Bowers, W.S.

de Oliveira Filho, A.M.

Pinchin, R.

Santos, C.E.

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545 TRANSACTIONS OF THE ROYAL Socmv OF TROPICAL MEDICINE AND HYGIENE, VOL. 74, No. 4, 1980 Brief Communications Activity of precocenes on the Chagas’s disease vector, Panstrongylus megistus A. M. DE OLIVEIRA FILHO, R. PINCHIN, C. E. SANTOS Nzicleo de Pesquisas de Produtos Naturais, Centro de Ci&cias da Satide, Universidade Federal do Rio deyaneiro, Rio de Janeiro, Brasil AND W. S. BOWERS Dept. of Entomology, Entomology-Plant Pathology Laboratory, Cornell University, Geneva, N, Y. 14456, USA The continued use of the organochlorine insecti- cides, HCH and dieldrin, for triatomine control is at present under review. Several organophosphorus, carbamate and pyrethroid insecticides are currently being tested and one, malathion, has already been introduced in the national vector control campaign in Argentina. It also has been suggested that juvenile hormone analogues (JHAs) may be used as control agents (GILBERT, 1976). However, as triatominebugs a;e most susceptible to JHAs orily during a few weeks of the fifth instar, their oractical use is somewhat limited (PINCHIN -et aZ.,& 1978a). This may not be so for another class of compounds with insect growth regulatory activity-the plant- derived precocenes. In 1976, Bowers announced the discovery of such compounds with anti-juvenile hormone activity. Their principal effects on OncoDeltus fasciatus (Dallas) were the induction of prec&ious “metamorphosis,’ and anti-gonadotropic and ovicidal activitv (BOWERS 1976. 1977: BOWERS et al., 1976). As we have noted some sikilarity in the structure-activity relationships for JHAs for 0. fasciatus and Panstrongylus megistus (Burmeister), it was considered feasible that the precocenes might also exhibit anti-juvenile hormone activitv on P. megistus and other triatomine bugs of importance in the transmission of Chagas’s disease. We describe here--the results of initiz experiments with the precocenes and discuss their possibilities as future vector control agents. Materials and Methods The specimens of P. megistus used in these experiments were from a laboratory colony main- tained at 27 +2”C and 65 i 15 y0 R.H. and given a weekly opportunity to feed on chickens. Acetone solutions of precocenes I and II (7-methoxy- and 6,7-dimethoxy-2,2-dimethylchromene, respectively) were applied topically, using a microsyringe, to the dorsal abdomen of each of the five nymphal instars, so that each insect received 100 pg of the compound. Insects in the control groups were treated with acetone (1~1) only. The insects were then main- tained in clean glass jars (700 ml capacity) fitted with filter paper roosts and closed with nylon netting. They were given a weekly opportunity to feed on chickens and their development was observed daily. Similarly, third and fourth instars, one to seven days after ecdysis, were treated with precocene I, orecocene II and urecocene II D~US i%(2. 5- hichlorophenyl)S, 7-himethyl-2, 6-dctadienyiamine a JHA effective in inducing juvenilization in P. megistus at 1 .O pg/fifth instar (PINCHIN et al., 1978b) and 10 pg/fourth instar (Oliveira F., unpublished results), at the dosage given in Table II, and observed as before. The mortality , time taken to ecdysis and morphological characteristics of the subsequent instars were recorded. Results and Discussion Table I shows the results of topical application of precocene I and II to all the nymphal instars of P. megistus. Treatment of the third and fourth instars with precocene II resulted in high percent- ages of precocious adultoids at the next ecdysis. Precocene I, however, showed only weak activity on the fourth instar. The percentages, corrected (Abbott’s formula) with respect to control group mortality, of apparently normal insects emerging upon the first moult after treatment, demonstrated the toxic effect of precocene II on the first and second instars and, to a lesser degree, of precocene I on the first instar. Furthermore, of all the insects which underwent ecdysis after treatment with precocene II, only one, a third instar which moulted to give an apparently normal fourth instar, moulted a second time. The resulting fifth instar died the day after ecdysis. Thus precocene II was 100 % effective in blocking normal development when applied to the first to fourth instars. For precocene-treated P. megistus, a marked delay in moulting was observed in those insects which underwent precocious metamorphosis. The precocious adultoids obtained had the appearance of adults in miniature, with the character- istic red and black coloration on the conexivium. The wings, though membranous and veined, were only partially developed, being small and curled up. The tarsi were, in general, two-segmented although some of the adultoids resulting from treated fourth instars had three-segmented tarsi. It had been Table I-Effects of topical application on all nymphal instars on P. megistus treated with precocene I or II at 100 ,ug/insect Compound Treated No. insects instar treated After 1st mouh After 2nd moult No. precocious adultoids % precocious adultoids % apparently normal /insects moulted [insects treated % apparently normal /insects treated 1st 90 0 0.0 2nd 20.0 65 22.2 (39*3)1 : 0.0 70.8 Precocene I 3rd 50 (92.7) (43.6)l 60.0 0.0 82.0 (92.9) (94.6) 4th 66.0 5th z: : 9.3 78~0(100~0) (82.5) 0.0 70~0(100~0) 68*0(100*0) - - 1st 30 2nd 25 : 0.0 10.0 (17.7) o-0 0.0 8.0 Precocene II 3rd 20 (10.5) (0.0) :: 83.3 4th 15 10-o (11.3) 84.6 13.3 5th 10 (18.9) 0 0.0 50-o (83.3) -- 1 Numbers in parentheses correspond to the percentages corrected (using Abbott’s formula) with respect to control group mortality. Table II-Effects of treatment of 3rd and 4th instar P. megktus (1 to 7 days old) with precocene I, II or II plus the Jm N-(2, 5-dichloro- phenyl)3,7-dimethyl-2,6-octadienylamine Treated Dose No. insects After 1st moult After 2nd moult instar Compound pg/nymph treated 56 precocious “/C apparently No. precocious adultoids/insects normal/insects No. precocious % precocious % apparently adultoids moulted treated adultoids adultoids/ normal/insects insects moulted treated Precocene I 100 30 0 o-o 73.3 (84.4)X 0 0.0 Precocene II 100 30 19 100.0 0.0 (0.0) 70*0(86*6)l 3rd Precocene 11 10 30 9 47.4 33.3 (38.2) -ii 2272 Precocene II 31 Rec. II -k JHA loo-:10 30 ii 0.0 71.0 (81.4) 0 2c3(28:8) 0.0 46.7 (53.5) 12 1OE 58*1(71.9) 0.0 (0.0) Precocene I 100 20 0 0.0 Precocene II 90~0(100*0) 0 o-0 100 20 19 100.0 o-0 (O-0) 55*0(64.9) 4th Precocene II Prec. II + JHA 100 $0 ii ?I i:; 85.0 (94.0) -i OT;; 70.0 (77 *8) 6 42.8” 7;TO(885) 40*0(47*2) 1 Numbers in parentheses correspond to the percentages corrected (using Abbott’s formula) with respect to control group mortality. 2 Adultoids not precocious because they moulted from the 5th instar. ACTIVITY OF PRECOCENES ON P. megistus 547 noted previously that if precocious metamorphosis occurs from the fourth instar, either with precocene application to third-instar 0. fasciatus (BOWERS \ ei hl., 1976) or to fourth instar Rhodnius prolixus (Stal), as described by TARRANT & Cum (1978), the tarsi in the resultant fifth instar adultoid are three- segmented, as in a normal adult. The adultoids were unable to copulate with each other or with normal adults and oviposition did not occur. They were also apparently unable to feed. Life expec&cy was short, averaging 20.9f4.2 davs for fourth-instar adultoids that moulted from treated third instars; adult female P. megistus can live under laboratory conditions for five months and males for even longer (SZUMLEWICZ, 1976). Conse- quently these fourth and fifth instar adultoids can be considered to have very low vector potential. Table II shows the results of the treatment of third and fourth instars with the precocenes at various dosages, and with precocene II plus a potent JHA for this insect. The slight difference in response to the precocenes for the insects aged up to one week (Table II) and those of undefined age (Table I) can be attributed to the fact that some individuals of the latter group had several opportuni- ties to feed before treatment, and hence the physio- logical processes involved in development and ecdysis would already have been initiated in some insects. Precocene II, applied to the third instar at 10 pg/insect, caused the emergence of precocious adultoids upon both the first and second moults after treatment. As expected, concurrent treatment with a JHA counteracted the effects caused by precocene. The third and fourth instars, treated with precocene II and a JHA (100 and 10 pg/insect, respectiv- ely), after passing through an intercalary instar, gave rise to fifth and sixth instar adultoids res- pectively All the fifth and 50”, of the sixth instar adultoids were similar in appearance to the precocious adultoids described above, although the sixth instar adultoids cannot strictly be termed precocious. However, the other sixth instars were characteristic of the partially juvenilized adultoids (PINCHIN et al., 1978b) which result from the application of JHA alone. Thus precocene ap- parently does not block the JH-receptor centres and probably inhibits JH production. This agrees with the results obtained for 0. fusciatus (BOWERS, 1977), in which females sterilized by precocene, upon receiving a dose of JH III, resumed oocyte development. Evidently compounds with anti-juvenile hormone activity may have a future as control agents. Contrary to JHAs, they have the advantages of shortening the life-cycle of triatomine bugs and inducing the appearance of sterile adultoids with negligible vector potential. At present it seems unlikely, however, that either JHAs or precocene analogues will completely replace insecticides in control campaigns, although they could well be useful as complementary pesticides in integrated control programmes. Acknowledgements We are indebted to the Ministry of Planning (FINEP), the National Research Council of Brazil (CNPq), and the Research Council of this University (CEPG/UFRJ) for financial assistance. References Bowers, W. S. (1976). Discovery of insect anti- allatotropins. In : The Juvenile Hormones. Gilbert. L. I. (Editor). New York: Plenum Press, pp. 394-40‘8. ’ Bowers; W. S., Ohta, T., Cleere, J. S. & Marsella, I’. A. (1976‘1. Discoverv of insect anti-iuvenile hormones in’ plants. Sckce, 193, 542-547. Bowers, W. S. (1977). Anti-juvenile hormones from plants : chemistry and biological activity. In: Produits Naturels et la Protection des Plantes. Citta de1 Vaticano: Pontifica Academia Scienti- arum, pp. 129-156. Gilbert, B. (1976). Possible use of insect hormone mimics in vector control. In: New approaches in American trypanosomiasis research. Proc. Int. Symp., Belo Horizonte, 1975. Pan American Health Organization, Washington, D.C., pp. 252- 288. Pinchin, R., Oliveira Filho, A. M. de, Figueiredo, M. J., Muller, C. A. & Gilbert, B. (1978a). Slow-release juvenile hormone formulations for triatomine control. Transactions of the Royal Society of Tropical Medicine and Hygiene, 72, 322-323. Pinchin, R., Oliveira Filho, A. M. de, Figueiredo, M. J., Muller, C. A., Gilbert, B., Szumlewicz, A. I’. & Benson, W. W. (1978b). Screening and structure-activity relationships for juvenile hormone analogues for Panstrongylus megistus, a primary vector of Chagas’ disease in Brazil. journal of Economic Entomology, 71,950-955. Szumlewicz. A. P. (1976). Laboratorv colonies of Triatominae, biology and population dynamics. In: New approaches in American trypanosomiasis research. Proc. Int. Symp., Belo Horizonte, 1975. Pan American Health Organization, Washington. DC., pp. 63-82. - - - . Tarrant, C. A. & Cupp, E. W. (1978). Morpho- genetic effects of nrecocene II on the immature stages of Rhodniu; prolixus. Transactions of the Royal Society of Tropical Medicine and Hygiene, 72, 666-668. Accepted for publication 31st August, 1979. 

Activity of precocenes on the Chagas's disease vector, Panstrongylus megistus

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