest during the Valedictory session.

Training Programmes on ‘AquacultureTechnologies and ProductivityEnhancement’

The NBFGR, at its Aquaculture Research& Training Unit (ARTU), Chinhat, organizeda series of short-term training programmessponsored by Uttar Pradesh DiversifiedAgriculture Support Project (UP-DASP),ATMA, Bihar and National FisheriesDevelopment Board (NFDB), Hyderabad. A

total of nine trainingprogrammes were conductedduring the period under report.A total of 208 progressive fishfarmers from different districtsof U.P. were trained in thesetraining programmes, as perdetails below:

The above trainingprogrammes were residentialand field oriented hands-ontrainings with practicaldemonstrations. Apart fromtheory classes, laboratorydemonstrations and exerciseswere made. Field visit to the fishfarms of the Institute were madeto expose the trainees withvarious fisheries activities.Participants of the training with the Director and staff of NBFGR

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A fish farmer receiving training certificate fromDr. J.K. Jena, Director, NBFGR

Batch Period No. of trainees Sponsored by First Batch July 13-17, 2010 23 UP-DASP Second Batch July 26-30, 2010 25 NFDB Third Batch August 03-07, 2010 14 UP-DASP Fourth Batch August 17-21, 2010 23 UP-DASP Fifth Batch September 06-10, 2010 35 UP-DASP Sixth Batch November 22-26, 2010 14 UP-DASP Seventh Batch December 08-12, 2010 27 NFDB Eighth Batch December 20-24, 2010 30 NFDB Ninth Batch 07- 11 March, 2011 17 ATMA, Bihar

Participants of a farmers’ training programme withthe Director and staff of NBFGR

Dr. S. N. Dwivedi, Former Additional Secretary, Govt. of Indiareleasing the Training Manual, along with Dr. J.K. Jena, Director,NBFGR and Dr. P.K. Varshney, Incharge, ARTU Chinhat NBFGR

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EXTENSION ACTIVITIES

Participation in ExhibitionsThe Institute participated in the following

exhibitions related to fisheries and aquaticresources in different parts of the countryduringthe year 2010-2011 :

Exhibition organized on the occasion ofIndian Aqua-Invest Congress and Expo-2010 at CIFE, Mumbai during 26 -28 May,2010.

“Innovation for Industry Meet in Fisheries”at Visakhapatnam on 8 September, 2010.

‘National Fish Festival’ named “Benaqua2010” at Nalban Fisheries Food Park, SaltLake City, Kolkata during 1-4 October,2010.

“Wildlife Week Celebrations” conductedby Department of Forest, Govt. of Keralaat Mangalvanam, Kochi during 03-07October, 2010.

"ICAR North Zone Farmers Fair 2010" atIVRI, Bareilly during 1-3 November, 2010.

The “Science Expo 2011” organized byRegional Science City, Lucknow during19-23 January, 2011.

The X Agricultural Science Congress heldat National Bureau of Fish GeneticResources, Lucknow during 10-12February, 2011.

Exhibition- cum-Mango Goshthi organizedby CISH, Lucknow on 27 February, 2011at Dashehari Village, Kakori, Lucknow.

The exhibition on the occasion of theAsian-Pacific Aquaculture 2011 at Kochi,Kerala during 17-20 January, 2011.

Visits by Farmers and StudentsThe following batches of farmers and

students visited the Institute during the period:

A group of 90 aqua-farmers fromMohanlalganj and Barabanki, UP, visitedARTU, NBFGR, Chinhat, Lucknow duringthe period of July to December, 2010.

A group of 60 B.Sc.students from DBPGCollege, Bachhravanvisited the Institute onDecember 16, 2010.

A group of 30 B.Sc.students from Sri JagdambaSaran Singh EducationInstitute, Gonda visited theInstitute on 4 February,2011.

A group of 40progressive farmers fromDistrict Faizabad, U.P.visited the Institute on 16February, 2011.Shri B.L. Joshi, His Excellency, Governor of UP and Shri Harish Rawat, Hon'ble Minister

of State for Agriculture and Dr. R.B. Singh, President, NAAS, being apprised about theNBFGR's achievements at exhibition pavillian during X Agril. Science Congress

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Technical Lectures/ talks delivered Dr. P. K. Varshney, Senior Scientist

delivered the following technical lectures/talks in different training programmes forfish farmers at different places:

Talks on “Scope of Aquaculture inUttar Pradesh” and “FreshwaterPrawn Farming” on 14 and 15

January, 2011, respectively, at FFDA,Aligarh.

Lectures on “FreshwaterAquaculture” and “FreshwaterPrawn Farming” on 30 and 31January, 2011, respectively, at FFDA,Aligarh.

A talk on “Freshwater PrawnFarming” on 6 October, 2010 atFFDA, Barabanki.

Lectures on “FreshwaterAquaculture” and “FreshwaterPrawn Farming” on 4 September,2010 at FFDA, Barabanki.

Dr. A.K. Singh, PrincipalScientist delivered an invited talk on

“Pineal melatonin and Fish Reproduction”at Department of Zoology, University ofLucknow under UGC-SAP Programme on30 March, 2011.

Shri Karna Veer Singh, Scientist (SS),delivered a lecture and conducted trainingas guest speaker for Computer Aided DrugDesigning at BCS- Insilico Biology, GomtiNagar, Lucknow on 15 May, 2010.

Dr. P. Sorgeloos along with Dr. J.K. Jena, Director and other dignitaries at NBFGRpavaillian during Asian Pacific Aquaculture 2011 at Kochi

Dr. B. Meenakumari, DDG (Fy), ICAR and Dr. Bengali Baboo, NationalDirector, NAIP, New Delhi at NBFGR pavillian during Innovation forIndustry Meet in Fisheries at Vishakhapatnam

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LIST OF PROJECTS

Institutional Projects

Sl. No. Project Title Personnel Period

Molecular Biology & Biotechnology Division

1 Cytogenetic characterization of prioritized marine and freshwater fish species.

B. Kushwaha, N.S. Nagpure and Ravindra Kumar

April, 2008 - March, 2011

2 Assessment of genotoxicity due to heavy metal pollution and heat stress in fishes

N. S. Nagpure, Ravindra Kumar, Basdeo Kushwaha and Poonam J. Singh

April, 2008 - March, 2011

3 Genetic characterization of commercially important fishes from the Western Ghats and marine habitat.

A. Gopalakrishnan, V.S. Basheer, T. Rajaswaminathan and P.R. Divya

April, 2009 - March, 2012

4 Cytogenetic and molecular characterization of freshwater catfishes using selected nuclear DNA region.

Ravindra Kumar, N. S. Nagpure, Basdeo Kushwaha, Mukunda Goswami and Akhilesh K. Mishra

April, 2009 - March, 2012

5 Protein profiling of Indian major carps based on Mass Spectrometric Analysis (Electrospray Ionization).

M Goswami, N. S. Nagpure, Ravindra Kumar and A. Srinivasan, Associate Professor (All India Institute of Medical Sciences, New Delhi)

April, 2010 – March, 2013

6 DNA Barcoding of Indian fishes.

Mahender Singh, A. Gopalakrishanan, K.K. Lal, Peyush Punia, V. Mohindra, U.K. Sarkar, M. Goswami and K.V. Singh

October, 2005 - March, 2012

7 Genetic stock structure elucidation in Tenuolosa ilisha and Channa striatus by microsatellites, mtDNA and molecular cytogenetic markers.

M. Singh, N.S. Nagpure, A. Gopalakrishnan and Ravindra Kumar

April, 2010 - March, 2013

Fish Conservation Division

8 Outreach Activity on Fish Genetic Stocks.

Kuldeep K. Lal, A. Gopalakrishnan, P. Punia, V. Mohindra, U.K. Sarkar, Mukunda Goswami and Rajeev K. Singh

April, 2008 – March, 2012

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S. N. Project Title Personnel Period

9 Genetic Approaches for conservation of prioritized Indian fish species.

Sub project 1:

Documentation of Genetic diversity in prioritized fish species belongings to the group featherback, carps, murrels and catfishes.

Kuldeep K. Lal, Vindhya Mohindra, Peyush Punia and Rajeev K. Singh

April, 2009 - March, 2012

10 Sub-project 2:

Development of breeding and sperm banking protocols for prioritized Indian finfishes belonging to groups catfishes, featherbacks, murrels and carps.

Sudhir Raizada, K.K. Lal, P.K. Varshney, A.K. Yadav and Sanjay Singh

April, 2009 - March, 2012

11 Sub-project 3: Documentation of mitochondrial genomes of fishes from important taxonomic groups.

Rajeev K. Singh and Vindhya Mohindra

April, 2010 - March, 2012

12 Neuroendocrine regulation of reproduction in threatened fishes.

A.K. Pandey, P. K. Varshney, R.S. Patiyal and A.K. Yadav

April, 2008 – March, 2011

13 Conservation of highly endemic freshwater fishes of the Western Ghats through milt cryopreservation and captive breeding.

V.S. Basheer, A. Gopalakrishnan, T. Raja Swaminathan , P.R. Divya, and K. Dinesh, Associate Professor, College of Fisheries, Kochi

April, 2009 - March, 2012

14 Role and potential of fisher folk organization and NGOs in Co-management of aquatic resources.

L.K. Tyagi, S.P. Singh, P.R. Divya, Amar Pal and Amit Singh Bisht

April, 2008 - March, 2011

15 AFLP fingerprinting and development of Microsatellite Markers for endangered Golden Mahseer.

Poonam Jayant Singh and

M. Goswami

April, 2008 - March, 2011

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S. N. Project Title Personnel Period

16 Quality seed production of selected carps, catfish and endangered species.

P. K. Varshney, A. K. Pandey, A. K. Yadav and Sanjay Singh

April, 2001 – March, 2014

Fish Health Management Division

17 Exploration of protozoan & monogenean parasites among carps and catfishes.

Rehana Abidi, Neeraj Sood and S.M. Srivastava

April, 2010 - March, 2013

18 Production of monosex/sterile common carp Cyprinus carpio using Aromatase Inhibitore.

A. K. Singh and P.P. Srivastava

April, 2009 - March, 2012

19 Development of monoclonal antibody based marker for monitoring humoral immune response in Catla catla.

Neeraj Sood, Gaurav Rathore, P.K. Pradhan and Peyush Punia

April, 2010 - March, 2012

20 Development of in vitro cellular model for assessment of innate immunity parameters of Labeo rohita.

Gaurav Rathore, Neeraj Sood, P.K. Pradhan and P. Punia

April, 2010 - March, 2012

21 Molecular assay for detection of Aphanomyces invadans for surveillance and prediction of epizootic Ulcerative Syndrome outbreaks.

P.K. Pradhan, Rehana Abidi, Gaurav Rathore, Neeraj Sood and T.Raja Swaminathan

April, 2009 - March, 2012

22 Disease surveillance of endemic and commercially important ornamental fishes of the Western Ghats.

T. Raja Swaminathan, A. Gopalakrishnan, V.S. Basheer and P.R. Divya

April, 2009 - March, 2012

Fish Taxonomy and Resources Unit

23 Development of digital repository of fishes of India. Sub-Project-I: Finfish Diversity Database. Sub Project-II: Taxonomic information system for selected fishfishes.

S.P. Singh, U.K. Sarkar, A.K. Pathak, A.Kathirvelpandian, R.Dayal, Reeta Chaturvedi and Ravi Kumar

April, 2007 - March, 2012

24 Fish Germplasm Exploration, Assessment, Cataloguing and Conservation for North Eastern Region.

U.K. Sarkar, Mukunda Goswami, M.S. Verma, Ravindra Kumar and A. Kathirvelpandian

April, 2007 - March, 2011

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Externally funded ProjectsS.N. Project Title Personnel Funding agency Period

1 Microsatellite markers for genetic variability studies in Penaeus (Fenneropenaeus) indicus.

A. Gopalakrishnan DBT, Govt. of India.

August, 2007 - August, 2010

2 Isolation and characterization of Flavobacterium sp. from fish and aquatic environmental samples.

Gaurav Rathore NBAIM, ICAR, New Delhi

August, 2006 - August, 2012

3 Bioprospecting of genes and allele mining for abiotic stress tolerance.

Vindhya Mohindra and Ravindra Kumar

NAIP, ICAR May, 2009- March, 2012

4 Genetic characterization and conservation biology of economically important Siluroid fish Ompok pabda of Tripura.

U.K. Sarkar and S. Banik, Tripura University

DBT, Govt. of India

March, 2011-February, 2014

5 Studies on fish biodiversity and aquatic environment of the Ken-Betwa River: An assessment prior to river interlinking for conservation of aquatic bio-resources.

U.K. Sarkar DBT, Govt. of India

September, 2007-August,

2010

6 Development and characterization of cell lines from Schizothorax richardsonii and Puntius chelynoides.

M. Goswamy and S.N. Bahuguna, HNB Garhwal Uni., Srinagar, Uttarakhand

DBT, Govt. of India

November, 2009-

November, 2012

7 Production of monoclonal antibodies to serum immunoglobulins of Ophiocephalus striatus and Labeo rohita and their application in immunoassays.

Neeraj Sood and Gaurav Rathore

DBT, Govt. of India

July, 2007-July, 2010

8 Ontogeny of the digestive system during larval development of Indian butter fish, Ompok bimaculatus

P.K. Pradhan International Foundation for Sciences (IFS), Swedan

November, 2008-

November, 2011

9 Assessment of genetoxic effect of tannery effluents in fishes of River Ganga.

N. S. Nagpure and Ravindra Kumar

UPCST, Lucknow

September, 2009-

September, 2011

10 Establishment of National Agricultural Bioinformatics Grid in ICAR

N.S. Nagpure, S.P. Singh, A.K. Pathak, U.K. Sarkar and Mahender Singh

NAIP ICAR (Component I)

April, 2010-March, 2012

11 Genetic characterization of common HAB species in the Indian EEZ.

A. Gopalakrishnan MOES, Govt. of India

April, 2009-March, 2012

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S.N. Project Title Personnel Funding agency

Period

12 Establishment of a National Repository at NBFGR, Lucknow for Conservation and Characterization of Fish Cell Lines.

M. Goswami and G. Rathore

DBT, Govt. of India

November, 2010-November, 2013

13 Genetic diversity in the natural populations of Cobia, (Rachycentron canadan) and silver pompret (Pampus argentius), candidate species for mariculture in India.

P.R. Divya, A. Gopalakrishnan and V.S. Basheer

DBT, Govt. of India

November, 2010-November, 2013

14 Harmonizing biodiversity conservation and agricultural intensification through integration of plant, animal and fish genetic resources for livelihood security in fragile ecosystems.

K.K. Lal, Peyush Punia, Vindhya Mohindra, U.K. Sarkar, L.K. Tyagi and Rajeev K. Singh

NAIP, ICAR August, 2010-August, 2014

15 Mass seed production of Indian Catfish Heteropneustus fossilis using sperm cryopreservation technology and its economic perspective for aquaculture

K.K. Lal and Peyush Punia

UPCAR, Lucknow

October, 2008-October, 2011

16 Development of cryopreservation protocol for breeding and grow out technology for gaint snakehead Channa marulius.

Peyush Punia and S. Raizada

UPCAR, Lucknow

April, 2010-April, 2013

17 Ecological impact assessment of African catfish Clarias gariepinus: Disease risks and potential for resource competition.

A.K. Singh, Rehana Abidi and A.K. Pathak

UPCAR, Lucknow

January, 2010-December, 2012

Externally funded ProjectsS.N. Project Title Personnel Funding agency Period

1 Evaluation of Butachlor commercial formulation for acute toxicity test and effect on growth performance of Cyprinus carpio

P.P. Srivastava and Rajesh Dayal

M/s Monsento India Ltd.

August, 2010- August, 2011

2. Evaluation of pertimethalian commercial formulation for acute toxicity test and effects on growth performance of Cyprinus carpio

P.P. Srivastava, N.S. Nagpure and A.K. Yadav

M/s Rallis India Ltd.

October, 2011-September, 2011

3. Evaluation of Atrazine commercial formulation for acute toxicity test and effects on growth performance and multilocation field studies to assess the impact on Cyprinus carpio

P.P. Srivastava, N.S. Nagpure, A.K. Yadav and Rajesh Dayal

M/s Syngenta India Ltd.

February, 2011-January, 2012

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Abroad:Dr. Vindhya Mohindra, Principal Scientistparticipated in the Open door workshop:Working with human genome sequence,organized by the Welcome Trust, U.K. at theAmerican University, Dubai during 19-21March, 2011.

Mrs. Poonam Jayant Singh, Scientist (SG)participated in the following:

NAIP sponsored International InternshipProgramme on IPR and TechnologyTransfer at University of Michigan, USAduring 10-24 July, 2010

Workshop on Exploring the Need forSpecific Measures for Access and Benefit-Sharing of Animal Genetic Resources forFood and Agriculture at Centre for GeneticResources, The Netherlands duringDecember 7-10, 2010 as a Panelist.

In India:Dr. W.S. Lakra, Director participated in thefollowing meetings and conferences during theperiod under report:

Director’s Conference at NASC Complex,New Delhi during 15-16 July, 2010.

Review Meeting of ICAR BiodiversityWork plan at NASC Complex, New Delhion 27 July, 2010.

Dr. J.K. Jena, Director participated in thefollowing meetings and conferences:

ICAR Regional Committee Meeting atBAU, Ranchi during 7-9 October, 2010.

International Consultation on DNABarcoding during 6-7 November, 2010 atNASC, New Delhi.

COML Meeting at Kochi and deliveredLead Lecture on DNA Barcoding of Finfishand Shellfish on 1 Dec. 2010.

National Seminar on Marine LivingResources at Kochi on 2 December, 2010.

5th National Conference on KVK atMPUAT, Udaipur during 22-24December, 2010 and delivered a LeadLecture on Fish Genetic Resources of Indiaand Health Management.

First Meeting of the Expert Group on Short-term and long-term measures for creatingan appropriate and effective legal andinstitutional framework for managementand control of aquatic animal diseases on18 February, 2011 at DAHDAF, NewDelhi.

ICAR Directors’ Conference at NASCComplex, New Delhi during February 23-24, 2011.

International Conference on TropicalIsland Ecosystem: Issues related toLivelihood, Sustainable Development andClimate Change during 23-26 March, 2011at CARI, Port Blair.

Review meeting of ICAR FisheriesResearch Institutes with regard to researchprogramme of XII plan with RAC Chairsconvened by Secretary, DARE & DG,ICAR, New Delhi on 17 March, 2011.

Dr. J.K. Jena, Director; Dr. K.K. Lal and Dr.Peyush Punia, Head of the Divisions and Dr.A.K. Pandey, Principal Scientist participated inthe Agricultural and Farmers Congress atBioved Research Society, Allahabad during 23-24 February, 2011.

Dr. J.K. Jena, Director; Dr. A. Gopalakrishnanand Dr. A.K. Singh, Principal Scientists; Dr .V.

PARTICIPATION IN SEMINARS/SYMPOSIA/WORKSHOPS/TRAININGS/MEETINGS

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S. Basheer, Sr. Scientist; Dr. T. RajaSwaminathan, Scientist (SS) and Dr. P. R.Divya, Scientist participated in the Asia PacificAquaculture 2011 (APA 2011), organized bythe World Aquaculture Society during 17-20January, 2011 at Kochi.

Dr. Pyush Punia, Head of Division and Shri A.S.Bisht, Technical Officer (T-5) participated in theIndian Aqua-Invest Congress and Expo-2010at CIFE, Mumbai during 26 -28 May, 2010.

Dr. N.S. Nagpure, Head of Divisionparticipated in the following meetings andconferences:

National Consultation on ‘Agro-Biodiversity’ at NASC Complex New Delhiduring 26-27 May, 2010 as a ‘Rapporteur’.

ZTM-BPD sponsored workshop onEntrepreneurship Development andBusiness Opportunity Workshop/Camp atIVRI, Izatnagar, Bareilly during 8-9 July,2010.

Workshop of NAIP project entitled“Establishment of National AgriculturalBioinformatics Grid (NABG) in ICAR” atIASRI, New Delhi during 16-17 September,2010 as a CCPI (Fisheries Domain).

Dr. A. Gopalakrishnan, Principal Scientistparticipated in the following meetings andconferences:

As a Member, Kerala State BiodiversityBoard, participated in three meetings of theBoard held during April – December, 2010.

Delivered a lecture on “DNA Barcodingof Aquatic Organisms” in the short-termtraining programme on ‘Tools andTechniques in Molecular Biology andBioinformatics’ on 18 February, 2011 atDepartment of Biotechnology, CochinUniversity of Science & Technology, Kochi.

Dr. A. Gopalakrishnan, Dr. V.S. Basheer andDr. P. R. Divya participated one day seminaron “Joining hands to conserve Kerala’s

biodiversity” as a part of InternationalBiodiversity day celebrations of Kerala Forestsand Wildlife Department and Kerala StateBiodiversity Board held at CMFRI during 22May, 2010.

Dr. A.K. Pandey, Principal Scientist participatedin the following meetings and conferences:

21st All India Congress of Zoology &National Seminar on BiodiversityConservation with Special Reference toFisheries and its Management for Food,Livelihood and Environmental Security,organized jointly by the Zoological Societyof India, Bodh-Gaya and Inland FisheriesSociety of India, Barrackpore at CentralInland Fisheries Research Institute (CIFRI),Barrackpore during 21-23 December, 2010.

National Seminar on Climate Change andits Impact on Biological Communities,organized jointly by Department ofEnvironmental Sciences. Dr. R.M. LohiaAvadh University, Faizabad and IndianAcademy of Environmental Sciences,Haridwar at Department ofEnvironmental Sciences, Dr. R.M. LohiaAvadh University, Faizabad during 12-13February, 2011.

Dr. U.K. Sarkar, Sr. Scientist participated in thefollowing meetings and conferences:

3rd DBT Task Force meeting onAquaculture and Marine Biotechnologyheld at New Delhi from 23-24 March, 2011and presented a research project proposal.

PAC Animal Science Meeting of the DSTheld at Central University, Patna on 14March, 2011 and presented a researchproject proposal.

Task Force meeting of the Uttar PradeshState Biodiversity Board, Lucknow held atLucknow on 9 March, 2011.

Meeting of the Central Pollution ControlBoard on Assessment of Fisheries with

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regard to Water Quality of River Gangaand Yamuna held at Parivesh Bhawan,New Delhi on 3 September, 2010.

A brainstorming meeting on BiodiversityConservation and Management organizedby the Ministry of Environment and Forestsat Paryavaran Bhawan, New Delhi on 9December, 2010.

Interface Meeting of the National FisheriesDevelopment Board at Hyderabad during15-16 December, 2010.

The Task Force Meeting of Uttar PradeshState Biodiversity Board held at Lucknowon 29 November, 2010.

Dr A. Gopalakrishnan, Principal Scientist andDr V. S. Basheer, Sr. Scientist, NBFGR CochinUnit participated in the IUCN Western GhatsFreshwater Biodiversity Assessment EvaluationWorkshop to assess the conservation status offreshwater teleosts during 7-11 October, 2010at Coimbatore, Tamil Nadu organized by theZoo Outreach Organization (ZOO),Coimbatore.

Dr. N.S. Nagpure, Head of Division; Dr. S.P.Singh, Principal Scientist; Dr. U.K. Sarkar, Sr.Scientist and Dr. Mahender Singh, Scientist (SS)participated in the Launch Workshop of NABGunder NAIP at IASRI, New Delhi during 17-18September, 2010 and the Partners Meet ofNABG under NAIP at NBAIM, Mau on 29October, 2010.

Dr. A.K. Singh, Principal Scientist participatedin the following meetings and conferences:

National Consultation on Biodiversity ofHigh Altitude Aquatic Resources,Conservation and Utilization during 29-30 September, 2010 organised byDirectorate of Coldwater FisheriesResearch, Bhimtal.

The XX Regional Committee Meeting IVat BAU, Ranchi during 7-9 Oct, 2010.

National Training Workshop onstrengthening CITES, implementationcapacity to ensure sustainable wild lifemanagement and non-detrimental trade inIndia organized by Wildlife Institute ofIndia at Dehradun during 20-21December, 2010.

Meeting of Expert Group to work outpossible arrangements and to formulateguidelines for Litopenaeus vannamei inInland freshwater held at NFDB,Hyderabad on 25 March, 2011.

Dr. U.K. Sarkar, Sr. Scientist and Dr. L.K. Tyagi,Scientist (SS) attended a Workshop onConservation of Fisheries and Other FaunalResources of River Ghagra, UP organized atSitapur by District Administration, Sitapur andKatraniaghat Foundation, Lucknow on 23December, 2010.

Dr. P.K. Varshney, Sr. Scientist deliveredlectures on “Freshwater Aquaculture” and“Freshwater Prawn Farming” on 30-31January, 2011, respectively, in a trainingprogramme organized by the FFDA, Aligah,UP.

Dr. L.K. Tyagi, Scientist (SS) and Shri A.S. Bisht,Technical Officer (T-5) attended “Science Expo2011” during 19-23 January, 2011 at RegionalScience City, Lucknow.

Dr. Rajeev Kr. Singh, Sr. Scientist participatedin 15h ADNAT Convention during 23-25,February, 2011 at CCMB, Hyderabad.

Mrs. Poonam Jayant Singh, Scientist (SG)participated in the Symposium on Genomicsand Biodiversity and presented a poster entitled“Legal perspectives in genetic resourceprotection : Implications of international regimefrom common heritage of mankind to theNagoya protocol during 23-25, February, 2011at CCMB, Hyderabad.

Dr. M. Goswami attended the followingtraining programmes and conferences:

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Symposium on AgriculturalBiotechnological Research at NASC, NewDelhi during 26-27 July, 2010.

International Training on Biosystematics atCMDE, Delhi University (Sponsored byDST) during 6-15 September, 2010

NAIP sponsored hands on training on“Stem Cell Research for Quality AnimalProduction” at NDRI, Karnal during 17-30 March, 2011.

Sh. S.V. Chaudhary, Sr. Library Assistant(T-4) participated in the following conferences

National Conference on Knowledge

Management in the Globalized Era during21-23, April, 2010 at Association of Agril.Librarians and Documentatists of India,New Delhi.

24 th National Seminar of IndianAssociation of Special Libraries andInformation Centres during 18-21,December, 2010 at DD University,Gorakhpur.

Shri Akhilesh Kr. Mishra, Technical Officer(T-5) participated in Akihil Bhartiya RajbhashaSammelan during 25-27, November, 2010 atGoa organized by Bhartiya Rajbhasa Parishad,Goa.

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Research papersAgwuocha, S., B.G. Kulkarni and A.K.

Pandey, 2011. Histopathological changesin hepatopancreas of intertidal clam,Gafrarium divaricatum, exposed to xylene,benzene and gear oil-WSF. J. Environ. Biol.,32: 35-38.

Bhoir, K.K., S.A. Suryawansghi and A.K.Pandey, 2011. Effect of sublethal heroinadministration on liver histo-morphologyof albino Rattus norvegicus. J. Appl. Biosci.,37: 38-41.

Chaturvedi, Anshumala, Vindhya Mohindra,Rajeev K. Singh, Kuldeep K. Lal, PeyushPunia, Ranjana, Anup Mandal, LalitNarain and W.S. Lakra, 2010. Populationgenetic structure and phylogeography ofCyprinid fish, Labeo dero inferred fromallozyme and microsatellite DNA markeranalysis. Mol. Biol. Rep. 38(5): 3513-3529.

Divya, P.R., P.C. Thomas, Vindhya Mohindra,K.K. Lal, R.K. Singh, A. Gopalakrishnan,Peyush Punia and W.S. Lakra, 2010. Amolecular approach to reveal the geneticidentity of parrot mussel and othersympatric mussel species distributed alongthe Kerala coast. J. Mar. Biol. Asso. India.52(1): 35-41.

Diwan, A.D., S Ayyappan, K.K. Lal and W.S.Lakra, 2011. Cryopreservation of fishgametes and embryos. Indian J. Anim. Sci.88(4), Suppl 1: 109-124.

Habib, Maria, W.S. Lakra, Vindhya Mohindra,Praveen Khare, A.S. Burman, AkanshaSingh, Kuldeep K. Lal, Peyush Punia andAsif A. Khan, 2010. Evaluation ofcytochrome b mtDNA sequencing geneticdiversity studies of Channa marulius(Channidae: Perciformes). Mol. Biol. Rep.38(2): 841-846.

Kumar, R, N.S. Nagpure, B. Kushwaha, S.K.Srivastava and W.S. Lakra, 2010.Investigation of the genotoxicity ofMalathion to freshwater teleost fish Channapunctatus (Bloch) using micronucleus testand comet assay. Arch. Environ. Contam.Toxicol., 58(1): 123-30.

Kumari, V., G. Rathore, U.K. Chauhan, A.K.Pandey and W.S. Lakra, 2011. Seasonalvariations in abundance of nitrifyingbacteria and nitrification in fish pond. J.Environ. Biol., 32:153-159.

Kupekar, Sandhya, B.G. Kulkarni and A.K.Pandey, 2010. Harmonal control ofMelanophores in the marine crab,Charybdis lucifera (Fabricius). J. Appl. Biosci.,36(1): 108-110.

Kushwaha, B., Ravindra Kumar, N.SNagpure, S.K. Srivastava, V.S. Basheer,M.K. Anil and W.S. Lakra, 2010.Chromosomal studies of three vulnerablemarine fishes from West Coast of India.Indian J. Geo-Marine Sci., 40(1): 62-66.

Lakra, W.S. and A.K. Singh, 2010. Risk analysisand sustainability of Pangasianodonhypophthalmus culture in India. Aqua. Asia,15(1): 34-37.

Lakra, W.S., U.K. Sarkar, R.S. Kumar, A.Pandey, V.K. Dubey and O.P. Gusain,2010. Fish diversity, habitat ecology andtheir conservation and management issuesof a tropical River in Ganga basin, India.Environmentalist, 30: 306- 319.

Lakra, W.S., M. Goswami and U.K. Sarkar,2011. Conservation biology of IndianMahseers. Indian J. Anim. Sci. 88(4), Suppl1: 98-108.

Lakra, W.S., M.S. Verma, M. Goswami, K.K.Lal, V. Mohindra, P. Punia, A.Gopalakrishnan, K.V. Singh, R.D. Ward

PUBLICATIONS

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and Paul Hebert, 2011. DNA barcodingIndian Marine Fishes. Mar. Ecol. Res., 11(1):60-71.

Lakra, W. S., T. R. Swaminathan and K. P.Joy, 2010. Development, characterization,conservation and storage of fish cell lines:a review. Fish Physiol. and Biochem., DOI10.1007/s10695-010-9411-x.

Lakra, W. S., M. Goswami, A.Gopalakrishnan, D.P. Singh, A. Singhand N.S. Nagpure, 2010. Geneticrelatedness among the fish species of genusChanna using mitochondrial DNA genes.Biochem. Systema. and Ecol., 38: 1212-1219.

Lakra, W. S., T.R. Swaminathan, G. Rathore,M. Goswami, K. Yadav and S. Kapoor,2010. Development and characterizationof three new diploid cell lines from Labeorohita (Ham.). Biotech. Prog. , DOI.10.1002/btpr.418.

Mani, I., R. Kumar, M. Singh, N.S. Nagpure,B. Kushwaha, P.K. Srivastava, D.S. Raoand W.S. Lakra, 2010. Nucleotide variationand physical mapping of ribosomal genesusing FISH in genus Tor (Pisces,Cyprinidae). Mol. Biol. Rep., 38: 2637–2647.

Mishra, Akhilesh K., W.S. Lakra, J.P. Bhatt,M. Goswami, N.S. Nagpure and VineetK. Dubey, 2010. Genetic diversity and itsimplications in the conservation ofthreatened fish species Barilius bendelisis(Hamilton). J. Ecophysiol. Occup. Health, 10:149-155.

Mishra, R.K., A.K. Yadav, P.K. Varshney,A.K. Pandey and W.S. Lakra, 2011.Comparative effects of different hormonalpreparations in induced spawning of thefreshwater catfish, Heteropneustes fossilis.J. Appl. Biosci., 3: 59-62.

Muneer, P. M. A., Remya Sivanandan, A.Gopalakrishnan, V.S. Basheer, K.K.

Musammilu and A.G. Ponniah, 2010.Development and characterization ofRAPD and microsatellite markers forgenetic variation analysis in the criticallyendangered yellow catfish Horabagrusnigricollaris (Teleostei: Horabagridae).Biochem. Genet., DOI 10.1007/s10528-010-9389-1.

Muneer, P. M. A, A. Gopalakrishnan, RemyaSivanandan, V.S. Basheer and A.G.Ponniah, 2010. Genetic variation andphylogenetic relationship between twospecies of yellow catfish, Horabagrusbrachysoma and H. nigricollaris (Teleostei:Horabagridae) based on microsatellite andRAPD markers. Mol. Biol. Rep. , DOI10.1007/s11033-010-0352-3.

Nwani, C.D., N.S. Nagpure, R. Kumar, B.Kushwaha, P. Kumar and W.S. Lakra,2010. Lethal concentration and toxicitystress of Carbosulfan, Glyphosate andAtrazine to freshwater fish Channapunctatus (Bloch). Int. Aquat. Res. 2: 105-111.

Nwani, C.D., N.S. Nagpure, R. Kumar, B.Kushwaha, P. Kumar and W.S. Lakra,2011. Mutagenic and genotoxic assessmentof atrazine-based herbicide to freshwaterfish Channa punctatus (Bloch) usingmicronucleus test and single cell gelelectrophoresis. Environ. Toxicol.Pharmacol., 31: 314–322.

Nwani, C.D., W.S. Lakra, N.S. Nagpure, R.Kumar, B. Kushwaha and S.K.Srivastava, 2010. Mutagenic andgenotoxic effects of carbosulfan infreshwater fish Channa punctatus (Bloch)using micronucleus assay and alkalinesingle-cell gel electrophoresis. Food Chem.Toxicol., 48: 202-208.

Nwani, C.D., W.S. Lakra, N.S. Nagpure, R.Kumar, B. Kushwaha and S.K.Srivastava, 2010. Toxicity of herbicide

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Atrazine: Effects on lipid peroxidation andactivities of antioxidant enzymes infreshwater fish Channa punctatus (Bloch).Int. J. Environ. Res. Public Health, 7: 3298-3312.

Padmakumar, K. G., L. Bindu, V.S. Basheer,A. Gopalakrishnan and W.S. Lakra, 2010.Threatened fishes of the World: Clariasdussumieri dussumieri Valenciennes, 1840(Clariidae). Environ. Biol. Fish., 87: 297-298.

Pandey, A.K., 2010. Hypothalamo-neurosecretory system of the marineteleost, Decapterus resselli (Ruppell 1830).J. Ecophysiol. Occup. Health, 10(1-2): 71-76.

Pandey A., V. K. Dubey, R. S. Kumar, U.K.Sarkar and W.S. Lakra, 2010. New recordof Glyptothorax brevipinnis (Hora, 1923)(Pisces, Siluriformes, Sisoridae) in the RiverKen, Central India. J. . Appl. Ichthyol., DOI:10.1111/j.1439-0426.2010.01590.x

Patiyal, R.S., R.C. Sharma, P. Punia, M.Goswami and W.S. Lakra, 2010. Length-weight relationship of Tor putitora(Hamilton, 1822) from the Ladhiya River,Uttarakhand, India. J. Appl. Ichthyol. ,26(3): 472-473.

Sani, R., B.K. Gupta, U.K. Sarkar, A. Pandey,V.K. Dubey and W.S. Lakra, 2010.Length-weight relationships of 14 Indianfreshwater fish species from the Betwa(Yamuna River tributary) and Gomti(Ganga River tributary) rivers. J. Appl.Ichthyol., 26(3): 456-459.

Sarkar, U.K. and W.S. Lakra, 2011. Life historytraits of freshwater fish population andimplications on aquatic biodiversityconservation: A review. Indian J. Anim. Sci.88(4), Suppl 1: 85-97.

Shukla, D., N.S. Nagpure, R. Kumar and P.J.Singh, 2010. Assessment of genotoxicity ofdichlorvos to Mystus vittatus (Bloch) bycomet assay. Indian J. Fish., 57(2): 39-44.

Singh, A.K., A.K. Pathak and W.S. Lakra,2010. Invasion of an exotic fish- commoncarp, Cyprinus carpio L. (Actinopterygii:Cypriniformes: Cyprinidae) in the GangaRiver, India and its impacts. ActaIchthyologica., 40(1):11-19.

Singh A. K. and W S. Lakra, 2011. Risk andbenefit assessment of alien fish species ofthe aquaculture and aquarium trade intoIndia. Reviews Aquac., 3(1): 3–18.

Singh, Rajeev K., Kuldeep K. Lal, VindhyaMohindra, Peyush Punia, Rama S. Sah,Akhilesh K. Mishra, Rajesh Kumar, B.N.Mishra and W.S. Lakra, 2010. Assessinggenetic differentiation in geographicpopulations of Labeo calbasu (Hamilton,1822) using allozyme markers. Biochem.Gen., 48: 760-778.

Sinha, R.K., R.K. Sinha, U.K. Sarkar and W.S.Lakra, 2010. First record of the southernsailfin catfish, Pterogoplichthys anisitsiEigenmann & Kennedy, 1963 (Teleostei:Loricariidae), Indian J. Appl. Ichthyol., 26,606– 608.

Sood, Neeraj, Dharmendra K. Chaudhary,Gaurav Rathore, Akhilesh Singh, W.S.Lakra, 2011. Monoclonal antibodies tosnakehead, Channa striatasimmunoglobulins: detection andquantification of immunoglobulin-positivecells in blood and lymphoid organs. FishShellfish Immunol., 30: 569-575.

Srivastava, S.M., P.P. Srivastava, R. Dayaland A.K. Pandey, 2010. ThreatenedBronze Featherback, Notopterus notopterus.Fishing Chimes., 30(6): 62-63.

Srivastava, S.M., P.P. Srivastava, R. Dayal,A.K. Pandey and S.P. Singh, 2010.Induced spawning of captive stock ofthreatened Bronze Featherback Notopterusnotopterus for stock improvement andconservation. J. Appl. Biosci. 36(2):144-147.

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Swaminathan, T. R., W.S. Lakra, A.Gopalakrishnan, V.S. Basheer, B.Kushwaha and K.A. Sajeela, 2010.Development and characterization of anew epithelial cell line PSF from caudal finof Green chromide, Etroplus suratensis(Bloch, 1790). In Vitro Cellular Develop.Biol.—Animal. DOI 10.1007/s11626-010-9326-y.

Swaminathan, T.R., W.S. Lakra, A.Gopalakrishnan, V.S. Basheer, B.Khushwaha and K.A. Sajeela, 2011 .Development and characterization of afibroblastic-like cell line from caudal fin ofthe red-line torpedo, Puntius denisonii(Day) (Teleostei: Cyprinidae). Aquac. Res.,DOI number DOI: 10.1111/j.1365-2109.2011.02854.x

Tiwari, M, N.S. Nagpure, D.N. Saksena, R.Kumar., S.P. Singh, B. Kushwaha, W.S.Lakra, 2011. Evaluation of acute toxicitylevels and ethological responses underheavy metal cadmium exposure infreshwater teleost, Channa punctata (Bloch).Int. J. Aqu. Sci., 2(1): 36-47.

Tiwari, M., N.S. Nagpure, D.N. Saksena andW.S. Lakra, 2011. Metallothionein mRNAexpression in freshwater teleost, Channapunctata (Bloch) under the influence ofheavy metal, cadmium–a dose kineticstudy. Int. Aquat Res., 3: 21-29.

Tiwari, Mohit, N.S. Nagpure, D.N. Saksenaand W.S. Lakra, 2010. Time kinetic studyof metallothionein mRNA expression dueto cadmium exposure in freshwatermurrel, Channa punctata (Bloch). J.Ecophysiol. Occup. Health., 10(1-2): 85-96.

Varshney, P.K., R.K. Agrihari, S.K. Singh andA.K. Yadav, 2011. Zooplankton diversityin changing environment of River Gomtiin Lucknow. Aquacult., 11(2): 141-153.

Yadav, A.K., R.K. Mishra, S.K. Singh, P.K.Varshney, A.K. Pandey and W.S. Lakra,

2011. Induced spawning of Asian catfish,Clarias batrachus, with different doses ofsGnRH based drugs. J. Exp. Zool., India,14(1): 199-202.

Books/Book Chapters:Bhoir, K.K., S.A. Suryawanshi and A.K.

Pandey, 2011. Light microscopic as wellas ultrastructural changes in adrenalcortex of Mus norvegicus induced by sub-lethal heroin administration. In: V.K.Gupta, A.K. Verma and G.D. Singh (Eds.)Perspectives in Animal Ecology andReproduction. Vol. 7. Daya PublishingHouse, New Delhi. pp. 315-330.

Bhoir, K.K., S.A. Suryawanshi and A.K.Pandey, 2011. Light microscopic as wellas ultrastructural changes in thyroid glandof Mus norvegicus induced by sublethalheroin administration. In: V.K. Gupta,A.K. Verma and G.D. Singh (Eds.)Perspectives in Animal Ecology andReproduction. Vol. 7. Daya PublishingHouse, New Delhi. pp. 487-502.

Kulkarni, B.G., F.A. Mohamed, S.S. Kupekarand A.K. Pandey, 2011. Effects of sublethalcypermethrin, carbaryl andmonocrotophos administration on certainbiochemical parameters in muscles andnerve cord of Periplanata americana. In: V.K.Gupta, A.K. Verma and G.D. Singh (Eds.)Perspectives in Animal Ecology andReproduction. Vol. 7. Daya PublishingHouse, New Delhi. pp. 116-127.

Lakra, W.S. and U.K. Sarkar, 2010.Conservation and management of aquaticgenetic resources of India. In: Kumar Dilip,K.V. Rajendran and S. Jahageerdar, (Eds.).Bioresource management and climatechange: Studium Press (India) Pvt. Ltd. pp1-16.

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Silas, E.G., Gopalakrishnan, A.,Ramachandran, A., Anna Mercy, T.V.,Kripan Sarkar, Pushpangadan, K.R., AnilKumar, P., Ram Mohan, M.K. andAnikuttan, K.K., 2011. Guidelines forgreen certification of freshwaterornamental fish. The Marine ProductsExport Development Authority, Kochi,India. xii + 106 p.

Singh, A.K., A. K. Pathak and W. S. Lakra,2010. Mapping of invasive exotic fishspecies of the Yamuna River system inUttar Pradesh, India. In: Nishida Tom andA.E. Caton (Eds.). GIS/spatial analysis infishery and aquatic sciences. InternationalFishery GIS Society, Saitama, Japan. 4:523-534.

Popular Articles:Jena, J. K. and U. K. Sarkar, 2010. Biodiversity

conservation and management of fishgenetic resources: Current scenario andpriorities. Souvenir. 21st All India congressof Zoology and National Seminar. 21-23December, 2010, CIFRI, Barrackpore,pp.37-43.

Jena, J.K., A. Gopalakrishnan and K.K. Lal,2011. Towards conservation andsustainable utilization of aquatic geneticresources: NBFGR’s perspective and efforts.In: Souvenir of X Agricultural ScienceCongress on Soil, Plant and Animal Healthfor Enhanced and Sustainable AgriculturalProductivity, 10-12 February, 2011,NBFGR, Lucknow, P. 454.

Kupekar, S.S, B.G. Kulkarni and A.K. Pandey,2011. Hormonal control of melanophoresin mangrove crab, Charybdis lucifera.Fishing Chimes, 31(1): 106-108.

Lakra, W.S. and U.K. Sarkar, 2010. NBFGR-Marching ahead in cataloguing andconserving fish genetic resources of India.Fishing Chimes. 30(1): 102-107.

Mishra, R.K., A.K. Yadav, P.K. Varshney andA.K. Pandey, 2011. Jalkrishi vividhikaranhetu youswanshi machhalion ka prajananavom palan (in Hindi). Vigyan, 97(1): 12-18.

Sarkar, U.K. and W. S. Lakra, 2010. Smallindigenous freshwater fish species of India:Significance, conservation and utilization.Aquaculture Asia, Vol. XV(3): 34-35.

Institute PublicationsThe library facilitated in bringing out the

following publications of the Bureau:

Training Manuals:1. Bioinformatics Tools & Resources in

Fisheries Research, 2011, NBFGR/NAIP/NABG Publication.

2. Jalkrishi taknikiyan evam utpadaktavriddhi (in Hindi). 2010. NBFGR,Lucknow.

Extension Bulletins:1. Innovative approach to conservation of

fish biodiversity: The concept of state fish,2010.

2. NBFGR-NEH Region collaborations andaccomplishments, 2010.

3. Outreach activity on fish genetic resources,2010.

4. Fish cell and tissue culture, 2010.

5. Guidelines for the Import of OrnamentalFishes into India/ W. S. Lakra, RehanaAbidi, G. Rathore, A. K. Singh, Neeraj Soodand T Rajaswaminathan. 2010. 6p.

6. Guidelines for Import and Culture ofTilapia/ W. S. Lakra , A. K. Singh, G.Rathore, Rehana Abidi, TRajaswaminathan and Neeraj Sood. 2010.20p.

7. Guidelines for Culture and Breeding of

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Litopenaeus vannamei/ W.S. Lakra and A.K.Singh, G. Rathore, Rehana Abidi, T.Rajaswaminathan and Neeraj Sood. 2010.7p.

8. Guidelines on Culture of Pangasius sutchiin India/ W.S. Lakra and A.K. Singh 2010.7p.

9. Sajeev Matsya Gene Bank (in Hindi)/ R.S.Patiyal, K.K. Lal, Peyush Punia and L.K.Tyagi. 2010. p.

10. Chromosomal diversity in Indian fishes,2010.

11. Genotoxicity studies in fishes, 2010.

Newsletters and others1. NBFGR News, Vol. 8 (1-2), January-June,

2010 (Bilingual).

2. Abstract book: X Agricultural ScienceCongress on Soil, Plant and Animal Healthfor Enhanced and Sustainable AgriculturalProductivity, 10-12, February, 2011,organized at NBFGR, Lucknow.

3. Souvenir: X Agricultural Science Congresson Soil, Plant and Animal Health forEnhanced and Sustainable AgriculturalProductivity, 10-12 February, 2011,organized at NBFGR, Lucknow.

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LINKAGES

The NBFGR worked in collaboration withthe following international and nationalorganizations and agencies during the periodunder report.

National Organization/Agencies

ICAR Institutes Central Institute of Freshwater

Aquaculture, Bhubaneswar, Orissa.

Central Inland Fisheries Research Institute,Barrackpore, West Bengal.

Central Marine Fisheries ResearchInstitute, Kochi, Kerala.

Central Institute of BrackishwaterAquaculture, Chennai, T.N.

Central Institute of Fisheries Education,Mumbai, Maharastra.

Directorate of Coldwater FisheriesResearch, Bhimtal, Uttarakhand.

National Bureau of Animal GeneticResources, Karnal.

National Bureau of Plant GeneticResources, New Delhi.

National Bureau of AgriculturallyImportant Microorganisms, Mau, UP.

ICAR Complex for NEH Region, Barapani,Shillong, Meghalaya.

International Organizations/Agencies The World Fish Centre, Penang, Malaysia.

Network of Aquaculture Centers in AsiaPacific (NACA), Bangkok, Thailand.

CSIRO Marine and Atmospheric Research,Hobart, Australia.

Institute of Biodiversity, University ofGuelph, Canada.

Universities and Colleges College of Fisheries, Kerala Agricultural

University, Panangad, Cochin, Kerala.

Regional Agricultural Research Station,Kerala Agricultural University,Kumarakom, Kottayam, Kerala.

School of Industrial Fisheries, CochinUniversity of Science & Technology, Kochi.

School of Life Sciences, NEHU, Shillong,Meghalaya.

Department of Zoology, GauhatiUniversity, Guwahati, Assam.

School of Life Sciences, DibrugarhUniversity, Dibrugarh, Assam.

Department of Life Sciences, ManipurUniversity, Manipur.

School of Life Sciences, R.G. University,Itanagar, Arunachal Pradesh.

College of Fisheries, Central AgricultureUniversity, Lambuchara, Agartala,Tripura.

St. Anthony College, Shillong, Meghalaya.

University of Delhi, Delhi.

HNB Grahwal University, Srinagar-Garhwal, Uttrakhand.

Dr. B.B. Ambedkar University, Lucknow.

University of Lucknow, Lucknow.

M.P. University of Agril. and Technology,Udaipur, Rajasthan.

C.S.K. Himachal Pradesh KrishiVishwavidhyalaya, Palampur, H.P.

Central Ministries/ Departments Ministry of Agriculture, New Delhi.

Ministry of Earth Sciences, New Delhi

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Ministry of Environment and Forests, NewDelhi.

Department of Biotechnology, New Delhi

State Ministries/ Departments Department of Fisheries, Govt. of U.P.,

Lucknow.

Department of Fisheries, Govt. of Kerala.

Department of Fisheries, Govt. of AndhraPradesh.

Department of Fisheries, Govt. of Punjab.

Department of Fisheries, Govt. of Haryana.

Department of Fisheries, Govt. of M.P.,Bhopal

Department of Fisheries, Govt. of HimachalPradesh, Bilaspur.

Department of Fisheries, Govt. of Orissa.

Department of Fisheries, Govt. of Assam,Guwahati.

Department of Fisheries, Govt. ofMeghalaya, Shillong.

Department of Fisheries, Govt. ofArunachal Pradesh, Itanagar.

Department of Fisheries, Govt. of Tripura,Agartala.

Department of Fisheries, Govt. ofNagaland, Kohima.

Department of Fisheries, Govt. of Manipur,Imphal.

Department of Fisheries, Govt. of Sikkim,Gangtok.

Department of Fisheries, Govt. of Mizoram,Aizwal.

Department of Forest, Govt. of U.P.

Zoological Survey of India, Dehradun.

M.P. State Biodiversity Board, Bhopal

U.P. State Biodiversity Board, Lucknow

A.P. State Biodiversity Board, Hyderabad

National Informatics Centre, Delhi andLucknow.

U.P. Remote Sensing Application Centre,Lucknow.

Non-Government Organizations Aquatic Biodiversity Conservation Society,

Lucknow.

Other Organizations National Institute of Oceanography,

Panaji, Goa.

Central Drug Research Institute,Lucknow.

Central Institute of Medicinal andAromatic Plants, Lucknow.

Kerala Forest Research Institute, Trichur,Kerala.

Indian Institute of Remote Sensing,Dehradun.

Sanjay Gandhi Post-Graduate Institute ofMedical Sciences, Lucknow.

Marine Products Export DevelopmentAuthority, Kochi.

Indian Institute of Toxicology Research,Lucknow.

Centre for Cellular & Molecular Biology,Hyderabad.

Wildlife Institute of India, Dehradun.

Fisheries Survey of India, Mumbai.

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LIBRARY AND INFORMATION SERVICES

using Internet were searched and arranged tosuit the requirements of users. Contents list ofthe selected journals were brought out andcirculated to all divisions and also sent to otherfisheries institutions. List of the books added tothe library has also been brought out onquarterly basis. The users of the libraryextensively used the Consortium of E-Resourceson Agriculture (CERA), to access the journalsrelated to agriculture and allied sciences.

Technical Reports and ReprographyServices

The library unit provided technical supportto bring out other departmental publications.The unit also attended to questionnaires onBureau’s infrastructure and other facilities. Theunit continued active reprography services.Comb binding, spiral binding, electro-databinding and lamination facilities fordepartmental reports were also provided.

Exchange ServicesThe Library continued exchange

relationship and resource sharing with leadingNational and International Research Institutesand Development organizations. To keepabreast of the activities of the Bureau, the librarysent the NBFGR Annual Report 2009-2010 andNBFGR Newsletters to various institutions andorganizations including, Universities, StateFisheries Departments, FFDAs, Krishi VigyanKendras, Entrepreneurs and Fish Farmers.

The NBFGR Library and DocumentationUnit acts as a repository of literature andinformation and provides latest information inthe field of fish diversity conservation, fishgenetics, fisheries and related aspects.

Resource DevelopmentThe library added a total of 339 documents

comprising 139 books, 149 serials and 51 annualreports. Now, the library has the total collectionof 5750 books, 2254 bound volumes of journals,3192 serials and 2261 reprints. The library hassubscribed 32 international journals and 46Indian journals. In addition to these, 49 currentjournals were received on gratis/exchangebasis. The total expenditure incurred by thelibrary during the year under report was Rs.17,99,131.

Library AutomationThe NBFGR library is operating in fully

automated environment. The various activitiesof library have been computerized usingintegrated library management software Libsys.The record of books, journals, maps, etc. wereentered in the database. Barcoding of books,periodicals and maps for automated circulationis under active process. Online Public Accesscatalogue is made available for the library users.

Information and Reference ServicesThe references from different databases

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STAFF ACTIVITIES

JoiningDr. J.K. Jena, Head, Aquaculture Production

and Environment Division, CentralInstitute of Freshwater Aquaculture,Bhubaneswar joined as the Director,NBFGR on 6.10.2010.

Shri Sunil Kumar, Administrative Officer, IISR,Lucknow joined as Administrative Officer,NBFGR, Lucknow on 4.02.2011.

Dr. Rajeev Kr. Singh, Scientist (Sr. Scale),NBFGR was appointed as Sr. Scientist,NBFGR. He joined on 11.03.2011.

Shri Sandeep was appointed as Jr.Stenographer. He joined on 23.06.2010.

Dr. Vikas Sahu was appointed as LaboratoryTechnician (T-3). He joined on 9.07.2010.

The NBFGR staff members welcome thenew members in the family.

RelievingDr. W.S. Lakra, Director, NBFGR was relieved

on 30 August, 2010 to enable him to joinas Director, Central Institute of FisheriesEducation, Mumbai.

Dr. R.S. Patiyal, Technical Officer (T-6),NBFGR, Lucknow was relieved on16.03.2011 to enable him to join as SeniorScientist at Champawat Field Centre ofDirectorate of Cold Water FisheriesResearch, Bhimtal, Nainital.

ResignationDr. Sanjeev Kr. Srivastava, Scientist (Sr. Scale)

resigned from the service and hisresignation was accepted by the ICARw.e.f. 15.05.2006.

PromotionsThe following staff members were

promoted to the next higher grade:

Shri Satyavir Chaudhary was promoted fromT-3 to T-4 w.e.f. 26.12.2005.

Dr. S.K. Srivastava, Former Technical Officerwas promoted from T-6 to TechnicalOfficer T (7-8) w.e.f. 12.12.2008.

Shri Ved Prakash was promoted from T-4 toTechnical Officer (T-5) w.e.f. 1.01.2009.

Shri Akhilesh Kr. Mishra was promoted fromT-4 to Technical Officer (T-5) w.e.f.16.07.2009.

Dr. (Mrs.) Ranjana Srivastava (Sinha) waspromoted from T-4 to Technical Officer (T-5) w.e.f. 27.07.2009.

Shri Ravi Kumar was promoted from T-4 toTechnical Officer (T-5) w.e.f. 04.04.2010.

Shri Sanjay Kr. Singh was promoted from T-4to Technical Officer (T-5) w.e.f. 12.06.2010.

Shri Amit Singh Bisht was promoted from T-4to Technical Officer (T-5) w.e.f. 26.06.2010.

Shri B.N. Pathak was promoted from T-3 to T-4 w.e.f. 12.04.2010.

Shri R.K. Shukla was promoted from T-3 to T-4 w.e.f. 1.01.2009.

Shri Om Prakash was promoted from T-2 to T-3 w.e.f. 1.05.2010.

Shri Ajay Kr. Singh was promoted from T-5 toTechnical Officer (T-6) w.e.f. 26.09.2010.

Shri Navin Kumar was promoted fromAssistant to Asstt. Administrative Officerw.e.f. 29.12.2010.

Shri Swapan Debnath was promoted from Sr.Clerk to Assistant w.e.f. 29.12.2010.

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Shri S.N. Srivastava was promotedfrom Sr. Clerk to Assistant w.e.f.29.12.2010.

Shri P.K. Awasthi was promoted from Sr. Clerkto Assistant w.e.f. 29.12.2010.

Shri Shreelal Prasad was promoted from Jr.Clerk to Sr. Clerk w.e.f. 29.12.2010.

Shri Vinay Kr. Srivastava was promoted fromJr. Clerk to Sr. Clerk w.e.f. 10.01.2011.

Financial up-gradationShri P.C. Verma, Jr. Clerk in the Pay Band –I

`5200-20200+Grade Pay of `2400/- wasgranted 2nd financial up-gradation of PB-Iin the grade pay of `2800/- under MACPScheme w.e.f. 01.08.2010.

The NBFGR family extends heartycongratulations to all the above staff membersfor their success.

Staff Welfare Activities

Institute Joint Staff CouncilThe Institute Joint Staff Council with the members mentioned below, was operative at the

Bureau during the period under report and considered the matters of common interest.

Official side1. Director, NBFGR - Chairman

2. Dr. N.S. NagpureHead of Division and HoO - Member

3. Dr. (Mrs) Vindhya Mohindra, Principal Scientist - Member

4. Dr. P.P. Srivastava, Senior Scientist - Member

5. Sh. Ashish Srivastava, AF&AO - Member

6. Sh. P. Lal, Assistant Administrative Officer - Member

7. Sh. Ajay Kumar Singh, T-6 - Member

Staff side1. Sh. Indrajit Singh, SSS - Secretary

2. Sh. S. N. Srivastava, Assistant - Member representative of CJSC

3. Sh. Amit Singh Bisht, T-5 - Member

4. Sh. Akhilesh Kumar Mishra, T-5 - Member

5. Sh. Santosh Kumar Singh, Jr. Clerk - Member

6. Shi. Anil Kumar, SSS - Member

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Staff Welfare Fund SchemeThe Staff Welfare Fund Scheme with the following members was at the Bureau during the

period under report and considered the matter for welfare of the staff.

1. Dr. N.S. Nagpure - ChairmanHead of Division

2. Dr. Sudhir Raizada - MemberPr. Scientist

3. Dr. L.K. Tyagi - MemberScientist (Senior Scale)

4. Shri Ashish Srivastava - MemberAF & AO

5. Mrs. Mamta Chakraborty - MemberPersonal Assistant (Lady Representative)

6. Shri Indrajit Singh, SSS - Member(Secretary, IJSC)

7. Mr. Laxchman Prasad - MemberSSS

8. Shri Panchoo Lal - Member-SecretaryAssistant Administrative Officer

Women’s CellThe Women’s Cell has been constituted at NBFGR, Lucknow with the following members:

1. Dr. (Mrs) Rehana Abidi - Head of the CellPrincipal Scientist

2. Dr. (Mrs.) Vindhya Mohindra - Member-Secretary Principal Scientist

3. Mrs. Reeta Chaturvedi - Member T-5

4. Mrs. Mamta Chakraborty - MemberPersonal Assistant

5. Shri Anil Kumar - MemberSSS

Grievance CellThe Grievance Cell with the following members has been constituted at NBFGR, Lucknow

under report period for addressing grievances in regard to service matters, etc., of staff:

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Nominated members (Official side)Dr. J.K. Jena, Director, NBFGR : Chairman

Dr. N.S. Nagpure, Head of Division, NBFGR : Member

Dr. A.K. Singh, Pr. Scientist, NBFGR : Member

Shri Ashish Srivastava, AF & AO, NBFGR : Member

Shri P. Lal, Asst. Adm. Officer, NBFGR : Member-Secretary

Elected members (Staff side)Dr. G. Rathore, Sr. Scientist, NBFGR : Member (Scientific)

Shri S.N. Srivastava, Assistant : Member (Administrative)

Shri Satyavir Choudhary, T-4 : Member (Technical)

Shri B.B. Bajpai, SSS : Member (Supporting)

Management CommitteeThe Institute Management Committee was represented by the following members nominated byDirector General, ICAR, New Delhi.

1. Director, NBFGR : Chairman

2. Dr. Madan Mohan, ADG (Marine Fy.), ICAR, New Delhi : Member (ICAR)

3. Dr. V.V. Singh, Principal Scientist, CMFRI, Centre, Mumbai : Member

4. Dr. T. Mahapatra, Principal Scientist, NRCPB, New Delhi : Member

5. Dr. M.S. Tantia, Principal Scientist, NBAGR, Karnal : Member

6. Dr. P.C. Agarwal, Principal Scientist, NBPGR, New Delhi : Member

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DISTINGUISHED VISITORS

The following distinguished visitors visitedthe Institute during the year:

Shri B.L. Joshi, His Excellency, Governor ofUttar Pradesh.

Shri Harish Rawat, Hon’ble Minister of Statefor Agriculture, Government of India.

Dr. S. Ayyappan, Secretary, DARE, Ministryof Agriculture, Govt. of India and DirectorGeneral, ICAR, New Delhi.

Dr. R.S. Paroda, Former Director General,ICAR, New Delhi.

Dr. Panjab Singh, Former Director General,ICAR, New Delhi.

Dr. Mangala Rai, Former Director General,ICAR, New Delhi.

Dr. S. N. Dwivedi, Former Additional Secretary,Govt. of India.

Dr. P.V. Dehadrai, Former DDG (Fisheries),ICAR, New Delhi.

Dr. C.D. Mayee, Chairman, ASRB, New Delhi.Dr. R.B. Singh, President, NASS, New Delhi.Shri G.B. Patnaik, Principal Secretary, H.E.,

Governor of UP.Dr. Kirti Singh, Former Chairman, ASRB, New

Delhi.Dr. K. Gopakumar, Former DDG (Fisheries),

ICAR, New Delhi.Dr. B. Meenakumari, DDG (Fisheries), ICAR,

New Delhi.Dr. H. P. Singh, DDG (Horticulture), ICAR,

New Delhi.Dr. S.A.H. Abidi, Former Member, ASRB, New

Delhi.Dr. N. K. Tyagi, Member, ASRB, New Delhi.Dr. S.L. Mehta, Former Vice-Chancellor,

MPUAT, Udaipur.Dr. A.N. Mukhopadhyaya, Former Vice-

Chancellor, Assam Agriculture University.Dr. B. Hanumaiah, Vice-Chancellor, B. R.

Ambedkar University, Lucknow.Dr. B. S. Bisht, Vice-Chancellor, G.B. Pant

University of Ag. & Tech., Pantnagar.Dr. Lalji Singh, Former Director, CCMB,

Hyderabad.Dr. Dilip Kumar, Former Director, CIFE,

Mumbai.Dr. M. C. Sharma, Director, IVRI, Izatnagar,

Bareilly.Prof. T.J. Pandian, Former National Professor,

ICAR.Dr. Anwar Alam, Secretary, NASS.Dr Bengali Baboo, ND, NAIP, ICAR, New

Delhi.Dr. A.D. Diwan, Former ADG (Marine Fishery),

ICAR, New Delhi.Dr. Madan Mohan, ADG (Marine Fishery),

ICAR, New Delhi.Dr. P.K. Seth, Chief Executive, Biotech Park,

Lucknow.Dr. C.S. Nautiyal, Director, NBRI, Lucknow.Dr. P.K. Joshi, Director, NAARM, Hyderabad.Dr. H. Ravishankar, Director, CISH,

Rehmankheda, Lucknow.Dr. R.L. Yadav, Director, IISR, Lucknow.Dr. P. Das, Former Director, NBFGR.Dr. K.K. Vaas, Former Director, CIFRI,

BarracporeDr. D. K. Arora, Director, NBAIM, Mau.Dr. P.S. Pathak, Former Director, IGFRI, Jhansi.Dr. A.P. Sharma, Director, CIFRI, Barrackpore.Dr. A. G. Ponniah, Director, CIBA, Chennai.Dr. P.C. Mahanta, Director, DCFR, Bhimtal.Dr. Mathura Rai, Former Director, IIVR,

Varansi.

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LIST OF PERSONNEL

Research ManagementDr. J.K. Jena - Director

Scientific Staff1. Dr. N.S. Nagpure - Head of Division2. Dr. K.K. Lal - Head of Division3. Dr. Peyush Punia - Head of Division4. Dr. (Mrs.) Rehana Abidi - Principal Scientist5. Dr. A. Gopalakrishnan - Principal Scientist (NBFGR Cochin Unit)6. Dr. A.K. Pandey - Principal Scientist7. Dr. Sudhir Raizada - Principal Scientist8. Dr. S.P. Singh - Principal Scientist9. Dr. A.K. Singh - Principal Scientist10. Dr. (Mrs) Vindhya Mohindra - Principal Scientist11. Dr. P.K. Varshney - Sr. Scientist12. Dr. Ravindra Kumar - Sr. Scientist13. Dr. Basdeo Kushwaha - Sr. Scientist14. Dr. U.K. Sarkar - Sr. Scientist15. Dr. Neeraj Sood - Sr. Scientist16. Dr. P.P. Srivastava - Sr. Scientist17. Dr. V.S. Basheer - Sr. Scientist (NBFGR Cochin Unit)18. Dr. Gaurav Rathore - Sr. Scientist19. Dr. Mukunda Goswami - Sr. Scientist20. Dr. Parvata Kumar Pradhan - Sr. Scientist21. Dr. Rajeev Kumar Singh - Sr. Scientist22. Mrs. Poonam Jayant Singh - Scientist (Selection Grade)23. Shri Ajey Kumar Pathak - Scientist (Selection Grade) (on study leave)24. Dr. Lalit Kumar Tyagi - Scientist (Sr. Scale)25. Dr. Mahender Singh - Scientist (Sr. Scale)26. Dr. T. Rajaswaminathan - Scientist (Sr. Scale) (NBFGR Cochin Unit)27. Shri Karan Veer Singh - Scientist (Sr. Scale)28. Dr. (Mrs.) Divya P.R. - Scientist (NBFGR Cochin Unit)29. Shri A. Kathirvelpandian - Scientist (on study leave at NBFGR

Cochin Unit)

(As on 31 March, 2011)

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Technical Staff1. Shri Rajesh Dayal - Field Officer T(7-8)2. Shri S.M. Srivastava - Field Officer T(7-8)3. Shri A.K. Yadav - Technical Officer (T-7-8)4. Shri A.K. Mishra - Electrical Foreman (T-6)5. Shri S.P. Singh - T-66. Shri Amar Pal - Technical Officer (T-6)7. Shri Babu Ram - T-68. Shri Ajay Kumar Singh - Field Surveyor (T-6)9. Shri Mohd. Gyas - Driver (T-5)10. Mrs. Reeta Chaturvedi - Computer Operator (T-5)11. Shri Ramashankar Sah - Technical Officer (T-5)12. Shri Subhash Chandra - Technical Officer (T-5)13. Shri Ved Prakash - Technical Officer (T-5)14. Shri Akhilesh Kr. Mishra - Technical Officer (T-5)15. Dr. (Mrs.) Ranjana Srivastava - Technical Officer T-516. Shri Ravi Kumar - Technical Officer T-517. Shri S.K. Singh - Technical Officer T-518. Shri Amit Singh Bisht - Technical Officer T-519. Shri S.K. Upadhyay - T-420. Shri Satyavir Chaudhary - Senior Library Asst. (T-4)21. Shri R.K. Shukla - Sample Sorter (T-4)22. Shri B.N. Pathak - Gestetner Operator (T-4)23. Shri B.K Rao - Sample Sorter (T-II-3)24. Shri Samarjit Singh - Driver (T-3)25. Shri Om Prakash - Driver (T-3)26. Shri Rajesh Kumar - Laboratory Asst. (T-3)27. Shri Om Prakash-II - Driver (T-3)28. Dr. Vikash Sahu - Laboratory Technician (T-3)29. Shri Madan Lal - Farm Technician (T-2)30. Shri Raj Bahadur - Lab. Technician (T-2)31. Shri Gulab Chandra - Electrician (T-2)32. Shri K.K Singh - Jr. Field Asst. (T-2)33. Shri Sree Ram - Laboratory Asst. (T-2)34. Shri P.C. Jaiswar - T-235. Shri Ram Bharose - T-2

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Administrative Staff1. Shri Sunil Kumar - Administrative Officer2. Shri Panchoo Lal - Asstt. Administrative Officer3. Shri Navin Kumar - Asstt. Administrative Officer4. Smt. Mamta Chakraborty - Personal Assistant5. Shri Tej Singh Seepal - Assistant6. Shri Jogendra Singh - Assistant7. Smt. Kaneez Fatima - Assistant8. Shri Swapan Debnath - Assistant9. Shri S.N. Srivastava - Assistant10. Shri P.K. Awasthi - Assistant11. Shri Sajivan Lal - Senior Clerk12. Shri Vinay Kumar Srivastava - Senior Clerk13. Shri Sreelal Prasad - Senior Clerk14. Shri Ram Sakal - Jr. Stenographer15. Shri Sandeep - Jr. Stenographer16. Shri Santosh Kumar Singh - Jr. Clerk17. Shri Ram Baran - Jr. Clerk18. Shri P.C. Verma - Jr. Clerk19. Shri Rajan Kr. Malhotra - Jr. Clerk

Skilled Supporting Staff1. Shri Laxman Prasad - Skilled Support Staff2. Shri Dukhi Shyam Deo - Skilled Support Staff3. Shri Anil Kumar - Skilled Support Staff4. Shri Indrajit Singh - Skilled Support Staff5. Shri Prahalad Kumar - Skilled Support Staff6. Shri Chhote Lal - Skilled Support Staff7. Shri Dinesh Kumar - Skilled Support Staff8. Shri Balram Babu Bajpai - Skilled Support Staff9. Shri Ashok Kumar Awasthi - Skilled Support Staff10. Shri Sidhnath - Skilled Support Staff11. Smt. Sabita Devi - Skilled Support Staff12. Shri Ram Lakhan - Skilled Support Staff13. Shri Sunit Kumar - Skilled Support Staff14. Shri Jai Narain Tiwari - Skilled Support Staff15. Shri Mahesh Chandra - Skilled Support Staff16. Shri Anwar - Skilled Support Staff17. Shri Sanjay Kumar - Skilled Support Staff18. Smt. Seema Devi - Skilled Support Staff19. Shri Ashok Kumar - Skilled Support Staff

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NBFGR Cochin Unit

A Research Unit of the Bureau is functioning in the campus of Central MarineFisheries Research Institute (CMFRI), Cochin, Kerala. This unit is carrying outresearch activities pertaining to genetic characterization, conservation andcataloguing of the vast fish genetic resources of marine and brackishwater ecosystemsof the country, as well as, of endemic freshwater fish species from the Western Ghats– the megabiodiversity ‘hotspot’.

Address : Scientist–in-ChargeNBFGR Cochin UnitCMFRI CampusPost Box No. 1603Ernakulam North P.O.Kochi – 682 018, Kerala.Telefax : 0484-2395570E-mail : nbfgrcochin@vsnl.net nbfgrcochin@eth.net

APPENDIX-I

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Aquaculture Research & Training Unit, Chinhat

An Aquaculture Research & Training Unit of the Bureau is functioning at Chinhat,Lucknow. This unit is carrying out human resource development activities includingpractical training programmes and fishery advisory services pertaining to fish culture,induced breeding, quality fish seed production, hatchery management and nursery pondmanagement.

Address : Scientist–in-ChargeNBFGR Aquaculture Research & Training UnitMalhore Road, ChinhatLucknow-227 105, U.P.Telefax : 0522-2815848E-mail : director@nbfgr.res.in

APPENDIX-II