journal of indian veterinary association december 2012, 10 (3)

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Editorial Board

INDEXED IN CAB INTERNATIONAL, EBSCO, HINARI AND INDEX COPERNICUS INTERNATIONAL

Journal ofIndian Veterinary AssociationKerala

Journal ofIndian Veterinary AssociationKerala

Vol. 10 Issue 3 December 2012

ISSN 0975-5195

JIVA

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< < J. Ind. Vet. Assoc., Kerala. 10 (3)02

CONTENTS

Vol. 10 Issue 3 December 2012 < < JIVA 03

RESEARCH ARTICLES 5 - 51

CLINICAL REPORTS 52 - 60

1. BETA CASEIN A1A2 POLYMORPHISM AND MILK YIELD IN VECHUR, KASARGODE DWARF AND CROSSBRED CATTLEE. M. Muhammed and M. Stephen 5

2. A HISTOLOGICAL STUDY OF THE STRUCTURAL CHANGESIN THE PANCREAS OF DIABETIC RATS B. Dhanush Krishna and Suguna Rao

3. THERAPEUTIC POTENTIAL OF AFANIL® AGAINSTBLOAT AND FOR EARLY RESTORATION OF RUMENFUNCTION IN BOVINESS. U. Digraskar, V. D. Muley, K. Ravikanth, M. Dandale and S. Maini

4. OPTIMIZATION OF PCR THROUGH MANIPULATIONOF CYCLE TIMES AND INCLUSION OF FORMAMIDES. Pramod, A. P. Usha, T. Venkatachalapathy and K. C. Raghavan

5. MILK FEEDING STRATEGIES FOR IMPROVING GROWTH PERFORMANCE IN CROSSBRED DAIRY CALVESA. J. Flamy and Joseph Mathew

6. MORPHOGENESIS OF MEDULLA OBLONGATAIN GOAT FOETUSESK. M. Lucy, K. R. Harshan, J. J. Chungath and N. Ashok

7. ROLE OF FECUNDITY GENES IN PROLIFICACYOF SMALL RUMINANTSAsha Abraham and Naicy Thomas

8. ECONOMIC ANALYSES OF THREE DIFFERENT METHODSOF PREGNANCY DIAGNOSIS IN DAIRY CATTLE OF KERALAArun Kurian, Josephine Francis and Jiss Job

9. PARTICIPATION OF SELF HELP GROUP MEMBERS ANDNON- MEMBERS IN PANCHAYATI RAJ SYSTEM -A COMPARATIVE STUDYAnu George, P. J. Rajkamal and R.S. Jiji

10. THERMAL STRESS IN DAIRY CATTLEA. Prasad, E.M. Muhammed, A. Kannan and T.V. Aravindakshan

10

15

19

24

28

34

38

42

45

11. SUCCESSFUL MANAGEMENT OF CHYLOTHORAXIN A DOG- A CASE REPORTG. Vijayakumar, S. Sivaraman, E. Venkatesakumar and M. Subramanian 52

CONTENTS

12. SURGICAL MANAGEMENT OF TESTICULAR SEMINOMA -A CASE REPORTLaiju. M. Philip and M. Ranjith Mohan 55

13. A REPORT ON THE OCCURANCE OF Hymenolepis anatineIN DUCKS UNDER BACKYARD SYSTEMG. Jyothimol, K. Syamala, M.N. Priya, C.K. Deepa, K.G. Ajithkumar,

Ajith Jacob George and Reghu Ravindran 57

14. MANAGEMENT OF SUBCUTANEOUSEMPHYSEMA IN A HORSEMir Aamir Ali and H.S. Mahesha 59

15. IMPACTS OF CLIMATIC CHANGE INANIMAL HUSBANDRY- ARE WE PREPARED?S.S. Rani 61

16. VACCINES FROM OUR GARDENTincy Mary John, N.M. Shah, B.S. Chandel and H.C. Chauhan 66

17. BIODIESEL PRODUCTION FROM ANIMAL FATS-AN EVER GREEN TECHNOLOGY FORTHE FUTURE ENERGY SECURITYJohn Abraham and Ramesh Saravana Kumar 72

GENERAL ARTICLES 61 - 77

ASSOCIATION NEWS 78 - 80

The editor/editorial board and referees are in no way responsible individually orcollectively for the views, data and technical details presented in the contributed papers

J. Ind. Vet. Assoc., Kerala. 10 (3) < <04


RESEARCH ARTICLE

ABSTRACT

â-Casein (â-CN) is the second most

abundant protein in cow's milk that contains

209 amino acids. Among the twelve â-CN

variants identified A1 and A2 are the common

types and others are very rare. A1 variant has

histidine at position 67 of the amino acid

sequence while A2 possess proline at this

position- this polymorphism has attracted much

public health attention, since consumption of

A1 milk is reported to cause various illness. The

high producing Bos taurus cattle are found to

possess more A1 allele. The present study was

undertaken to assess the relationship between

â-CN A1/A2 polymorphism with milk yield in

Vechur, Kasargode and Crossbred cattle. The

average daily milk yield was 1.27±0.05 kg in

Vechur cattle and the peak yield of crossbred

cattle was 10.71±1.046 kg. The average daily

milk yield for A1A2 genotype in Vechur cattle

was 1.52±0.08 kg and 1.14±0.04 kg for A2A2

genotype and the difference in the milk yield of

the two genotypes was significantly different

1 *PhD scholar, Email:[email protected] 2Professor at Department of Animal Breeding, Genetics and

Biostatistics, College of Veterinary and Animal Sciences,

Mannuthy, Thrissur

BETA CASEIN A1A2 POLYMORPHISM AND

MILK YIELD IN VECHUR, KASARGODE DWARF

AND CROSSBRED CATTLE*

1 2E. M. Muhammed and M. Stephen

College of Veterinary and Animal Sciences, Mannuthy, Thrissur

*Part of MVSc thesis of the first author submitted at Kerala Agricultural University

(p=0.05). In crossbred cattle the peak yield (kg)

of A1A1 genotype was 14.64±3.181,

8.54±0.194 for A1A2 genotype and

9.09±0.125 for A2A2 genotype. The A1A1

genotype in crossbred cattle showed

significantly higher average peak yield

compared to other genotypes and difference

between A1A2 and A2A2 genotypes were not

significantly different (p=0.05). Thus it can be

concluded that selection for enhancing milk

production may increase the frequency of

harmful A1 allele in bovine population.

Key words: Beta casein gene A1and A2,

Vechur cattle, AS-PCR, Milk yield.

INTRODUCTION

India has vast resources of livestock. It

plays a vital role in the Indian economy and in

improving the socio economic conditions of rural

masses. Agriculture and livestock sector share

16.74 percent and 4.36 percent in the economy of

India. As per Livestock census 2003, India has

total livestock population of 485 million with

185.2 million cattle and 97.2 million buffalo which

are 65 percent and 34 percent respectively of total

bovine population (283.1million). In Kerala 96

percent of livestock population is cattle producing

98.50 percent of total milk (Anon, 2009). More

than 82 percent of the cattle in Kerala are

crossbreds except indigenous breeds like Vechur.

Vechur cattle once thought to be extinct due to

extensive crossbreeding are the first native cattle of

Kerala to be approved as a distinct breed and the

smallest breed in the world. Vechur cattle are

maintained by a few farmers and at Kerala

Veterinary and Animal Sciences University.

Milk is a common source of animal protein

and associated micro elements for vegetarians.

Cow's milk contains two major protein groups:

caseins and whey proteins and out of which

caseins account for 80 percent of milk proteins.

Bovine milk contains four caseins: alpha S1

(CSN1S1 39-46 percent of total casein), alpha S2

(CSN1 S2 8-11 percent), beta (CSN2, 25-35

percent), and kappa (CSN3 8-15 percent). â-

Casein (â-CN) is the second most abundant protein

in cow's milk that contains 209 amino acids.

Bovine â-CN gene belongs to the cluster of four

casein genes located on chromosome 6. There are

12 genetic variants of â-CN: A1, A2, A3, B, C, D, F,

H1, H2, I and G out of which A1 and A2 are the

most common. So the present study was

undertaken, realising the importance of â-CN gene

polymorphism in discovery of markers linked to

economically important traits such as milk

production.

MATERIALS AND METHODS

Blood samples were collected from 72

Vechur cattle (60 from KVASU farm and 10 from

Vechur Conservation Trust, Vechur), 14 Kasargode

dwarf cattle (KVASU farm) and 100 crossbreds

from different parts of Kerala viz, Kozhikode (22),

Kannur (15), Malappuram (14), Wayanad (15),

Thrissur (24) and Kottayam (10).

Collection of Samples

About 5 ml of blood was collected from each

animal in a sterile 15 ml polypropylene centrifuge

tube (vacutainer) containing Ethylene Diamine

Tetra Acetic acid (EDTA) as anticoagulant (1 mg/ml

of blood). The samples were brought to the 0 0

laboratory at 4 C and stored at -20 C until DNA

extraction.

AS-PCR amplification

There are 12 alleles (A1, A2, A3, B, C, D, F,

G, H1, H2, I and G) of â-CN in bovines. Of all the

12 â-CN variants all except A2 produce BCM-7

upon enzymatic digestion in the gut. Among

BCM-7 yielding variants A1 is the predominant

type and other alleles are very rare. Therefore in

this study alleles are classified as A1 and A2 only.

Based on this, two allele specific reverse primers

each one matched to one of two alleles at 3' end (G

for â-CN A2 and T for â-CN A1) along with a

common forward primer were used (Keating et.

al., 2008). The ASPCR products were checked

electrophoretically using 2 percent agarose.

Statistical analysis

The allelic and genotype frequencies at

A1/A2 locus were calculated by direct counting

method and the variations of the allelic

frequencies among the three groups were

analyzed by the Chi-square test of significance as

described by Snedecor and Cochran (1994)

considering the allelic frequencies in a 2×2 table.

Collection of data

The average daily milk yield of Vechur

cattle was collected from the records maintained


in the farm. For crossbred animals, the peak yield

was obtained from the farmers. The milk yield of

Kasargode dwarf cattle were not recorded as the

number of observations was very less. There were

only two genotypes (A1A2 and A2A2) in Vechur

cattle and three genotypes- A1A1, A1A2 and A2A2

in crossbred cattle. The difference in milk yield of

the genotypes was compared by t-test and

ANOVA.

RESULTS AND DISCUSSION

Genotype and Allele Frequencies of â-CN

The allele and genotype frequencies of â-

CN A1A2 in Vechur, crossbred and Kasargode

dwarf cattle are presented in Table 4.1. In Vechur

cattle the genotype frequencies of A1A1, A1A2

and A2A2 were observed as 0, 0.34 and 0.66

whereas in crossbred cattle the corresponding

frequencies were 0.32, 0.28 and 0.40 respectively.

Of the 72 Vechur cattle typed, none of the animals

were of A1A1 genotype. No A1A1 genotypes were

found in Kasargode dwarf animals also and the

corresponding frequencies in Kasargode dwarf

cattle was recorded as 0.79 (A1A2) and 0.21

(A2A2). The frequencies for A1 and A2 alleles in

Vechur cattle were noted as 0.2 and 0.80

respectively and for crossbreds the frequencies

were recorded as 0.46 and 0.54, respectively. In

the Kasargode dwarf cattle A1 and A2 allele

frequencies were obtained as 0.39 and 0.61

respectively.

â- CN Polymorphism and Milk Yield

The average daily milk yield of 50 Vechur

cows were found to be 1.27±0.07 kg and the

average peak yield of 100 crossbred cattle were

found to be 10.71±1.05 kg. â-CN A1/A2

polymorphism had significant effect on average

daily milk yield in the Vechur cattle and peak yield

in crossbred cattle (Table 4.2). In t test, Vechur

cattle carrying A1A2 genotype showed a

significantly higher average daily milk yield

(1.52±0.08 kg) compared to A2A2 genotype

(1.14±0.04 kg). ANOVA showed significant

difference between two genotypes in crossbreds.

Crossbred cattle carrying A1A1 genotype showed

significantly higher average peak yield

(14.64±3.181 kg) whereas A1A2 and A2A2

genotypes had lower average peak yield

(8.54±0.019 kg and 9.09±0.125 kg respectively).

The peak yields of A1A2 and A2A2 were not

significantly different while the peak yield of A1A1

genotype was significantly higher than the other

two genotypes.

Where a, b, c are allelic frequencies in different groups


Means with different superscript in a row

differ significantly (p=0.05) Figures in parenthesis

denotes number of observations (2008) in Italian

Holstein Friesian and Nilsen et.al. (2009) in

Norwegian Red cattle.A higher milk yield was

reported for heterozygous A1A2 animals by Ng-

Kwai-Hang et.al. (1986) and Ojala et.al. (1997) in

Holstein Friesian cattle. In contrast, milk yield was

not associated with â-CN A1/A2 polymorphism as

reported by Sarbour et.al. (1996) in Canadian

Holstein and Boettcher et.al. (2004) in Holstein

Friesian cattle.

As observed in the present study, selection

for increasing milk yield may contribute for the

higher proportion of undesirable A1 alleles in the

population (Ron et.al. 1994 and Velmala et.al.

1995). Considering the public health implication,

adequate weightage should be given to select bulls

with A2A2 genotype while making selection for

increasing milk yield of crossbreds. The proportion

of A2 allele is high in Vechur cattle. Based on the

present results, careful breeding strategies can be

adopted to develop a Vechur herd/breed with

A2A2 genotype and its milk can be marketed at a

premium price.

ACKNOWLEDGEMENTS

The authors are thankful to the Dean,

College of Veterinary and Animal Sciences,

Mannuthy, Director, Department of Animal

Breeding, Genetics and Biostatistics, College of

Veterinary and Animal Sciences, Mannuthy,

Thrissur, Kerala and Vechur Conservation Trust,

Vechur, Kerala, for providing facilities for the

study.

REFERENCES

Anand, L. F. 2009. Leptin gene polymorphism in

Vechur and Crossbred cattle of Kerala.

MVSc. Thesis, Kerala Agricultural University,

Thrissur, p. 79.

Anon. 2009. Annual report, Department of Animal

Husbandry and Dairying, Government of

India, New Delhi. Available:http://

www.dahd.nic.in.[15 Feb 2010].

Boettcher, P.J., Caroli, A Stella, A., Chessa, S.,

Budelli, E., Canavesi, F., Ghiroldi, S. and

Pagnacco, G. 2004. Effects of Casein

Haplotypes on Milk Production Traits in

Italian Holstein and Brown Swiss Cattle.

J. Dairy Sci. 87:4311-4317.

Chandran, R. R and Stephen, M. 2010. Evaluation

of lactation milk yield in crossbred cattle.

Ind. Vet. J. 87:363-364.

Comin, A., Cassandro, M., Chessa, S., Ojala, M.,

Dal Zotto, R., De Marchi, M., Carnier, P. and

The recorded average daily milk yield of

Vechur cattle was 1.27±0.07 kg in the present

study against 2.17 kg reported by Girija (1994), 2.2

kg reported by Iype and Venketachelapathy

(2001). In crossbred cattle the peak yield was

recorded as 10.71±1.046 kg. The 305 day milk

yield of crossbred cattle of Kerala is reported as

2406 kg by Chandran and Stephen (2007) and

2106 kg by Anand (2009).

Significantly increased level of milk yield

obtained in A1A1 crossbred cattle is in line with

findings of Ron et.al. (1994) in Israeli Holstein cows

and Velmala et.al. (1995) in Finnish Ayrshire cattle.

Higher milk production from A2 allele is reported

by Ng-Kwai-Hang et.al.(1990) in Holstein Friesian,

Ikonen et.al. (1999) in Finnish Ayrshire, Freyer

et.al. (1999) in Holstein Friesian, Comin et.al.


Gallo, L. 2008. Effects of composite â-casein

genotypes on milk coagulation, quality and

yield traits in Italian Holstein cows. J. Dairy

Sci. 91:4022-4027

Freyer, G., Konig, S., Fischer, B., Bergfeld, U. and

Cassell, B.G. 2008. Crossbreeding in dairy

cattle from a German perspective of the past

and today. J. Dairy. Sci. 91:3725-3743.

Freyer, G., Liu, Z., Erhard, G.T. and Panicke, L.

1999. Casein polymorphism and relation

between milk production traits. J. Anim.

Breed. Genet. 116: 87-97.

Girija, C.R. 1994. Characterization and evaluation

of dwarf cattle of Kerala, Ph.D. Thesis, Kerala

Agricultural University, Thrissur. p. 112.

Ikonen, T., Ojala, M. and Ruottinen, O. 1999.

Associations between milk protein

polymorphism and first lactation milk

production traits in Finnish Ayrshire cows.

J. Dairy Sci. 82:1026-1033.

Iype, S. and Venketachelapathy, R.T. 2001. Vechur

cattle of Kerala, Kerala Agricultural

University, Thrissur, p 37.

Keating, F., Smith, T.J, Ross, R.P. and Cairns, M.T.

2008. A note on the evaluation of a beta-

casein variant in bovine breeds by allele-

specific PCR and relevance to â-

casomorphin. Irish J. Agric. Fd Res. 47: 99-

104.

Ng-Kwai-Hang, K.F., Hayes, J.F., Moxley, J.E. and

Monardes, H.G. 1986. Relationships

between milk protein polymorphisms and

major milk constituents in Holstein-Friesian

cows. J. Dairy Sci. 69:22-26.

Ng-kwai-Hang, K.F., Monardes, H.G. and Hayes,

J.F. 1990. Association between genetic

polymorphism of milk proteins and

production traits during three lactations.

J. Dairy Sci. 73:3414-3420.

Nilsen, H., Oslen, H.G., Hayes, B., Sehesred, E.,

Svendsen, M., Nom, T., Meuwissen, T. and

Lien, S. 2009. Casein haplotypes and their

association with milk production traits in

Norwegian Red cattle. Genet. Selection

Evolution 41:1-12.

Ojala, M., Famula, T.R. and Medrano, J.F. 1997.

Effects of milk protein genotypes on the

variation for milk production traits of

Holstein and Jersey cows in California.

J. Dairy Sci. 80:1776-1785.

Sarbour, M. P., Lin, C.Y., Lee, A. J. and. Mcalllster,

A. J. 1996. Association between milk protein

genetic variants and genetic values of

Canadian Holstein bulls for milk yield traits.

J. Dairy Sci. 79:1050-1056.

Velmala, R., Vikki, J., Elo, K. and Tanila, A.M. 1995.

Casein haplotypes and their association with

milk production traits in the Finnish Ireshire

cattle. Anim. Genet. 26:419-425.


RESEARCH ARTICLE

ABSTRACT

The present study was aimed to

demonstrate the histological changes in the

pancreas of streptozotocin (STZ) induced

diabetes in a rat model. Twenty rats were

randomly separated into two groups of ten

rats each. Group I served as normal control

and group II served as diabetic. The diabetic

condition was induced in group II by

streptozotocin. Light microscopic evaluation

of islets showed highly swollen â-cells with

loss of cytoplasmic granularity, vacuolations,

necrosis, elongated and fusiform â-cells and

sparse cellularity in diabetic rats. The present

study showed that induction of diabetes using

STZ results in the alteration of the morphology

of endocrine part of pancreas in rats.

Keywords: â-cells, Diabetes Mellitus, Necrosis,

Streptozotocin

1PhD Scholar, Department of Pathology, College of Veterinary

and Animal Sciences, KVASU, Mannuthy, Email ID:

[email protected]

2Professor, Department of Pathology, Veterinary College,

Bangalore

A HISTOLOGICAL STUDY OF THE STRUCTURAL

CHANGES IN THE PANCREAS OF DIABETIC RATS*

1 2B. Dhanush Krishna and Suguna Rao

Department of Pathology, Veterinary College, KVAFSU, Hebbal, Bangalore

*Part of MVSc. thesis submitted by the first author to KVAFSU, Bidar

INTRODUCTION

Diabetes mellitus (DM) is a major health

problem worldwide. The economic impacts of

diabetes with its complications and associated

diseases are large (Sarah et al., 2004). According to

recent estimates, the prevalence of diabetes

mellitus is 4 percent worldwide and that indicates

143 million persons are affected which will

increase to 300 million by the year 2025 (Analava

et al., 2007).

Animal models in diabetes research are

very common where rats are the first choice of use,

comprising over 85 percent of these models (US

Department of Agriculture, 1989). It may have

been because of the pathogenesis of diabetes in

animal models is most likely similar to the

pathogenesis in humans. Streptozotocin (STZ) has

been extensively used to induce diabetes for

various diabetes studies in laboratory animals. STZ

has been reported to be capable of generating

reactive oxygen species resulting in oxidative stress

and cell death. Also, STZ has been found to be a

better chemical inducer for diabetes than alloxan

(Szkudelski, 2001).

The present study was designed to

demonstrate the histological changes on the

endocrine component of pancreas in STZ induced

diabetes in a rat model.




Care and management of experimental animals

The present study was carried on approval

from Institutional Animal Ethics Committee of

Veterinary College, Bangalore. Adult male Albino

Wistar rats weighing 220±40g were maintained

under standard laboratory conditions. The animals

were randomly separated into two groups, group I

as normal control, while group II as diabetic.

Induction of experimental diabetes

After a two-week acclimatization period,

DM was induced experimentally in group II by a

single intraperitoneal injection of a freshly

prepared STZ solution (Sigma Chemicals, USA)

dissolved in 0.1 M ice-cold citrate buffer (pH 4.5)

at a dose of 45 mg/kg to overnight-fasted rats (Babu

and Prince, 2004). Control rats received an

intraperitoneal injection of citrate buffer alone.

After three days of STZ administration, blood

glucose levels of each rat were determined. Rats

with a blood glucose level above 300mg/dL were

considered diabetic and included in this study. The

blood glucose level was measured using

commercially available biochemical kits (Span

Diagnostics, Bangalore) (Tietz, 1976).

Histological Evaluation

Two animals from each group were th th theuthanized on 15 , 30 and 45 day of the

experiment using an overdose of ether for studying

the progressive effects of STZ. Animals were

immediately dissected and pancreas was observed

for evidence of gross pathology. For the light

microscopic examinations, samples from tail

portions of the pancreas were fixed in 10 percent

neutral buffered formalin and embedded in

paraffin. The paraffin blocks were cut into pieces

with a thickness of 4 µm. The sections were stained

with Hematoxylin and Eosin (H&E) (Bancroft and

Gamble, 2008) for the evaluation of islets injury.

Statistical Analysis

Data were analyzed using a commercially

available statistics software package (Graph Pad

Prism for Windows Version 5.0). All values are

presented as Mean ± Standard Error (SE). The data

were analyzed using paired sample student t test

(p < 0.001; two tailed).


A significant (p < 0.001) decrease in the

body weight was observed in the diabetic group

when compared to the control. The blood glucose

level was significantly increased (p < 0.001) in the

diabetic group when compared to the control.

The diabetic control rats appeared grossly

emaciated. The pancreas showed slight congestion

and progressive decrease in the size which became th

appreciable from 15 day of the present study. On th

45 day, the pancreas was atrophied and appeared

as a thin gelatinous strip.

Histopathology of islets of Langerhan's of

pancreas of control animals revealed normal

architecture with compact arrangement of â- cells

and non â- cells throughout the study. In the

present study, post STZ injection, the pancreas

showed numerous lobules with normal appearing

exocrine component in diabetic control rats on th

15 day. However, pancreatic islets were reduced

in number per lobule and showed loss of normal

architecture. The normal distribution of non

â-cells and â-cells was altered with much

reduction in the number of â-cells and disorderly

arranged non â- cells. The â-cells were swollen,

vacuolated, necrotic or elongated and fusiform

with condensed nucleus and showed loss of

cytoplasmic granularity.

Fig 1A. Islet of control animal showing normal

architecture with compact arrangement ofth

â- cells and non â- cells on 15 day (H&E X 200)

Fig 1B. Islet of diabetic control showing atrophiedth

and vacuolated â- cells on 15 day (H&E X 200)

thOn 30 day post STZ injection, there was

further decrease in the number of islets which

appeared shrunken and showed increased severity thin those lesions that appeared on 15 day in â-cells.

thOn 45 day post STZ injection, occasional small

sized islets were appreciated which were difficult

to locate. There was severe reduction in the

number of â-cells. Occasional persisting â-cells

were highly swollen, vacuolated and showed loss

of cytoplasmic granularity. This was in agreement

with earlier reports. (Papaccio et al., 2000 and Mir

et al., 2008). They appeared irregular in shape,

reduced in the size, with some assuming 'star fish'

appearance. Besides, there was an increase in the

number of non â- cells and mild degrees of fibrosis

with infiltration of a few inflammatory cells were

also observed. The histopathology of islets in

diabetic rats indicated progressive destruction of th th

â-cells from 15 to 45 day of the investigation.

Fig 1C. Islet showing extensive vacuolation andth

necrosis of â- cells on 30 day (H&E X 200)

Fig 1D. Islet showing atrophied and “starfish”thappearance on 45 day (H&E X 200)


According to the American Diabetes

Association (ADA), most common symptoms of

diabetes include polydipsia, weight loss and

polyphagia those were evidently present in the

diabetic groups in the present experiment. The rdelevation of blood sugar level on 3 day confirmed

the establishment of diabetes mellitus in rats which

is attributed to the selective cytotoxicity of STZ on

â- cells (Bedoya et.al., 1996).

The decrease in cellularity within islets

observed in the study reflects the cytotoxity of STZ.

The reduction in the number of â- cells was also

noticed. STZ possesses diabetogenic effect

mediated through pancreatic â-cell destruction.

STZ appears to cause cytotoxicity by a number of

mechanisms. STZ on entry into the â- cells via a

glucose transporter (GLUT2) gets spontaneously

decomposed to form an isocyanate compound

and a methyldiazohydroxide which cause intra

molecular carboxylation and alkylation of cellular

components respectively especially that of DNA of

â-cells (Varva, 1960). However, the DNA damage

of â-cells of pancreas is mainly by alkylation with

carbonium ion produced by methyldiazohyd-

roxide (Elsner et.al., 2000).

The DNA damage induces activation of

poly ADP-ribosylation, a process that is more

important for the diabetogenicity of STZ than just

DNA damage itself. Poly ADP-ribosylation leads to +

depletion of cellular NAD and ATP. Enhanced ATP

dephosphorylation after STZ treatment supplies a

substrate for xanthine oxidase resulting in the

formation of super oxide radicals. Consequently,

hydrogen peroxide and hydroxyl radicals are

generated. Further, STZ liberates toxic amounts of

nitric oxide that inhibits aconitase activity and

participates in DNA damage. As a result of the STZ

action, â-cells undergo destruction by necrosis.

STZ selectively destroys the pancreatic insulin

secreting â- cells, leaving less active cells and result

in a diabetic state (Szkudelski, 2001).

In the present study, â- cells in some islets

were found to be fusiform. The change in the

shape of cells can be attributed to the partial

damage of STZ to some â- cells. Aybar et.al. (2001)

have reported that use of lower dose of STZ

produced an incomplete destruction of pancreatic

â-cells even though rats became permanently

diabetic.

Brownlee (2001) postulated a concept

that link hyperglycaemia-induced damage by

different mechanisms that finally leads to cellular

stress. Firstly, hyperglycemia increase movement

of glucose through polyol pathway and sorbitols

are produced which in turn causes osmotic stress

to cells and dihydronicotine amide adenine

dinuleotide phosphate (NADPH) is consumed,

depleting intracellular glutathione. Secondly,

hyperglycemia increases concentrations of

advanced glycation end products. These

glycosylated proteins are formed by non-

enzymatic reactions and changes in protein

structure may alter their cellular functions. Thirdly,

glucose activates various isomers of protein kinase

C which in turn affects the expression of nitric

oxide, endothelin, nuclear factor kappa B (NF-kB)

and plasminogen activator inhibitor. Finally,

hyperglycemia increases the flux of glucose

through the hexasomine pathway effecting

inflammatory mediators and insulin resistance.

The combined effect of these mechanisms results

in over-production of superoxides by the

mitochondrial electron-transport chain, causing

cellular stress and damage that was clearly

appreciated in the form of islets injury in the

present study. STZ induced hyperglycaemia has


been described as a useful experimental model to

study the activity of hypoglycaemic agents. So the

result of the present study is useful to evaluate the

hypoglycaemic agents in pre-clinical trials.

REFERENCES

American Diabetes Association: Report of the

Expert Committee on the diagnosis and

classification of diabetes mellitus. 2007,

Diabetes Care, 30, S42-47.

Analava, M., Bhattacharya, D and Roy,S. 2007.

Dietary influence on type 2 Diabetes

(NIDDM). J. Hum. Ecol., 1:139-147.

Aybar, M. J., Sanchez Riera, A. N., Grau, A and

Sanchez, S. S. 2001. Hypoglycaemic effect

of the water extract of Smallanthus

soncifolius (yacon) leaves in normal and

diabetic rats. J. Ethnopharmacol. 74: 125-

132.

Babu, P. S and Prince, P. S. M. 2004. Antihyper-

glycaemic and antioxidant effect of

hyponidd, an ayurvedic herbomineral

formulation in streptozotocin-induced

diabetic rats. J. Pharm. and Pharmacol., 56:

1435-1442.

Bancroft, J. D and Gamble, M. 2008. Theory and th

practice of histological techniques. 6 Ed.

Churchill Livingstone, United States of

America, 121 p.

Bedoya, F. J., F. Solano and Lucas, M.1996.

Nmonomethyl-arginine and nicotinamide

prevent streptozotocin induced double

strand DNA break formation in pancreatic

rat islets. Experientia, 52:344-347.

Brownlee, M. 2001. Biochemistry and molecular

cell biology of diabetic complications.

Nature, 414: 813-820.

Elsner, M., Guldbakke, B., Tiedge, M., Munday, R

and Lenzen, S., 2000. Relative importance

of transport and alkylation for pancreatic -

ce l l tox i c i t y o f s t rep tozo toc in .

Diabetologia. 43: 1528-1533.

Mir, S. H., Baqui, A., Bhagat, R.C., Darzi, M.M and

Shah, A.W. 2008. Biochemical and

histomorphological study of streptozotocin

- induced diabetes mellitus in rabbits. Pak. J.

Nutr., 7: 359- 364

Papaccio, G., Pisanti, F.A., Latronico, M.V.,

Ammendola, E and Galdieri, M.2000.

Multiple low dose and single high dose

treatments with streptozotocin do not

generate nitric oxide. J. Cell Biochem., 77:

82-91.

Sarah, W.,Gojka, R.,Anders, G.,Richard, S and

Hilary, K., 2004. Global prevalence of

diabetes. Diabetes Care.2004; 27: 1047-

1053.

Szkudelski, T. 2001. The mechanism of alloxan

and streptozotocin action in cells of the rat

pancreas. Physiol. Res., 50: 536-546.

Tietz. 1976. Fundamentals of clinical chemistry.

W.B.Saunders Co., Philadephia.

US Department of Agriculture: Animal welfare

enforcement report fiscal year 1988.

Washington, DC, 1989.

Varva, J.J., Deboer, C. and Dietz, A. 1960.

Streptozotocin, a new antibacterial

antibiotic. Angtibiot. Ann. 230-235.


ABSTRACT

Efficacy of herbal oral formulation ® Afanil (Supplied by M/s Ayurvet Ltd, Baddi, HP;

India) was evaluated against bloat condition in

bovines on the basis of improvement in clinical,

hematological and rumen fluid analysis. For this

a clinical study was undertaken on 20 cases of

bovines suffering from bloat presented to

Department of Veterinary Medicine, College of

Veterinary and Animal Sciences, Parbhani,

Maharashtra. Total cases were divided in two

treatment groups T and T of 10 animals each. T 1 2 1

was administered conventional antizymotic

mixture of vegetable oil 750 ml-1.5 liter mixed

with 20-30 ml of turpentine oil orally. T was 2

treated with herbal anti bloat oral formulation

®(Afanil ) @ 100 ml twice daily till recovery.

Supportive and fluid therapy was also given to

the animals from both the groups. Time period

required in hours for resumption of distended

rumen and rumination process was significantly ®

lesser (P < 0.01) in animals treated with Afanil

(6.08±0.37 and 6.95±0.39) as compared to

conventional anti bloat therapy in T (9.65±0.71 1

and 10.75±0.71). 2.50± 0.17 and 1.70± 0.15

days was the mean time period required for

complete clinical recovery after conventional ® and Afanil anti-bloat therapy respectively.

®THERAPEUTIC POTENTIAL OF AFANIL

AGAINST BLOAT AND FOR EARLY RESTORATION

OF RUMEN FUNCTION IN BOVINES

S. U. Digraskar, V. D. Muley, K. Ravikanth, M. Dandale and S. Maini

College of Veterinary and Animal Science, MAFSU, Parbhani, Maharastra and

Ayurvet Limited, Baddi, Himachal Pradesh.

INTRODUCTION

Bloat is an over distention of the rumen

and reticulum with the gases of fermentation

either in the form of a persistent foam mixed with

the ruminal contents called primary or frothy bloat,

or in the form of free gas separated from the ingesta

called secondary or free-gas bloat (Merck

Veterinary Manual, 2011). It occurs due to feed

rations containing a high proportion of grains or

due to feeding of fresh green legumes. In acute

tympany or bloat, due to progressive distension of

rumen there is respiratory and cardiac distress

which if not relieved, may rapidly lead to asphyxia,

cardiac failure and death (Radostits et. al, 2003).

®Afanil efficiently normalized the altered

hematological as well as rumen pH conditions as

compared to that of T . In present clinico-1

®therapeutic trial, Afanil herbal anti bloat

preparation was found to be effective to treat

bloat cases of bovines in comparison to

conventional therapy. It contains herbal extracts

and carminative volatile oils which ensure quick

and prompt relief in bloat cases hence may be

successfully employed in the treatment.

®Keywords: Afanil , bloat, antizymotic, tympany,

herbal


RESEARCH ARTICLE

® Comparative efficacy of the Afanil herbal

anti bloat therapy with that of conventional

therapy was evaluated in the present clinical study.


The present investigation was undertaken

at Department of Veterinary Medicine, College of

Veterinary and Animal Sciences, Parbhani,

MAFSU, Maharashtra. Twenty bovine cases

presented with clinical features of frothy as well as

free gas bloat formed the basis of the experiment.

The clinical cases were divided in two treatment

groups T and T of 10 animals each. Feeding and 1 2

clinical history was recorded in every case which

followed blood and ruminal fluid sample

collection for further analysis.

Animals in group T (n=10) suffering from 1

bloat of dietary origin were administered

conventional antizymotic mixture of vegetable oil

750 ml-1.5 liter mixed with 20 -30 ml of turpentine

oil orally. Group T (n=10) was treated with herbal 2

®anti bloat oral formulation Afanil 100 ml twice

orally daily till recovery (not more than 2 days in

this experiment). Supportive therapy with

antihistaminic, IV fluids (RL/DNS) along with

sodium bicarbonate or ascorbic acid was also

instituted depending upon the cause of bloat in

affected bovine. Clinical signs were recorded after

treatment.

The efficacy of the treatment in both

groups was assessed on the basis of improvement

in, a) Clinical parameters e.g. time period required

for resumption of distended rumen, rumination

and other physiological processes. b)

Hematological- TLC, DLC and c) Rumen fluid

analysis- pH and consistency.


Diagnosis was made on basis of the

correlating the feeding history with that of clinical

signs exhibited by the animals. Most of the animals

had clinical features comprised of over distended

abdomen with bulging of upper left flank,

anorexia, absence of rumination and retarded

ruminal motility, dehydration, dyspnoea, dullness

and depression. Tympanic or drum-like sound was

heard on percussion of left paralumbar fossa. Milk

yield was greatly reduced or absent in lactating

animals. All of the animals were having the feeding

history of highly succulent leguminous fodder,

flour or grains.

A prompt response to treatment with ® Afanil was noticed. Over distended rumen started

getting rid of gases and froth within minutes and

minimum time period required for rumen to

completely assume its normal shape was only 4.50

hrs after the initiation of therapy (Table 1). While

that for conventional treatment required 6.5 hrs.

Mean time period required for resumption of ® distended rumen after treatment in Afanil treated

group was 6.08±0.37 hrs against 9.65±0.71 hrs

required for conventional therapy. Similar findings

were also noted by Gahlaut (1998).

Mean time period required for resumption ® of rumination process (Table 1) after Afanil

therapy was 6.95±0.39 hrs therapy. In

comparison to this 10.75±0.71 hrs were required

with conventional therapy. A complete clinical

recovery occured in mean time period of ®

1.70±0.15 days after Afanil therapy while

conventional therapy required more time for

complete recovery i.e. 2.50±0.17 days.

Blood parameters with reference to TLC

and DLC of affected bovines before and after


® Afanil therapy were studied. Differential

Leukocyte Count (DLC) showed eosinophilia to

the extent of 5.70±0.37 in affected cases which

were improved to 3.0±0.26 and 2.70±0.21 in

group T and T animals respectively. As ruminal 1 2

and blood plasma histamine concentrations

increase during ruminal lactic acidosis (Asenbach

and Gabel, 2000) eosinophilia may be because of

histamine upsurge in bloat cases which was ®significantly brought down in Afanil treated

group.

Analysis of ruminal fluid before and after

treatment in both the groups was done (Table 2). ®

Results obtained after analysis showed that Afanil

efficiently brought the altered levels of the ruminal

pH to normal level required for optimum ruminal

fermentation and fiber digestion.

The carminative, antispasmodic, anti-

flatulent, stomachic, alterative and laxative ®properties of Afanil are contributed by its

constituent herbs for which these activities have

been well documented (Nadkarni, 1954; Chopra

et. al, 1982; Kapoor, 1990; Hussain et. al 1992).

A Period required for resumption of distention ofa brumen in hours after treatment (in hours) 9.65± 0.71 6.08±0.37

B Period required for resumption of ruminationa bprocess in hours after therapy (in hours) 10.75± 0.71 6.95± 0.39

C Period required for overall clinical recovery e.g.

normal appetite, physiological parametersa band milk (in days) 2.50± 0.17 1.70± 0.15

Means bearing different superscript in a row differ significantly at P=0.05 leve

Sr.No Clinical parameters

Group T1

Conventionaltherapy

Group T2®

Afanil therapy

Table 1. Mean values of clinical parameters in different treatment regimen against bloat in bovines.

Table 2. Evaluation of pH of ruminal fluid in clinical cases of bloat before and after treatment.

® Group T (Conventional therapy) Group T (Afanil therapy)1 2

Case no. BT AT BT AT

Mean ± SE 7.87± 0.43 7.15± 0.23 7.47± 0.58 6.77± 0.15

BT: Before treatment AT: After treatment


In the present clinico-therapeutic trial, the ®

product Afanil herbal anti bloat preparation was ,

found to be safe, effective, non-toxic and

promising therapy to treat frothy as well as free gas

bloat cases of bovines in comparison with

conventional therapy. Results in the present

experiment corroborates well with earlier findings

of Sharma (1996), Gahlaut (1998) and Bhardwaj

(1998).

®Thus, it was concluded that Afanil , a

liquid emulsion containing herbal extracts and

carminative volatile oils ensures quick and prompt

relief in bloat cases and may be successively used

in field conditions to prevent the losses from bloat

origin.

ACKNOWLEDGEMENT

Authors are thankful to Associate Dean,

COVAS, Parbhani for providing necessary facilities

required for the present Clinico-therapeutic trial

and Ayurvet Limited, Baddi for providing

necessary samples for carrying out present

research experiment.

REFERENCES

Aschenbach, J.R and Gabel, G.J. 2000. Effect and

absorption of histamine in sheep rumen:

Significance of acidotic epithelial damage

J. Anim. Sci., 78:464-470

Bhardwaj, U. 1998. Therapeutic efficacy of herbal

anti-tympanitic agents for recurrent

tympany in buffaloes. Ind. Vet. Med. J. 22:

327-328

Chopra, R.N., Chopra, I.C., Handa, K.L and

Kapoor, L.D. 1982. Indigenous drugs of ndIndia. 2 edn. Academic Publishers,

Calcutta.

® Gahlaut, K.S.1998. Therapeutic efficacy of Afanil

for ruminal tympany. Dairy Guide, April-

June, 57-59.

Hussain, A. 1992. Dictionary of Indian Medicinal

Plants. Central Institute of Medicinal and

Aromatic Plants, Lucknow.

Kapoor, L.D. 1990. CRC Handbook of Ayurvedic

medicinal plants. CRC Press, Boca Raton,

Florida.

Merck Veterinary Manual. 2011. http://www.

merckvetmanual.com/mvm/index.js?

cfile=htm/bc/ Accessed 22/7/2012

Nadkarni, A.D. 1954. Dr. K.M. Nadkarni's Indian

Materia Medica. Popular Book Depot,

Bombay.

Radostits,M., Gay,C.C., Blood,D.C and Hunchcliff,

K.W. 2003. Veterinary Medicine. 10th Ed.

W.B. Saunders Company Ltd., 493-500.

Sharma, V and Sharma, S.D. 1996. Clinical efficacy ®

of Afanil in treatment of acute tympany and

bloat. The Veterinarian, 20:18-19.


OPTIMIZATION OF PCR THROUGH MANIPULATION

OF CYCLE TIMES AND INCLUSION OF FORMAMIDE

1 2 3 4S. Pramod , A. P. Usha , T. Venkatachalapathy and K.C. Raghavan

Kerala Veterinary and Animal Sciences University

ABSTRACT

Polymerase chain reaction (PCR) is an in

vitro technique to produce million fold copies of

a particular segment of DNA. PCR should

optimally yield a unique product unless

designed otherwise. Artefacts appear many

times, which affect the success of downstream

applications. Prevention of non specific

ampl i f i ca t ion us ing fo rmamide and

manipulation of cycling time is discussed in this

article.

1 2PhD Scholar, CASAGB, Mannuthy, Professor & Head, CPPR, 3Mannuthy, Associate Professor & Head, University Goat

4Farm, Mannuthy, Director, CASAGB, Mannuthy

INTRODUCTION

Polymerase chain reaction (PCR) is an in

vitro cell free technique to produce a large number

of copies of a specific segment of DNA. The

technique of PCR (Mullis, 1983) evolved during

four decades after the establishment of double

stranded helix structure of DNA by Watson and

Crick (1953). Kleppe (1971), an associate of

Hargobind Khorana probably introduced the

concept of PCR; with two primers, and cycles of

repair synthesis after addition of DNA polymerase

in each round. Earlier PCR relied upon DNA

polymerase I enzyme of Escherichia coli or rather

its 'Klenow fragment', which was thermolabile and

needs to be added afresh before each cycle. Chien

et.al. (1976) purified DNA polymerase from

Thermus aquaticus (EC 2.7.7.7), which later

became famous as 'Taq' polymerase. The molecule

had a weight in the range of 63-68 KDa with an

optimum temperature of activity at 80°C in a

buffer of pH of 8.0. Ever since Taq polymerase was

included in PCR eliminating the need to add DNA

polymerase before initiation of each cycle, it

gained acceptance and was used for a myriad of

downstream applications.

Even though the PCR optimally should

yield a unique product (unless designed otherwise)

many times unintended artefacts also appear. It

becomes important to avoid such spurious

amplifications so as to improve the success rate of

downstream applications like molecular cloning,

PCR-RFLP etc. This article attempts to throw light

into the optimisation of PCR, when multiple

spurious amplicons appear along with the

expected product.


PCR reactions were set up so as to contain

20-100 ng of template DNA, 5 nM each of forward

and reverse primers (Sigma), 200 mM of each

dNTP's (Fermentas), 2.5 mM of MgCl (Sigma), 2


RESEARCH ARTICLE

0.75 U Taq polymerase (Sigma), polymerase buffer

(Sigma) to a final concentration of 1X, diluted with

autoclaved Millipore water in a volume of 10 µL

which was amplified in a Bio-Rad thermal cycler.

'Gradient' is an option in most thermal

cyclers for optimization of cycling conditions. The

positions for reaction tubes are arranged rows and

columns. The positions in a row can be

programmed to maintain one particular

temperature and is called a block. When gradient

option is set, the temperature remain constant in a

row and varies along a column of tube positions.

The programme used for gradient PCR was as

follows; initial denaturation at 94°C for three

minutes, followed by 35 cycles of denaturation at

94°C for one minute, annealing temperature over

the gradient for 30 seconds and extension at 72°C

for 1 minute with a final extension time of 2

minutes at 72°C. The gradient used in this

experiment is presented in table (1).

Sl No Block Temperature (°C)

FUT1 HSD 17â

1 A 65.0 59.0

2 B 64.3 58.4

3 C 63.2 57.4

4 D 61.4 55.8

5 E 59.3 53.9

6 F 57.7 52.5

7 G 56.6 51.5

8 H 56.0

Table 1: Block temperatures in gradient PCR

The loci under study were segments from

Fucosyl transferase 1 (FUT1) and 17â Hydroxy

steroid dehydrogenase (HSD17â) genes, which are

candidate genes (Spotter and Distl., 2006) for litter

traits in pigs. The primer pair FUT1 F: 5'-

GCCGCCACCTCTGTCTGACC-3' and FUT1 R: 5'-

TACCCCCTGGGCCTCTTGCC-3' were designed

from Genbank accession no. L50534.1 using

Primer-Blast tool (available at http://www.ncbi.

nlm.nih.gov/tools/primer-blast/ index.cgi?

LINK_LOC=BlastHome). The forward and reverse

primers had a length of 20 bases each with a GC

content of 70 percent. The melting temperature

(T ) of forward primer was 65.23°C and that of m

reverse primer was 66.0°C. The T of an m

oligonucleotide is the temperature at which half of

the molecules remain free of secondary structures.

The expected product size for this amplicon was

578 bp.

A segment of HSD 17â gene was amplified

using the primers described by Jacobs et.al. (2002).

The nucleotide sequence of forward primer was

5'- CTCCCACCCCACCTGTTC-3' and that of the

reverse primer was 5' CCGTTCACCACCCCTCCTC-

3', with an expected product size of 273 bp. The

forward primer was 18 nucleotides long while the

reverse primer had a length of 19 nucleotides. The

GC content was 66.67percent and 68.42 percent

respectively for the forward and reverse primers.

The melting temperature (T ) of forward primer m

was 59.24°C while that of the reverse primer was

61.96°C.

Gradient PCR was done across a block

temperature of 56°C and 65°C for FUT1 locus and

50 - 59°C in the case of HSD17â locus. The

amplified products were loaded into 2 percent

agarose (SRL labs) gel with pre incorporated

ethidium bromide (SRL labs) and electrophoresed

in Tris acetate EDTA buffer at 85 volt for 45 minutes

before being photographed in Geldoc

(Pharmacia). The output of gradient PCR for FUT1


is presented in figure (1) and that of HSD 17â in

figure (2). Several non specific products could be

observed in both the loci. In order to suppress

spurious amplifications, formamide was

introduced into the reaction mix, along with the

reduction of extension time. The composition of

reaction mixture was modified by including 0. 75

percent V/V of formamide (USB) and was used to

amplify FUT1 (Fig: 3) and HSD 17 â loci (Fig: 4).

However mere addition of formamide did not

remove all non specific amplicons and a new PCR

programme was used in case of HSD 17 â gene

(Table 2).

Table 2: Modified PCR programme used for HSD

17â locus

RESULTS

The result from gradient PCR of FUT1 locus is

given in Fig (1) and that of HSD 17 â is provided in

Fig (2). Four non specific products lower in

molecular mass than the expected amplicon (578

bp) were observed in case of FUT1 locus. A

product in excess of 1 Kb also appeared when the

annealing temperature was below 59.3°C. The

larger product failed to amplify when the annealing

time was reduced from 30 seconds to 15 seconds,

with the simultaneous reduction of extension time

from one minute to 45 seconds. A second gradient

PCR was performed with formamide (0.75 percent

V/V) incorporated in the reaction mixture using the

modified programme. Problem of spurious

amplifications could be successfully controlled in

FUT1 locus (Fig: 3) and an annealing temperature

was selected for subsequent amplification of this

locus.

In case of HSD 17 â locus, four non specific

products, all above the intended product of 273

bp was observed. Addition of formamide and

reduction of extension and annealing times did not

quite well remove non specific amplifications (Fig:

5, lanes 5 to 7). Annealing time was reduced to 15

seconds and the extension time was reduced to 30

seconds. The PCR program was further modified

by introducing an additional step with three cycles,

at a higher annealing temperature and the

spurious amplification could be suppressed (Fig.4).

DISCUSSION

Efficiency of PCR reflects the capability of a

primer pair to produce faithful amplification,

which is close to the theoretical doubling of target

sequence per cycle. Ill designed primers allow co-

amp l i f i c a t i on o f un in tended r eg ion s

compromising both specificity and efficiency. The

principles of PCR primer design were reviewed by

Singh and Kumar (2001). However in many cases,

whole genome sequence is not available in public

domain which makes the researcher to choose

primers without the knowledge of other probable

targets.

The annealing temperature (T ) used for a

primer- template interaction is a function of T . m

The annealing temperature occurs within a range

of 4-10°C lower than T and increased T in PCR m a

results in poor or no amplification (Wu et.al.,

1991). In case of both FUT1 and HSD 17â, the


annealing temperature was observed to be within

this range. It implies that the nearest neighbour

thermodynamics (Santa Lucia.,1998) can efficiently

predict the T . Lorenz (2012) suggested that the m

melting temperature of the primers should range

between 45-65°C with the window between 52-

58°C being the optimum. However T for the m

primer pair should not differ by more than 5°C.

Dieffenbach et.al., (1993) were of the opinion that

PCR primers should maintain a reasonable GC

content. Oligonucleotides which were 20 bases

long with a 50percent G + C content generally had

Tm values in the range of 56-62°C. Primers for both

the loci were within the length limit and had

acceptable T . Even with GC content in excess of m

50 percent, spurious amplicons had appeared in

the PCR for FUT1 and HSD 17â loci. The high GC

content is reflected in the elevated T of the m

primers. The factor that the length of primers for

HSD 17â being slightly less (18 and 19 bases) than

the optimum, could also have favoured

mispriming.

The concentration of MgCl , dNTP's and 2

Taq polymerase also influence the occurrence of

spurious amplicons. Higher concentration of

MgCl , dNTP and Taq polymerase is known to 2

favour mispriming and generation of unintended

products. The recommended concentration of

MgCl varies between 0.5 to 5.0 mM. In this 2

experiment, a concentration of 2.5mM MgCl was 2

used which is well within this range.

Sarkar et.al. (1990) used formamide to

prevent spurious amplification in PCR, which

employed primers with GC content in excess of 55

percent. They had further observed that in case of

primers with high GC content, formamide added

to the reaction mixture at 1.5 to 5 percent levels

prevented spurious amplification. In the present

study, formamide included at 0.75 percent V/V,

was sufficient to banish non specificity.

Inclusion of a separate cycle with three

steps (steps 2 to 4, Table: 2) to amplify HSD 17â

comes from 'Touchdown' programme (Don et.al.,

1991). The touchdown programme is widely used

to amplify repetitive sequences, like microsatellite

markers which generate lot of noise. In order to

overcome spurious priming, few initial cycles are

run at higher annealing temperature thereby

enhancing the specificity of primer binding.

Addition of a high temperature step in this experiment

possibly increased the specificity of PCR.

Innis et.al. (1988) observed that Taq

polymerase added nucleotides at a rate of about

60 nucleotides per second. Some other reports

suggest the speed to be between 35 and 100

nucleotides per second. The rule of the thumb is to

provide extension time of one minute per Kb size

of the product. In current study, reduction of

extension time also played a part in blocking

spurious amplifications in both FUT1 and HSD

17â loci.

CONCLUSION

Non specific amplification in the case of

primers with high GC content could be averted by

using formamide and adjustment of annealing and

extension time and introduction of a high

temperature step so as to favour the formation of

the product of interest.

ACKNOWLEDGEMENTS

INSPIRE fel lowship provided by

Department of Science & Technology,


Government of India, funds and facilities provided

by KVASU and ICAR is thankfully acknowledged.

REFERENCES:

Chien, A., D.B. Edgar and J.M. Trela. 1976. DNA

polymerase from the extreme thermophile

Thermus aquaticus. J. Bact., 127 (3): 1550-

1557.

Dieffenbach, C. W., Lowe, T.M and Dveksler, G.S.

1993. General concepts for PCR primer

design. Genome Res.,3: S30-S37.

Don, R.H., P.T. Cox, B.J. Wainwright, K. Baker and

Mattick, J.S. 1991. 'Touchdown' PCR to

circumvent spurious priming during gene

amplification. Nucleic Acids Res. 19(14):

4008.

Innis, M.A., B. K.B. Myambo, D.H. Gelfand, and

Brow, M.A.D. 1998. DNA sequencing with

Thermus aquaticus DNA polymerase and

direct sequencing of polymerase chain

reaction-amplified DNA. Proc. Natl. Acad.

Sci. 85: 9436-9440.

Jacobs, K.., M. Mattheeuws, , M. Van Poucke, A.

Van zeveren and Peelman, l.J. 2002.

Characterization of the porcine FABGL

gene. Anim. Genet. 33: 220-223.

Kleppe, K., E. Ohtsuka, R. Kleppe, I. Molineux and

H.G. Khorana. 1971. Studies on

polynucleotides. XCVI. Repair replications

of short synthetic DNA's as catalyzed by DNA

polymerases. J. Molec. Biol. 56: 341-61.

Lorenz, T. C. 2012. Polymerase Chain Reaction:

Basic Protocol Plus Troubleshooting and

Optimization Strategies. J. Vis. Exp. (63),

e3998, DOI: 10.3791,3998.

SantaLucia, J. Jr. 1998. A unified view of polymer,

dumbbell, and oligonucleotide DNA

nearest-neighbor thermodynamics. Proc.

Natl. Acad. Sci. U. S. A. 95: 1460-1465.

Sarkar, G., S. Kapelner and Sommer, S.S.1990.

Formamide can dramatically improve the

specificity of PCR. Nucleic Acids Research.

18(24): 7465.

Singh, V.K. and Kumar. A. 2001. PCR primer

design. Molecular Biology Today, 2 (2): 27-32

Spotter, A. and Distl, O. 2006. Genetic approaches

to the improvement of fertility traits in the

pig. Vet. J., 172: 234-247

Watson, J.D and Crick, F.H.C. 1953. A structure for

deoxyribose nucleic acid. Nature. 171: 737-

738

Wu, D., L. Ugozzoli., B.K. Pal., J. Qian and Wallace,

R.B. 1991. The effect of temperature and

oligonucleotide primer length on the

specificity and efficiency of amplification by

the polymerase. DNA and cell biology, 10

(3): 233-238


ABSTRACT

The success of dairy industry depends

to a large extent on rearing of calves to a

breedable age at a faster rate and with

minimum mortality. The higher the plane of

nutrition the earlier the onset of puberty and

thus quicker the returns. In this context, a study

was conducted to assess the effect of challenged

milk feeding on growth performance in dairy

calves. Eighteen calves born in Kerala

Agricultural University Livestock Farm,

Mannuthy were selected at random and were

divided into three groups of six animals each.

The calves in the first group were fed with milk in

normal regime as per package of practices

recommendations (Control group). Animals in

the second and third group were fed 25 percent

and 50 percent extra milk as that of normal

regime. The body weight and average daily th

weight gain of all animals were recorded till 4

month of age and analyzed. The overall result

indicated that there was a significant increase

(P<0.05) in daily average weight gain in animals

fed with 25 percent and 50 percent extra milk

when compared to animals in the control

group. It was also observed that the daily

1 2Research Assistant ([email protected]), Prof. and

head, University Livestock Farm, Mannuthy, College Of

Veterinary and Animal Sciences, Mannuthy, Thrissur

average weight gains recorded in animals in the

second and third group were significantly higher

(P<0.05) than that of the control during the

fourth month of age indicative of the carry over

effect of the feeding system. Based on the result

it is recommended that a higher level of milk

feeding regime during pre-weaning state will

not only influence the growth rate during the

period but also beneficially contribute to the

growth performance in later stages.

Key words: Challenged milk feeding, extra

milk allowance, milk feeding strategy.

INTRODUCTION

An efficient calf feeding system is critical

because it determines the future income and

sustainability of dairy farms (Tozer et.al., 2001).

The primary goal of most liquid feeding programs

for dairy calves is to double the birth body weight

within 8 weeks of age and minimize the morbidity

primarily diarrhea and respiratory diseases and

mortality ( Jasper et.al., 2000). The higher the

plane of nutrition the earlier the onset of puberty

and thus quicker the returns ( Von et.al., 2006).

Nature's way of feeding calves includes free access,

nursing until satiated, frequent meals per day and

suckling. Conventional rearing systems usually

limit access, restrict milk intake per meal,

MILK FEEDING STRATEGIES FOR IMPROVING GROWTH

PERFORMANCE IN CROSSBRED DAIRY CALVES

1 2A.J. Flamy and Joseph Mathew

College Of Veterinary and Animal Sciences, Mannuthy, Thrissur

RESEARCH ARTICLE


encourage rapid feeding or gorging, restrict meals

per day or provide milk in pails( Davis et.al.,2011 ).

Feeding less milk results in poor growth due to lack

of needed nutrients( Huber et.al., 1984). In many

animal models, it is well documented that the

amount of nutrients consumed early in life has

long-term effects on future performance

(Pettersson. 2000 ). In this context, a study was

conducted to assess the effect of challenged milk

feeding on growth performance in dairy calves

reared in Kerala Agricultural University Cattle

Farm.


Eighteen calves born in University

Livestock Farm and Fodder Research and

Development Scheme, Mannuthy were selected

at random and were divided into three groups of

six animals each. The calves in the first group were

fed with milk in normal regime as per package of

practices recommendations (Control group).

Animals in the second and third group were fed 25

percent and 50 percent extra milk as that of

normal regime. This schedule in all groups was

continued until the calves attained 3 months of age

at weaning. The body weight and average daily th

weight gain of all animals were recorded till 4

month of age and analyzed.

RESULT

The observations are summarized in the

following tables

1 D173 25% Extra 29 35 45 57 71

2 D175 25% Extra 26 34 40 54 70

3 D176 25% Extra 22 33 39 56 80

4 D185 25% Extra 19 27 39 51 64

5 D188 25% Extra 20 25 39 54 64

6 D191 25% Extra 20 24 32 39 56

7 D174 50% Extra 25 35 43 56 70

8 D177 50% Extra 21 32 41 55 78

9 D178 50% Extra 17 26 37 52 76

10 D183 50% Extra 26 35 47 60 72

11 D186 50% Extra 26 30 45 59 77

12 D192 50% Extra 25 29 35 52 54

13 D181 Control 24 31 38 47 55

14 D182 Control 23 28 37 46 60

15 D184 Control 22 28 37 47 54

16 D189 Control 31 38 43 55 61

17 D190 Control 25 33 43 51 56

18 D193 Control 23 26 39 46 55

Table 1 : Details of Weight recorded (Grams) during the first, second, third and fourth month

Experimental

group (Kg)

Birth

Weight (Kg)

Wt. in

30 days (Kg)

Wt. in

60 days (Kg)

Wt. in

90 days (Kg)

Wt. in

120 days (Kg)Sl.

No.Calf No


STATISTICAL ANALYSIS

The overall result indicated that there was

a significant increase (P<0.05) in daily average

weight gain in animals fed with 25 percent and 50

percent extra milk when compared to animals in

the control group. It was also observed that the

daily average weight gains recorded in animals in

the second and third group were significantly

higher (P<0.05) than that of the control during the

fourth month of age indicative of the carry over

effect of the feeding system.

DISCUSSION

Accelerated milk feeding adds economy

by reducing calf mortality rate and cutting calving

interval (Terre et al., 2006). Most dairy producers

feed restricted quantities of milk to calves because

of cost and the perception that increased milk

intake may lead to a higher incidence of diarrhoea,

reduced calf starter feed intake and reduced mass

gain. Results from several studies indicate that

feeding more milk or high-quality milk replacer

does not cause diarrhoea ( Jasper et.al., 2002).

The concept of accelerated feeding for

young milk-fed calves is now well-accepted as an

alternative to traditional restricted feeding.

Research and field experience have highlighted

many important aspects that are required for

successful implementation of accelerated milk

feeding systems. A step-down or gradual weaning

process facilitates a smoother transition to dry feed

( Flower and Weary, 2001). Colostrum-deprived

calves or calves that are undergoing transport stress

will not respond as well to increased amounts of

milk and may in fact be impacted negatively.

Benefits to accelerated milk-feeding programs

include: decreased age at first calving ,

improvements in health, and increased milk

production ( Kung et.al., 1997). Adequate milk

feeding during younger ages decreases stress in

calves and thus improves the performance (Fiems

et.al., 1982, Diaz et.al., 2001).

Based on the result of this study, it is

recommended that a higher level of milk feeding

regime during pre-weaning state will not only

influence the growth rate during the period but

also beneficially contribute to the growth

performance in later stages. Ongoing research will

provide the necessary input variables to model the

overall economic impact of accelerated milk

feeding programs


REFERENCES

Davis, R., VandeHaar, C. A., Wolf, J. S., Liesman, L.

T., Chapin and Weber, M.S. 2011. Effect of

intensified feeding of heifer calves on

growth, pubertal age, calving age, milk

yield, and economics. J. Dairy Sci.

94:3554-3567.

Diaz, M. C., Van, A. M. E., Smith, J. M., Kelsey, J. M

and Hutten, E. L .2001. Composition of

growth of Holstein calves fed milk replacer

from birth to 105 kilogram body weight.

J. Dairy Sci. 84: 830-842

Fiems, L.O., Boucque, C.V., Cottyn, B.G and

Buysse, F.X., 1982. Effect of feeding

techniques and age at weaning on the

performances of bucket-fed and suckling

reared calves. In: Signoret, J.P. (Ed.),

Welfare and Husbandry of Calves, Current

Topics in Veterinary Medicine and Animal

Science, vol. 19, pp. 149-167.

Flower, F. and Weary, D.M .2001. Effects of early

separation on the dairy cow and calf.

Separation at 1 day and 2 weeks after

birth. Appl. Anim. Behav. Sci. 70:275-284.

Jasper, J. and D. M. Weary. 2002. Effects of Ad

Libitum Milk Intake on Dairy Calves.

J. Dairy Sci. 85 :3054-3058.

Huber, J. T., Silva ,A. G., Campos, O. F and

Mathieu, C. M. 1984. Influence of feeding

different amounts of milk on performance,

health, and absorption capability of baby

calves. J. Dairy Sci. 67: 2957-2963

Kung, L. J., Demarco, S., Siebenson, L.N., Joyner,

E., Haenlein, G.F.W. and Morris, R.M.

1997.An evaluation of two management

systems for rearing calves fed milk

replacer. J. Dairy Sci., 80: 2529-2533

Pettersson, K., 2000. Results from a questionnaire:

housing, feeding and management of

dairy calves in Sweden. In: Tielen, M.J.M.,

Voets, M.T. (Eds.), Proceedings of the 10th

International Congress on Animal

Hygiene, vol. 1. Maastricht, The

Netherlands, pp. 421-425.

Terre,M. Bach, A., Geng, D and Devant, M. 2006.

Performance and behaviour of calves

reared in groups or individually following

an enhanced-growth feeding programme.

J. Dairy Res. 73: 480-486

Tozer, P. R and Heinrichs, A .J. 2001 .What affects

the costs of raising replacement dairy

heifers: A multiple-component analysis.

J. Dairy Sci. 84: 1836-1844

Von , A. G., Wolf, F. Hötzel, M and Weary, D. M..

2006 .Effects of continuous versus

periodic milk availability on behavior and

performance of dairy calves. J. Dairy Sci.

89: 2126-2131


ABSTRACT

Prenatal morphogenesis of medulla

oblongata (MO) was studied using 46 goat

foetuses ranging from 2.5 cm CRL (40 days of

gestation) to 41.5 cm CRL (full term). By 40

days of gestation, the roof plate region of the

rhombencephalon expanded enormously and

as a result, the entire alar and basal plates of the

neural tube were displaced laterally and

ventrally. Nuclei first appeared in medulla by

48 days (4.0 cm CRL) and nerve fibres crossing

in different directions broke up the gray

substance into a mixture of gray and white

mater, the reticular formation. Trapezoid body

started developing by 48 days. Medullary

pyramids appeared on the ventral surface by 81

days of age (13.0 cm CRL). Percentage

contribution of medulla oblongata to the total

brainstem weight increased progressively

during gestation (from 13.90 percent in second

month to 17.57 percent in the fifth month).

When compared to cerebrum and cerebellum,

the MO along with the other regions of the

brainstem was noted to be a slow growing

region. During second month, contribution of

¹Associate Professor, CV&AS, Mannuthy, ²Professor (Retired), 4³Professor and Head, CV&AS, Pookot, Professor and Head,

Department of Veterinary Anatomy and Histology, CV&AS,

Mannuthy.

MO to the total brain weight was 4.69 percent,

which gradually reduced to 3.46 percent in the

fifth month indicating rapid growth of the region

in the first half of gestation followed by a gradual

“cephalic shift” of function from phylogene-

tically older brainstem to the higher cerebral

and cerebellar cortices. Towards term, this

region was well developed and the relative

maturity of the MO in goats at birth justifies the

classification of goat as a prenatal brain

developer.

Key words: Medulla oblongata, Prenatal

development, Goat.

INTRODUCTION

Medulla oblongata is the caudal portion of

the brain, located between the pons rostrally and

spinal cord caudally, resting on the basioccipital

bone. Postnatal studies on the histomorphology of

the hindbrain have been made in different

domestic animals (Jenkins, 1978; King, 1987;

Rizzo, 2006 and Konig and Liebich, 2007).

However, prenatal developmental changes have

not been well documented in ruminants. Hence,

this study was planned to investigate prenatal

morphogenesis of medulla oblongata in goats.

MORPHOGENESIS OF MEDULLA OBLONGATA*IN GOAT FOETUSES

1 2 3 4K. M. Lucy , K. R. Harshan , J. J. Chungath and N. Ashok

College of Veterinary and Animal Sciences, Mannuthy.*Part of the PhD thesis submitted by the first author to Kerala Agricultural University.

RESEARCH ARTICLE



Prenatal morphogenesis of medulla

oblongata (MO) in goats was studied using 46 goat

foetuses ranging from 2.5 cm CRL (40 days of

gestation) to 41.5 cm CRL (full term). The material

available in the Department of Anatomy and those

collected from the farms and clinics were used for

the study. Body weight, body parameters and skull

parameters of the subjects were recorded. The

age of the foetuses was calculated from the 0.33

formula, W = 0.096 (t-30) derived by Singh

et.al. (1979) for the goat foetuses, where 'W' is the

body weight of the foetus in g and 't' is the age of the

foetus in days. Based on the age, the foetuses were

divided into four groups representing second,

third, fourth and fifth months of gestation. The

heads were separated at the occipito-atlantal

junction and the brain was then carefully dissected

out and fixed in 10 percent neutral buffered

formalin. After recording the whole brain

parameters, the MO was separated at the caudal

border of the pons (rostral boundary) and the

rostral limit of origin of first pair of cervical spinal

nerves (caudal boundary). Measurements were

taken and the data were analysed statistically

(Snedecor and Cochran, 1985). Standard

procedures were adopted for histoarchitectural

studies (Luna, 1968).


Development in the Second Month

Medulla oblongata, the caudal most

segment of the brainstem, extended from the level

of first pair of cervical spinal nerves to the caudal

edge of the pons (Fig. 1). It lay on the unossified

basioccipital and this cartilaginous skeleton

developed in the sixth week of gestation.

Measurements of medulla oblongata at different

stages of gestation are given in table 1. By 40 days,

the roof plate region of the embryonic

rhombencephalon expanded enormously. As a

result, the entire alar and basal plates of the neural

tube were displaced laterally and ventrally. Arey

(1957) compared this to an opened book whose

hinge was the floor plate. The lumen became the

fourth ventricle covered dorsally by the thin, single

layer of ependyma, the roof plate. This constituted

the anterior and posterior medullary vela (Figs. 2

and 3). These vela were continuous with the

cerebellum cranially and the roof of the central

canal of spinal cord caudally. The sulcus limitans

present on the ventrolateral wall of the fourth

ventricle provided the plane of division of the

medulla into a ventromedial basal plate and a

dorsolateral alar plate. Similar observations were

made in dog foetuses by Jenkins (1978). The

lumen was filled with CSF. Nuclei appeared by 48

days and nerve fibres crossing in different

directions broke up the gray substance into a

mixture of gray and white known as the reticular

formation. The trapezoid body started developing

at 48 days (4 cm CRL). The point of emergence of

the facial nerve from the medulla is illustrated in

figure 4. The endolymphatic duct also could be

seen within the petrous temporal.

Vascular mesenchyme occupied the

ependymal roof; the combined membrane, the

tela choroidea, infolded as vascular tufts into the

cavity of the myelencephalon constituting the

choroid plexus of the fourth ventricle. This

developed in the goat foetus in the sixth week of

gestation. Keith (1947) reported that the choroid

villi developed on the ventricular surface of the

caudal medullary velum at eight weeks in the

human foetus and CSF was being produced during


the third month. Later during the seventh week,

the ependymal roof plate became broad and thin.

The cavity of the rhombencephalon thus

expanded to the sides, flattened dorsoventrally

and was filled with CSF. The rhombic lip, ridge

where the tela joined the alar plate was made up of

three to four layers of cells. Harrison (1978)

reported that the cells of rhombic lip were actively

mitotic and provided large number of neuroblasts,

which migrated cephalad into the ventral aspect of

the hindbrain where they formed the pontine

nuclei and the olivary nuclear complex.

Caudally the medulla oblongata was

continuous with the spinal cord. The fourth

ventricle narrowed posteriorly to be continued as

the central canal of spinal cord. Dorsal wall of this

region showed thickened epithelium constituting

the circumventricular organ. Medulla oblongata

contributed 4.69 percent of the brain weight and

13.90 percent of the brainstem weight at this stage.

Fig. 1 Dorsal surface of brain (55 days)1. Cerebrum; 2. Corpora quadrigemina;

3. Cerebellum; 4. Medulla oblongata

Fig. 2 C.S. of the medulla oblongata at the levelof rostral medullary velum (48 days). H&E. x 1001. Medulla oblongata; 2. Nuclear aggregation;

3. Fourth ventricle with CSF; 4. Rostral medullary velum;5. Body wall; 6. Internal glial limiting membrane


Fig. 3 C.S. of the medulla oblongata at the level of

caudal medullary velum (48 days). H&E. x 100

1. Caudal medullary velum; 2. Choroid plexus;

3. Fourth ventricle with CSF; 4. Medulla oblongata;

5. Nuclear aggregation; 6. Median sulcus;

7. Sulcus limitans

Fig. 4 C.S. of the medulla oblongata showing thefacial nerve and endolymph duct (48 days). H&E.x100

1. Medulla oblongata; 2. Facial nerve;3. Endolymph duct; 4. Petrous temporal bone

Fig. 5 C.S. of the medulla oblongata showingforamen of Lushcka and caudal medullary velum.(76 days). H&E. x 100 1. Foramen of Lushcka;

2. Caudal medullary velum; 3. Choroid plexus;4. Sulcus limitans

Development In The Third Month

The medullary pyramids could be

distinguished from 81 days (13 cm CRL) and were

in the form of longitudinal ridges on either side of

the ventral median fissure but they were not

widely separated in the rostral portion. These

agree with the findings of Dellmann and Mc Clure

(1975) in small ruminants. However, in cattle the

pyramids were widely separated at the point of

emergence from the caudal aspect of pons.

The dorsal surface of the medulla formed

the floor of fourth ventricle as noted in the second

month. The caudal medullary velum projected

from the dorso-medial angle of medulla

oblongata. The fourth ventricle communicated

with the subarachnoid space by the foramen of

Lushcka (Fig. 5). Floor of the ventricle was marked

by a deep median sulcus, which became shallower

rostrally. On each side of the median sulcus was a

continuous ridge, the medial eminence, bounded

laterally by the sulcus limitans as observed by Truex

and Carpenter (1969) in man and Dyce et al.

(1996) in domestic animals. The medial

eminence, or trigonum hypoglossi was formed by

the nucleus of hypoglossal nerve. The lateral

eminence was occupied by the caudal poles of the

medial and inferior vestibular nuclei. In between

the medial and lateral eminences was the

intermediate eminence, the trigonum vagi.

Mean weight of medulla oblongata was

0.220 ± 0.032g during third month. Width of

medulla oblongata was more than its height

throughout gestation. Average length and width of

medullary pyramids were 0.805 ± 0.031cm and

0.230 ± 0.013cm, respectively. The trapezoid

body was clearly demarcated and the mean

rostrocaudal distance was 0.117 ± 0.005cm.


RESEARCH ARTICLE

Dellmann and Mc Clure (1975) reported that the

trapezoid body was more clearly demarcated in

small ruminants than in cattle.

Development In The Fourth Month

Medulla oblongata contributed 17.57

percent of the brainstem weight. Percentage


brainstem weight increased progressively during

gestation. During the fourth month, mean length,

width and thickness of the medulla oblongata were

1.183 ± 0.027cm, 0.855 ± 0.019cm and 0.682 ±

0.011cm, respectively. Unlike in the pons region,

maximum width of medulla oblongata exceeded

the maximum height. Morphological features did

not change much during the fourth month. Length

and width of medullary pyramids increased 15.03

and 93.04 percent, respectively from third to

fourth month. More increase in width

corresponded to growth of cerebrum since these

fibres have their origin in the cerebral cortex.

Development In The Fifth Month

Trapezoid body was clearly demarcated

from the pons. Cranio-caudal length of trapezoid

body was 0.298 ± 0.003cm. Mean weight of

medulla oblongata increased three-fold during

fifth month. Corresponding changes were also

noticed in the length, height and width of medulla

oblongata (Table. 1). During second month


brain weight was 4.69 percent which gradually

reduced to 3.46 percent in the fifth month

indicating rapid growth of the region in the first half

of gestation followed by a gradual “cephalic shift”

of function from phylogenetically older brainstem

to the higher cerebral and cerebellar cortices.

When compared to other divisions of brain, the

medulla oblongata along with other regions of

brainstem was noted to be a slow growing region.

Ventral median fissure was flanked by the

pyramids. Mean length and width of medullary

pyramids were 1.585 ± 0.088 cm and 0.656 ±

0.008 cm, respectively. Grossly, medulla oblongata

was adult-like during the fifth month (Fig. 6).

Fig. 6 Dorsal view of brainstem (124 days)1. Thalamus; 2. Pineal gland; 3. Rostral colliculus;

4. Caudal colliculus; 5. Fourth ventricle;6. Medulla oblongata

The medulla oblongata is a great

suprasegmental conveyor and co-ordinator for

pathways and nuclei involved with vital regulatory

and protective processes which affect the whole

body. Towards term, this region was well

developed and the relative maturity of the MO in

goats at birth justifies the classification of goat as a

prenatal brain developer. Foetal brain is most

vulnerable when it is growing rapidly and

nutritional deficiencies and diseases during the

growing period can cause permanent damage.

REFERENCES

th Arey, L.B. 1957. Developmental Anatomy. 6 edn,

W.B.Saunders Company, Philadelphia. pp.

454-501.


Dellmann, H.D and Mc Clure, R.G. 1975. Central

nervous system. Sisson and Grossman's thThe Anatomy of the Domestic Animals. 5

edn, (Ed.) Getty R. W.B. Saunders

Company, Philadelphia. pp.1065-80.

Dyce, K.M., Sack, W.O. and Wensing, C.J.G.

1996. Textbook of Veterinary Anatomy. nd

2 edn, W.B. Saunders Company,

Philadelphia. pp. 259-324.

Harrison, R.G. 1978. Clinical Embryology.

Academic Press, London. p. 250.

Jenkins, T.W. 1978. Functional Mammalian ndNeuroanatomy. 2 edn, Lea and Febiger,

Philadelphia. p. 480.

Keith, A. 1947. Human Embryology and th

Morphology. 6 edn, Williams and

Wilkins, Baltimore. pp: 117-53.

King, A.S. 1987. Physiological and Clinical

Anatomy of the Domestic Mammals.

Oxford University Press, New York. p.325.

Konig, H.E. and Liebich, H.G. 2007. Veterinary

Anatomy of Domestic Animals, Text Book rdand Colour Atlas. 3 edn, Schattauer, New

York. pp: 40-48.

Luna, L.G. 1968. Manual of Histological

Staining Methods of the Armed Forces rd

Institute of Pathology. 3 edn, Mc Graw-

Hill Book Company, New York. p.258.

Rizzo, D.C. 2006. Fundamentals of Anatomy and ndPhysiology. 2 edn, Thomson Delmar

Learning, Australia. pp: 244-69.

Singh, Y., Sharma, D.N. and Dhingra, L.D. 1979.

Morphogenesis of the testis in goat. Indian

J. Anim. Sci. 49: 925-31.

Snedecor, G.W. and Cochran, W.G. 1985. thStatistical Methods. 7 edn, The Iowa State

University Press, USA. p. 313.

Truex, R.C. and Carpenter, M.B. 1969. Human th

Neuroanatomy. 6 edn, The Williams and

Wilkins, Baltimore. p. 673.


ROLE OF FECUNDITY GENES IN PROLIFICACY

OF SMALL RUMINANTS

Asha Abraham¹ and Naicy Thomas²

College of Veterinary and Animal Sciences, Mannuthy

INTRODUCTION

The aim of every breeder is to get

maximum profit possible from the animal he is

rearing. This can be achieved by improving the

genetic worth of the stock by proper selection

methods. For the traits that are to be selected, their

relative economic value should be established.

Kidding/lambing percentage is the most important

factor affecting profitability in small ruminants.

Increasing prolificacy offers greater potential for

improving reproduction rate and production

efficiency. Only way to increase the numerical

productivity (no. of kids produced per goat per

year) is that those which are closely linked to

reproductive parameters. Improvement of

reproductive traits in livestock species has become

of increasing interest,especially in small ruminants,

where small increases in litter size can equal large

gains in profit.

PROLIFICACY

Prolificacy is measured as the ability of a

female animal to produce large number of young

ones in their life span through high ovulation rate

and high embryo survival. But, it is difficult to

obtain selection improvements in traits associated

with reproduction, since kidding/lambing

percentage is lowly heritable. Differences among

does/ewes in litter size (single, twin, triplet) are

largely due to non-genetic factors, such as

management and nutrition. Genetic change is

permanent but, nutrition and management vary

from year to year. Common strategy for increasing

prolificacy via genetic means is to select ewes that

are more likely to produce multiple births and to

select rams that are more likely to sire prolific

daughters. This can be accomplished through

simple selection based on birth type or by selecting

for a composite trait, such as weight of young ones

weaned. In the absence of any other information,

it is best to select males and replacement female

lambs/kids that are born as multiples from young

females.

FECUNDITY GENES

1. Booroola gene (FecB)

It is a single gene in chromosome 6 in

sheep which is the main reason for higher

prolificacy of certain breeds. This gene has effects

on granulosa cell maturation, oocyte development

and its function. The increase in prolificacy is due

to autosomal mutation that occurred in this gene

which causes increase in ovulation rate and litter

¹MVSc scholar, ² Assistant Professor, Centre for Advanced

Studies in Animal Genetics and Breeding, College of

Veterinary and Animal Sciences, Mannuthy

RESEARCH ARTICLE


size. Term “Booroola” was taken from the name of

the ranch in Australia, where the sheep carrying

single gene for prolificacy were first discovered.

Booroola gene can be transferred to any other

breed by crossbreeding. This FecB gene is

responsible for the higher prolificacy of Finn sheep,

Romanov, Booroola Merino and British Milk

Sheep. Up until now, the most numerous breed of

sheep containing the FecB gene is Booroola

Merino, the breed it was originally discovered in.

Feeding maintenance quality hay or pasture is

enough for lambing rates of 240-300 percent in

heterozygous Booroola ewes.

Ancestor and original source of Booroola

Merino sheep FecB gene is the Garole Sheep from

the Sunderban (Bengal) area of India. Garole is a

small ewe of 12-15 kg, with a mean litter size of 2.3

lambs, adapted to hot, humid, swampy conditions

of rice paddies. 10 ewes and 2 rams were originally

imported to Australia in 1792, from Calcutta,

India. The FecB gene mutation is found in the

Garole sheep. Genetic evidence supports the

historical records that the prolific Garole sheep,

when introduced to Australia, bred with the much

larger 35-40 kg Merino sheep. Garole sheep are

virtually all homozygous for the FecB gene (BB).

2. GDF9 / FecG gene

Growth differentiation factor 9 (GDF9) is

seen in chromosome 5 in sheep and goat. They are

expressed in oocytes and play an important role in

ovar ian fol l iculogenesis . S ingle s t rand

conformation polymorphism (SSCP) studies of

exon 1 and flanking of GDF9 gene reveals two

silent mutations (183A>C and 336C>T) in AA

genotype in comparison to BB genotype. Studies

show that allele A has correlation with prolificacy

in Jining Grey goat. (Chu, 2004).

3. BMP15 / FecX gene

Bone morphogenetic protein 15 (BMP15)

is an X linked gene seen in sheep and goat. BMP15

gene is essential for oocyte and follicular

development. Higher prolificacy of Inverdale,

Lacaune, Belclare, Small Tailed Han ewes and

Jining Grey goats is due to this gene. Single strand

Normal and FecB gene carriers kept

at Marshall Building in Edinburgh


conformation polymorphism (SSCP) studies show

SNPs in exon 1 and exon 2. Two point mutations

(G963A and G1050C) are found in AB genotype in

comparison to AA genotype (Vacca, 2010).

4. POU1F1 gene

POU domain, class 1, transcription factor 1

(POU1F1) is otherwise known as Pit1 and GH

factor 1. It is seen in chromosome 3 in sheep.

POU1F1 is an important transcription factor for

Growth Hormone. Single strand conformation

polymorphism studies reveals six mutations -

C256T in exon 3, C53T and T123G in intron,

G682T, T723G and C837T in exon (Feng, 2012).

5. Estrogen Receptor gene

Estrogen receptor gene in sheep has two

regions - ESR1 in chromosome 6 and ESR2 in

chromosome 14. Single strand conformation

polymorphism studies show SNP of exon 1 of

Estrogen Receptor (ESR) gene. AC"G mutation

was noticed at the 363bp of exon 1 in BB genotype

on comparison to AA genotype. Marked assisted

selection (MAS) of animals with estrogen receptor

gene polymorphisms can be done to increase litter

size and thus increase in economic value to mutton

producers (Xiao-Dan, 2005).

6. Prolactin receptor gene

Prolactin receptor gene is seen in

chromosome 5 of sheep. This gene mainly

interacts with prolactin and thus increases

proli f icacy. Single strand conformation

polymorphism studies of exon 10 of prolactin

receptor gene reveals two mutations (186G"A

and 220T"C) in AB genotype in comparison to AA

genotype. (Zhang, 2007).

7. FSH receptor gene

FSH receptor gene is seen in Chromosome

2 in sheep. It affects the follicular growth and

hence plays an important role in prolificacy.

Variations in aminoacid sequence of receptor

protein are due to point mutations. Single strand

conformation polymorphism studies shows that

polymorphisms at codon 307 and 680 influence

responsiveness to FSH and affects prolificacy

(Tisdall, 1995).

8. KiSS-1 gene

Kisspeptin (KiSS-1) gene is important for

proper GnRH function and thus affects prolificacy.

It is seen in chromosome 1 in sheep. Single strand

conformation polymorphism studies reveals that

polymorphism of intron 2 results in two SNPs

T2643C and 8bp base deletions (2677AGTT

CCCC) giving rise to four different genotypes CC,

TT, TG and TC (Hou, 2011).

9. INH gene

INH gene (Inhibin gene) is essential for

normal oocyte and follicular maturation and

affects prolificacy. Inhibin genes - INHA and

INH A are found in chromosome 2 in sheep.

Single strand conformation polymorphism studies

reveal a C865T silent mutation in exon of INHA

gene (Yuanqing He, 2010).

CONCLUSION

Genetic improvement of reproductive

â


traits has traditionally been restricted to use of

quantitative genetic methods but gain has been

limited when using these methods. Recent

improvement in molecular genetics provided that

the major genes associated with reproduction can

be utilized in breeding through marker-assisted

selection (MAS). Reproductive traits are often

suggested as prime targets for marker-assisted

selection for their low heritability and the fact that

the trait can be measured only in one sex.

The fecundity genes has posed the unique

and exciting opportunity to add a high level of

prolificacy to sheep and goat, that fit the

environment well, without having to add

undesirable traits of other breed. These genes have

proved to be trackable and persistent, after its

introduction to a breed. Fecundity traits offer a

new option that can allow certain breeds

producing lambs/kids to obtain a high level of

prolificacy by genetic introgression of desired allele

of the fecundity gene. The presence of fecundity

genes, Booroola and Inverdale has been proved as

a cause for high prolificacy for Malabari goats also.

So, by using genetic molecular markers we can

detect the mutations which results in high

prolificacy in certain breeds (Davis, 2004).

Breeders can select males from these breeds to

incorporate into crossbreeding and can attain high

level prolificacy. Speed and efficacy of selection is

expected to increase by the use of these molecular

markers in selection.

REFERENCES

Chu, M. X., Li, B. X., Wang, J. Y., Ye, S. C and Fang,

L. 2004. Association between PCR-SSCP of

growth differentiation factor 9 gene and

high prolificacy in Small Tail Han sheep.

Anim. Biotechnol. 15(2):111- 120

Davis, G. H. 2004. Fecundity genes in sheep.

Anim. Reprod. Sci. 82-83:247-253

Feng, T., Chu, M. X., Cao, G. L., Tang, Q. Q., Di, R.,

Fang, L., and Li, N. 2012. Polymorphisms of

caprinePOU1F1 gene and their association

with litter size in Jining Grey goats. Mol. Biol.

Rep. 39(4):4029-4038

Hou, J. X., An, X. P., Wang, J. G., Song, Y. X., Cui, Y.

H., Wang, Y. F., Chen, Q. J. and Cao, B. Y.

2011. New genetic polymorphisms of KiSS-

1gene and their association with litter size in

goats. Small Ruminant Research. 96 (23):

106-110.

Tisdall, D. J., Watanabe, K., Hudson, N. L., Smith,

P. and Mc Natty, K. P. 1995. FSH receptor

gene expression during ovarian follicle

development in sheep. J. Mol. Endocrinol.

15: 273-281.

Vacca, G. M., Dhaouadia, A., Rekikb, M.,

Carcangiua, V., Pazzolaa, M. and Dettoria,

M. L. 2010. Prolificacy genotypes at

BMPR1B, BMP15 and GDF9 genes in North

African sheep breeds. Small Ruminant

Research. 88(1): 67-71.

Xiao-Dan, B. I., Chu Ming-Xing, Jin Hai-Guo, Fang

Li and Ye Su-Cheng. 2005. Estrogen

Receptor as a Candidate Gene for

Prolificacy of Small Tail Han Sheep. Acta

Genetica Sinica. 2005 -2010

Yuanqing He, Xiaoke Ma, Xiaoyong Liu, Cunxia

Zhang and Jun Li. 2010. Candidate gene

polymorphism and its association in

Chinese goats. J. Agri. Sci. 2:1.

Zhang, G. X., Chu, M. X., Wang, J., Fang, L and Ye,

S. C. 2007. Polymorphism of exon 10 of

prolactin receptor gene and its relationship

with prolificacy of Jining Grey goats. Yi

Chuan. 29(3):329-336


ABSTRACT

Level of participation of livestock farmers

in Panchayati Raj system of democratic

decentralisation was analysed in the study. A

comparison was made between Self Help Group

(SHG) members and non members in terms of

level of participation. The results showed that the

level of participation was medium to high for

SHG members, whereas, it was low to medium

for most of the non members. The difference was

statistically significant.

Key words: Panchayati Raj, Democratic

decentralisation, Self Help Group, Participation

1 2 3PhD Scholar, Professor and Head and Associate Professor

Department of Veterinary and Animal Husbandry Extension

College of Veterinary and Animal Sciences, Mannuthy, Kerala

INTRODUCTION

Democratic decentralisation, often

referred as Panchayati Raj system in India, is meant

for the transfer of authority, responsibility and

accountability from central to local governments.

Any rural development programme in order to be

effective should ensure people's liberal

participation. Thus, in the Panchayati Raj system of

local self governance, the emphasis has shifted

towards the participation of people through

PARTICIPATION OF SELF HELP GROUP MEMBERS AND

NON- MEMBERS IN PANCHAYATI RAJ SYSTEM -*A COMPARATIVE STUDY

1 2 3Anu George , P. J. Rajkamal and R.S. Jiji

College of Veterinary and Animal Sciences, Mannuthy.*Part of M.V.Sc thesis submitted by the first author to Kerala Agricultural University

Panchayati Raj Institutions (PRIs), in planning,

formulation and execution of development

programmes. Decentralisation is effected in terms

of political, financial and administrative

dimensions.

Panchayati Raj system of democratic

decentralisation was revamped in Kerala state

since 1995. It has coordinated grass root level

planning. It is vested with the authority of

formulation and implementation of projects. Local

needs are found out through the gram sabha and

resources are allocated based on the priority of

needs. People participate in the governance,

decis ion making and implementat ion.

Transparency and accountability are the key

features of the system. Local self governance is

important because, weaker sections of the society

have greater chances of participation in decision

making which might affect their lives directly or

indirectly.

The common people, especially the

livestock farmers, have vested much hope in

Panchayati Raj believing that need-based

participatory planning at grass roots can take place

to their own benefit. They are known to participate

in the Panchayati Raj system at varying levels

hopeful of receiving benefits. Self Help Groups

(SHGs), neighbourhood groups (NHGs),

Gramsabha etc. are the venues of participation.

RESEARCH ARTICLE


Self Help Group (SHG) is a small group of

rural people, usually 10-20 in number, gathered

voluntarily to undertake some common income

generating activities through mutual trust and

mutual help (Arunkumar, 2005). In Kerala, SHGs

are gaining more impetus nowadays and many are

taking livestock rearing as the key activity.

Furthermore, the poverty eradication mission of

Kerala Government, Kudumbasree is also

organizing women SHGs including that of livestock

based SHGs.

The present study was carried out with the

objective of comparing the level of participation of

SHG members and non members in PRIs.

METHODOLOGY

The study was conducted in Thrissur

district of Kerala state. Two block panchayats, viz.,

Ollukkara and Irinjalakuda were selected

purposively, since one of the key activities of SHGs

in these blocks was livestock rearing. There were

65 viable livestock- based SHGs in these blocks.

The list of SHG members was prepared with the

help of office bearers of these SHGs. One hundred

members were selected from among the total of

675 members, and they formed the sample of SHG

members. The sample of non-members comprised

of 100 livestock owners selected randomly from

the list prepared with the help of secretaries of milk

co-operative societies and extension personnel

working in these blocks. Thus a total of 200

livestock farmers formed the sample of the study.

The level of participation in PRIs meant the

degree of respondents' participation in the various

Panchayati Raj bodies and activities concerned

with the planning and implementation of projects.

It was measured by personal interview method

using the schedule developed for the purpose. The

findings were expressed as percentage. Based on

the Delenius-Hodges cumulative f method, the

respondents were grouped into three groups as

low, medium and high for level of participation.


Panchayati Raj system of participatory

planning replaced the earlier Community

Development Programme to ensure people's

participation in community development.

According to Mishra (1994) participation means

co-operating or taking part in something and the

mere presence, even the silent presence of an

individual or a representative of an organisation at

different levels can be taken as participation.

1 Low < 8 00 60

2 Medium 8 - 31 21 38

3 High > 31 79 2

Total 100 100

Level of participation in PRIs

Table. 1. Distribution of respondents based on

the level of participation in PRIs n=200

SHGmembers

Non-members

Sl.

NoCategory Score

Frequency (Percentage)

Data in table 1 show that majority (79

percent) of the SHG members were having high

level of participation in PRIs. For the rest 21 per

cent, the level was medium. There was nobody in

the low category. But the situation is different as

far as non members are concerned. The level of

participation in PRIs was low for 60 percent of the

non SHG members, medium for 38 percent and

high for only 2 percent of the respondents. As for


non SHG member livestock owners, reporting

either a medium level or low key participation in

PRIs, was not at all the result normally expected.

This is a precarious situation and as such this

system does not seem to give any hope as far as

livestock owners are concerned. Remedial

measures are urgently needed on a war footing

before it collapses.

Z test on selected variables

Table 2. Z test on selected variables

1 Level of

participation 28.41±1.56 22.96±2.79 17.02**

** (p<0.01)

SHGmembers

Non-members

Sl.

NoVariables

Mean ± SE

Z value

Z test indicated that (table 2) there is

significant difference between the SHG members

and non members in terms of level of participation

in Panchayati Raj.

While studying the perception of

veterinary surgeons of Thrissur District of Kerala

about people's participation in PRIs, Tajne (2003)

confirmed that majority of them felt the extent of

people's participation to be only somewhat

satisfactory. David (1998) also reported that

participation of people in the planning process to

be not satisfactory.

There are apparent synergies between

SHGs and local politics also. Through membership

in SHGs rural people can gain experience in

regular meetings, taking decisions and allocating

money. Since SHG members have more public

contact than others, they can effectively

participate in campaigning too (Anon, 2006). SHG

membership generally contributes to women

getting nominated in election to local bodies.

CONCLUSION

Majority of livestock owners were

reporting either a medium level or low key

participation, where as majority of SHG members

level of participation in PRIs was reported to be

high which was statistically significant. Capacity

building through appropriate training programmes

would be the best strategy for better participation.

The results indicate the need to reconsider the

nature and intensity of SHG promotion. SHG

promotion cannot be considered as a one-shot,

simple input. It has to be more strategic, adaptive

and long-term.

REFERENCES

Anon. 2006. Self Help Groups in India-A study of

the lights and shades. Executive summary of

EDA Rural systems Pvt. Ltd in association

with APMAS for CRS, USAID, CARE,

GTZ/NABARD, p20.

Arunkumar, D. 2005. A critical analysis of Swa-

Shakti programme in Karnataka, M.Sc (Agri)

Thesis, University of Agricultural Sciences,

Dharwad, p112.

David, R.P. 1998. Decentralised planning in

Kerala-Case study of two panchayats. MA

Project Report, Calicut University, p85.

Tajne, S.B. 2003. Work environment of Veterinary

Surgeons of Thrissur District for extension

activities under Panchayati Raj. M.V.Sc.

Thesis, Kerala Agricultural University,

Thrissur, p124.

Mishra, Y. 1994. People's participation in

production process under watershed.

Kurukshetra 42:28-30.


THERMAL STRESS IN DAIRY CATTLE

1 2 3 4A. Prasad , E. M. Muhammed , A. Kannan and T. V. Aravindakshan

College of Veterinary and Animal Sciences, Mannuthy, Thrissur

1 3Assistant Professor, Associate Professor, Department of 2 4Livestock Production Management, PhD Scholar, Professor,

Department of Animal Breeding, Genetics and Biostatistics,

College of Veterinary and Animal Sciences, Mannuthy,

Thrissur.

ABSTRACT

Crosses of temperate cattle reared in

tropical ambience lapse in heat tolerance,

fertility and disease resistance. However, cross

breeding zebu with high yielding exotic cattle

seems necessary to meet the need for enhanced

composite milk production per unit in view of

food security. Thermal stress is considered as the

main factor responsible for reduction of milk

yield in tropical climate. Any model for the study

of thermal stress in dairy cattle should

encompass related effect of ambient parameters

of temperature, humidity, wind speed and solar

radiation as the stressor and behavioural,

autonomic, neuroendocrine and immunological

endpoints as responses. The responses reach

different response states depending on the

severity of the stressor. Response measurements

are at behavioural, physiological, biochemical

and cellular level, which also include hormone,

protein and gene expression assays. By fitting

these measurements to the described model we

can work out the biological and economic cost of

thermal stress and the level of adaptation of the

dairy animal in question. Such studies taking in

to consideration the diverse nature of climatic

factors is imperative for finding ameliorative

measures to reduce the thermal stress

experienced by the existing cattle population

and for the possible genetic and management

strategies for evolving and maintaining a

climatically adapted dairy stock in a state like

Kerala. This review analyses suitable model for

climatic adaptation studies in the hot and humid

climate especially in small holder production

systems.

INTRODUCTION

Challenge before the scientific community

of the tropical world is to find ways to enhance

milk production in the prevailing climatic

conditions. Historically the traditional livestock

production largely depended on heat tolerant

native breeds that produced less milk compared to

temperate exotic breeds. The dairy sector now

largely comprises of extensive and expanding

crossbred population in Kerala. For crossbreds,

increased air temperature, and humidity

measured as Temperature Humidity Index (THI)

above critical thresholds are related to low dry

matter intake (DMI) and to reduced efficiency of

milk production (West, 2003).


RESEARCH ARTICLE

The Zone of Thermoneutrality (Fig. 1) with

in which no additional energy above maintenance

is expended to heat or cool the body (for livestock 0it is between -0.5 to 20 C) and the upper critical

temperature (B on the right side ) may reach to 25-0

26 C (West, 2003). The ambient temperature of

hot humid region is above this critical temperature

during several months of each year. Effective

environmental temperature is a combined effect of

ambient temperature and humidity. The

combined effect is quantified as Temperature

Humidity Index (THI). The normal THI to maintain

production in dairy cattle is 72. In our state, during

most days in a year it is hot and humid and hence

the THI is high enough to cause significant heat

stress.

So the dairy cattle especially those with

exotic blood cannot express its optimum

production potential. Constant exposure of cattle

to high temperature causes a rise in its rectal

temperature, a decline in feed intake, increase in

water intake, a decrease in production of milk,

changes in milk composition, reduction in growth

and even a loss in body weight. This is the reason

for deterioration in the performance of temperate

dairy cattle when introduced into tropical

countries. (Sastry and Thomas, 2005).

The extent of reduction in feed intake is

proportional to the thermal stress i.e. how long and

how much the cow is subjected to temperature

beyond thermo neutral zone. VFA is reduced in

summer, especially acetic acid. Feed efficiency is

considerably reduced. Digestible energy o

utilization was 60 percent at 21 C but only 40 o

percent after 7 days exposure to 32 C and 31

percent after 14 days exposure (Jones and

Stallings, 1999). High ambient temperature and

humidity causes depression in yield of fat and SNF.

Lactose percentage is also depressed when cows

are exposed to high ambient temperature.

Genetic potential of well adapted local

breeds that can be maintained without special

feed concentrates or preventive health care are

being increasingly recognized in this context. The

prevailing harsh climate and the anticipated future

climatic changes are real threats to the

sustainability of the sector. An exact understanding

of the impact of thermal stress on dairy cattle and

possible ameliorative measures are thus extremely

important. Thermal stress alone does not act

separately in reducing the productivity of the

animal, but multiple stresses act on the animal

when nutrition is compromised (Sejian et.al. 2012).

Since, interpretation of the effect of multiple

stressors are difficult, multiple responses of the

thermal stress alone is discussed here.

Fig.1 Kleiber's law of metabolic heat production and

core body temperature as influenced by environment

temperature. C = lower critical temperature D= point

of reduction of metabolic heat. B on the left side =

lower point of zone of thermal neutrality below which

chemical regulation is needed to maintain

homeothermy. B on the right side = upper critical

temperature.


'Stress' conceptual clarity

The word 'stress' is perceived by different

people differently. Even among animal scientists,

different disciplines approach stress in different

angles. Here an attempt is made to elucidate the

word stress in the context of thermal stress in dairy

cattle. Stress is a part of life. All life forms have

evolved mechanism to cope with the stresses of

their lives. Moberg and Mench (2000) defined

stress as the biological response elicited when an

individual perceives a threat to its homeostasis.

The threat is the stressor. When stress response

truly threatens the animal's well-being then the

animal experience distress.

The stressor in our context is the effective

environmental temperature represented by the

temperature, solar radiation, humidity and air

movement in the immediate vicinity of the dairy

cow. Our challenge is to determine when their

stress becomes distress and to determine how to

measure stress and distress in animals.

Evolving a model for assessing thermal stress in

dairy cattle

Moberg (2000) divided stress response in

to three general stages

1. Recognition of a stressor

2. Biological defense against the stressor

and

3. Consequence of the stress response.

A stress response begins with the CNS

perceiving a potential threat to homeostasis. Then

the cow develops biological response or defense

that consists of four general biological defense

responses

1. Behavioural

2. Autonomatic nervous system

3. Neuro Endocrine

4. Immune response

Now let us attempt to evolve a model

forthermal stress in dairy cattle which better

clarifies our problem and will help in attempting to

assess it scientifically.

(Behavioural, autonomic,

neuro endocrine, immunological)


Challenges in measuring thermal stress

Scientists rely on a variety of endocrine,

behavioural, autonomic nervous system and

immunological end points to measure stress.

Unfortunately, none of these measures has proved

to be a litmus test for stress. Further, complicating is

inter animal variability in stress response.The

above model demonstrates that all the responses to

the perceived stressor can reach any one of the

response state depending the severity of the

stressor and the vulnerability of the animal. The

vulnerability depends upon the modifiers of the

responses. Modifiers which shape animal's

organization of its biological defenses are early

experience, genetics and age. The responses are

not mutually exclusive but need to be measured

independently for better assessment. After

assessing them independently measures related to

each response must be compared with normal

values to arrive at the respective response state. As

any one single response cannot be considered to

be exclusively due to stress, the non parametric

approach of considering the mode of response

states for the entire array of tests may be taken as

the animal's thermal stress level. Different

responses and possible measurements are

explained below and illustrated in Fig. 2.

Behavioural response

A cow seeking shade during direct solar

exposure is an example. But in case of dairy cattle

behavioural response options are limited by

confinement. When the severity of the stressor

increases, this response can manifest as altered

behaviour, pre pathological behaviour and even

pathological behaviour. But all the animals will not

show same degree of alterations in behaviour to

the same degree of stressor.

Autonomic nervous system response

During thermal stress the autonomic

nervous system affects a diverse number of

biological systems including the cardiovascular,

gastro intestinal, exocrine glands and adrenal

medulla. The results are altered heart rate, blood

pressure and gastrointestinal activity. But stress

activation of autonomic nervous system is of

relatively short duration. According to the grade of

the stressor and the animal's vulnerability each of

the autonomic nervous system responses also

differs.

Neuro endocrine

Hormones secreted from hypothalamic

pituitary system have a broad long lasting effect on

the changes induced by body stress. The secretions

of pituitary hormones have been implicated in

failed reproduction, altered metabolism, immune

competence and behaviour. The hypothalamic -

pituitary- adrenal (HPA) axis has been primary

neuroendocrine axis. However, the secretion of

prolactin and somatotropin has proven to be

equally sensitive to stress and has considerable

economic importance. Likewise, thyroid

stimulating hormone and gonadotropins (LH and

FSH) are either directly or indirectly modulated by

thermal stress.

Fig. 2. Extrapolation of Fig 1 to depict the response

states of the animal when subjected to effective

temperature above the upper critical temperature.

(The column 1 - altered, 2- pre pathological and. 3-

pathological response states.


Increased secretion of the adrenal

glucocorticoids and cortisol has been found to be

associated with stress and investigators frequently

cite an increase in circulating cortisol as a proof of

stress. Colborn et.al., (1991) found that stallions

secreted similar amounts of cortisol whether the

stallions were restrained, exercised or permitted to

mate with a mare. Serum cortisol concentrations

are often used to evaluate stress, but due to the

marked variability, faecal corticosterone has been

used to evaluate stress in cattle (Morrow et.al.,

2002). Dairy cattle secrets cortisol during restrained

and when they are approached by strangers. So it is

difficult to simply use secretion of cortisol or any

other hormone to differentiate between non

threatening stress and distress. Even among the

same breed, some cows may produce more

cortisol to the same amounts of thermal stressors.

Immune response

We have long attributed the increased

incidence of disease in cows suffering from thermal

stress. Immune system in its own right is one of the

major defense systems responding to stressor

(Dunn, 1988). Measurement of immune

competence offers us a potentially powerful tool

for evaluating the disease components of distress.

According to Kumar et.al., (2011) heat stress is one

of the wide varieties of factors which cause

oxidative stress in-vivo. Reactive oxygen species

(ROS), constantly generated in vivo as an integral

part of metabolism cause oxidative stress when

their level exceeds the threshold value. Superoxide

ions, hydrogen peroxide and hydroxyl ions (ROS)

produced as a result of heat stress are highly

reactive and they cause damage to the

polyunsaturated fatty acids of lipoprotein layer of

cell membrane. This lipid peroxidation leads to the

formation of lipid peroxides which are again

harmful to the cell components. The amount of

lipid peroxides is measurable and hence can be

used as reliable indicator of level of thermal stress.

Reduced glutathione present in the animal acts as

an anti oxidant by getting itself oxidized. So in a

distressed animal the amount of reduced

glutathione will be low.

The heat shock response is a highly

conserved cascade of altered protein and gene

expression in animals. The altered intracellular

proteins secretion is due to the concerted action of

physiological stress response which constitutes a

system wide gene network coordinated across a

variety of cells and tissues to minimize effects of

adverse environmental conditions. Endocrine and

metabolic responses are as result of gene

expression changes that include (1) activation of

heat shock transcription factors (HSFs), (2)

Increased expression of HSPs and deviated

expression and synthesis of extra proteins, (3)

Increased protein, glucose and amino acid

oxidation and reduced fatty acid metabolism, (4)

Endocrine system activation and (5) Immune

system activation (Richter et.al., 2007).

Diverse physiological stresses (thermo-

dynamics, mutant proteins and oxidative injury)

produce multiple changes in a cell that ultimately

affect protein structures and function. Cells from

different phyla initiate a cascade of events that

engage essential proteins, the molecular

chaperones, in decisions to repair or degrade

damaged proteins as a defense strategy to ensure

survival. Molecular chaperones such as the heat

shock family of stress proteins (HSPs) actively

participate in an array of cellular processes,

including cytoprotection. The versatility of the

ubiquitous HSP family is further enhanced by

stress-inducible regulatory networks.


Biological cost of thermal stress

Whether or not the stress altered functions

are beneficial in helping the cow to cope is not our

immediate concern. The changes in biological

function during stress result in a shift of biological

resources away from biological activities occurring

before the stressor. For example, energy originally

utilized for growth or reproduction might be

needed by animal to cope with stress. This change

in biological function during stress is the biological

cost of stress. During prolonged stress or when

stress is severe the biological cost is significant and

the work of stress becomes a significant burden to

the body. It is during such stress that the animal

enters the next stages of stress pre pathology and

pathology.

In a hot humid climate like the one in

Kerala, the dairy cows suffer from chronic distress

which is the sum total of all the compensations or

displacements caused by the thermal stress

applied. This chronic distress represents the totality

of discomfort, felt by the cow in a hot environment.

The cows under this chronic distress will have

reduced feed intake and increased water

consumption. The body temperature will rise often

stabilizing at a higher level. There will be shift in

body water from intra cellular to extra cellular and

extracellular to vascular space which will be

mobilized to effect evaporative thermal cooling

from skin and respiratory tract.

Increased respiratory activity leads to

excessive blowing out of carbon dioxide through

the expired air thus upsetting carbonic acid

bicarbonate buffer system. In order to maintain

acid base balance, bicarbonate, mainly in the form

of sodium bicarbonate or potassium bicarbonate is

excreted through urine. Excessive loss of sodium

from body triggers corticosteroid hormone.

Ultimately acid base and mineral balances are

disturbed. Excessive moisture through evaporative

channels can cause dehydration. Under hot

condition, when subjected to water scarcity,

energy utilization efficiency is reduced and

reproductive efficiency is progressively affected.

Acclimation and adaptability factor

Acclimation is within life time phenotypic

response to environmental stress and is a

homeothermic process driven by endocrine system,

whereas, adaptability involves evolutionary changes

that occur over time scales covering multiple

generations. Alterations in gene expression and

changes in cellular signaling are key components

of adaptability. The changes in gene expression

mainly include that of HSPs. The major symptom

in crossbred cattle is reduced feed intake, when

the animals are under heat stress. Even if they are

fed adlibitum, that too high quality feed, the

animals do not take feed. This may be due to

oxidative cellular stress that leads to cell starvation.

Cell starvation often leads to inability of the cells to

utilize glucose and other energy releasing

molecules. When the cells are under starvation,

growth, production and reproduction are

compromised first, and then the vital functions of

body. All these changes are associated with altered

nutrient partitioning that accounts for production

loss. In the acclimated state, metabolism is

adjusted to minimize detrimental effects of

increased thermal loads. We have to determine

the basis of altered energy metabolism during

thermal stress in exotic and indigenous animals

which may lead to opportunities for improved

animal performance via altered breeding

strategies.


What contributed more to the natural

selection are the fitness traits such as fertility and

viability. The cattle breeds evolved in temperate

and tropical climate are adapted to respective

climate in terms of these fitness traits. When they

are taken to alternate conditions fitness traits are

maintained where as production traits are

compromised. So it is not possible to get a high

production from these breeds in alternate

conditions (i.e. temperate breeds in tropical

climate and tropical breeds in temperate climate).

CONCLUSION

Thermal stress is a major threat to the

viability and sustainability of milk production

around the world, especially in tropical climate.

The Inter-governmental Panel on Climatic Change

(IPCC) has forecasted global warming leading to 0rise in average temperature by 1.8-4 C by the year

2100. Thus, the deteriorating prevailing condition

is going to be aggravated unless measures to

alleviate the thermal strain are addressed at

appropriate level. For suggesting genetic,

nutritional and management measures to

overcome this challenge, scientific operational

definition of thermal stress elucidated through

suitable models are imperative. The model

suggested in the present study is based on various

reviews on different aspects of the subject. The

model depicts the different levels of stress response

to the perceived stressor and various methods to

measure the responses. This model will also help in

determining the level of acclimation and

adaptability of cattle.

REFERENCES

Colborn, D. R., Thomson, D. L., Roth, T. L.,

Capehart, J. S. and White, K.L.1991.

Response of cortisol and prolactin to sexual

excitement and stress in stallions and

geldings J. Anim. Sci.69:2556-2562.

Hutcheson, D.P. and Cole, N.A. 1986.

Management of transit stress syndrome in

Cattle: Nutritional and Environmental

Effects. J. Anim. Sci. 62:555-560.

Jones, G.M. and Stallings, C.C. 1999. Reducing

Heat Stress for Dairy Cattle. Virginia

Cooperative Extension, Dairy Publication,

404-200.

Kumar, S.B.V., Ajeet, K and Meena, K. 2011. Effect

of heat stress in tropical livestock and

different strategies for its amelioration.

J. Stress Physiol. Biochem. 7 (1): 45-54.

Moberg, G. P and Mench, J. A. 2000. The Biology

of Animal Stress Basic Principles and

Implications for Animal Welfare, CABI

Publishing

Morrow, C. J., Kolver, E. S., Verkerk, G. A. and

Matthews,L.R. 2002. Faecal glucocorticoid

metabolites as a measure of adrenal activity

in dairy cattle. Gen. Comp. Endocrinol.

126: 229-241.

Rao, G.S.L.H.V.P. 2003. Agricultural Meteorology

Kerala agricultural University, Thrissur,

Kerala, India, p.231.

Richter, K., Haslbeck, M. and Buchner, J. 2010.

The heat shock proteins: Life on the verge

of death. Molecular cell. 40:253-266.

Sastry, N. S. R and Thomas, C. K. 2005. Livestock th

Production Management (4 ed.). Kalyani

Publishers, New Delhi, pp. 13-15.

Sejian, V., Naqvi, S. M. K., Ezeji, T., Lakritz, J and

Lal, R. 2012. Environmental stress and

amelioration in livestock production.

Springer, Newyork . pp 129-144.

West, J. W. 2003. Effects of heat stress on

production in dairy cattle J. Dairy Sci.

86:2131-2144


SUCCESSFUL MANAGEMENT OF CHYLOTHORAX

IN A DOG- A CASE REPORT

1 2 3 4G. Vijayakumar , S. Sivaraman , E. Venkatesakumar and M. Subramanian

Veterinary College and Research Institute, Namakkal

INTRODUCTION

Chylothorax is the accumulation of chyle

in the pleural cavity and has been reported in dogs,

cats, humans and other species (Ettinger and

Feldman, 2010). Any disease or process that

increases systemic venous pressures (i.e. right heart

failure, mediastinal neoplasia, cranial venacava

thrombi or granuloma) may cause chylothorax.

CASE HISTORY AND CLINICAL FINDINGS

A non descript male dog aged about 8

years was brought to Veterinary College and

Research Institute hospital with history of

dyspnoea and not taking food for five days. Clinical

examination of the dog revealed pale mucous

membrane, cough, abducted elbows, severe

dyspnoea, oral breathing / panting, lethargy,

exercise intolerance and tachycardia. Auscultation

revealed muffled heart sound and murmur.

Haemato -biochemical examination did not reveal

any abnormality. Thoracocentesis was done and

about 520ml of chyle was removed on the day of

presentation (Fig 1). Analysis of the pleural effusion

revealed chyle (Fig 2) white coloured, opaque with 3

2.5g/dl protein and WBC 5.23 x10 /cumm).

Ultrasonography of the thorax and heart revealed

pleural effusion and tricuspid value insufficiency.

Electrocardiography and radiography of the

animal in standing posture depicted reduced QRS

complex and fluid accumulation in the pleura with

loss in cardiac silhouette respectively.

Fig 1. Needle Thoracocentesis

Fig 2. Chyle- White coloured and opaque in nature

1Associate Professor (Corresponding author), Department of Veterinary Clinical Medicine, Veterinary College and Research

2Institute, Namakkal, [email protected]. Assistant Professor, TVCC, Veterinary College and Research Institute,

3 4Thanjavur. Assistant Professor and Professor and Head, Department of Veterinary Clinical Medicine, Veterinary College and Research Institute, Namakkal

CLINICAL REPORT


TREATMENT

The dog was administered with Ringers

lactate (10ml/kg body weight intravenous) to

counteract shock along with frusemide (4mg/kg

IM). Enalapril (0.5mg/kg PO bid), frusemide

(4mg/kg PO bid). Salt restricted diet were

advocated. it was suggested that the animal should

not be put to exercise. Amoxicillin-Cloxacillin

(20mg/kg PO bid) was prescribed for five days. The th

animal was presented again on 8 day for check

up. Dog showed good clinical improvement and

was able to take food and water by itself. The

degree of dyspnoea was reduced and animal was

able to lie down without any respiratory distress.

The dog had uneventful recovery from pleural

effusion and is presently under treatment with

enalapril and frusemide for valvular insufficiency.

DISCUSSION

Chylothorax is more commonly caused by

transmural leakage of chyle that occurred through

intact but dilated lymphatic vessels. Underlying

diseases that have been reported to cause

chylothorax include heart disease (cardiomy-

opathy, pericardial effusion, heartworm disease,

tetralogy of Fallot, tricuspid dysplasia,

cortriatriatum dexter), lymphatic or mediastinal

neoplasia, fungal granuloma, venacaval

thrombosis, peritoneal pericardial diaphragmatic

hernia, lung lobe torsion, and congenital

abnormalities of thoracic duct (Birchard et. al.,

1998). In most small animal patients, chylothorax

is considered to be idiopathic because the specific

aetiology remains unknown (Ettinger and

Feldman, 2010). In the present case, chylothorax is

due to heart disease caused by tricuspid valve

insufficiency. Sturgess (2001) reported that the

common presenting signs in chylothorax were

restrictive breathing pattern, dyspnoea,

tachpnoea, coughing, weight loss, muffled heart

sound, and decreased lung sound on ventral area.

The clinical signs noticed in the present study are

similar to the reports of the above author. Chyle

appeared white milky to pink coloured, opaque.

(Fossum, 2007). Radiography was helpful in

identifying masses, neoplasm, cardiac disease or

lung lobe torsion. Echocardiography identified

cardiac diseases and effusion (Ettinger and

Feldman, 2010). Emergency procedure that has to

be adopted in patients with pleural effusion is

therapeutic needle thoraco-centesis. Needle th

thoracocentesis is done at 7 intercostal space on

the ventral one third of the thorax and removal of

fluid at the rate of 10ml/kg body weight is sufficient

to result in significant improvement in respiration

(Sturgess, 2001). In the present study, clinical signs,

radiography and ultrasonography were useful in

diagnosing chylothorax. Other medical

management strategies included use of low-fat

diets, benzopyrone octreotide in addition to

treatment of underlying cause. Benzopyrone may

increase the number and function of macrophages

to remove protein from lymph and promote fluid

reabsorption. Octreotide, a somatostatin analog

that inhibits gastric, pancreatic and biliary

sectretions, promote gastrointestinal water

absorption can also be used (Rasiah et.al.,2003). If

chylothorax is idiopathic, surgical management

with ligation of thoracic duct with pericardiectomy,

pleuro peritoneal shunting, pleuro venous shunting

or pleurodesis could be done (Fossum, 2007).

SUMMARY

A successful management of chyloth-orax

due to heart disease in a dog with thoracocentesis,

frusemide, and enalapril is placed on record.


ACKNOWLEDGEMENT

The authors are thankful to the Dean,

Veterinary College and Research Institute,

Namakkal for the facilities provided.

REFERENCES

Birchard, S.J., Smeak,D.D and McLoughlin, M.A.

1998. Treatment of idiopathic chylothorax

in dogs and cats. J. Am. Vet. Med. Assoc.,

212:652.

Ettinger, S.J and Feldman, E.C. 2010. A textbook of th Veterinary Internal Medicine, 7 edition,

Saunders Elsevier, Philadelphia.

rdFossum, T.W. 2007. Small Animal Surgery. 3

Edition. Mosby Elsevier, St. Louis, Missouri.

Sturgess, K. 2001. Diagnosis and Management of

chylothorax in dogs and cats. In Practice,

23:506.

Rasiah, S.V., Oei, J and Lui ,K. 2003. Octreotide

in the treatment of congenital chylothorax.

J. Paed. Child Health, 40:585.

The architect of 'white revolution', Varghese Kurien, who led 'Operation Flood' to

transform India from a milk-deficient country to the world's biggest milk producer died on 9-9-

2012 at the age of 90. Hailed as the undisputed 'Milkman of India', who created the billion

dollar brand Amul, he is credited with laying the foundation of the nation's co-operative dairy

model.

The Indian Government had conferred on him the Padma Vibhushan. He was also the

recipient of World Food Price, Ramon Magsaysay award for Community Leadership, Carnegie-

Wateler World Peace Prize and International Person of the Year award from US.

Born in Kozhikode, Kerala, on November 26, 1921, Kurien had graduated in science from Loyola College

in Chennai (1940) and obtained his degree in engineering from the Guindy College of Engineering in Chennai.

After a stint at TISCO, Jamshedpur, Kurien got the Government of India's scholarship to study dairy engineering.

Following specialised training at Imperial Institute of Animal Husbandry and Dairying in Bangalore, Kurien went to

the USA where he completed his masters' degree in mechanical engineering with dairy engineering as a minor

subject from Michigan State University in 1948. On his return to India, Dr. Kurien was assigned to join

Government Creamery located at Anand in Gujarat to serve his bond period. Kurien then joined Kaira District

Cooperative Milk Producers' Union Limited in 1949 on the request of Tribhuvandas Patel, the then Dairy

Chairman. The dairy was formed at the initiative of Sardar Vallabhabhai Patel. Later, Patel asked Kurien to help set

up a dairy processing plant which saw the birth of Amul.

Amul's co-operative model became a success and it was replicated throughout Gujarat. The different

dairy unions were later brought under the banner of Gujarat Co-operative Milk Marketing Federation (GCMMF).

Dedicating his professional life to empowering the Indian farmers through co-operatives, Kurien, served the

GCMMF from 1973 to 2006, and the Institute of Rural Management (IRMA) from 1979 to 2006. Kurien's tenure at

Anand changed the destiny of Indian dairy industry. The first dairy co-operative union in Gujarat was formed in

1946 with two village dairy co-operative societies as its members.

By 1955, Kurien led to the development of the iconic Amul brand for selling the milk of the co-operative.

In 1965, Kurien's leadership caught the attention of the Prime Minister Lal Bhadur Shashtri. He asked Kurien to

lead the National Dairy Development Board (NDDB) and replicate the Co-operative success story of Amul across

the country. In 1970, with the help of the World Bank, the NDDB started “Operation Flood” which, over the next

26 years, transformed India from a milk importer to world's top most milk producing country. Kurien came to be

known as the “Milkman of India” and the “Father of White Revolution".

HOMAGE TO Dr. VARGHESE KURIEN: INDIA'S WHITE KNIGHT


SURGICAL MANAGEMENT OF TESTICULAR

SEMINOMA - A CASE REPORT

1 2Laiju. M. Philip and M. Ranjith Mohan

Animal Husbandry Department, Kerala

INTRODUCTION

Testicular tumors are common and

account for 4 to 7 percent of all tumors in male

dogs. Most affected dogs are over 5 years of age,

with a median age of 10. The majority of tumors

occur in undescended testicles located in the

inguinal canal or abdominal cavity. Tumors in

descended testicles are less common (Grieco, V.

et.al, 2008). The affected testicle is often larger and

firmer than its neighbour and has an irregular,

nodular surface. At times the testicle is normal size

but feels hard. In this paper a case of Testicular

Seminoma and its surgical management in a

German Shepherd dog is discussed.

CASE PRESENTATION

A five year old German Shepherd dog was

presented to Veterinary Polyclinic, Mannarkkad

ABSTRACT

Testicular Seminoma was diagnosed in a

five year old monorchid German Shepherd dog.

After Ultrasoundscanning and laboratory

examination, scrotal ablation was done to repair

the condition.

with a complaint of scrotal enlargement (Fig. 1).

On detailed examination, the animal was found to

be monorchid and it was confirmed by

Ultrasonography. The scrotal sac was hard in

consistency. Ultrasound scanning revealed

thickened testicular wall and a homogeneous

testicular mass of low echogenicity (Fig. 2).

Cytological examination of Fine needle aspirate

revealed confirmation of seminoma of testis.

Hence decided to perform scrotal ablation.

The surgical site was prepared aseptically

after restraining the animal. The dog was

premedicated with Atropine Sulphate at the rate

1Veterinary Surgeon, Veterinary Polyclinic, Mannarkkad2Veterinary Surgeon, Veterinary Dispensary, Karakurissi

Fig. 1 Dog with Enlarged Scrotum

0.45mg/kg body weight intra muscularly, followed

by an intravenous bolus injection of Xylazine

Hydrochloride at the rate 1 mg/kg body weight and

Ketamine Hydrochloride at the rate 5 mg/ kg body


CLINICAL REPORTS

weight. An elliptical incision was made around the

base of the scrotum. Separated the scrotal skin and

fascia to expose the spermatic cord. The testis was

removed along with the spermatic cord and

scrotal sac, after ligating the testicular artery and

testicular vein separately. Apposed the skin incision

in a horizontal mattress pattern. Parenteral

medication was done using Inj. Amoxicillin-

Dicloxacillin 500 mg for three days. The animal

had an uneventful recovery.

Fig. 2 Ultrasound scanning revealing homogeneous

testicular mass of low echogenicity

Fig. 3. Surgically Removed Testicular Tumour

DISCUSSION

Testicular tumours affect mostly the Boxer,

German shepherd, Afghan hound, Weimaraner

and Shetland sheepdog type of breeds.

There are three common types of

testicular tumours. Sertoli cell tumours,

seminomas and Leydig (interstitial) cell tumours.

Seminoma is a unilateral, single, often benign

tumour of the testis; however, malignant forms of

the tumor have been reported in rare cases. It is the

second most common tumor of the testis in male

dogs, typically affecting older dogs (over the age of

four). Seminomas develop due to cryptorchidism,

a fetal abnormality which occurs when one or both

testes fail to descend into the scrotum from where

they develop in the abdomen (Hayes, H.M. Jr and

Pendergrass, T.W. 1976). There is 16-times risk of

developing seminoma in the cryptorchid testicle

compared to descended testicle in dogs with

unilateral cryptorchidism (Reif, J. S et.al, 1979).

Seminoma in descended testicle found in younger

dogs and associated with contralateral cryptorchid

testicle.Ultrasound examination is a sensitive and

relatively specific technique for the diagnosis of

testicular tumors.

REFERENCES

Grieco, V., Riccardi, E. and Greppi, G.F. 2008.

Canine testicular tumours a study on 232

dogs. J. Comp. Path. 138: 86-89.

Hayes, H. M. Jr and Pendergrass, T. W. 1976.

Canine testicular tumours: Epidemologic

Features 410 dogs. Int. J. Cancer. 18 (4) :

482 - 487

Reif, J. S., Maguire, T.G., Kenney, R.M and Brodey,

R.S. 1979. A. Cohort study of canine

testicular neoplasia. J. Am. Vet. Assoc. 175

(7) : 719-723.


A REPORT ON THE OCCURANCE OF Hymenolepis anatina

IN DUCKS UNDER BACKYARD SYSTEM

1 2 2 2 2G. Jyothimol , K. Syamala , M.N. Priya , C.K. Deepa , K.G. Ajithkumar ,

3 2Ajith Jacob George and Reghu Ravindran

College of Veterinary and Animal Sciences, Pookot

Hymenolepis anatina

INTRODUCTION

Reports on the occurrence of helminth

infections in anseriform birds affecting their health

and well-being are scant (Schiller,1951).

Tapeworms of the genus Hymenolepis have been

frequently encountered in large numbers in water

fowls reared under intensive conditions (Islam

et.al., 1988) . The present communication reports

the occurrence of a cestode parasite, Hymenolepis

anatina in domestic ducks from Wayanad, Kerala,

and its pathogenic effects.

ABSTRACT

Two different cases of mortality in ducks

reared under backyard system due to severe

cestode parasitism were reported. On

postmortem examination varying degrees of

enteritis was also observed. The parasites were

collected and identified based on morphology

as Hymenolepis anatina. The present

communication reports the occurrence of H.

anatina for the first time in domestic ducks of

Kerala.

1 2MVSc Scholar, Assistant Professor, Department of Veterinary 3Parasitology, Associate professor, Department of Veterinary

Pathology, College of Veterinary and Animal Sciences, Pookot,

Wayanad


Two cases of mortality in domestic ducks

reared under backyard system were reported.

Case number 1:- Forty out of 100 ducks of 6 month

age reared by a private owner from Mananthavady

died. The owner reported that the birds were

brought one month before from a poultry market

at Thrissur.

Case number 2: Three out of 10 ducks

owned by an agricultural farmer from Kalpetta

showed mortality.

In both cases, a representative number of

birds were brought to the Department of

Veterinary Pathology, College of at Veterinary and

Animal Sciences, Pookot, Wayanad for conducting

postmortem examination. Parasites were collected

for routine processing and identification. Parasites

were identified based on (Singh, 2003).

RESULT

It was observed that the cestode parasites

were present in large numbers as to occlude the

lumen of intestine which has resulted in enteritis

of varying degree. The parasites were identified as

Hymenolepis anatina based on morphological

features. Each proglottid showed three testis and

one bilobbed ovary within a single segment (Fig.1)


CLINICAL REPORT


Vitelline gland was clearly seen behind the ovary.

Genital pore was unilateral in each segment. The

scolex revealed the presence of ten hooks on the

rostellum, which was typical for H. anatine (Fig.2)

Fig.1: Hymenolepis anatina segments

Fig.2: Hymenolepis anatina hooks on rostellum

DISCUSSION

Cestode parasitism in wild ducks usually

maintain a balance, where the host is not affected

with serious consequences. Among the helminthic

infections of anseriform birds, the most commonly

observed parasites were of Hymenolepidae family

(Schiller,1951). Kharchenko (1960) observed that

hymenolepids are most common in summer and

spring season.

Usually most of the Hymenolepis sp. were

considered to be non-pathogenic but severe

infections may cause death. In the present study,

H. anatina is reported for the first time from

domestic ducks of Kerala. There were no available

reports on the pathogenicity of these parasites. The

mortality observed in ducks could be attributed to

the severe cestode parasitism.

REFERENCES

Islam, M. R., Shaikh, H and Baki, M. A. 1988.

Prevalence and pathology of helminth

parasites in ducks of Bangladesh.

Veterinary Parasitology. 29:73-77

Kharchenko, O. N. 1960. The problem of

helminth - carriers among ducks with

hymenolepid infection. J. Helminthologia.

4: 249-253

Schiller, E. L.1951. The cestoda of anseriformes of

the North Central states. The American

Midland Naturalist. 46: 444-461

Singh, K. R. S. 2003. Veterinary Helminthology.

Di rec tora te o f in format ion and

publication, Indian Council of Agricultural

Research, New Delhi. pp: 221

ABSTRACT

A kathiawari horse aged three years was

presented with swelling throughout the body.

Clinical examination revealed a penetrating

wound in the axillary region with subcutaneous

emphysema. The wound was closed with

antibiotic impregnated gauze with minimal

debridement an uneventful recovery occurred.

Keywords: Axillary wound, subcutaneous

emphysema

1 2Assistant Veterinary Officer, Veterinary officer, Bangalore Turf

Club Limited, No.52, Racecourse road, Bangalore.

MANAGEMENT OF SUBCUTANEOUS

EMPHYSEMA IN A HORSE

1 2Mir Aamir Ali and H.S. Mahesha

Bangalore Turf Club Limited, Racecourse road, Bangalore.

INTRODUCTION

Axillary wounds are caused by penetration

of a sharp object like trees, fences, gates and

usually do not involve the thorax (Laverty et.al.,

1996). Marked subcutaneous emphysema

develops because the wound tissue becomes a one

way channel for air and as the horse moves the air

progressively accumulates in soft tissues. The

trapped air may migrate through fascial planes with

potential to cause pneumothorax, (Hance et.al.,

1992).

CASE HISTORY AND TREATMENT

A kathiawari horse aged about 3 years was

presented with a complaint of swelling throughout

the body and also involving the face. Clinical

examination revealed a penetrated wound in the

maxillary region with diffuse accumulation of air

with crepitation and the skin pitting on pressure,

with congested mucus membranes and anorexia

were the other associated symptoms. The wound

was cleaned with normal saline and the wound

was closed with antibiotic ointment and sterile

guaze. Fig. (1and 2).

Fig.1. Diffusely swollen head and body


CLINICAL REPORT

Fig. 2. Penetrating wound in the axilla

Tetanus toxoid was given initially and

gentamicin was given intravenous at 6.6mg per Kg

body weight once daily along with metronidazole

at 25mg per Kg orally four times daily and penicillin

sodium at 22000 I.U. per Kg body weight

intravenously twice daily. Phenyl butazone was

given at the dose rate of 2.2 mg per Kg

intravenously once daily was administered for ten

days.

RESULT AND DISCUSSION

The clinical symptoms as observed in the

current case were also reported by Laverty et.al.,

(1996), Hassle (2007). Axillary wounds were found

associated with subcutaneous emphysema and

predisposing to pneumothorax that was not

observed in this case. Stall rest was advised to avoid

further accumulation of air and continued until

recovery. Surgical closure of the wound was not

considered as reported by Marble et.al., (1996),

Joellugo (2006) and Florent David et.al., (2008).

Antibiotic impregnated gauze was well tolerated

by the horse and following which the horse

recovered without any respiratory complication.

REFERENCES

Florent David and Sheila Laverty, 2008. Thoracic

Trauma. In:Five minute Veterinary Consult

Equine, Jean pierre Lavoie and Kenneth

Hinchcliff (Ed. 2), Elsevier., pp.756-757.

Hance, S.R and Robertson, J.T. 1992. Subcutaneous

Emphysema from an axillary wound that

resulted in pneumomediastinum and

bilateral pneumothorax in a horse. J. Am.

Vet. Med. Assoc., 200:pp.1107.

Hassel, D.M. 2007. Thoracic Trauma in horses.

Vet. Clin. Equine., 23:67-80.

Joel Lugo. 2006. Thoracic disorders. In Equine

Surgery Auer and Stick (Ed. 3), Saunders

Elsevier St. Louis USA., pp. 620-621.

Laverty, S., Lavoie, J. P., Pascoe, J.R and Ducharme,

N. 1996. Penetrating wounds of the thorax

in horse. Equine. Vet. J., 28:pp.220.

Marble, S.L., Edens, L.M and Shiroma, J.T. 1996.

Subcutaneous emphysema in a neonatal

foal. J. Am. Vet. Med. Assoc. 208:pp.97.

BEST SCIENTIFIC PAPER AWARD OF INDIAN VETERINARY ASSOCIATION, KERALA

Indian Veterinary Association, Kerala has decided to give best scientific paper awards annually

for the best papers published in Journal of Indian Veterinary Association, Kerala from the year 2012.

The awards will be for a best Clinical paper and a best Research paper. The award consists of a citation

and a cash award which will be distributed in the Veterinarians Annual Convention.


ABSTRACT

Climatic change and food security are

two important emerging global problems. The

increasing animal population to meet the food

security is also considered as a major catalyst

for climatic change and emerging zoonoses.

Developing countries are likely to be worse

affected by climatic change and difficult to

cope with the adaptation or mitigation

measures for want of economical and

technological support. Species and region

specific research are essential to formulate

ideal but affordable mitigation measures to

meet the climatic stresses. Sensitization of

farmers about its importance and capacity

bui lding are the immediate needs.

Collaboration of multidisciplinary experts and

agencies is also important for desired results.

IMPACTS OF CLIMATIC CHANGE IN

ANIMAL HUSBANDRY- ARE WE PREPARED?

S. S. Rani

Assistant Director, Livestock Management Training Centre,

Kudappanakkunnu, Thiruvananthapuram

CLIMATIC CHANGE AND GLOBAL WARMING

Extreme weather events and its damaging

effects are already experienced all over the world.

Climatic change is now a reality and the most

obvious manifestation is the rising average

atmospheric temperature otherwise known as

global warming. The mean global annual

temperature increased between 0.4 to 0.7° C.The

impacts could be multifarious like rainfall, melting

glaciers, rising sea level, recurrent droughts and

floods, threat to biodiversity, increased plant and

animal diseases and infinite number of challenges

on public health (Sarkar, S. and Padaria, R. N.

2010). The Intergovernmental Panel on Climatic

Change (IPCC) predicts the increase in global

average surface temperature between 1.8°C and

4.0° C by 2100 and extinction of approximately 20

to 30 percent of plant and animal species with

severe consequences for food security in

developing countries (FAO).

Effect of Green House Gases (GHG) like

Carbon dioxide, Methane and Nitrous Oxide

towards global climatic change is already

established. At smaller levels these gases have a

protective role of life on earth. Carbon dioxide is

widely considered the most important human-

induced GHG. Estimated GHG emission from

animals is 18 percent, mainly Methane, which got

25 times the Global Warming Potential (GWP) of

Carbon dioxide (FAO). Climate-changing gases

are released into the atmosphere at every step of

meat, egg, and milk production and potentially

disrupting weather, temperature, and ecosystem

health. Globally emissions from pig manure alone

account for almost half of all GHG emissions from

farm animal manure. (Koneswaran, G and

Nierenberg, D, 2008).


GENERAL ARTICLE

DIRECT IMPACTS ON ANIMAL HEALTH

India is an agrarian country with the largest

cattle population in the world. Livestock

population have a crucial role in developing

countries by its contribution to food security and

poverty alleviation. It is also noted that the global

livestock sector is growing faster than any other

agricultural subsector. But Methane emission from

cattle population through enteric fermentation

and manure management is a matter for concern

in climatic change.

Impact of climatic changes could be in four

ways: heat-related diseases and stress, extreme

weather events, feed grain forage availability and

price and adaptation of animal production systems

to new environments and emergence or re-

emergence of infectious diseases, especially

vector-borne diseases critically dependent on

environmental and climatic conditions.

Most important direct effect could be heat

stress and reduced nutrient intake affecting

production performance (Mariara, J.K., 2008). The

results of impact assessment study in Africa

indicate that large livestock farms are more

vulnerable to climate change and are likely to lose

net revenue while small farms are much less

vulnerable and will probably get advantage, atleast

against the risk of dryness, livestock offer a good

substitute for crops. (Seo, S.N. and Mendelsohn.

R.2007). It is a requisite to mitigate the climatic

stresses, if a high producing and less tolerant

animal to be reared in harsh environmental

situation. (Naqvi, S.M.K and Sejian, V. 2011).

Mitigation to overcome heat stress could be

Physical modification of environment

Genetic development of less sensitive breeds

Improved nutritional management schemes

=

=

=

Some current economic practices such as

shades, sprinklers and ventilation will be suitable

for adapting to reduce heat stress in dairy cows.

Shades are the simplest method to reduce the

impact of high solar radiation. Open sided

construction for sheds will increase natural

ventilation and if not, providing specially designed

fans will be ideal. An effective way of cooling cattle

is evaporative spray-mist, fog and sprinkling

systems.

Climate change will have a substantial

effect on global water availability in the future. Not

only will this affect livestock drinking water

sources, but it will also have a bearing on livestock

feed production systems. Reduction of livestock

numbers to more productive animals leads to

more efficient milk/ meat production and lower

GHG emissions. Different animal feeding and

manure management (collection, storage,

spreading, biogas production), management of

feed crop production are other mitigation

measures. (IFAD, 2007).

Emergence/ Re-emergence of diseases:

Predictions on all the impacts are difficult

but a change in climate can result in changes in

species composition of vectors, pathogens and

augmenting its spread and even the emergence of

new pests and diseases. New transmission

modalities and host ranges complicate the

epidemiology of some diseases. (FAO, 2008).

Temperate countries will be more prone to such

diseases. Changes in the spatial distribution of

vectors/pathogens, animal populations with little

or no immunity would be exposed and suffer

major disease impacts (FAO, 2008).Diseases such

as Bluetongue have expanded their range due to


global warming. Extreme weather events such as

heavy rainfall or droughts often trigger disease

outbreaks.

PUBLIC HEALTH IMPACTS

Climate change is having far-reaching

consequences on human health, perhaps most

starting signals could be growing conflicts, hunger

and then to diseases. (Koneswaran, G and

Nierenberg, D, 2008). Climate change may result

in food-borne zoonoses. Additional /inappropriate

use of pesticides and veterinary drugs could be

another concern. Mycotoxins in food is going to be

a growing problem in the coming era.

Vulnerability of the impact could be more

on developing countries where animals and

human are exposed directly to natural resources

for food and less capacity for adapting mitigation

measures. Rural poor communities rely greatly on

agriculture and livestock for their survival, at the

same time these are amongst the most climate-

sensitive economic sectors. So climatic change can

aggravate poverty.

NEED FOR A 'ONE HEALTH' APPROACH

It is estimated that up to 75 percent of

newly recognised infectious diseases of humans

are from zoonotic pathogens- Bovine Spongiform

Encephalitis, Nipah virus, Monkey Pox, H5N1,

West Nile Virus, Rift Valley Fever and many more to

add. It is a clear realisation that infective agents

circulate between wildlife, domestic animals and

humans. It is also proved that no one discipline or

sector of society has enough knowledge and

resources to prevent the emergence or resurgence

of diseases in today's globalized world. The 'One

Health' approach needs contributions from

multidisciplinary experts like agricultural scientists,

veterinarians, anthropologists, economists,

educa to r s , eng inee r s , en tomo log i s t s ,

epidemiologists, microbiologists, nutritionists,

physicians, public health professionals, sociologists

and local communities (Black, P. F. et.al. 2008)

Responses to Climatic changes and future

strategies

1. Adaptation to reduce the vulnerability of

people and ecosystems

2. Mitigation to reduce the magnitude of impact

in the long term.

The Conference of the signatories under

the United Nations Framework Convention on

Climate Change (UNFCCC) have been

unsuccessful till date in negotiating consensus on

emission reductions. The chances for global

agreement in preventing or mitigating climate

change are increasingly decreasing. Therefore we

need to prepare people for adjusting to the

impacts of climate change.

Across the globe, governments are

increasingly adopting national programmes to deal

with the threat of climate change. According to the

national action plan on climatic change by the

Prime Minister's council, possible risks were

analysed with observed impacts and eight

National Missions were formulated in strategic

areas including a National Mission for sustainable

agriculture. Under the aegis of these missions

implementation of various activities are in

progress. Some of them are conservation of water

through practices like rain water harvesting, risk

financing like crop/animal insurances, disaster

management and proactive programmes on

capacity building, surveillance and control of

vector borne diseases like Japanese Encephalitis,


Dengue fever, Malaria,Filariasis, energy audits in

large energy consuming units, promotion of

biofuels.( GOI., 2009)

Government of Karnataka invited related

departments to prepare their action plans,

strategies or vision documents for evaluation but

only half the agencies could prepare the requisite

documents in time. Government of Kerala have

the department of Environment and Climatic

Change to co-ordinate reduction, adaptation or

mitigation measures.

METHANE EMISSION REDUCTION STRATEGIES

This is very important in India with huge

livestock population. Also Methane is the GHG

with the shortest life span,10-12 years when

compared to CO with 120 years. So any effort to 2

reduce methane emission will be beneficial to slow

global warming. It could be managemental,

nutritional or advanced biotechnological

strategies. (Naqvi, S.M.K and Sejian, V. 2011).

Methane emission from exotic/crossbred cows

with high productivity is proved to be less. Amount

of feed consumed and its digestibility are two

important factors that determine total Methane

production.

WHAT NEXT?

Identifying task force to formulate policies

in relevant sectors, research to develop

vulnerability indices on various parameters in the

sector and to formulate adaptation strategies that

best suited for the farming situation of our state are

crucial to meet the hardships of climatic change.

Capacity building of staff at various levels to cope

up with the mitigation measures and sensitisation

of farming community about this phenomenon are

other immediate interventions required.

CONCLUSION

Climate change is an ongoing process.

Starving in both animals and human can be worse

in the developing world. There is a two-way

relationship between livestock production and

environmental health. On the one hand, livestock

contribute to climate change and other

environmental problems, and at the same time

livestock health and productivity can be adversely

affected by these same environmental upsets.

(Sherman, D.M.2010).

Research should be initiated by

responsible forum to formulate policy for

sustainable livestock at one end. Developing

farmer friendly technologies are important

especially in the quality of feed to improve milk

production and decrease emission of GHG.

Control of emerging diseases need concerted

efforts in research, control measures for vector and

pathogen including their transboundary

movement in the era of enhanced global trade,

travel and tourism. Climate determines ecosystem

health over time, but weather drives immediate

outbreaks and disasters. Hence implications of

climatic change on different fields connected with

plant, animal and human life need to be studied in

depth at different levels for early mitigation. The

environmental impacts of animal production

require more focussed attention from

international organizations, governments,

producers, and consumers on meat and dairy

production.


REFERENCES

Black, P. F., Murray, J. G. and Nunn, M. J. 2008.

Managing animal disease risk in Australia:

the impact of climate change. Rev. sci. tech.

Off. int. Epiz., 27 (2), 563-580

FAO. 2008. Climate related Transboundary pests

and Diseases, Technical background

document from the Expert Consultation th th

held on 25 to 27 February 2008 at FAO,

Rome. Available at ftp://ftp.fao.org/docrep/

fao/meeting/013/ai785e.pdf. Accessed on

19-3-2012

GOI. 2009, National Action Plan on Climate

Change. Available at india.gov.in/

allimpfrms/alldocs/15651.doc

IFAD 2007. Livestock and climatic change. The

International Fund for Agricultural

Development (IFAD) Strategic Framework

2007-2010. Available at www.ifad.org/sf/

Accessed on 21-3-2012

Koneswaran, G and Nierenberg, D 2008. Global

Farm Animal Production and Global

Warming: Impacting and Mitigating Climate

Change. Environ Health Perspect 116:578-

582.

Mariara, J.K. 2008. The Economic Impact of Global

Warming on Livestock Husbandry in Kenya:

A Ricardian Analysis. African Economic

Conference on Globalization, Institutions

and Economic Development of Africa.

Tunis, 12-14th November 2008. Available at

www.afdb.org .Accessed on 19-3-2012

Naqvi, S.M.K and Sejian,V. 2011. Global climate

change: Ro le o f L i ves tock .As ian

J.Agric.Sci,3(1):19-25

Report prepared for Department of Ecology and

Environment, Karnataka.Available at

www.empri.kar.nic.in/Karnataka. Accessed

on 17-3-2012

Sarkar, S and Padaria, R. N. 2010. Farmers'

Awareness and Risk Perception about

Climate Change in Coastal Ecosystem of

West Bengal. Indian Res. J. Ext. Edu. 10 (2)

SAT eJournal | ejournal.icrisat.org December. 4

(1). Available at www.icrisat.org/journal/pdf.

Accessed on 17-3-2012

Seo, S.N. and Mendelsohn. R. 2007. Climate

Change Impacts on Animal Husbandry in

Africa: A Ricardian Analysis. World Bank

Policy Research Working Paper 4261, June

2007. Available at www.worldbank.org/pdf.

Accessed on 20-3-2012.

Sherman, D.M. 2010. A Global Veterinary medical

Perspective on the concept of one health:

Focus on livestock. ILAR Journal. 51(3): 281-

287.

Slenning, B.D. 2010. Global Climate Change and

Implications for Disease Emergence.

Veterinary Pathology. 47(1) 28-33

State Action Plan on Climate Change Karnataka

2010. Rapid Assessment of Sectoral Actions

Thornton, P, Herrero, M, Freeman, A and Mwai, O.

2007. Vulnerability, Climate change and

Livestock Research Opportunities and

Challenges for Poverty Alleviation


VACCINES FROM OUR GARDEN

1 2 3 4Tincy Mary John , N.M. Shah , B. S. Chandel and H.C.Chauhan

College of Veterinary Science and Animal Husbandry, SDAU, Gujarat

INTRODUCTION

There is a saying that, “Prevention is better

than Cure”. The best way to prevent a disease is to

be immunized and the most effective way to get

immunized is by vaccination. Thus, Vaccines can

said to be a boon in the field of medical science.

The world of vaccines can be classified mainly into

two groups Classical Vaccines and New

Generation Vaccines. Classical vaccines are made

up of killed /inactivated or live attenuated

microbial agents or bacterial toxoids. With the

advancement in the field of biotechnology and

molecular biology, we were able to introduce New

Generation Vaccines like naked nucleic acid

vaccines, vectored vaccines, anti-idiotypic

vaccine, recombinant DNA vaccine etc. Still,

according to WHO reports, the mortality rate due

to illness are increasing every year especially in the

third world countries. This may be due to

constraints on vaccine production, distribution

and delivery. The search by scientists to solve these

limitations ultimately results in the development of

“EDIBLE VACCINES”.

EDIBLE VACCINES

hey are simply sub-unit vaccines that are

edible in nature. Here, the gene of interest is

introduced into plants and then these altered

plants are induced to manufacture the

corresponding proteins. This process is known as

transformation and the altered plants are called

transgenic plants. (Sibila, J. et.al.2005)

The first report of edible vaccine appeared

in 1990 in the form of patent application

published under the international patent

cooperation treaty. (Curtiss, R. I and Cardineau,

C.A. 1990) It was regarding the successful

expression of Streptococcus mutans surface

protein A in tobacco. As this bacterium causes

dental caries, it was envisaged that the stimulation

of a mucosal immune response would prevent the

bacteria from colonizing the teeth and there by

protect against tooth decay. (Mason, H. S and

Arntzen, C. J. 1995) Later in 1992, Charles Arntzen

and his coworkers successfully expressed

hepatitis B surface Antigen in tobacco plants in a

cost effective manner. This paved the way for

worldwide acceptance to the concept of edible

Edible vaccines are those vaccines based

on genetically engineered expression of an

antigenic protein by an edible plant. In simpler

words, t

1 2M.V.Sc Scholar, Professor & Head, Department of Veterinary 3 4Microbiology. Professor & Head, Associate professor,

Department of Animal Biotechnology, College of Veterinary

Science & A.H.,S. D. Agricultural University

GENERAL ARTICLE


vaccine. (Mason, H. S. et.al.1992) Till then, various

attempts have been made to develop edible

vaccines against human and animal diseases

including Norwalk virus particles, (Mason, H. S

et.al.1996) Rabies (Hooper, D.C. et.al., 1994)

Gastroenteritis, Cholera (Arakawa, T. 1997).

PRODUCTION OF EDIBLE VACCINE

1. Appropriate plant virus is genetically

engineered to express the desired peptides/

proteins. The recombinant virus is then inoculated

into the plant. Large numbers of new plants are

grown and chimeric virions are extracted and

purified. The resultant plant edible vaccines are

utilized for immunological purpose.

2. In another approach, the gene of interest is

integrated with plant vector by transformation. A

variety of techniques have been used to introduce

transgene into plant cell; these could be grouped

into following categories:

v Agrobacterium mediated gene transfer: The

appropriate gene construct is inserted into

the T-region of a disarmed Ti plasmid of

Agrobacterium. The recombinant DNA is

placed into Agrobacterium; a plant

pathogen which is co-cultured with the

plant cells or tissues to be transformed. The

drawback of this method is that it gives low

yield and the process is slow. This method

worked especially well for dicotelydenous

plants like potato, tomato and tobacco.

Studies have also proved that the genes are

For producing Edible Vaccines, any of the

two methods can be followed: (Shah, P.C et.al.

2011)

expressed by this method in experimental

animals and plants.

v Biolistic method: The gene containing DNA

coated metal (e.g. gold, tungsten) particles

are fired at the plant cells using gene gun.

Those plant cells that take up the DNA are

then allowed to grow in new plants, and are

cloned to produce large number of

genetically identical crop. This method is

quite attractive because DNA can be

delivered into cells of plant which makes

gene transfer independent of regeneration

ability of the species. But the chief limitation

is that the gene gun is highly expensive.

v Electroporation: Here there is introduction

of DNA into cells by exposing them for brief

period to high voltage electrical pulse which

is thought to induce transient pores in the

plasma lemma. The cell wall presents an

effective barrier to DNA therefore, it has to

be weakened by mild enzymatic treatment

so as to allow the entry of DNA into cell

cytoplasm.

“SECOND GENERATION”EDIBLE VACCINES

Scientific community developed the

second generation edible vaccine which provides

protection against several pathogens. These

multicomponent edible vaccines can be obtained

by crossing two plant lines harboring different

antigens. Yu and Langridge (Jie, Yu and William

H.R. Langridge. 2001) fused Cholera toxin (CT) B

and A2 subunit complementary DNAs (cDNAs) to

a rotavirus enterotoxin and enterotoxigenic


Escherichia coli fimbrial antigen genes and

transferred into potato. It was found that this

trivalent edible vaccine could elicit humoral

responses, as well as immune memory B cells and T

helper cell responses which are hallmarks of

successful immunization.

MECHANISM OF ACTION

Mucosal Immune System (MIS) is the first line

of defense mechanism of body and thereby the

most effective site for vaccination. The most

effective route of mucosal immunization is oral

route. (Korban, S. S. et.al. 2002). The transgenic

plant parts with desired gene are fed directly since

the outer tough wall of plant cells acts to protect

near payer's patches

(Rudzik, R. et.al.1975).These released

antigens are taken up by the M-cells and is

presented to B cells with the help of antigen

presenting cells (APC). The activated B-cells get

differentiated into plasma cells and secrete IgA

class of antibody and elicit mucosal immunity and

humoral immunity. Another important component

of mucosal immunity is T-cell mediated immune

response where the T-cells specifically recognize

pathogens and directly kill the invader themselves.

It also helps indirectly to the antibodies to clear

the

antigens against attack by enzymes, gastric and

intestinal secretions. This method is known as bio-

encapsulation. The plant cell wall breaks in the

intestines and antigens are

released.

infection. T-cells produced in the mucous are

capable of travelling the mucosal tissues through

special 'homing' receptors on their membranes.

This means that if an immune response is

generated in gastrointestinal lining, T-cells

produced there can travel to other mucosal sites,

(e.g. the lungs, nasal cavity) providing protection

over a large surface area

Figure showing mechanism of action in the

intestinal mucosa

CLINICAL TRIALS

Edible vaccines have been checked for

their efficacy in humans. The results of human

trials that were tested for different transgenic plants


have showed positive responses and no major

safety concerns. ( Streatfield, S.J. 2001). Transgenic

plant-made vaccines are also being used in

animals. The first edible vaccine to demonstrate

efficacy in animal trials was against Transmissible

gastroenteritis virus (TGEV) in pigs. Livestock

animals are fed with transgenic plants, like

Arabidopsis thaliana, alfalfa and potato with

antigens to protect them from pathogens;

including FMDV, BRV and bovine viral diarrhea

virus (BVDV)( Streatfield, S.J. 2001)

ADVANTAGES

1. Trigger mucosal immunity (traditional

vaccines may bypass this) which is the body's first

line of defense.

2. Adjuvant to enhance immune response is not

necessary.

3. Cost-effective in terms of manufacturing,

storage and transportation. Edible vaccines are

made in molecular farms, and not in multimillion

dollar machines. Since most countries have soil-

rich land, it provides them the convenience in

obtaining edible vaccines, and long distance

transportation is not required. Maintenance of

cold chain not required because plant tissues can

be dried or, as when the seeds are targeted, have

low moisture content. (Pascual, D. W.2007). Thus

the amount spent yearly to preserve vaccine can be

saved.

4. Their production is highly efficient and can be

easily scaled up. For example, hepatitis-B antigen

required to vaccinate whole of China annually,

could be grown on a 40-acre plot and all babies in

the world each year on just 200 acres of land!

5. Easily available since produced from plants. If

we run out, we can simply plant more. Moreover,

we can select the local/native crop of a particular

area and can engineered it to produce the vaccine.

6. Giving an oral vaccine would require little or

no training at all, which reduces the requirement

of trained professionals.( Streatfield, S.J. 2001)

7. Eliminates the need of syringes & needles for

administration. Needle administered vaccines are

plagued with problems of re-use, mis-use and an

occasional lack of sterilization.

8. Plants cannot host most human pathogens, so

the vaccines will not pose a danger to humans.

9. Fear of contamination with animal viruses like

the mad cow disease, which is a threat in vaccines

manufactured from cultured mammalian cells, can

be eliminated.

10. They are subunit preparations containing

only the antigen and not any pathogenic genes.

Thus it enhances the safety of individual.

11. Multi-component ability of vaccine allows to

harbour more than one antigen to prevent many

diseases in same individual. (Jie, Yu and William

H.R. Langridge. 2001)

12. Antigen can be protected by means of

bioencapsulation.

13. They can seroconvert even in presence of

maternal antibodies, thus having a potential role in

protecting infants against like Group-B

Streptococcus, Respiratory Syncytial Virus etc.

which are under investigation.

FUTURE CHALLENGES/CONCERNS

1. There is need of proper distinguishing

characters to identify between 'vaccine fruit' and

'normal fruit' to avoid misadministration of vaccine


which could lead to tolerance.

2. Antigen selection involves safety concerns of

whether or not selected antigens are compatible

enough with the selected plant type to be

expressed (Lal, P. et.al. 2007)

3. How can the vaccine dose be controlled? This

remains the most difficult task. Consistency of

dosage form differs from fruit to fruit, plant to plant

and generation to generation. It is determined by

protein content, patient's age, weight, ripeness of

the fruit and quantity of the food eaten. If low

doses are consumed then the production of

antibodies are less likely to occur, and if high doses

are consumed it may cause tolerance. (Lal, P. et.al.

2007)

4. Glycosylation patterns in plants differ from

those in humans and could affect the functionality

of the vaccines.

5. Allergic reactions to the fruit or vegetable

expressing the foreign antigen may be an issue.

7. Certain foods are not eaten raw (e.g.

potato) and needs cooking which will denature or

weaken the protein present in it. (Moss, W. J.

et.al.1999)

8. Variable storage conditions for edible

vaccine are also a major problem. Potatoes

containing vaccine could be stored for longer time

while a tomato does not last long. Thus these

6. The doubt still exists about whether the

antigens be able to survive the hostile, acidic

conditions of the stomach and even if they did, will

they be able to trigger the immune system in the

right way. Although initial trials have shown

promising results in human subjects, it is not clear

what will happen when the person comes in

contact with the actual virus.

vaccines need to be properly stored to avoid

infection through microbial spoilage.

9. Another concern is if the transgenic plants

are mass produced, they may have an inconsistent

expression caused by the small interfering RNAS.

(Tonks, A. 2007)

10. There is chance of plant/crops (food)

contamination through cross pollination and of

vaccine itself in plant debris spreading dust and

other pollutant in surfaces and ground waters. The

vaccine antigen may affect browsing animals and

humans living in the area drinking vaccine polluted

water or breathing vaccine polluted dust. The

cultivation and production should be limited to

facilities like greenhouse, or in plant tissue culture

that prevent their environmental release.

CONCLUSION

Transforming plants to carry vaccines is

one of the latest innovations of medical technology

and promises greatest hope for the floundering

biotech industry. It can prove to be very effective, if

rightly implemented, in providing accessibility to

developing and underdeveloped countries where

rates of diseases are relatively high. Edible vaccines

are prominent over typical traditional vaccines due

to its positive aspects like they are cost-effective,

safe, easy to administer and can store at the site of

production. But it has to overcome the above

mentioned concerns and technical obstacles to

become a reality. Future research is also required

to demonstrate whether these vaccines meet the

standards of quality (purity, potency, safety,

efficacy and durability) defined for vaccines by the

World Health Organization. It can give us new and

dramatic hope for improved life. Although in the

initial stages of development, a day is not away


when we will be able to pluck a fruit from the

garden, munch on it and get protected from

diseases… making needles needless…

As Hippocrates said, “Let thy food be thy

medicine”

REFFERENCES

Arakawa, T., Chong, D. K and Merritt, J. L. 1997.

Expression of cholera toxin B subunit

oligomers in transgenic potato plants.

Transgenic Res. 6:403-13.

Hooper, D .C ., Pierarrd, L., Modelska , A., Otvos,

L. J and Fu, Z. F .1994. Rabies ribonucleo-

capsid as an oral immunogen and immun-

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Jie, Yu and William, H.R. Langridge. 2001. A plant-

based multicomponent vaccine protects

mice from enteric diseases. Nature

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Korban, S. S., Krasnyanski, S. F and Buetow, D. E.

2002. Food as production and delivery

vehicle for human vaccine, J. Am. Coll.

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Lal, P., Ramachandran, V. G., Goyal, R and Sharma,

R. 2007: Edible vaccines: Current status and

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Mason, H. S., Ball, J. M and Shi, J. J. 1996.

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Implications of human immunodeficiency

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29:4452-4455

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Streatfield, S.J. 2001: Mucosal immunization using

recombinant plant-based oral vaccines.

Methods, 38:150-157.

Tonks, A. 2007: Oral vaccines: Spoonful of

antigens. Br. Med. J, 335:180-182.


1Assistant Professor, Department of L.P.M, College of

Veterinary and Animal Sciences, Pookode, Wayanad. 2Professor and Head, Department of L.P.M, Veterinary College

and Research Institute, Namakkal, Tamil Nadu.

“BIODIESEL PRODUCTION FROM ANIMAL FATS”

AN EVER GREEN TECHNOLOGY FOR THE FUTURE ENERGY SECURITY

1 2John Abraham and Ramesh Saravana Kumar

College of Veterinary and Animal Sciences, Pookode, Wayand.

INTRODUCTION

The world is on a quest for energy, the

premier source of which is now petroleum. Energy

is the most fundamental requirement of every

nation as it progress through the ladder of

development. India with 16 percent of the world

population could boast of only 0.5 percent of the

world oil reserves. The current level of production

barely caters to 26 percent of the petroleum

demand. India imports 75 percent of its crude oil

requirements. The estimated crude oil import cost

comes to about 10 percent of the country's G.D.P.

Besides the billions spent on importing crude oil,

we are also importing billions of tonnes of CO and 2

other green house gases causing climatic changes

as evidenced by changing rainfall patterns, rising

sea levels and temperatures. The demand for

diesel is five times higher than the demand for

petrol in India and any increase in diesel price

immediately pushes up inflation. Therefore, the

government is still incurring huge cost of

subsidising diesel. With the Indian economy

poised for a robust growth of 9 to 9.5 percent for ththe 12 plan period, energy security has become

the key issue in policy formulation and planning.

India's energy security would remain

vulnerable until alternative fuels to substitute/

supplement petro-based fuels are developed

based on indigenously produced renewable

feedstocks. In biofuels, the country has a ray of

hope. Biofuels are non-polluting and virtually in

exhaustible. Biofuels can increasingly satisfy these

energy needs in an environmentally benign and

cost-effective manner while reducing dependence

on import of fossil fuels and thereby providing a higher

degree of National Energy Security (Anon. 2009).

The national biofuel policy 2009,

proposed 20 percent blending of biodiesel by

2017. This huge demand cannot be met from non-

edible vegetable oil feedstock alone. Therefore,

development and utilization of new indigenous

biomass feedstock for production of bio fuel and

development of next generation of more efficient

bio fuel conversion technologies are the need of

the hour.

In this context, biodiesel production from

animal fats offers new scope as a potential means

to stimulate rural development, lower emission of

harmful pollutants and decrease green house gas

emission, while contributing to national energy

security by reducing dependence on oil imports

and mitigation of climatic changes vis-à-vis

providing good fuel properties for the diesel

engine.

GENERAL ARTICLE


Definition of Biodiesel

Biodiesel is defined as “the mono alkyl

ester of long chain fatty acids derived from

renewable lipid feed stock such as vegetable oil or

animal fats, for use in compression ignition (diesel)

engines” (National Biodiesel Board, 1996)

Blends

Blends of biodiesel and conventional

hydrocarbon based diesel are products most

commonly distributed for use. Much of the world

uses a system known as the "B" factor to state the

amount of biodiesel in any fuel mix:

100 percent biodiesel is referred to as

B100, while

20 percent biodiesel is labelled B20

5 percent biodiesel is labelled B5

Blends of 20 per cent biodiesel with 80 per

cent petroleum diesel (B20) can generally be used

in all diesel engines without any modification.

Biodiesel in its pure form (B100), may require

certain engine modifications to avoid maintenance

and performance problem.

Importance of Biodiesel

Biodiesel has good fuel properties,

comparable to or even better than petroleum

diesel. It has 10 percent built-in oxygen content

that helps it to burn fully. Its cetane number (an

indication of its fuel burning efficiency) is 72 for

biodiesel derived from tallow and 72.5 for

biodiesel derived from chicken oil, higher than

54.4, the cetane number of most petroleum

diesels. The esters of the long-chain fatty acids of

biodiesel are excellent lubricants for the fuel

injection system. It has a higher flash point than

diesel, making it a safer fuel. Other advantages are

=

=

=

the almost zero sulphur content and the reduced

amount of carbon monoxide, unburned

hydrocarbons and particulate matter in the

exhaust.

BIODIESEL PRODUCTION TECHNOLOGY

Biodiesel Feedstock

Biodiesel is typically made from vegetable

oil though animal fat can also be used. Rapeseed

oil has 82 percent of the share of the world's

biodiesel feedstock, followed by sunflower oil (10

percent), soy bean (5 percent) and palm oil (3

percent). The choice of feed is country specific and

depends on availability. Other feedstocks include

waste vegetable oil (WVO), algae which can be

grown using waste material and oil from

halophytes such as Slicornia bigelovii.

Biodiesel Production Process

The major steps required to synthesise

biodiesel are as follows

1. Feed stock pre-treatment : If waste

vegetable oil (WVO) is used, it is filtered

to remove dirt, charred food, and other

non-oil material often found.

2. Determination and treatment of free fatty

acids (FFA) : A sample of the cleaned

feedstock oil is titrated with a

standardised base solution in order to

determine the concentration of free fatty

acids (carboxylic acid) present in the oil

sample.

3. Transesterification: Transesterification

(also called alcoholysis) is the reaction of

fat or oil with an alcohol to form esters

and glycerol. The reaction is as shown in

the equation below.


Where R , R and R are long hydrocarbon chains 1 2 3

sometimes called fatty chain.

Alcohol

Alcohols are primary and secondary

monohydric aliphatic alcohols having 1-8 carbon

atoms (Sprules and Price 1950). Among the

alcohols that can be used are methanol, ethanol,

propanol, butanol and amyl alcohol. Methanol is

most frequently used because of its low cost and its

physical and chemical advantages (polar and

shortest chain alcohol).

Alkali/ Base catalyst

The alkalis include Sodium hydroxide,

Potass ium hydroxide, Carbonates and

corresponding Sodium and Potassium alkoxide

such as Sodium methoxide, Sodium ethoxide,

Sodium propoxide and Sodium butoxide.

Acid Catalyst

Sulphuric acid, Sulfonic acids and

Hydrochloric acid are used as acid catalyst.

Lipase Enzyme

Lipases can also be used as bio-catalysts.

Alkali Catalysed Process

Alkali-catalyzed transesterification is

much faster than acid-catalyzed transesterification

and is most often used commercially. For an alkali-

catalyzed transesterification, the glycerides and

alcohol must be substantially anhydrous (Wright

et.al., 1944) because water makes the reaction

partially change to saponication, which produces

soap. Low free fatty acid content in triglycerides is

required for alkali-catalyzed transesterification.

Acid Catalysed Process

Acid catalysed process are used for direct

esterification of free fatty acids in a high free fatty

acid (FFA) feed stock, or to make esters from soap

stock (Keim, 1945).

Multiple Step Process

Canakci and Van Gerpen (2001)

investigated an acid catalyzed pre-treatment step

followed by a base catalyzed step as an effective

conversion method for low cost raw materials.

They found that by using methanol and sulfuric

acid and a reaction temperature of 60°C the free

fatty acid content of a feedstock could be

significantly reduced.

The preferred method for high FFA feed

stock is acid catalysis followed by base catalysis.

Ultrasonic-Reactor Method

In the ultrasonic reactor method, the

ultrasonic waves cause the reaction mixture to

produce and collapse bubbles constantly. This

cavitation provides simultaneously the mixing and

heating required to carry out the transesterification


process. Thus using an ultrasonic reactor can

drastically reduces the reaction time, reaction

temperatures, and energy input.

Micro-Wave Method

Current research is being directed into

using commercial microwave ovens to provide the

heat needed in the transesterification process. The

microwaves provide intense localized heating that

may be higher than the recorded temperature of

the reaction vessel.

ALTERNATE PROCESSING TECHNIQUES

Enzymes

Enzymes have shown good tolerence for

the free fatty acid levels of the feedstock, but the

enzymes are expensive and unable to proceed the

reaction to completion. Immobilisation of enzymes

and use of multiple enzymes in sequence may

provide future opportunities in this area. (Nelson

et.al., 1996)

Co-Solvent

Boocok et.al., (1998) developed a novel

technique for accelerating the transesterification

reaction rate. He proposed the addition of co-

solvent to create a single phase and this accelerates

the reaction. The commonly proposed co-solvents

are Tetra hydro furan and Methyl tertiary butyl

ether. The major draw back of this method are the

hazard level associated with the co-solvent and

recovering and recycling the co-solvent.

Supercritical Methanol System

By increasing both the temperature and

the pressure, a critical point is obtained at which

gasses and liquids are indistinguishable fluids.

Matter that exists in this new phase is called a super

critical fluid (SCF). Methanol at supercritical

conditions becomes an excellent solvent and

dissolves the feedstock and reacts readily without a

distinct catalyst, but energy costs of production are

similar or less than catalytic production routes.

(Saka and Kudsina, 2001).

Animal Fats for Biodiesel Production

The amount of animal fat used by the bio-

diesel industry has more than doubled from 2006

to 2008. According to a report by the Informa

Economics, approximately 20 percent of the bio-

diesel manufactured in the United States in 2008

was produced from animal fats greases and

recycled cooking oils.

Whole dead poultry carcasses and poultry

slaughter wastes can be used to recover fats and

oils. The dead poultry birds having an average

body weight of 1.25 kg have a fat content of 14.55

percent and by solvent extraction method 96.1

percent of this fat can be recovered. A two step

processing reaction, acid catalysed esterification of

FFA followed by base catalysed transesterification

of triglycerides could convert 97.62 percent of this

fat to good quality biodiesel. A litre of biodiesel can

be produced from chicken fat at the cost of 22.00

rupees. The blending of biodiesel to commercial

diesel at 20 percent can substantially reduce the

smoke (47.14 percent) compared to that of diesel

in a CRDI engine (John Abraham, 2012). Thus the

blending of biodiesel at 20 percent to commercial

diesel can reduce the import of costly crude oil and

simultaneously, substantially reduce the engine

emission. The total weight of the dead birds


available in India per year was estimated at 2.4

lakh tons as per the standard mortality of the

industrial average (Chandrasekaran, 2009). Which

can be converted to 8500 tonnes of biodiesel. Thus

biodiesel production technology provides

opportunity to produce highly valued biofuel from

dead animal and birds. (Wealth from waste). At the

same time, this concept can also solve the major

problem of unscientific disposal of dead birds and

slaughter waste. Currently a 5 million dollar plant is

being built in the USA, with the intent of producing

11.4 million litres of biodiesel from an estimated

one billion Kg of chicken fat produced annually at

the Tyson poultry plant.

Rearing animals for meat and biodiesel can

be the production objectives in days to come. This

concept can revolutionise animal production

sector. Especially pigs and poultry can contribute in

a big way because of their excellent attributes such

as prolificacy, short generation interval and quick

body weight gain for slaughter. Fat less pork at a

premium price is gaining consumer acceptance

phenomenally. This provides the separated fat to

be converted to biodiesel economically.

Conclusion

The biofuel industry is poised to make

important contribution to meeting India's energy

needs by supplying clean domestic fuel.

Simultaneously it also provides other advantages

like

4 It provides a market for excess production

of vegetable oils and animal fats.

4 It decreases, although will not eliminate

the country's dependence on imported

crude oil.

4 Biodiesel in renewable and does not

contribute to global warming due to its

closed carbon cycle.

4 By biodiesel blending the overall carbon

dioxide emission can be reduced by 78

percent compared with petroleum based

diesel fuel.

4 The exhaust emissions of carbon

monoxide, unburned hydrocarbons and

particulate emissions from bio-diesel are

lower than with regular diesel fuel.

Unfortunately there might be a slight

increase in oxides of nitrogen (NO )x

4 When added to regular diesel fuel it can

convert fuel with poor lubricating

properties, such as modern ultra-low

sulphur diesel fuel into an acceptable fuel.

4 Provide good fuel properties for the diesel

engines.

Biodiesel is not a replacement technology;

it is a transition technology to help clean

up our existing fuel and stream line

distribution by keeping the manufacture

and consumer of fuel as local as possible,

all the while keeping the revenue in the

national economy. It is not about

replacing all of our petroleum imports; it is

about not importing so much.

REFERENCES

Anon. 2009. National Policy on Bio-fuel.

Government of India, Ministry of New and

Renewable Energy.

Boocock, S.K., S.K. Konar, V. Mao, C. Lee and S.

Buligan. 1998. Fast formation of high purity

methyl esters from vegetable oils, JAOCS, 75:

1167-1172.


Canakci, M and Van Gerpen, J. 2001. Biodiesel

Production from Oils and Fats with High Free

Fatty Acids. Transaction of the Am. Soc. Agric.

Eng., 44 (6): 1429-1436.

Chandrasekaran, D. 2009. Poultry waste

management- Situation in India. Paper

presented at the national seminar on wealth

from livestock waste, 12-13 November

2009. Veterinary College and Research

Institute, Namakkal, Tamil Nadu.

John Abraham. 2012. Utilization of dead poultry

birds for biodiesel production. Ph.D thesis

submitted to the Tamil Nadu Veterinary and

Animal Science University, Chennai.

Keim, G. I.1945. Testing fat and fatty oils, U. S.

Patent No. 2,383,601.

Nelson, L.A., Foglia, T.A and Marmer, W.N. 1996.

Lipase catalyszed production of bio-diesel.

JAOCS, Vol. 73, no.8.

Saka, S. K. Kusdiana. 2001. Biodiesel fuel from

rapeseed oil as prepared in supercritical

methanol, Fuel 80, (2001), 225-231.

Sprules, F. J and Price, D. 1950. Production of fatty

esters. U. S patent 2, 366-494.

Wright, H. J., Clark, J.B and Coburn, H.V. 1944. A

report on ester interchange. Oil and Soap

21,145-148.


KERALA VETERINARY SCIENCE CONGRESS 2012

ASSOCIATION NEWS


The Kerala Veterinary Science Congress 2012 was

organised by Indian Veterinary Association, Kerala

in association with Dept. of Annual Husbandry,

Kerala State Veterinary Council, KLD Board, KSPDC

and HLL Life Care Ltd on 10th and 11th Nov 2012 at

Trivandrum. The Science congress provided and

effective platform for veterinary practitioners,

researchers and academicians to present their works

and to interact among themselves as well as with the

peers of the noble profession. Over 200 delegates

from across the country participated in the science

Congress. The Science congress was inaugurated by

Dr Renukaprasad, Hon’ble Vice Chancellor of

Karnataka Veterinary, Animal & Fisheries Sciences

University. Dr. Radhakrishna Pillai, Director, Rajiv

Gandhi Centre for Biotechnology, Trivandrum

released the compendium of the science congress

which was received by Dr. K G Suma, Director of

Animal Husbandry. Dr. Arunkumar, President, IVA

Kerala presided over the function. Dr. K

Udayavarman, President, KSVC & Director of

Museum & Zoo, Dr. Ani S Das, MD, KFL Ltd.

addressed the gathering. Dr. K C Prasath, President

KVSSA, and Dr. E K Easwaran, President AHOAK

offered felicitations. The participants were

welcomed by Dr. Theodore John, General Convenor.

In the Scientific session that followed papers

were presented in 3 session viz. Animal Health

Science, Animal Production and management and

Basic Veterinary subjects and field veterinary

experience. Dr. Harikrishna Kumar, SVS, VH,

Aryanad, Dr. Aravind, SVS, DVC, Kollam and Dr.

Geetharani, VS, Kollam were winners of best paper/

poster presented in session 3 comprising of field

veterinary experience. The lead papers in each

session was given by Dr. Kumanan, Director of

Reserach TANUVAS, Dr. Sreenivasan, Director of

Distance Education, TANUVAS and Dr. Ani S Das,

MD, KFL Ltd. The valedictory Address was given by

Dr. Sreenivasan, Director of Distance Education,

TANUAS and Dr. N N Sasi, Registrar of Kerala State

Veterinary Council gave away the prizes to the

winners.

thKVSSA leadership training camp was held on 6 and th

7 October 2012 at N C Gardens Beach Resort,

Parappanangadi, Malappuram, Dr. K. R. Arun Kumar, IVA

President inaugurated the training camp at a function

presided by Dr. K. C. Prasath, KVSSA President. Dr. Joby

George, General Secretary, KVSSA welcomed the participants

and Dr. K. A. Sajeeev Kumar, Dr. Ajaykumar felicitated. Dr.

Sivadas K. T proposed the vote of thanks. The faculty members

for the camp were Mr. M. N. Chandran Nair and Mr.

Subramanian (International trainers of JCI). 30 Veterinarians

participated in the camp.

KVSSA LEADERSHIP TRAINING CAMP 2012


WORLD RABIES DAY COMMEMORATION

Indian Veterinary Association, Kerala

organized a technical seminar on rabies eradication

at Hotel National Park, Ettumanoor on 28-09-2012.

The function was inaugurated by the Hon’ble

Finance Minister, Sri. K M Mani, Dr. Arunkumar,

IVA, Kerala State President presided over the

function. Dr. C Sreekumar, General Secretary, IVA

Kerala welcomed the gathering. The delegates were

veterinary surgeons deputed from all the 14 district

of Kerala. Dr. Ilona Otter, DVM, Director, World

Wide Veterinary Services India (WVS) handled the

highly informative technical session. Sri. K M Mani in

his inaugural address stressed the need for humane

treatment and handling of animals. He noted that

Animal Birth Control (ABC) of dogs and Rabies

vaccination are the correct and most humane way to

control human rabies cases. He said killing of dogs is

not an accepted method to control rabies anywhere

in the world. The fear of rabies is what makes society

so critical of stray dogs. He lauded the efforts of IVA,

Kerala in addressing this critical issue by conducting

awareness campaigns, implementation of ABC

programme in institutions cases throughout the

district of Kottayam and sponsoring vaccines free of

cost for pet dogs and cats in the run up to the

commemoration programme. The Finance Minister

accepted a memorandum and acknowledged the

high risk veterinary doctors face and announced that

risk allowance will be sanctioned to all cadres of

doctors working in AHD.

As part of the World Rabies Day commemoration,

IVA Kottayam unit has conducted nearly 60 animal

birth control surgeries (spaying) for free in all taluks

of the kottaym district. The association also

conducted free anti-rabies vaccination

programmes in all veterinary hospitals with the

cooperation of its members. IVA state unit

congratulates the office bearers and the members

of Kottayam unit for taking up such an ethical

initiative which is first of its kind in the country.

ABC SURGICAL CAMP

Dr. Jacob Alexander, Veterinarian of Thiruvananthapuram zoo and his team was

honoured for the timely diagnosis and successful treatment of Trypanosomiasis by

awarding “Ponnada” on 26/07/ 2012 at a function held at the Zoo. Ministers K.B. Ganesh

Kumar, P. K. Jayalakshmi, K. Muraleedharan, MLA, and District Governor of Lions Club

International P. Surendran were among the dignitaries present on the occasion.

Dr. P. Rajeev, District Epidemiologist, ADCP, Alappuzha (1989 batch, COVAS,

Mannuthy), received ''1st DAN BLACK BELT “ in “ TAEKWONDO” from “ KUKKIWON”

( World Taekwondo Headquarters ), South Korea. Taekwondo is a Korean martial art and

a recognized Olympic game. He got training at the Masters Taekwondo Federation,

Pathirappally, Alappuzha, Kerala and spent many years for this marvellous achievement.

The black belt test was conducted by Asia’s one and only Olympic referee Mr Maradi

Chandrasekhar who holds 7th DAN Blackbelt in Taekwondo

SIGN POST & ACHIEVEMENTS

OBITUARY

Dr. V. Sathianesan (75), Former Professor and Head, Department of Veterinary

Parasitology, CVAS, Mannuthy left for his heavenly abode on 02.12.12. Dr. Sathianesan

was an authority in Helminthology, especially gastro intestinal nematodes in ruminants.

He was always keen in the activities of the Indian Association for the Advancement of

Veterinary Parasitology (IAAVP) and had been the member of its Executive committee.

He is survived by his wife, Smt. Saralamani and two daughters, Anila and Anupa.May his

soul rest in peace.

Dr H. Sivasubrahmaniam Addl. Director (Rtd) passed away on 21/10/2012 at

Trivandrum. He was regarded as scholar of the profession and was a well wisher of the

Veterinary Community. He contributed his personal collection of books to IVA Library.

May his soul rest in peace.

Dr. C.N. Ramachandran Nair (71), Addl. Director (Rtd) passed away on 25/09/ 2012 .

He served the Animal Husbandry Department in various offices and got retired from

service as Addl. Director. May his soul rest in peace


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journal of indian veterinary association december 2012, 10 (3)

Documents

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