Avaliação biológica de machos esterilizados por radiação gama, para utilização em programas de controle populacional de Aedes aegypti (DIPTERA: CULICIDAE) / Biological evaluation of sterilized males by gamma radiation for use in the Aedes aegypti (Diptera: Culicidae) population control programs

Este estudo avaliou o desempenho biológico e reprodutivo de machos de Aedes aegypti esterilizados por radiação gama e seu potencial para uso no controle da espécie, através da técnica do inseto estéril (TIE). A eficácia de esterilização de diferentes doses de radiação gama, 30, 40 e 50 Gy, foi avaliada em machos irradiados (MI) na fase de pupa. O seu efeito sobre a longevidade e competitividade de acasalamento dos MI, sobre a fecundidade e fertilidade das fêmeas acasaladas com estes machos, bem como de fêmeas também irradiadas (FI), foram parâmetros analisados. Os testes de competitividade foram conduzidos em gaiolas teladas (2 X 2 m) em condições simuladas de campo, onde foram liberados machos irradiados e machos não irradiados (MNI), nas seguintes proporções: 5:1, 10:1 e 15:1. Os resultados revelaram que o fitness biológico e reprodutivo dos mosquitos são afetados pela radiação, de modo que os efeitos são diretamente proporcionais a dose de radiação utilizada. O parâmetro de longevidade apresentou diferenças significativas, tanto quando os MI foram analisados de forma individual quanto em grupo. Para a maior dose, MI e FI viveram em média 19,6 ± 2,2 dias e 25,8 ± 2,2 dias, respectivamente, enquanto que no grupo controle, estes valores foram de 24,9 ± 2,5 dias e 30,4 ± 2,5 dias...

This study evaluated the biological and reproductiv e performance of Aedes aegyptimales sterilized by gamma radiation and its potential for use in controlling the species through the Sterile Insect Technique (SIT). The sterilization efficacy of different doses of gamma radiation, 30, 40 and 50 Gy, was assessed in irradiated males (IM) in the pupal stage. The direct effects of gama radiation on the longevity and competitiveness of IM, as well as the effect on the fecundity and fertility of females mated with these males, and also irradiated females (IF), were analyzed. The competitiveness was examined by tests conducted in large cage (2 x 2 meters) in semi field conditions where irradiated males (IM) and non-irradiated males (NIM) were released in the following proportions: 5:1, 10:1 and 15:1. The results revealed that the biological and reproductive fitness of the mosquitoes are affected by radiation, so that the effects are directly proportional to the radiation dose. The longevity parameter showed significant differences...

Participation of irradiated Anopheles arabiensis males in swarms following field release in Sudan.

BACKGROUND: The success of the sterile insect technique (SIT) depends the release of large numbers of sterile males, which are able to compete for mates with the wild male population within the target area. Unfortunately, the processes of colonisation, mass production and irradiation may reduce the competitiveness of sterile males through genetic selection, loss of natural traits and somatic damage. In this context, the capacity of released sterile Anopheles arabiensis males to survive, disperse and participate in swarms at occurring at varying distances from the release site was studied using mark-release-recapture (MRR) techniques. METHODS: In order to assess their participation in swarms, irradiated and marked laboratory-reared male mosquitoes were released 50, 100 or 200 m from the known site of a large swarm on three consecutive nights. Males were collected from this large swarm on subsequent nights. Over the three days a total of 8,100 males were released. Mean distance travelled (MDT), daily probability of survival and estimated population size were calculated from the recapture data. An effect of male age at the time of release on these parameters was observed. RESULTS: Five per cent of the males released over three days were recaptured. In two-, three- and four-day-old males, MDT was 118, 178 and 170 m, and the daily survival probability 0.95, 0.90 and 0.75, respectively. From the recapture data on the first day following each release, the Lincoln index gives an estimation of 32,546 males in the natural population. DISCUSSION: Sterile An. arabiensis males released into the field were able to find and participate in existing swarms, and possibly even initiate swarms. The survival probability decreased with the age of male on release but the swarm participation and the distance travelled by older males seemed higher than for younger males. The inclusion of a pre-release period may thus be beneficial to male competitiveness and increase the attractiveness of adult sexing techniques, such as blood spiking.

Mating competitiveness of sterile male Anopheles coluzzii in large cages.

BACKGROUND: Understanding the factors that account for male mating competitiveness is critical to the development of the sterile insect technique (SIT). Here, the effects of partial sterilization with 90 Gy of radiation on sexual competitiveness of Anopheles coluzzii allowed to mate in different ratios of sterile to untreated males have been assessed. Moreover, competitiveness was compared between males allowed one versus two days of contact with females. METHODS: Sterile and untreated males four to six days of age were released in large cages (~1.75 sq m) with females of similar age at the following ratios of sterile males: untreated males: untreated virgin females: 100:100:100, 300:100:100, 500:100:100 (three replicates of each) and left for two days. Competitiveness was determined by assessing the egg hatch rate and the insemination rate, determined by dissecting recaptured females. An additional experiment was conducted with a ratio of 500:100:100 and a mating period of either one or two days. Two controls of 0:100:100 (untreated control) and 100:0:100 (sterile control) were used in each experiment. RESULTS: When males and females consort for two days with different ratios, a significant difference in insemination rate was observed between ratio treatments. The competitiveness index (C) of sterile males compared to controls was 0.53. The number of days of exposure to mates significantly increased the insemination rate, as did the increased number of males present in the untreated: sterile male ratio treatments, but the number of days of exposure did not have any effect on the hatch rate. DISCUSSION: The comparability of the hatch rates between experiments suggest that An. coluzzii mating competitiveness experiments in large cages could be run for one instead of two days, shortening the required length of the experiment. Sterilized males were half as competitive as untreated males, but an effective release ratio of at least five sterile for one untreated male has the potential to impact the fertility of a wild female population. However, further trials in field conditions with wild males and females should be undertaken to estimate the ratio of sterile males to wild males required to produce an effect on wild populations.

Aedes albopictus (Skuse) males in laboratory and semi-field cages: release ratios and mating competitiveness.

To control the container-breeding mosquito and major vector of dengue and chikungunya Aedes albopictus, the sterile insect technique (SIT) is proposed as a component of integrated vector management programs in endemic areas. For the technique to be successful, released males, sterilized with 35 Gy of ionizing radiation during the pupal stage, must be able to compete for mating opportunities with wild counterparts and successfully copulate with wild females to induce sterility in the population. Any reduction in competitiveness can be compensated for by increasing the ratio of released sterile to wild males, a ratio which must be optimized for effectiveness and efficiency. Fruit fly SIT programs use field enclosures to test the competitiveness of sterile males to monitor the quality of the colony and adjust release ratios. This is laborious and time consuming, and for mosquito programs it would be advantageous if similarly useful results could be obtained by smaller scale laboratory tests, conducted on a more regular basis. In the present study we compared the competitiveness, as measured by hatching rate of resulting egg batches, of irradiated males measured in small and large laboratory cages and semi-field enclosures in a greenhouse setting, when competing in a 1:1, 3:1, and 5:1 ratio with fertile males. The sterile males were found to be equally competitive when compared to unirradiated counterparts, and a 5:1 ratio was sufficient to reduce, but not eliminate, the fertility of the female populations, irrespective of cage size. Variability in hatch rate in eggs laid by individual females and so-called indeterminate matings, when we could not be certain whether a female had mated a fertile or a sterile male, could be investigated by closer investigation of mating status and the frequency of multiple matings in Ae. albopictus. The laboratory results are encouraging for the effectiveness of the SIT using irradiated males of this species, and we support further assessment in the field.

Irradiated male tsetse from a 40-year-old colony are still competitive in a Riparian forest in Burkina Faso.

BACKGROUND: Tsetse flies are the cyclical vectors of African trypanosomosis that constitute a major constraint to development in Africa. Their control is an important component of the integrated management of these diseases, and among the techniques available, the sterile insect technique (SIT) is the sole that is efficient at low densities. The government of Burkina Faso has embarked on a tsetse eradication programme in the framework of the PATTEC, where SIT is an important component. The project plans to use flies from a Glossina palpalis gambiensis colony that has been maintained for about 40 years at the Centre International de Recherche-Développement sur l'Elevage en zone Subhumide (CIRDES). It was thus necessary to test the competitiveness of the sterile males originating from this colony. METHODOLOGY/PRINCIPAL FINDINGS: During the period January-February 2010, 16,000 sterile male G. p. gambiensis were released along a tributary of the Mouhoun river. The study revealed that with a mean sterile to wild male ratio of 1.16 (s.d. 0.38), the abortion rate of the wild female flies was significantly higher than before (p = 0.026) and after (p = 0.019) the release period. The estimated competitiveness of the sterile males (Fried index) was 0.07 (s.d. 0.02), indicating that a sterile to wild male ratio of 14.4 would be necessary to obtain nearly complete induced sterility in the female population. The aggregation patterns of sterile and wild male flies were similar. The survival rate of the released sterile male flies was similar to that observed in 1983-1985 for the same colony. CONCLUSIONS/SIGNIFICANCE: We conclude that gamma sterilised male G. p. gambiensis derived from the CIRDES colony have a competitiveness that is comparable to their competitiveness obtained 35 years ago and can still be used for an area-wide integrated pest management campaign with a sterile insect component in Burkina Faso.

Radiation biology of mosquitoes.

There is currently renewed interest in assessing the feasibility of the sterile insect technique (SIT) to control African malaria vectors in designated areas. The SIT relies on the sterilization of males before mass release, with sterilization currently being achieved through the use of ionizing radiation. This paper reviews previous work on radiation sterilization of Anopheles mosquitoes. In general, the pupal stage was irradiated due to ease of handling compared to the adult stage. The dose-response curve between the induced sterility and log (dose) was shown to be sigmoid, and there was a marked species difference in radiation sensitivity. Mating competitiveness studies have generally been performed under laboratory conditions. The competitiveness of males irradiated at high doses was relatively poor, but with increasing ratios of sterile males, egg hatch could be lowered effectively. Males irradiated as pupae had a lower competitiveness compared to males irradiated as adults, but the use of partially-sterilizing doses has not been studied extensively. Methods to reduce somatic damage during the irradiation process as well as the use of other agents or techniques to induce sterility are discussed. It is concluded that the optimal radiation dose chosen for insects that are to be released during an SIT programme should ensure a balance between induced sterility of males and their field competitiveness, with competitiveness being determined under (semi-) field conditions. Self-contained 60Co research irradiators remain the most practical irradiators but these are likely to be replaced in the future by a new generation of high output X ray irradiators.

The influence of late-stage pupal irradiation and increased irradiated: un-irradiated male ratio on mating competitiveness of the malaria mosquito Anopheles arabiensis Patton.

Competitiveness of released males in genetic control programmes is of critical importance. In this paper, we explored two scenarios to compensate for the loss of mating competitiveness after pupal stage irradiation in males of the malaria mosquito Anopheles arabiensis. First, competition experiments with a higher ratio of irradiated versus un-irradiated males were performed. Second, pupae were irradiated just prior to emergence and male mating competitiveness was determined. Males were irradiated in the pupal stage with a partially or fully-sterilizing dose of 70 or 120 Gy, respectively. Pupae were irradiated aged 20-26 h (young) as routinely performed, or the pupal stage was artificially prolonged by cooling and pupae were irradiated aged 42-48 h (old). Irradiated males competed at a ratio of 3:1:1 to un-irradiated males for mates in a large cage design. At the 3:1 ratio, the number of females inseminated by males irradiated with 70 Gy as young pupae was similar to the number inseminated by un-irradiated males for the majority of the replicates. At 120 Gy, significantly fewer females were inseminated by irradiated than by un-irradiated males. The irradiation of older pupae did not result in a significantly improved male mating competitiveness compared to the irradiation of young pupae. Our findings indicate that the loss of competitiveness after pupal stage irradiation can be compensated for by a threefold increase of irradiated males, but only for the partially-sterilizing dose. In addition, cooling might be a useful tool to facilitate handling processes of large numbers of mosquitoes in genetic control programmes.

Mating competitiveness of male Anopheles arabiensis mosquitoes irradiated with a partially or fully sterilizing dose in small and large laboratory cages.

Male mating competitiveness is a crucial parameter in many genetic control programs including the sterile insect technique (SIT). We evaluated competitiveness of male Anopheles arabiensis Patton as a function of three experimental variables: (1) small or large cages for mating, (2) the effects of either a partially sterilizing (70 Gy) or fully sterilizing (120 Gy) dose, and (3) pupal or adult irradiation. Irradiated males competed for females with an equal number of unirradiated males. Competitiveness was determined by measuring hatch rates of individually laid egg batches. In small cages, pupal irradiation with the high dose resulted in the lowest competitiveness, whereas adult irradiation with the low dose gave the highest, with the latter males being equal in competitiveness to unirradiated males. In the large cage, reduced competitiveness of males irradiated in the pupal stage was more pronounced compared with the small cage; the males irradiated as adults at both doses performed similarly to unirradiated males. Unexpectedly, males irradiated with the high dose performed better in a large cage than in a small one. A high proportion of intermediate hatch rates was observed for eggs collected in the large cage experiments with males irradiated at the pupal stage. It is concluded that irradiation of adult An. arabiensis with the partially sterilizing dose results in the highest competitiveness for both cage designs. Cage size affected competitiveness for some treatments; therefore, competitiveness determined in laboratory experiments must be confirmed by releases into simulated field conditions. The protocols described are readily transferable to evaluate male competitiveness for other genetic control techniques.

[Response of interspecific competition between Ulva pertusa and Grateloupia filicina to UV-B irradiation enhancement].

The interspecific competition between Ulva pertusa and Grateloupia filicina and it's response to the UV-B irradiation enhancement were analyzed using mono-culture and co-culture methods. The study adopted reasonable experimental design and took biomass as the main examined index. Results showed that the relation of interspecific competition included both allelopathy effect and nutrient competition. Specific growth rates of U. pertusa under treatment with abundant nutrition and limited nutrition was 2.54 and 2.47 times of those of G. filicina. Thus, compared to U. pertusa, G. filicina was in inferior position. UV-B irradiation could inhibit the growth of U. pertusa and G. filicina under the condition of mono-culture. The higher the dosage and the longer exposure of UV-B irradiation were, the more significant the inhibitive effect was. When they were cultured together, low dosage [1.6 kJ x (m2 x d)(-1)] and medium dosage [4.8 kJ x(m2 x d)(-1)] of UV-B irradiation reduced the competitive ability of U. pertusa, and weights of U. pertusa and G. filicina declined 6.81% and 3.88% in low dosage, and 10.47% and 6.98% in medium dosage, respectively. So the relation of interspecific competition tended to be at a balanced level even though U. pertusa was still the dominant algae. However, on the 12th day, weight of U. pertusa decreased by 13.09%, but the value of G. filicina was 14.72%, which was higher than that of U. pertusa. Therefore, high dosage [9.6 kJ x (m2 x d)(-1)] of UV-B irradiation had more serious inhibitive effect on G. filicina, and competitive dominant position of U. pertusa tended to be more obvious. Thus, UV-B changed the relation of competitive balance of U. pertusa and G. filicina, which changed along with the dosage of UV-B. Moreover, UV-B irradiation might influence the metabolism of the allelochemicals produced by U. pertusa and G. filicina in a long time.