The developmental pattern of leg paralysis in Lucilia sericata (Meig.) induced by 0.3 pi Risella 17 oil

(applied topically) was observed to differ according to whether the appendages belong to the fore-, mid-, or

hindlegs. In general the forelegs were least susceptible and the number of paralytic legs increased with time.

By 3 days after treatment most of the affected flies had 3-4 legs paralysed. Also evident is a strong association

between the type of paralytic symptoms and the kind ofappendages

Lim, G. S.

Universiti Putra Malaysia Institutional Repository

Pertanika 3(1), 56-60 (1980)Notes on the Developmental Pattern of Leg Paralysis Inducedby Risella 17 oil on Lucilia sericata (Meigen) 1G. S. LIMCrop Protection Research Branch, Malaysian Agricultural Research and Development Institute (MARDI),Serdang, Selangor, Malaysia.Key words: Risella 17 oil; blowfly; Lucilia sericata (Meig.); leg paralysis developmental pattern.RINGKASANCorak pembentukan lumpuh kaki pada Lucilia sericata (Meig.) yang disebabkan oleh 0.3 pi minyakRisella 17 (yang digunakan secara topikal) didapati berbeza menurut 'appendages' samaada yang terdapatpada kaki hadapan, tengah atau belakang.Biasanya kaki hadapan adalah paling kurang kelemahannya dan jumlah kaki yang lumpuh makinbertambah dengan masa. Tiga hari selepas rawatan didapati kebanyakkan lalat-lalat yang menerima rawatanmempunyai 3-4 kaki yang lumpuh. Hubungan yang kuat di antara jenis tanda-tanda lumpuh dan jenis-jenis"appendages' juga diperhatikan.SUMMARYThe developmental pattern of leg paralysis in Lucilia sericata (Meig.) induced by 0.3 pi Risella 17 oil(applied topically) was observed to differ according to whether the appendages belong to the fore-, mid-, orhindlegs. In general the forelegs were least susceptible and the number of paralytic legs increased with time.By 3 days after treatment most of the affected flies had 3-4 legs paralysed. Also evident is a strong associa-tion between the type of paralytic symptoms and the kind ofappendages.INTRODUCTIONAmong the various kinds of oils, Risella 17 oilis commonly used in insecticide formulations. Onseveral occasions, it has been used alone or withinsecticides in blowfly studies (Bard, 1961; Lewis,1962, 1963; Busvine, 1962, 1971). These treat-ments included varying dosages applied topically,by injections, or as surface residues. Although noinducement of leg paralysis was noted in theseearlier investigations, Risella 17 oil was found toinduce leg paralysis in a later study (Lim, 1972,1976) with the blowfly, Lucilia sericata (Meig.)Following this, studies were conducted to investi-gate further into the effect of the oil, particularlyon the pattern of paralytic development. It ishoped that these investigations would pro¥ide notonly a deeper insight into their nature and relation-ships, but also a better. understanding on someaspects of the mortality process since the condi-tion of paralysis is noted to be an extended stateof moribund condition (Lim, 1972, 1976).MATERIAL AND METHODSThroughout the studies, the temperature wasmaintained at 26 ± 1DC and the relative humidity65 ± 5%. All the flies used were unmated andof the same age (4 days old) and brood. Within 24hours of emergence, the males were segregated onthe basis of the distance between the eyes, whichin the female is approximately more than one-third the total width of the head (Aubertin, 1933).This early separation enabled the flies to remainunmated and their age known throughout thestudies.lTaken in part from a thesis for theD.I.C. and M.Sc. degree of the University of London. The study wasconducted at the Field Station, Silwood Park (Berkshire), Imperial College of Science and Technology,England.56G. S. LIMDuring treatment, the flies were temporarilyimmobilized by chilling. Application was madewith a foot-operated Burkard micro-applicatorfitted with an ordinary 1 ml glass syringe thatcarried a bent needle with a blunt tip. Each fly wastopically applied with a nominal dose of 0.3 ill asdelivered by the applicator. The site of applicationwas on the anterior margin of the second lastabdominal segment and on the ventral surface.After treatment, the flies were kept in plasticcontainers (10 cm diameter x 3.7 cm high) ingroups of 10 flies/container. Water and food in theform of granulated sugar were provided. At regularintervals, the flies were observed for symptoms ofparalysis; each fly being considered paralytic whenit had at least two legs paralysed.As a control check, a parallel set of untreatedflies was used.RESULTS AND DISCUSSIONParalytic legs induced by Risella 17 oil werecharacteristics; and had distinctive positions.Altogether, four major types of effects have beenidentified (Lim 1972, 1976), these being: Type I,paralytic legs pointing backward; Type II, paralyticlegs pointing sideways; Type III, affected legsfolded; and Type IV, paralytic legs breaking away.The developmental pattern of these forms isobserved to differ according to whether the legsaffected belong to the fore-, mid-, or hindlegs.100ForelegsThe forelegs were observed to be the leastsusceptible to paralysis under the conditionsstudied. At 1 week after treatment, not more than12% of the treated flies exhibited paralysis in theforelegs (Fig. 1). Among these, no individual hadboth the forelegs affected.Paralysis in the forelegs may be of Type II,Type III or Type IV. All these effects occurred inapproximately equal proportion.MidlegsUnlike the case in the forelegs, paralysis inthe midlegs was readily induced by Risella 17.Even at 2 hours after treatment, 20% of the treatedflies had already developed paralysis in at leastone of the midlegs. This condition rapidly in-creased with time, more than 90% being affectedfrom 2 days after treatment (Fig. 1).Initially, most of the affected flies had onlyone of the midlegs affected. However, by 2 daysafter treatment, the majority had developedparalysis in both legs.In the affected midlegs, all the three maintypes of paralytic symptoms were observed. Themost common was Type III; more than 50% of theflies exhibiting it by 2 days after treatment. Type Isymptom was relatively less frequent, being notmore than 22% throughout the seven-day observa-tional period, while the frequency of Type II80 /~a60 o---<l ForeleO"~ Midleo€--Hindeo.,j!-40 /1>i 200 3 ?~2 4 2 3 4 5 6 7HOURS DAYSPeriod After TreatmentFig. 1. Development of leg paralysis on male Lucilia sericata (Meig.) at varying periods after treatmentwith Risella 17 oil, applied topically at 0.3 ill.57DEVELOPMENTAL PATTERN OF LEG PARALYSIS INDUCED BY RISELLA 17 OILsymptom was intermediate. The latter was ex-hibited by 27-32% of the treated flies in 1-3 daysafter treatment. However, all the main symptomswere obvious, with significant numbers possessingthem by 1 day after treatment.Type IV symptom was also a common feature.The number suffering this effect increased withtime from 23% to 71 % at, respectively, 1 and 7days after treatment.HindlegsAs in the midlegs, paralysis inducement byRisella 17 in the hindlegs was rapid and common.A similar pattern of development was observed i.e.the numbers affected increased rapidly with time(Fig. 1). For instance 26%, 50%, 88%, 91 % and 94%of the treated flies showed hindleg paralysis at2 hrs., 4 hrs., 3 days, 5 days and 7 days aftertreatment, respectively.The major paralytic symptom exhibited bythe hindlegs was Type I. This symptom developedrapidly, increasing from 26% at 2 hrs. after treat-ment to more than 70% from 3 days onwardsafter treatment. None of the other conditions (i.e.Type II, Type III, and Type IV) appeared before 2days after treatment. Comparatively, very few fliessuffered these effects. At 7 days after treatment,only 24% developed Type II symptom while lessthan 6% exhibited Type III and Type IV symptoms.All The Legs Considered TogetherNumber of legs affected: The total number of80legs affected in a particular fly was observed tovary with the time interval from treatment (Fig. 2).The longer the interval period, the greater was thenumber of legs affected. For example, at 2 hrs.after treatment, the percentage number of flieshaving one, two, and three affected legs was verylow (8% - 14%). By the fourth hour, 20% - 24%of the flies had two and three legs affected. Thelatter increased to about 32% at 1 and 2 daysafter treatment.As more of the legs in a fly became paralytican obvious change in the number of flies having afew affected legs (one - three) to that havingfour - five affected legs was noticed. Thus, theformer condition gradually decreased with time.Among the latter, flies with four affected legswere predominant (53%) at 3 days after treatmentand this condition, with slight variation, was main-tained thereafter throughout the observationalperiod.Flies having five affected legs were noted only5 days and after from treatment. However, only afew showed this condition; there being 5% - 12%.Throughout the study no fly was observed tohave all its six legs affected. "Symptom development with time: Initially,up to about 4 days after treatment, all the. threemajor paralytic symptoms developed progressivelywith time. However, except for Type I and Type IIparalysis which remained more or less constant,j60u".,........o"'~402 HOURS 4 HOURS 1 DAY 3 DAYS 5 DAYS0123456 0123456012345601234560123456Number of legs affectedFig. 2. Paralytic effect of Risella 17 oil (0.3 J1l/fly) in relation to the number of legs affected in maleLucilia sericata (Meig.) at varying period after treatment.58G. S. LIMType III paralysis showed a sharp decline, fallingfrom 68% at 4 days after treatment to 35% at 7days after treatment (Fig. 3). As recovery of TypeIII symptom was not observed~ the decline wasprobably due to the increasing loss of the legsaffected in this manner. This appeared to be so asType III and Type IV (legs breaking away) wereboth common features of the midlegs.then indicated the maximum effect induced by theoil. This was based on the findings that the limitingeffect was reached by the third day from the oilapplication.Due to the large number of zero and smallfrequencies obtained (Table la), the data had tobe combined as a 2 x 2 contingency table (Table100 SYMPTOM:0---0 Type0---0 Type II~ Type IV....--. Type III80.~;0:::_40o"U~60.!-oo M;;.__~~;"-'_"""~_-_.....' ....__-'-__....L.__.....I.__....I.__....I.__.....l2 4HOURS2 3 4DAYS5 6 7Period after treatmentFig. 3. Development of different types of leg paralysis in male Lucilia sericata (Meig.) induced by Risella17 oil, applied topically at 0.3 pl//Zy.Association of leg position and types ofsymptom: it was noticed that certain paralyticsymptoms normally developed only on certain pairof legs in the affected fly. For example, Type Isymptom was most commo~ on the ~indlegs butcomparatively rare on the ffildlegs, while Type IIIwas predominantly on the midlegs. In the midlegs,symptoms of Type II, Type III and Type IV werenormally observed, with Type III being the mostcommon. These suggest that there may be anassociation between certain symptoms and thetypes of legs in an affected individual. This isclearly evident from Table 1 which shows thefrequencies of the types of legs and symptoms at3 days after treatment. The data at 3 days aftertreatment are used as the symptoms exhibited1b) and analysed by the Fisher exact probabilitytest (Siegel, 1956). In the analysis, it was foundthat P = 0.467 x 10-13 which shows conclusivelythat the types of paralytic symptoms developedin affected individual are highly dependent on thetypes oflegs affected.In general, it is noted that there is a strongassociation between certain symptoms and thetypes of legs in an affected individual. Also, eachof the major symptoms rarely occurred by itself.More often, an affected fly would exhibit com-bination of the different symptoms; many com-binations being possible and these varied withdifferent individuals.59DEVELOPMENTAL PATTERN OF LEG PARALYSIS INDUCED BY RISELLA 17 OILTABLE INumber of Lucilia sericata (Meig.) exhibiting the differenttypes of paralytic sypmtoms in relation to the differenttypes of legs. The paralytic symptoms were induced byRisella 17 oil applied topically at 0.3 f.1l on the male flies.a. Uncombined values recorded.Paralyticsymptoms Forelegs Midlegs HindlegsType I 0 0 24Type II 0 J J JType III 0 20 4Type IV J 11 Jb. Data from "a" combined as 2 x 2 contingency table.Paralytic Forelegssymptoms + Hindlegs TotalMidlegsType 11+Type III+ Type IV 43 6 49Type I 0 24 24Total 43 30 73Some of the symptoms appeared to be inter-changeable, at least for short period, e.g. Type IIto Type III symptoms and vice versa. This wasmore common in flies in the initial period ofsymptom development, suggesting that the diffe-rent symptoms observed were probably due todifferent degrees in the effect of the oil at varyingintervals from treatment. Also, once paralyticsymptoms have developed there is generally norecovery, i.e. the paralytic effects are permanentin the affected flies.60ACKNOWLEDGEMENTGrateful thanks are due to my supervisor,Dr. C.T. Lewis, for his constant interest andguidance.REFERENCESAUBEIlTIN, D. (1933): Revision of the genus LuciliaR-D. (Diptera, Calliphoridae). Linnean Society'sJ. Zoology. 38: 289-436.BARD, J. M. (1961): Influence of the soh'ent on thetoxicity of injected solutions of an insecticide.M.Sc. Thesis (London), Department of Zoologyand Applied Entomology, University of London.BlJSVINE, J. R. (1962): A laboratory technique formeasuring the susceptibility of houseflies andhlowflies to insecticides. Laboratory Practice, 11:464-468.BlJSVINE, J. R. (1971): A critical review of the tech-niques for testing insecticides. CommonwealthAgric. Bureaux, London.LEWIS, C. T. (1962): Diffusion of oil films overinsects. Nature (London) 193: 90~.LEWIS, C. T. (1963): Some applications of radio-isotopes to the study of the contamination ofinsects by insecticide solutions. Proc. Ser. illt.Atomic Ellergy Agency. 74: 135-146.LIM, G. S. (1972): The effect of chemicals appliedtopically on Llicilla sericata (Meig.) with specialreference to Risella 17 oil. M.Sc. Thesis (London).Department of Zoology and Applied Entomology.Uni"ersity of London.LIM, G. S. (1976): Inducement of leg paralysis inhlowfly (Lucilia sericata Meig.) hy Risella 17 oil.Mal. Agric. Res., 5: 19-23.SIEGEL, S. (1956): Nonparametric statistics for thehehavioUlal sciences. New York. McGraw-Hili.( Received 20 February 1980)

Notes on the Developmental Pattern of Leg Paralysis Induced

by Risella 17 oil on Lucilia sericata (Meigen)

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Notes on the Developmental Pattern of Leg Paralysis Induced by Risella 17 oil on Lucilia sericata (Meigen)

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