A comparative study has been earried out in order to investigate the effects of cytarabine and ftorafur on the larval development and sex segregation of the Mediterranean fruit fly.Peer reviewe

Burgos, R.

Muñiz, Mariano

Digital.CSIC

BURGOS R., MUNIZ M. INHIBITION OF LARVAL GROWTH OF CERAT/T/S CAPITATA WIED. BY ADDITION OF ANTIMETABOLITES TO THE LARVAL DIET Es/ratio da:	 Frustula Entomologica Nuova Serie, vol. IX (XXII) , pp . 129-133 - 1986 PACINI EDITaRE PISA R. BURGOS, M. MUNIZ (*) INHIBITION OF LARVAL GROWTH OF CERATITI5 CAPITATA WIED. BY ADDITION OF ANTIMETABOLITES TO THE LARVAL DIET INIBIZIONE DELLA CRESCITA LARVALE DI CEROT/ TI S CAP/TATA WIED. MEDIANTE ADDIZIONE DI ANTIMETABOLITI ALLA DIETA ARTIFICIALE ABSTRACT A cornpa ra tive study has been ea r ried ou t in order lO investigat e the effects of cyta ra­bin e (l -~-arabinofurano s ylc ylo sine ) and ftorafu r [s-flu oro-I-(lelrah ydro-2-furyl)-ura eil] on the larval dc velopment. r -T " -'-:: igh l and sex segre gat ion of the Mediterranean fru it fly. INTRODUCTION After Knipling proposed the insect eradication by release of laboratory reared individuals by ionizing radiation (KNIPLtNG, 1955), man y control programs using this technique have been carried out in the a ffec ted countries. In the last ten years, the effects of irradiation and chemicals on the reproduction of insects of medical and economic importance have be en evaluated. In certain cases, insects sterilized by chemicals have appeared to be more vigorous than those sterilized by ionizing radiation , but effective doses tended to reduce sexual competitiveness, mating frequency and longevity of treated males and oviposition and longevity of treated fem ales (CAVALLORO and DELRIO, 1981; EL-GAZZAR el al., 1983; GUERRA, 1975; PATTERSON e l al. , 1977; SANTlAGO-ALVAREZ and SARASUA, 1983, etc.). However, little work have be en done about the incidence of combination of chemical and physical treatments on their reproductive activity. CHANG and Hsu ha ve demonstrated that precocene 11 s ign ífican tly reduces sex attrancy in males of the Mediterranean fruit Ily (CHANG and Hsu , 1982; Hsu and CHANG, 1982); on the other hand, ovaries treated with the five most active benzyl-1.3-benzodioxoles showed a range of 28 to 78% of treated individuals ex­hibiting retardation in ovary growth (CHANG e l al.. 1984). GUERRA (1975) showed that combinations of two substerile treatments (antimetabolites and r-irradiation) were more effective than each of them separately. Ftorafur [(5-Fluoro-I-(tetrahydro-2-furyl)-uracil)] a nd cy ta ra b ine (I -p-D­Arabinofuranosylcytosine) have been the ch emicals used in this study. Ftorafur is a nu cleoside antimetabolite that acts by inhibiting the DNA synthesis (HILLERS e l al ., 1967) and cytarabine contains, in contrast to normal nucleosides, cytyd ine a nd desoxycytíd íne arabinose as synthetic nucleoside instead of ri bose and desox­yr ibose; it is a cytostatic from the range of pyrimidine antagonist. Its effect is att ributed prirnarly to the inhibition of the de soxy cytidine synthesis; however, in add ition , inhibition of the cytidilic acid and kin ase a lso takes place, as well as incorporation of the compound into nucleic acids (LAY et al., 1971). Our proposal in this paper is to s tudy comparatively the effects of two an­timetabolites (ftorafur and cytarabine) on the larval development parameters of Ceratitis capitata Wied. In a next work we will investigate the joint action of these chemicals and r-irradiation on the reproductive activity of this species in order to find useful resuIts for practical cont ro l programs. (*) Instituto de Edafologia y Biología Vegetal (C.S.J.C.) el Serrano 115 Dpdo. 28006 Madrid, Spainal. - 130 ­MATERIAL ANO METHOOS The antimetabolites were incorporated separately into a new larval diet (ANDRÉS and MUNIZ, 1984; MUNIZ and ANDRÉS, 1983) to give several concentrations ranging from 10 to 50 ppm with ftorafur and from 4 to 20 ppm with cytarabine. 80 neonata larvae were daily seeded in glass vials (2 cm dia. x 9 cm long) contain­ing 5 grams of the larval diet and provided with a porous stopper to get a good aeration. Pupae were daily removed and the emerged adults were recorded and sexed. The number of replica tes varied according to the dosages and treatments and no lower than 5 of them were carried out. Conditions during the experiments were 26 ± 1°C;. 65 ± 5% RH and a light: dark regime of 12: 12 hours (1900 lux). Statistical differences are indicated in the tables of results. Means followed by the same letter do not differ s ignificantly at the 0.05 level by the Student's t-test, TABLE 1 - Eiiect s 01 [torajur and cytarabinae on th e larval development of th e Mediterranean [ruit fly, Ceratitis capitata Wied. (i< ± S .E .) Doses Pupation time Pupation Pupal weight (mg) (ppm) (days) (%) (7 day-old pupae) O 7.39 ± 0.06 a 93 .0J ± 0.09 a 9.19 ± 0.09 a (n = 21) (n = 21) (n = 53) Ftorajur 10 8.40 ± 0.14 bcdh 87 .38 ± 0.13 ab 9.55 ± 0.08 b (n = 5) (n = 5) (n = 14) 15 8.26 ± 0.08 e 87.39 ± 0.15 ab 9,09 ± 0.30 ab (n = 6) (n = 6) (n = 19) 20 8.75 ± 0.14 d 89.87 ± 0.12 ab 8.99 ± 0.24 a (n = 6) (n = 6) (n = 22) 50 10.99 ± 0.11 e 84 .73 ± 0.11 b 8.78 ± 0.32 a (n = 6) (n = 6) (n = 29) Cylarabine 4 7.41 ± 0.05 ab 92.63 ± 0.17 a 7.95 ± 0.15 b (n = 6) (n = 6) (n = 20) 6 7.89 ± 0.13 cd 91 .02 ± 0.03 a 8.06 ± 0.17 be (n = 7) (n = 7) (n = 25) 8 8.04 ± 0.09 d 93.80 ± 0.08 a 7.84 ± 0.19 be (n = 7) (n = 7) (n = 25) 10 8.83 ± 0.19 efh 92 .29 ± 0.05 a 7.06 ± 0.20 ed (n = 9) (n = 9) (n = 42) 15 8.87 ± 0.10 f 85.69 ± 0.08 b 6.88 ± 0.22 d (n = 6) (n = 6) (n = 38) 20 11.58 ± 0.11 g 65.42 ± 0.15 e 5.41 ± 0.25 f (n = 5) (n = 5) (n = 35) - 131 ­RESULTS AND DISCUSSION In table I is presented the action of ftorafur and cytarabine on different larval develop­ment parameters of Ceratitis capi ta ta Wied. It ca n observe that a sign ifica nt retard of 3 days in the pupation time with regard to control wa s obtained wh en the concent ra t ion of ftorafur in the larval diet was 50 ppm, but this effect was found with cyt arabine at the rate of 20 ppm. We ights of 7 day s old pupae were practically identical in cont ro l and ftorafur treat­ments and significantly different in the cont rol and cyt arabine ones; in this case pupal we ight s lowe r than 7 mg were o b ta ined a l dosage of 15 and 20 ppm. TABLE 2 - Effect s o] [toraiur and cy ta rabine orz the adult emergence, sex segregation and inhibition o i the larval migration o ] th e Mediterrarzearz [ruit jly, Ceratitis capita ta Wied. Data are presented as %. (x ± S.E.) Inhibition of Dose s Adult emergenee Sex segrega tion the larval migration (ppm) (referred to pupae) Males Females (Pupa t ion in the vials) o 98.90 ± 0.04 a 50.77 ± 0.02 a 49.23 ± 0.02 a 7.52 ± 0.08 a (n = 19) (n = (9) (n = 21) Ftoralur 10 98.77 ± 0.21 a 51.87 ± 0.22 a 48.13 ± 0.22 a 9.37 ± 0.05 ac (n = 5) (n = 5) (n = 5) 15 98.27 ± 0.11 a 49 .60 ± 0.03 a 50.40 ± 0.03 a 10.39 ± 0.17 ac (n = 6) (n = 6) (n = 6) 20 96.91 ± 0.20 ac 47 .91 ± 0.04 a 52.10 ± 0.04 a 9.91 ± 0.31 ac (n = 6) (n = 6) (n = 6) 50 95.14 ± 0.06 be 55 .50 ± 0.07 a 44 .50 ± 0.07 a 21.38 ± 0.47 e (n = 6) (n = 6) (n = 6) Cytarabin e 4 94.88 ± 0.37 abe 52.37 ± 0.04 a 46.63 ± 0.04 a 9.19 ± 0.03 a (n = 6) (n = 6) (n = 6) 6 97.58 ± 0.32 ab 57.46 ± 0.14 a 42.54 ± 0.14 a 11.75 ± 0.27 ac (n = 5) (n = 5) (n = 7) 8 99.51 ± 0.22 a 50.47 ± 0.13 a 49.53 ± 0.13 a 8.60 ± 0.09 a (n = 5) (n = 5) (n = 7) 10 92.36 ± 0.07 b 50 .00 ± 0.01 a 50.00 ± 0.01 a 18.64 ± 0.33 bcd (n = 7) (n = 7) (n = 9) 15 85.91 ± 0.14 e 51.27 ± 0.09 a 48.73 ± 0.09 a 12.54 ± 0.43 ad (n = 6) (n = 6) (n = 6) 20 46.25 ± 0.63 d 40.73 ± 0.20 a 59.27 ± 0.20 a 47.51 ± 0.92 e (n = 5) (n = 5) (n = 5) 132 In a former study, the pupation time of larvae reared in a diet including 5-fluorouracil at 10 ppm wa s about 11 day s, 3.5 da ys later than the one obta ined in the standard diet (BURGOS and MUNIZ, 1981). On the other hand, yield of pupae with cyta rab ine at 20 ppm was lower than the one found with ftorafur at 50 ppm. With respect to adults emergence, referred to pupae, the same effect can be ob served in table 11. The larval diet including cytarabine at the rate of 10 ppm showed a toxicity higher than the one obtained with ftorafur at the highest concentration. Sex ratio was practically 1:1 in control, ftorafur and cyta rabine treatments, but a tendency to decrease this proportion was found when cyta ra­bine was included in the larval diet at the rate of 20 ppm (40.73% for males and 59.27 % for females). On the other hand, larval migration was also studied ; a significative de crease in the ability to leave the artificial medium was found in larvae reared in diets with ftorafur at 50 ppm with regard to the control; however, this effect was ob­tained with cytarabine at 10 ppm. In this case, the inhibition of the larval migra­tion at 20 ppm was almost twice highe r than the one obtained with ftorafur at 50 pprn. From these results it can be concluded that the toxic effects in larvae reared in diets that include cytarabine were higher than the ones observed when ftorafur is incorporated to the larval medium. For practical purposes, do ses greater than 20 ppm a nd 8 ppm with ftorafur and cytarabine respectively, are not ad visable. ACKNOWLEDGEMENTS The au thors wish lo express their gratitud to Mrs. C. González Godino for her co llabo­rat ion . SUMMARY A compa ra live s tudy has been ca r ried out in order to inve stiga le the effec ts of cytara­bine (I -~-D-arabin ofu r anos ylcytosin e) and ftorafur [5-fluoro-I-(lelrahydro-2-furyl)-uracil) on the larval development , pupal weight and sex segrega tion of the Medite rranean fruit fly, when th ese ant imetabolites were included in the larval diet a t concentrations ranged from 4 ppm to 20 ppm (Cytarabine) and from 10 ppm to 50 ppm (Ftorafu r). The results obta ined showed that, in gene ral, the tox ic effec t was higher with cyt ara­bine than with Fto rafur. An increase of 4 days in the pupation time was observed when cytarabine was included in the larval rearing medium at a dosage of 20 ppm but the sa me effecl was found with ftorafur a t 50 ppm. On the other hand, a decrease in the wei ghts of 7 day-old pupae was obtained with cytarabine al 4 ppm. However, only a lendency lo this eff ect was found with ftorafur al 15 ppm. Al the rare of 15 ppm in both cases , the yield of pupae wa s lower than the one obtained with the laboratory st andard diet . RIASSUNTO Sono riportat i i risultati degli stud i su gli effe tt i dell a cytarabina ( I -~-a ra b i no fu rano­sylcytosina) e ftorafur [5-fluoro-I-(tetrahyd ro-2-furil)-uracilJ sullo sviluppo larvale, pes o pu ­pal e e segregazione sessuale della mosca mediterranea della frutta . GIi antimetaboliti somministrati nella dieta larvale in concentrazioni comprese tra 4 e 20 ppm per la cy ta rabina e tr a lO e 50 ppm per il ftorafu r hanno dim ostrato una maggio­re tossi cita del primo prodotto. E stato inolt re evidenzia lO un incremento di 4 giorni della durara pupale qu ando la - 133 ­cytarabina veniva inclusa nel substrato artificiale in quant ita di 20 ppm; lo stesso risultato e stato ottenuto con lo ftorafur in dose di 50 ppm. É st ata ugualmente registrata una diminuzione del peso delle pupe ottenute da larve nutrite con medium con tenente 4 ppm di cy ta ra bina; lo stesso tipo di tendenza e stato registrato con ftorafur a dosi di 15 ppm. REFERENCES ANDRÉS M.P., MUNIZ M., 1984 . Desarrollo de un a nueva d ieta larvaria para Ceratitis capi ta ta Wied. Bol. S ervo Plagas., 10 (1): 85-116. BURGOS R., 1981 - Alteraciones morfológicas en Ceratitis capitata Wied. por tratamiento con 5-fluoruracilo en la dieta larvaria . Graellsia , 37: 77-96. BURGOS R., MUNIZ M., 1982 - Efe ctos del 5-fluor u racilo sobre el desarrollo larvario de Ceratitis capitata Wied . Graellsia, XXXVII: 97-121. CAVALLORO R., 1981 - Establishment of continuously in vitro growing cell lines of Med -fly. Revu e Canadienne de Biologie , 40 (2): 181-186. CAVALLORO R., DELRIO G., 1974 - Sterilizzazione di Dacus oleae e Ceratitis cap itata con radiazioni gamma e neutroni veloci . Redia, 55: 373-392. CAVALLORO R., DELRIO G., 1981 . Radiost erilization of male a nd femal e Sitophilus ory z.ae (L.). Redia , 44: 157-164. CAVALLORO R., MUNIZ M., POZZI G., 1984 . Comparative radiosensitivity of Medfly cells and embryos. Redia, 67: 421-433 . CHANG F., HSU c.L., 1982 - Effect of precocene Ir on sex a tt ra ncy in the Medit erranean fru it fly. Ceratitis capitata (Wied.). Ann. Entomol . Soco Am., 75: 38-42. CHANG F. et al. , 1984 - Effect of precocene and benzyl-I-3-benzodioxole derivates on sex attrancy in the Mediterranean fruit fly. Ibid., 77: 47-50. EL-GAZZAR L.M., DAME DA, SMITTLE B.J ., 1983 - Fertility and cornpet it iveness of Culex qu inquelasciatus males irradiated in nitrogen . J. Econ. Entomol., 76: 821-823. GUERRA A.A., 1975 - Sexual ste riliza tion of Tobacco budworms with combinations of oral chemosterilants and y.irradiation . Ent. Soco Am. , 68 (1), 1-3. HILLERS S. et al. , 1967 - Analogs of pyrimidine-nucleosides. I.N-(a-Furanidyl) derivates of natural pyrimidine bases on their antimetabolites. Dokl Akad USSR , 176 (2): 332-335 . LAY H.L. et al ., 1971 - Experiences with cy tosine arabinoside in childhood leukaemia and Iymphoma. The Medical Journal 01 AUSlralia, 2: 187. MELLADO M. et a l., 1972 . Ensayos de lucha autocida contra Ceratitis capita ta Wied . An. IN lA/Ser. Pral . Veg., 2: 421-433 . MUNIZ M., ANDRÉS M.P. , 1983 - Investigaciones bási cas para la inclusión de Hansenula an omala como aporte proteico en la dieta larvaria de Ceratitis capitata Wied . Graellsia , XXXIX: 16 5-174. OHINATA K. el a l., 1977 . Mediterranean fruit Fly : Sterility and sexual competitiveness in laboratory after treatments with y-irradiation in air, carbon dioxide. Helium, nitro­gen or partial vacuurn . J. Econ . Entomol ., 70 (2): 165-168. PATTERSON R.S. et al. , 1977 - Release of radiosteril ized males to contro l Culex pipien s quinqueiasciatus. J. Med. Entomol., 14: 299-304 . SANTIAGO-ALVAREZ c., SARASUA M.J. , 1983 - Repense de Ceratitis capitata Wied. au diflubenzuron . In Proc. of the CEC/IOBC International Symposium Fruit Flies of Eco­nomic Importance. Athens/Greece/16-19 November 1982. (Ed . R. Cava lloro): 219-225 . SHARMA V.P. et a l., 1973 . Chemosterilization of the tropical home mosquito Culex pipiens quinquelasciat us Wied .: Laboratory a nd field ca ge studies. Bull. W.H.O., 48: 45-48. manoscritto presentato il 28 agosto 1987 

Inhibition of larval growth of Ceratitis capitata Wied. by addition of antimetabolites to the larval diet.

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