Vector competence of sterile male Glossina fuscipes fuscipes for Trypanosoma brucei brucei: implications for the implementation of the sterile insect technique in a sleeping sickness focus in Chad.

BACKGROUND: Human African trypanosomiasis (HAT) is a neglected tropical disease caused by Trypanosoma brucei gambiense transmitted by tsetse flies in sub-Saharan West Africa. In southern Chad the most active and persistent focus is the Mandoul focus, with 98% of the reported human cases, and where African animal trypanosomosis (AAT) is also present. Recently, a control project to eliminate tsetse flies (Glossina fuscipes fuscipes) in this focus using the sterile insect technique (SIT) was initiated. However, the release of large numbers of sterile males of G. f. fuscipes might result in a potential temporary increase in transmission of trypanosomes since male tsetse flies are also able to transmit the parasite. The objective of this work was therefore to experimentally assess the vector competence of sterile males treated with isometamidium for Trypanosoma brucei brucei. METHODS: An experimental infection was set up in the laboratory, mimicking field conditions: the same tsetse species that is present in Mandoul was used. A T. b. brucei strain close to T. b. gambiense was used, and the ability of the sterile male tsetse flies fed on blood with and without a trypanocide to acquire and transmit trypanosomes was measured. RESULTS: Only 2% of the experimentally infected flies developed an immature infection (midgut) while none of the flies developed a metacyclic infection of T. b. brucei in the salivary glands. We did not observe any effect of the trypanocide used (isometamidium chloride at 100 mg/l) on the development of infection in the flies. CONCLUSIONS: Our results indicate that sterile males of the tested strain of G. f. fuscipes were unable to cyclically transmit T. b. brucei and might even be refractory to the infection. The data of the research indicate that the risk of cyclical transmission of T. brucei by sterile male G. f. fuscipes of the strain colonized at IAEA for almost 40 years appears to be small.

Mosquito mass rearing: who's eating the eggs?

For the sterile insect technique, and other related biological control methods where large numbers of the target mosquito are reared artificially, production efficiency is key for the economic viability of the technique. Rearing success begins with high quality eggs. Excess eggs are often stockpiled and stored for longer periods of time. Any pests that prey on these eggs are detrimental to stockpiles and need to be avoided. Psocids of the genus Liposcelis (Psocoptera, Liposcelididae) are common scavengers consuming various types of organic material that are distributed globally and thrive in warm damp environments, making insectaries ideal habitats. In this short report, we investigated the species that has been found scavenging stored mosquito eggs in our insectary and identified it to be Liposcelis bostrychophila Badonnel, 1931. Additional observations were made to determine whether these predators indeed feed on mosquito eggs, and to suggest simple, effective ways of avoiding infestation.

TITLE: Élevage de masse de moustiques : mais qui mange les œufs ? ABSTRACT: Pour la technique des insectes stériles et les autres méthodes de lutte biologique associées, dans lesquelles un grand nombre de moustiques cibles sont élevés artificiellement, l'efficacité de la production est essentielle pour la viabilité économique de la technique. Le succès de l'élevage commence par des œufs de bonne qualité. Les œufs excédentaires sont souvent stockés pendant de longues périodes. Tous les organismes nuisibles qui exploitent ces œufs nuisent à ces stocks et doivent être évités. Les psoques du genre Liposcelis (Psocoptera, Liposcelididae) sont des charognards répandus qui consomment diverses matières organiques, sont répartis dans le monde entier et prospèrent dans des environnements chauds et humides, ce qui rend les insectariums des habitats idéaux pour eux. Dans ce court rapport, nous avons étudié l'espèce qui mangeait des œufs de moustiques stockés dans notre insectarium et nous avons déterminé qu'il s'agissait de Liposcelis bostrychophila Badonnel, 1931. D'autres observations ont été faites pour déterminer si ces prédateurs se nourrissent effectivement des œufs de moustiques et suggérer des moyens simples et efficaces pour éviter l'infestation.

Identification of critical factors that significantly affect the dose-response in mosquitoes irradiated as pupae.

BACKGROUND: The sterile insect technique (SIT) for use against mosquitoes consists of several steps including the production of the target species in large numbers, the separation of males and females, the sterilization of the males, and the packing, transport and release of the sterile males at the target site. The sterility of the males is the basis of the technique; for this, efficient and standardized irradiation methods are needed to ensure that the required level of sterility is reliably and reproducibly achieved. While several reports have found that certain biological factors, handling methods and varying irradiation procedures can alter the level of induced sterility in insects, few studies exist in which the methodologies are adequately described and discussed for the reproductive sterilization of mosquitoes. Numerous irradiation studies on mosquito pupae have resulted in varying levels of sterility. Therefore, we initiated a series of small-scale experiments to first investigate variable parameters that may influence dose-response in mosquito pupae, and secondly, identify those factors that potentially have a significantly large effect and need further attention. METHODS: In this study, we compiled the results of a series of experiments investigating variable parameters such as pupal age (Aedes aegypti), pupal size (Ae. aegypti), geographical origin of mosquito strains (Ae. aegypti and Ae. albopictus), exposure methods (in wet versus dry conditions, Ae. albopictus) and subsequently in low versus high oxygen environments [submerged in water (low O2 (< 5 %)] and in air [high O2 (~ 21 %)] on the radiosensitivity of male pupae (Ae. aegypti, Ae. albopictus and Anopheles arabiensis). RESULTS: Results indicate that radiosensitvity of Ae. aegypti decreases with increasing pupal age (99% induced sterility in youngest pupae, compared to 93% in oldest pupae), but does not change with differences in pupal size (P = 0.94). Differing geographical origin of the same mosquito species did not result in variations in radiosensitivity in Ae. aegypti pupae [Brazil, Indonesia, France (La Reunion), Thailand] or Ae. albopictus [Italy, France (La Reunion)]. Differences in induced sterility were seen following irradiation of pupae that were in wet versus dry conditions, which led to further tests showing significant radioprotective effects of oxygen depletion during irradiation procedures in three tested mosquito species, as seen in other insects. CONCLUSIONS: These findings infer the necessity to further evaluate significant factors and reassess dose-response for mosquitoes with controlled variables to be able to formulate protocols to achieve reliable and reproducible levels of sterility for application in the frame of the SIT.

Chilling, irradiation and transport of male Glossina palpalis gambiensis pupae: Effect on the emergence, flight ability and survival.

BACKGROUND: The sterile insect technique (SIT) requires mass-rearing of the target species, irradiation to induce sexual sterility and transportation from the mass-rearing facility to the target site. Those treatments require several steps that may affect the biological quality of sterile males. This study has been carried out to evaluate the relative impact of chilling, irradiation and transport on emergence rate, flight ability and survival of sterile male Glossina palpalis gambiensis. RESULTS: Chilling, irradiation and transport all affected the quality control parameters studied. The emergence rate was significantly reduced by long chilling periods and transport, i.e. from 92% at the source insectary in Burkina Faso to 78% upon arrival in Senegal. Flight ability was affected by all three parameters with 31% operational flies lost between the production facility and the destination site. Only survival under stress was not affected by any of the treatments. CONCLUSION: The chilling period and transport were the main factors that impacted significantly the quality of sterile male pupae. Therefore, in the operational programme, the delivery of sterile male pupae was divided over two shipments per week to reduce the chilling time and improve the quality of the sterile males. Quality of the male pupae may further be improved by reducing the transport time and vibrations during transport.

Symbiotic microbes affect the expression of male reproductive genes in Glossina m. morsitans.

BACKGROUND: Tsetse flies (Diptera, Glossinidae) display unique reproductive biology traits. Females reproduce through adenotrophic viviparity, nourishing the growing larva into their modified uterus until parturition. Males transfer their sperm and seminal fluid, produced by both testes and male accessory glands, in a spermatophore capsule transiently formed within the female reproductive tract upon mating. Both sexes are obligate blood feeders and have evolved tight relationships with endosymbionts, already shown to provide essential nutrients lacking in their diet. However, the partnership between tsetse and its symbionts has so far been investigated, at the molecular, genomic and metabolomics level, only in females, whereas the roles of microbiota in male reproduction are still unexplored. RESULTS: Here we begin unravelling the impact of microbiota on Glossina m. morsitans (G. morsitans) male reproductive biology by generating transcriptomes from the reproductive tissues of males deprived of their endosymbionts (aposymbiotic) via maternal antibiotic treatment and dietary supplementation. We then compared the transcriptional profiles of genes expressed in the male reproductive tract of normal and these aposymbiotic flies. We showed that microbiota removal impacts several male reproductive genes by depressing the activity of genes in the male accessory glands (MAGs), including sequences encoding seminal fluid proteins, and increasing expression of genes in the testes. In the MAGs, in particular, the expression of genes related to mating, immunity and seminal fluid components' synthesis is reduced. In the testes, the absence of symbionts activates genes involved in the metabolic apparatus at the basis of male reproduction, including sperm production, motility and function. CONCLUSIONS: Our findings mirrored the complementary roles male accessory glands and testes play in supporting male reproduction and open new avenues for disentangling the interplay between male insects and endosymbionts. From an applied perspective, unravelling the metabolic and functional relationships between tsetse symbionts and male reproductive physiology will provide fundamental information useful to understanding the biology underlying improved male reproductive success in tsetse. This information is of particular importance in the context of tsetse population control via Sterile Insect Technique (SIT) and its impact on trypanosomiasis transmission.

Tolerance of low temperature and sterilizing irradiation in males of Glossina pallidipes (Diptera: Glossinidae).

Investigations into the possibility of using the chilled adult release system are continuing as an alternative method to the release of sterile tsetse flies, Glossina pallidipes Austen (Diptera: Glossinidae) in cardboard boxes. Exposing tsetse flies to 4°C for 6 h caused negligible mortality. A combination of chilling and irradiation resulted in reduced quantities of seminal contents being transferred to females. Mortality of flies after bulk irradiation was lower when a thermos flask was used than expanded polystyrene. Mortality after removal from cold storage increased with age. Flies that did not have a blood meal for 3 d prior to exposure to cold had a lower overnight survival than flies that were deprived of a blood meal for 1 or 2 d. Exposure of adult male tsetse flies to low temperature should be for as short a duration as is practical, so that the fitness of the released sterile flies is not unduly compromised. It is also necessary to ensure that losses are minimized during bulk irradiation of adult flies. It would be desirable to have minimal losses after the combined effects of irradiation, cold, and transportation, such that a sufficient number of sterile male flies will still be available to successfully compete for mating opportunities with wild females.