2013 International Nuclear Atlantic Conference - INAC 2013 Recife, PE, Brazil, November 24-29, 2013 ASSOCIAÇÃO BRASILEIRA DE ENERGIA NUCLEAR - ABEN ISBN: 978-85-99141-05-2

INCREASE OF ACCEPTABILITY PERIOD OF Musca domestic L., 1758 (DIPTERA: MUSCIDAE) PUPAE, IRRADIATED BY GAMMA RADIATION AS HOST OF THE PUPAL Spalangia endius, Walker, 1839 (: PTEROMALIDAE)

Natanael M. Itepan1, Sara E. D. Z. Itepan2 and Valter Arthur3

1 Instituto Federal de São Paulo – IFSP - Área de Física Rua Diácono Jair de Oliveira, 1005 - Santa Rosa. 13423204 Piracicaba, SP [email protected]

2 Faculdade de Filosofia Ciências e Letras de Ribeirão Preto – FFCLRP - USP Av. Bandeirantes, 3900 05508000 Ribeirão Preto, SP. [email protected]

3Centro de Energia Nuclear na Agricultura – CENA - USP LIARE Av. Centenário, 303 13400970 Piracicaba, SP [email protected]

ABSTRACT

This experiment was carried out in Biological Control of Domestic "Eduardo Hiroshi Mizumoto" Laboratory at Entomology and Acarology Department (LEA/ESALQ/USP) and in Food Irradiation and Radioentomology Laboratory (LIARE/CENA/USP). The gamma radiation source used was a Co-60 irradiator model Gammabeam-650 of the Atomic Energy of Canada Ltd. whose activity in the beginning of the experiments was 9.8x1013 Bq. (2,644 Ci). The lots of pupae of Musca domestic L., 1758 and the parasitoid Spalangia endius Walker, 1839 had been kept in acclimatized room with 25 ± 2°C of temperature and 70 ± 5% of relative humidity. This experiment was designed to investigate if the non-viabilization M. domestic pupae, using gamma radiation, could build up the acceptability period as host of the pupal parasitoid S. endius. At these age intervals, the dose to prevent adult emergence was 25, 220, 360 and 520 Gy respectively. The 1, 2, 3 and 4 days old pupae were irradiated (dose rate: 1,510 Gy/hr) and exposed to the parasitoid S. endius at a proportion of one female parasitoid to five housefly pupae, during different periods after the irradiation. The results allow us to conclude that irradiation increase the acceptability period of the housefly pupae by the parasitoid. The best age to irradiate the housefly pupae was one day.

1. INTRODUTION

The pressing necessity of increasing the animal protein availability in rational and economic way, gave origin to the modern systems of animal production. These systems cause a great manure accumulation, with enormous potential for the proliferation of synantropic . The flies are of great importance in doctor-veterinary medicine, therefore they are transmitters of some pathogenic agents (virus, rickettsia, bacteria and protozoa), act in the egg dissemination of helminthes [1,2]. The Family Pteromalidae (Hymenoptera: Cahlcidoidea) includes great number of species parasites, many with a very important paper as agents in the control of synanthropic flies [3]. Although they are gifts in the field, it does not have an adjusted number to exert good control [4]. However, with the mass creation of these in laboratory [5], and repeated releases, they can thus, to win the great potential reproductive of these plagues, and to get a satisfactory control level [4,5,6,7,8]. Leading experiments the field level had observed that Muscidifurax uniraptor and Pachycrepoideus vindemiae acted in the surface and until 3 cm of depth, in manure poultry, already Spalangia spp attacked the hosts located between 5 to 10 cm depth [9]. The success achieved with the release was facilitated by extensive laboratory studies that determined the increased efficiency control of parasitoid S. endius [10]. These studies were used to develop a computer model to simulate the control of M. S. domestica endius [11]. Figure 1 shows the relationship between host and parasitoids.

Figura1. Diagram of the model of interaction between Musca domestica and pupal parasitoid Spalangia endius [11].

The irradiations cause its deleterious effect in the muscles that help the adult to emerge, provoking partial or complete insufficiency of this process. However, pupas not viable, in the majority of the cases, contain completely imago [13]. The technique of the irradiation in the creation of parasitoids is used for the inhibition of the emergency of the flies and the elimination of the multiparasitism for the maintenance of pure colonies of parasitoids [12]. Pupae of M. domestica with: 0-1, 1-2, 2-3, 3-4, 4-5 days radiating them with rate of dose of 2020 Gy/hour, got LD100 respectively: 100, 250, 450, 850 and 1350Gy [14]. One of the important factors in the radio sensitivity of some insects is the dose rate employed [15]. The objective of this work was to test the possibility to increase the period of acceptability of pupae, M. domestica, as hostess of the parasitoid S. endius, with the irradiation process.

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2. MATERIALS AND METHODS

This experiment was carried out in Biological Control of Domestic Fly "Eduardo Hiroshi Mizumoto" Laboratory at Entomology and Acarology Department (LEA/ESALQ/USP) and in Food Irradiation and Radioentomology Laboratory (LIARE/CENA/USP). The used species, S. endius and M. domestica had been gotten of colonies kept for the Laboratory of Biological Control. The radiation source used gamma was a irradiator of Co-60 type Gammabeam 650, of the Atomic Energy of Canada - Ltd., whose activity in the beginning of the experiments was of 9,8x1013 Bq. (2644Ci). The lots of pupae of M. domestica had been kept in room acclimatized with temperature of 25 ± 2°C and 70 relative humidity of ± 5%, until reaching, the desired ages. To evaluate the acceptability of pupae radiated by the parasitoid, the doses for pupae of 1, 2, 3 and 4 days had been respectively: 25, 220, 360 and 540 Gy, with rate of dose of 1510 Gy/hour. Each treatment had 5 repetitions, with 20 pupae each. The ratio host: parasitoid was: 5:1. Each repetition was conditioned in a polystyrene cup, with capacity of 250 ml, whose cover the central part was extracted, and glues a screen to allow the gaseous exchanges. In each repetition one was placed of cotton piece absorbed in water and another one in honey solution 25%. After the exposition of pupas to the parasitoids, by an interval of 1 day, each repetition was individualized in a glass pipe of 2,3 cm of diameter for 9 cm of height, closing with a peace of cotton water-repellent, where if it gave to the emergency of the parasitoids and flies. They had been carried through the counting of the emerged parasitoids of each repetition. The experiment followed the casualization project entirely contends the following factor: * Irradiation, with the following levels: pupas radiated with 1, 2, 3 and 4 days, and pupas not radiated, * Interval of age where pupa was displayed to the with the following levels: 1 to 2, 2 to 3, 3 to 4, 4 to 5, 5 to 6, 6 to 7, 7 to 8, 8 to 9, and 9 to 10 days. For pupas radiated the interval of exposition to the parasitoid it was not up to 4 the 5 days; therefore after this period occurs the emergency of the adults of the house fly. For the analysis the level of 1 factor was fixed in relation to the levels of the other factor. Statistics distribution free was used, by means of the test Qui-square [12].

3. RESULTS AND DISCUSSION

The lineage of the parasitoid S. endius is gifts Table 1. The analysis of the results, for the test Qui-square, to a level of significance of 5%, is gifts in Table 2. Analyzing pupae radiated one does not perceive that it does not have difference in the lineage of S. endius in the intervals of the age of pupa. It inside did not have difference of the 4 tested intervals, between radiated pupae of not irradiates [15,16]. Pupae radiated with 2, 3 and 4 days had provided an increase of acceptability in 1 day; already radiated with the 1 day this period drew out in 2 days. The irradiation obtained to draw out the period of acceptability of pupae for the parasitoids. Pupae when radiated with 1 day, they had been the ones that had presented greater period of acceptability (with 5% of significance). Dissecting pupae made impracticable, noticed that the irradiation does not affect histogenesis, therefore all the imago had presented completely [12].

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Table 1 Lineage of gotten S. endius of pupae of M. domestica submitted to the treatments: not radiated, radiated with 1, 2, 3 and 4 days of age and displayed to the parasitoids in different intervals of the age of pupas (of 1 the 2 up to 9 the 10 days).

Intervals of Repetitions TREATMENTS exposition to No Irradiated Irradiated Irradiated Irradiated the parasitoid irradiated with 1 day with 2 day with 3 day with 4 day 1 15 14 ------2 16 15 ------1 to 2 days 3 12 17 ------4 15 14 ------5 9 16 ------Total-Average 67 (13,4) 76 (15,2) 1 15 11 16 - - - - 2 14 12 12 - - - - 2 to 3 days 3 9 14 15 - - - - 4 17 14 14 - - - - 5 8 9 14 - - - - Total-Average 63 (12,6) 60 (12,0) 71 (14,2) 1 13 11 8 5 - - 2 10 10 15 10 - - 3 to 4 days 3 8 12 11 4 - - 4 14 12 11 8 - - 5 15 11 12 6 - - Total-Average 65 (13,0) 56 (11,2) 57 (11,4) 33 (6,6) 1 8 10 11 8 10 2 5 12 14 5 8 4 to 5 days 3 8 12 9 8 6 4 6 9 11 8 15 5 8 8 10 6 9 Total-Average 35 ( 7,0) 51 (10,2) 55 (11,0) 35 (7,0) 48 (9,6) 1 - - 2 1 2 0 2 - - 5 0 1 0 5 to 6 days 3 - - 3 1 0 1 4 - - 5 1 2 0 5 - - 5 1 0 0 Total-Average 20 (4,0) 4 (0,8) 5 (1,0) 2 (0,4) 1 - - 1 0 0 0 2 - - 1 0 0 0 6 to 7 days 3 - - 0 0 0 0 4 - - 2 0 0 0 5 - - 0 0 0 0 Total-Average 4 (0,8) 0 (0,0) 0 (0,0) 0 (0,0) 1 - - 0 0 0 0 2 - - 0 0 0 0 7 to 8 days 3 - - 0 0 0 0 4 - - 0 0 0 0 5 - - 0 0 0 0 Total-Average 0 (0,0) 0 (0,0) 0 (0,0) 0 (0,0) 1 - - 0 0 0 0 2 - - 0 0 0 0 8 to 9 days 3 - - 0 0 0 0 4 - - 0 0 0 0 5 - - 0 0 0 0 Total-Average 0 (0,0) 0 (0,0) 0 (0,0) 0 (0,0)

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1 - - 0 0 0 0 2 - - 0 0 0 0 9 to 10 days 3 - - 0 0 0 0 4 - - 0 0 0 0 5 - - 0 0 0 0 Total-Average 0 (0,0) 0 (0,0) 0 (0,0) 0 (0,0)

Table 2. Analysis of the averages of the lineage of gotten S. endius of pupas of M. domestica submitted to the treatments: not radiated, radiated with 1, 2, 3 and 4 days of age displayed to the parasitoid in different intervals of the age of pupas (1 the 2 days up to 9 the 10 days).

Intervals of Treatments exposition to No Irradiated Irradiated Irradiated Irradiated the parasitoid Irradiated with 1 day with 2 day with 3 day with 4 day 1 to 2 days A 13,4 A A 15,2 A ------2 to 3 days AB 12,6 A A 12,0 A A 14,2 A -- -- 3 to 4 days AB 13,0 A AB 11,2 AB AB 11,4 AB A 5,6 B -- 4 to 5 days B 7,0 A AB 11,2 A AB 11,0 A AB 7,0 A A 9,6 A 5 to 6 days -- AB 4,0 A B 0,8 AB B 1,0 AB B 0,4 B 6 to 7 days -- B 0,8 A C 0,0 B C 0,0 B C 0,0 B 7 to 8 days -- C 0,0 A C 0,0 A C 0,0 A C 0,0 A 8 to 9 days -- C 0,0 A C 0,0 A C 0,0 A C 0,0 A 9 to 10 days -- C 0,0 A C 0,0 A C 0,0 A C 0,0 A

Letters to the right of the average, comparisons in the direction of the lines and to the left, in the direction of the columns to a level: 5% of significance (x2).

4. CONCLUSIONS

On the basis of the gotten results, I concluded that: - The non-viable pupae of M. domestica with gamma draws out acceptability for S.endius pupal parasitoid; - Radiated Pupae with 1 day had been the ones that had provided the biggest period of acceptability for the parasitoid S. endius.

6. REFERENCES

1. ANÔNIMO. 1986. Faxina semanal mantém baixa a população de moscas. Gado Holandês, São Paulo, 50(127):25-27. 2. ANÔNIMO. 1984. Importância das moscas na criação de animais, especialmente bovinos. Gado Holandês, São Paulo, 45(115):54-8. 3. RUEDA, L.M. & AXTELL. Guide to common species of pupal parasites (Hymenoptera: Pteromalidae) of the house fly and other muscoid flies associated with poultry and livestock manure. Raleigh, North Carolina University Agricultural Research Service, 1985a 88p. (Technical Bulletin, 278). 4. MORGAN, P.B. & PATTERSON, R.S. Sustained realeases of Spalangia endius to parasite field populations of three species of filth breeding flies. Journal of Economic Entomology. College Park, 70(4): 450-2, 1977.

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5. MORGAN, P.B.; LABRECQUE, G.C. & PATTERSON, R.S. Mass Culturing the Microhymenopteran parasite Spalangia endius. Journal of Medical Entomology, Lanham. 14(6): 671-3. 1978. 6. MORGAN, P.B.; PATERSON, R.S.; LABRECQUE, G.C. WEIDHAAS, D.E.; & BENTON, A. Supression of field population of houseflies with Spalangia endius. Science, Washington, 189:388-9, 1975. 7. OLTON, G.S. & LENGER, E.F. Winter inoculative releases of parasitoids to reduce house flies in poultry manure. Journal of Economic Entomology, College Park, 68(1):35-8, 1975. 8. RUTZ, D. & AXTEEL, R.C. Sustained releases of Muscidifurax raptor (Hymenoptera: Pteromalidae) for house fly (Musca domestica) control in two types of caged-layer poultry houses. Environmental Entomology, Maryland, 8(6): 1105-10, 1979. 9. RUEDA, L.M. & AXTELL, R.C. Effect of depth of house fly house fly pupae in poultry manure on parasitism by six species of Pteromalidae (Hymenoptera). Journal of Entomological Sciences, Tifton, 20(4):444-9, Oct., 1985. 10. MORGAN, P.B.; PATTERSON, R.S. & LABRECQUE, Q.C. Host-parasitoide relationship of the house fly, Musca domestica L., and the protelean parasitoid, Spalangia endius Walker (Hymenoptera Pteromalidae and Diptera: Muscidae). Journal of the Kansas Entomological Society, Lawrence, 49, 483-488, 1976. 11. WEIDHAAS, D.B.; HAILE, D.E.; MORGAN, P.B.; LABRECQUE, G.C. A model to simulate control of House Flies with a pupal parasite Spalangia endius. Environmental Entomology, Lanham, 6(4): 489-500, 1977 12. ITEPAN, N.M.; BERTI FILHO, E.; COSTA, V.A.; ARTHUR, V. & COSTA, N. Utilização de pupas de Musca domestica L (Diptera: Muscidae) irradiadas com raios gama (60 Co) na criação dos parasitos Spalangia endius Walker, Muscidifurax raptor Girault & Sanders e Pachycrepoides vindemiae (Rondani). In: CONGRESSO BRASILEIRO DE ENTOMOLOGIA, 11, Campinas, 1987. Resumos, Campinas, SEB, 1987, p. 234. 13. GRECCHI, M.A. Alterações biológicas em Musca domestica L., 1758 (Diptera: Muscidae) induzidas pela radiação gama do cobalto-60. Piracicaba, 1989. 170p. (Mestrado - ESALQ/USP). 14. DOMARCO, R.E. Influência da taxa de radiação gama sobre a radiosensibilidade de cinco espécies de insetos, Piracicaba, 1977. 68p. (Mestrado - ESALQ/USP). 15. NEGRILLO, B.J. 1992. Métodos não paramétricos uni e multivariados. Piracicaba, (Primeira edição), 215 p. 16. MORGAN, P.B.; SMITTLE, B.J.; & PATTERSON, R.S. Use of irradiated pupae to mass culture the microhymenopterus pupal parasitoide Spalangia endius Walker (Hymenoptera: Pteromalidae): I. Musca domestica L. (Diptera: Mucidae). Journal of Entomological Science, Tifon, 21(3): 22-7, 1986. 17. ITEPAN, S.E.Z. Progênie de Spalangia endius Walker, 1839 (Hymenoptera: Pteromalidae) em pupas de Musca domestica L., 1758 (Diptera: Muscidae) que foram inviabilizadas com a radiação gama do Cobalto-60 e resfriadas. Ribeirão Preto, 1992. 60p. (Mestrado - Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto/USP).

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