STUDIES ON THE SUSCEPTIBILITY

Level Of Tlie Housefly Musca Domestica Nebulo To DDT IN ALIGARH

PROJECT REPORT SUBMITTED IN THE PARTIAL FULFILMENT

OF THE REQUIREMENTS FOR THE AWARD OF THE

DEGREE OF MASTER OF SCIENCE IN ZOOLOGY (PARASITOLOGY).

Course No. 5-Zy-72

1975—76

BY

JOSEPHINE PONDAYI-ZESAGULI (MRS) M.Sc. (Fiaal)

ROLL i. 0. 71 ZYM—32 ENROLMENT No, L 4111

DB?ART.:.;EN^ir OF ZOOLOGY ALIGAEH MUSLIM UNIVERSITY

ALIGARK

JUNH, IZ15

r / i^'

' /)S~ r '

Z^3/ _ '/

DS2831

STUDIES ON THE SUSCEPTIBILITY

LEWa OF THE HOUSEav WUgCA. DOWESTICf NEBULO TO DOT

IN M.IGARH

Project Report Submitted in the Par t ia l Fulfilnant of the Requiranenta

for the Award of the Degree of Raater of Science in Zoology (PARASITXOGY} •

Courae No* S«fl ->72

1975 - 76

Bv

JOSEPHINE PONDIIVI^ESCULI (nRS)

n.Sc* (Final)

Roll No. 7l2irn-32

CNROLnENT No. L4111

ZOOLOGY OEPARTPIENT

ALIGARH PIUSLIPl UNIVERSITY, (^IGARH.

JUNE, 1976

STUDIES ON THE SUSCEPTIBILITY

LEV/a OF THE HOUSEFLY WUSCj DOnESTICA NEBULO TO DDT

IN M.IGARH

» C K « O M L E O G £ i n E N T S

I an grateful to Or* ^ O B I I A* Ansari for superulBing ay project* I alao

wLah to thank Ptofaseor S« n« Ham, Head of the Zoology Oepartoianty Mlgarh

nualiai Univeraity, ULigazh for providing the nacaaaary laboratory f a c i l i t i a a .

l-aatly, but not the least^ I wiah to expraaa ay gratitude to the Covemaant

of India, Cultural Scholarahip thich financed mjf atudiea*

JOSEPHINE PONDWUESAGULI (RRSi

Section of Paraaitology,

Zoology Department,

^ igarh Ruslin Uniueraity,

HL I C A R H-

C O N T E N T S

£fl6£Jift.

**• INTfOOUCTION 1

2 . REUIEIil OF LITERATURE 3

3 . nATERIM. AND TECHNIQUE 6

4 . RESULTS AND DISCUSSION 8

TABLE 8 (ai

GRAPH 8 (b>

5. simum 10

6 . REFERENCES 11

I N T R O D U C T I O N

The houaefly, liUftSB JUtttila i s a oosn»polltan species and has great medical

inportaioB* The f ly serves as a direct agent and vector of nany pathogens

especially those causing enteric dieeases* These enteric diseases, for

example^ dysentry are due to the fact that f l i e s frequent to i l e ts , dumps

and even faeces for egg-laying. Here, they pick parasitic protozoans e.g.

Cnt ^anoeba, ^iardla etc* , and helminth ova on their hairy and sticky feet or

actually swallow t h a i . Latsr, they depoeit the organlama In Kitchens, dining

tablee and exposed edible substences. Que to their regurgitation habit, the f l i es

vomit the germe from their food reservoir with the saliva to dissolve the eugar

end alao via their faecse ( f ly specke)* Qiandler reports that such specks are

said to be produced by a well-fed f ly every 5 nlnutee a l l day. So, f l i es in

this way transmit bacteria causing anthrax, typhoid, dysentry and food poisoning*

They also harbour the virus for polioinyelltis and may produce myiasis* Spirochaetes

cauelng yawe are also transmitted by f l i e s .

Obviously, the potential danger of f l l e e oennot be minimised. The denger

described above l i ee mainly in their feeding habits. I t i s alao attributable to

their high fecundity, f e r t i l i t y and the short duration of their developmental

cycle. The fecundity i s better appreciated I f one raelises that a female, within

i t e l i f e time yiich averages from 6 to 8 weeks, w i l l have depoeited up to

2,000 egge* Hence, the danger of f l i e s i s multiplied in geometric progression.

Back home in f^odeeia, there are s t i l l many deaths and diseases among man

and his livestock caused by parasltee* Typhoid and dysentry are ooromon among

dilldren Wiose l ives are nipped in the bud and thus their potential i s a great

loss to the country's progress*

- 2 -

Han has sines bscome awars of tha imminent danger of f l i es and has t d e d to

e}cbarroinate than maChanlcally and biologically* But the most affective measure l a

the control tsy chemicala eo called aa insecticides. The chemicals may include

chlorinated hydrocarbonsf organophosphorous con^unda and plant products as

pyrsthroids* Among tha chlorinated hydrocarbona, the most oommonly used

inescticidas against f l i es i s DOT,

The praeent project on suaceptltilllty of houseflias to ODT in Aligarh waa

takan up ao that I gat a )^sed to the method of spproach in tha control of

inaacta* Thia wi l l make me better equipped to go back and help al leviate

the problem of these peats htiich are of medical and veterinary importance

together with other reeearchea related to toxicology*

- 3 -

R E t f l E M O F L I T E R A T U R E

DOT waa th» f i r s t insocticlda of thtt dtlocinabed hydEocartwn gcoup to to used

on a Mocld Mido scdLs againot houaefliaa and othar pasta* This waa alao tha f i r s t

GhsKLcidi to induce taaiatanca in inaacta« Maturally i t drew graabar raaaarch effort

than any other ceapound*

OOT Mae f i ra t amJa in 1874* I t e inaackicidal pzopectiea uera dlacouarad by

Hullar in tha Baala LaboratetiaA of tha SuLaa Caapany of 3«R. Gaigy S.A« in 1939«

Tha tMM inaacticLdo quickly psauad axcaptionaLly affactive by checking the plague

of ColotadB b ^ l e in t94t tftiloh thseatanad the SMlaa potato crop at a time iriisn

a l l food ciopa uare of utnaat inportanca« The inaact idda M M thoroughly teated

againat a kdda ranga of inaacta inclriddtng •aaquitaaa and houaafliaa. I t took a

year to aatabliah i t s inaatit iddal adtion*

The dLfficultiea of ooaaunication during l^srld liiar I I reatrictatt tha apraad

of thia inforaaitioR to tha f raa uorld* The cheodcal was ultinataly introduced

into the United Kingdoa and tha U«S.A* toMarda the end of 1942» and by that time

much waa kno«n concerning the coapound*

Rullar (l9S9) reported thifc coapounding DOT into 5 or 10^ duate and spraya

proved to have a laating affect againat f l i e s and noaquitoas in rooaaf barracka

and f era buildinga.

Miiaaan <1944) ahowad that DOT had affective reaidual action and acted aa a

narva poiaon on tha f l i aa *

Bukbon (l945) deacribad tha work carried out by Buavina (l94S) ahoudlng that

the lethal doaa of DOT for •"*>•»*• '*innn rhimntfrn waa 1 ag par aq* cm. of surface,

librk haa also been dona on the toxic aanifaatatitm of DOT in other aniaale

including aan.

Lindqulat and co*Herkara (1944) reported that DOT applied aa a apciy in a

auitahla eolvent raaliined aa a nearly inviaibla depoait after tha l iquid had

volati l iaad and acted aa a residual contact inaacticide against housefllss.

Coaparisons of the insacticidal reaulta of DOT in varieua ooapounds as paints.

- 4 -

waxes etc* have also been done*

•ueetaan (l946) reooMunded spot treatnent of DOT hhers f l i es csllect for

feeding aid teeting* In 1947 SiMetnan got excellent f ly control with DOT in tMo

daily herds and reepective bacns*

Kellsr (l95S) tested the effectiveneee of QOT in baita*

Ae onre and aore DOT uae used i t kias noticed that there uaa gradual decre^e in

i t e effectlvenees* This was because jcttiijyumsa **^^ developed in the f l i ee hhich

nuet not be confused Mitb jfjjiBs J^SJJUCiasi (Hoskins and (cordon, l95&/« Thsre are t«o

types of resistance yiich heve bean differentiated as t ft f>||l,Qif and phyV g 10^ T"^

(gsnstic) resistances*

Hess (1952) was the f i r s t to describs bshavioral resistance in mosquitoes

folloy.ng contact with DOT, Since than i t hae bean noticed in f l i ee alao*

Busvins (l963) observed that vigor tolersnca snd behavioral reeiatonce ware

not l ikely to preeent eerious obstacles aa physiological reeietance to najor

inaact control achenes*

PhY8tBtetti.Bal. .WBtrtanCl i s due to the preaenca of an enzyme OPT^YdratfttHttiLniae

litich converts DOT to OOC Perry and Hoekins ( l9Sl) found this detoxification of

DOT as a factor in ths resitance of housefliee* Linquist jj^La-flL* ( l 9 5 l ) reported

that thie conv^nion of DOT to OOC i s mainly concentrated in the fat body and

gut region or in the cuticle hypodermis*

lAiereaa DOT i s toxic, OOC i s non-toxic to the insect* Hence, physiological

rssistance can develop to the point of immunity*

MLeaman (l957) obssrvsd that OOTi^ssistant f l i ss had « MnniJrf bai^i^ar nhidn

dissolvBS the DOT. Hance, DOT becomes insffective to such DOT-resistant f l i e s *

Brown ( l96 l ) recorded 36 inetances of DOT«ceaiatance*

- 5 -

Synthei^s of anzyn^ i s ganabically conttollsd* Hsncs, 00T-««ai9tant f l i a s

Mi l l also produce DOTHrasistant psogeny* The inheritance mechanism or OOT-

rasiatanca haa bean found to be aanof actorial* insari (l969) agreea with thia

aingla gana hypothesis but found i t to be not oonplately dondnant.

Khan and ihaaci (l964) noted that l i i i l a lUi iai JHHOUifli JtaOO^iiAft llsd

davalopad raaistance for (K)T, ths Indian houasfly (Buece dniaetiga ^itauia) had

not attained any aignificant degree of realatence* They alao fomd that HuSSS

do—ti,q« *f*in*if1^ 99^ from C^cutta waa l iab le to become more reaitant than the

n ' HiP aub««pecie8. •

Psi ( l96 l ) obtained only 1*6 - 2.0 t l M a DOT •—(itf-anca in iSyttGUdBOaiifcifiA

jUtBllA f n > s Delhi vil lage*

ScLnca i t Mae f i r s t discovered in Swden in ( lM6)» resistance has been

giotdng at an alacning taite and yet insect ic ide are s t i l l being effective*

Buewine (l963) e j^a ina thia obearvation aa being due to the Iscalieation of

resistance in nany instsncea and alao beceuee a l i e i t e d number of apeciee ittov.

reaiatance to both groups of diiesinatad ineaot ic i iM*

However i f e jec t ion preeeure continuee, resistance mill ult ieately spread

throu^ the range of a apeciea* Thie» I think, i e a very important and crucial

point to bear in nind in any toxLoological undertaking since the rate of growth of

resitwice i s wsll corrslatsd with selectiva prsssurs*

M U T E R i a L S a N O T E C H N I Q U E

ISXl LLJJL§«-

Flies u»re collactad from various loca&itlsa in Aligarh \dz> ShaMhad HaMMt,

S«N« Hall nsBS and tha City* Thaaa f l i aa uara pooled togabher to get a homoganaous

stock*

The r i iea waca nainfcainad in k&ra nai ci^aa in tha laboratocy at rooca tanpara-

tura* Thaaa were fed on buffalo nLlk which Mas diluted wLth wetar. Tha nilk was put

in pet t i (tt.ahee with aoaked cotton* The patr l diahea ware changed daily*

g fc H R I N G»,

The f l i ea readily ouipoaitad in the p i t r i diahea* i hatch of eggea was

aapamtaly aabedded in glaaa jars 4 X 8 " under the pads of nilk-aoakad-ootton*

Cara waa taken to embed a aingla batch of eggs i n a Jar, otheriAaa owarcxoMding

would result in wary aaall pupae and f l i ea with a high aorta l i ty* The feapa of the

jars ware oovared with aualin dotha aecured with a rubber band to pre\Mnt ovlpoaition

froiB outaids f l i e s and at the aana t ins allow constant aeration* Evsrydaf fresh

aoaked cotton waa added to the J are and a dry layer of cotton uaa placed on the top

oT the pad on tha f i f t h day* Larvae migrate to this dry layer and pupate in i t .

The pupae were ieolated from the dry cotton and put in petr i dlehea aeparetely

acoorc^Lng to the embedding date* Theee ware allowed to hatch in muslin cloth cages*

I t took approximately 12 •> 14 tteya from the i n i t i a l atage of the egg to the emergence

of adi l ta* i ien theee f l i e a become four-day old» they are fu l ly mature and are ready

for testing*

- 7 -

T E S T I N G .

Pure CX3T uas taken ae the teet cheaical* i atodc aoiution of 1^ concentration

was nads* M least f ive fonaulations ranging fron 0»3125^ to 1^ were prepared.

Oose of the insec t idde vaB maintained as Q«Q01B nl / f iy*

Testing was done by topical application of the insecticide through a odcro-

applicator outfit* The f l i e s were anaaethebized uith carbon dLoxida* A drop of the

insecticide wee a^ilied on the doreiMi of the f l iee*

Treated fl leevere dropped into tieaua papas cages* Sugar cubes were provided

ae Bource of food end the jspening of the cege was ca|>ped with dampened cotton*

The cages were kept in the laboratory at room temperature*

After 24 hours of testing* the number of dead and l i v e f l i e s were counted in

each cage* nortality percantagaa wen calculated for each DOT concentration using

the formula I->

Wtimber of dead f l i e s X lOO

Totml number of f l i e s tested 1

Live f l i e s were those liiich could s t i l l fly and walk properly, the rest were considered

as dsad.

- 8 -

R E S U L T S fcWD D I S C U S S I O N

Fxoa the Tabl» th» nalas seen to bs nace auacaptihle to DOT trsatoMrtt than ,

the fanales since there were ante demiiRales (664) than fenalee (384i* This glv/ )»

a ration of females dead to maksa dead of 1 > 1»78.

I t appears that femalee nay be havdng a better inherent cc^jadty for QOT

detoxification than the males* But actually the higher percentage in malee i e

becauBO of thiHLr incrsaeed metabolic activity* Qbvlouely t lp id diffuaion end is

abeorption of thia ineect idde taking place in males as compared to females*

The reeults are very consistent and i t ma be concluded that males are more

susceptible than females*

^%0 i * e . that concentration of OOT uhich i s capable of ki l l ing 50^ of the

fly population aa calculated from the graph idea found to be 0*135* This i s a lower

value than uhat Khan and Aneari (l964) got for HUgarh f l i e s* They found LC g for

Wuaca t^qifai^qa »fH»^n to be 0*32 and that "^ ' IITBH (ttWUlbi litB y *? * was 0*29

against DOT dissolved in A ^ ^ n s *

Sen ( l959 | obtained §00 mortality with 49 DOT in Huifia dai—at^ga uictna from

Calcutta* lihile developing resistance to DOT in these f l i e s . Khan and Aieari raiied

the value of the LCgQ to 1*44 in the atMtia. ^^5 and to 14.5 in the y idna " 2

generationa*

From thia comparieon even though i t i s adsittedly not extensive, i t can be

concluded that Ai.igaih f l i e e are e t i l l very susceptible to DOT treatment* This

means that OOT could be used to control these f l i e s* In fact Allgarh f l i e s are

Buch a menace that chamo-^reatment should be tried with OOT but only for a short

duration before the f l i e s become recant* A campaign must be launched to educate

the people a l l around about the dangers of houseflies and how each individual

can asalet by pepper eanitation and deecrindn^e disposal of garbage and other

refuse* This will get rid of the potential breeding places*

- 8 (a>

T It 8 L £

PERCENTUGE WQRTi nV OF HOUSEFLIES HT DIFFERENT ODNCENTRATlOttOF OPT

(oncentration No of Np.of fiJBs dead

V^IIA " " l " P-ialea Total ^ nortality

0,03125 316 40 27 67 21,2

0*0625

0,125

0.25

0*5

342 1

332

313

316

68

117

121

164

51

57

72

76

119

174

193

240

34,7

52.4

61,68

75.3

1.0 320 174 98 272 85.0

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PR03ITS

. 9 .

100! •octal i ty in the f i l e s could be obtained by application of 7% DOT as

calculatad frem the graph* This would, houever, be impractical since acetone

can diaaolve only up to 6^ O^T, Beyond th i s dosage the DOT uLU remain

undissolved* So, another batter soltiwnt UDUld have to be substituted for ths

acetone A th is X%V9X at DOT concentration.

Therefore, females showed less susceptibility to OOT than males* LCgQ

Msa found to be only 0*135 iJiich indicates that the f l i e s have net yet developed

resistance against CXJT Md 7^ DOT i e i^at i a needed to give IQO t mortality diasolved

in eome s u i t ^ l s solvent*

- 10 -

s u n n a R Y

Aiigach housaflles WBCS collected fron vedoue l e c e l l t l M and pooled to get (x

hoeogaReouB etock« Ftom this etock F<t ganerad^lon was otibeined* Four-day old

f l i e s uere tested by topical application on the docsua with various concentra­

tions of DOT diseolued in acetone* The eueceptibility level of theee f l i e s Mas

assessed from a plottMi probability curve*

It was found that LCg^ was 0.135 of DOT and that 7j( DOT can give IQQ^

nortality* Ralee were found to be nere eusceptible to DOT then featalss*

- 1 1 -

R E F E R E N C E S

1 * KNSARI, 3«A«l Studies on the i s o l a t i o n and genetic nature of spec i f ic

insect ic ide resistance i n Housefl ies*

•BBtYy*^aqaKtf ^ t 70 ( i 9 6 9 ) .

2 . BELDING, OU-.t Text Book of f i n i c a l Paraa i to loov . %)pleton-Century-Crofts,

Inc« N.Y* PS53 - 855 ( l936)«

3 . BROiiN, IUli<*A«t nechanien of resistance against inssc t ic idas* annual, Rauiau

f^f Er fci M lntw 5^ 301 - 326 ( l 9 6 0 ) .

4 . aUSVINE, 3«R«t The present status of insec t ic ide raatatanca^ B u l l . >ihrld Health

qgQaniaatian.^g Suppl . , 31 > 40 ( l 9 6 3 ) ,

5 . CMIPBELL, G*A« and ^Sl, T # F . . I OPT and Wauar Infaefcl,<^da«. Chapman &

Hiall L t d . , London ( l 9 5 0 ) ,

6 . CHANOLER, ««C« and READ, C ^ . , i IntroAicfcityi nf Paraa l to lnov. hfLlay Eastern

Ptf t . L t d . , New D e l h i . ( 1 * 1 ) .

7 . HESS, A.O.f ftWELcan ?aUU Trap* » t d . " Y B M S, 3 7 1 9 1 9 5 2 > .

B. HOSKINS, W. n . , ; & GORDON, H.T . I %thropod resisltance to chemicals.

jQUgt ^Onn. %lt«» A, 89 - 122 ( l 9 5 6 ) .

9 . KELLER, 3 . C , , I Poison Baits f o r the Cbntiol of Housefl ies on m i l i t a r y

paaei^uations, J . Eonn> Enfc. ^ , 528 - 529 ( l 9 5 5 ) .

10 . KHMi, N.H. & UNSARI, 3 . l l , t OOT-rssistance hazard i n the Indian housef l ies ,

nuasa rinaMigtlfin xiatiftUA and Vhiasa dBBtatAM

jtiasLoa. JaJboL^aBaku JSS, is ( i 9 6 4 ) .

1 1 . LINDQUIST, A , U , niM^OEN, A . H , , UILaON, H.G« and 30NES, h . l U , t

J> ^fiOnt ,£nt»> ,37, 132 ( l 9 4 4 ) . *

1 2 . niCKS, 0 . l i , , t Xnsecticide-Realstance. I Revieid of Oevelopments i n 195B and

1959. B u U , Ufartd Heaitih ftSO* -22» 519 - 529 (1960)

1 3 . P«L, R.I Trans, R. Soc. Troo. Had. Hyp. , j g , 125 - 126 (1951^^

1 4 . PERW, A«S« & HOSKINS, U.n«.} The de tox i f ica t ion of DOT sa a factor i n the

resistance of house f l i es . 3> Econ., Enfc. 44 (s )

850 - 857 ( l 9 5 l ) .

- 12 -

15. SWEE7WAN, H#L«t 3 . Eoan, Enfc. ^9, 380 (l946>.

16. SUCCmAN, B,L,t Comparative effectiuensss of OOT #<0 000 for control of f l i e s ,

3^ £cot^. Er t> jjQ ( 4 ) , 565 - 566 Cl947).

17. WEanAN, R, and FEN30S, P «. W.ittt SchMBJ^ fln1i> 888., JlS, 179 (l944>!

18. MIESnAN, R.I T««^^ PKyi><«i«gy. J , 1S7 - 197 (1967).

* Oitiginal kbrke not conaulted.