A bacterium against the tiger: further evidence of the potential of noninundative releases of males with manipulated Wolbachia infection in reducing fertility of Aedes albopictus field populations in Italy.

BACKGROUND: Incompatible insect technique (IIT) is a population suppression approach based on the release of males with manipulated Wolbachia infection inducing egg inviability in wild females. We here present results of multiple field releases of incompatible ARwP males carried out in 2019 in a 2.7-ha green area within urban Rome (Italy) to assess the effect on Aedes albopictus egg viability. Data are compared with results obtained in 2018, when the approach was tested for the first time in Europe. RESULTS: An average of 4674 ARwP males were released weekly for 7 weeks, resulting in a mean ARwP:wild male ratio of 1.1:1 (versus 0.7:1 in 2018). Egg-viability dynamics in ovitraps significantly varied between treated and control sites, with an estimated overall reduction of 35% (versus 15% in 2018). The estimated proportion of females classified as mated with ARwP males was 41.8% and the viability rate of eggs laid by these females (9.5%) was on average significantly lower than that of females only mated with wild males (87.8%); however, high variability in fertility was observed. Values of ARwP male competitiveness were 0.36 and 0.73 based on the overall viability rate of eggs in ovitraps and on female fertility, respectively; thus, well above the conventional 0.2 threshold for an effective suppressive impact in the field. CONCLUSIONS: Results further support the potential of IIT as a tool to contribute to Ae. albopictus control in the urban context, stressing the need for larger field trials to evaluate the cost-efficacy of the approach in temperate regions. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Design of a modular controlled unit for the study of bioprocesses: Towards solutions for Bioregenerative Life Support Systems in space.

Space exploration beyond the Low Earth Orbit requires the establishment of Bioregenerative Life Support Systems (BLSSs), which, through bioprocesses for primary resource recycling, ensure crew survival. However, the introduction of new organisms in confined space habitats must be carefully evaluated in advance to avoid unforeseen events that could compromise the mission. In this work, we have designed and built an experimental chamber, named Growing/Rearing Module (GRM), completely isolated and equipped with micro-environmental monitoring and control systems. This unit is specially intended for the study of single bioprocesses, which can be composed to design functional BLSSs. GRM can be implemented with specific devices for the biological system under study and the control of environmental parameters such as temperature, humidity, lighting and if required, pressure of gaseous components. GRM was validated in experiments of both microgreen cultivation, as a source of fresh food for astronauts, and rearing of the decomposer insect Hermetia illucens for bioconversion of organic waste. During the study of each bioprocess, the environmental and biological data were recorded, allowing to make preliminary assessments of the system efficiency. The GRM, as a completely confined environment, represents the first self-consistent unit that allows to fine-tune the optimal parameters for the operability of different bioprocesses. Furthermore, the upgradability according to the mission needs and the functional integrability of modules differently equipped are the keys to GRM versatility, representing a valuable tool for BLSSs' design.

Increased biting rate and decreased Wolbachia density in irradiated Aedes mosquitoes.

BACKGROUND: Releasing considerable numbers of radiation-sterilized males is a promising strategy to suppress mosquito vectors. However, releases may also include small percentages of biting females, which translate to non-negligible numbers when releases are large. Currently, the effects of irradiation on host-seeking and host-biting behaviors have not been exhaustively investigated. Information is also lacking regarding the effects of sterilizing treatment on the endosymbiotic bacterium Wolbachia, which is known to affect the vector competence of infected mosquitos. METHODS: To ascertain the effects of irradiation on females, the pupae of two Aedes albopictus strains, differing in their natural or artificial Wolbachia infection type, and Aedes aegypti-which is not infected by Wolbachia-were treated with various doses of X-rays and monitored for key fitness parameters and biting behavior over a period of 2 weeks. The effect of radiation on Wolbachia was investigated by quantitative polymerase chain reaction (qPCR) and fluorescence in situ hybridization (FISH) analysis. RESULTS: Partial Aedes albopictus female sterility was achieved at 28 Gy, but the number of weekly bites more than doubled compared to that of the controls. Radiation doses of 35 and 45 Gy completely inhibited progeny production but did not significantly affect the survival or flight ability of Ae. albopictus females and caused a tripling of the number of bites per female per week (compared to untreated controls). These results were also confirmed in Ae. aegypti after treatment at 50 Gy. Wolbachia density decreased significantly in 45-Gy-irradiated females, with the greatest decreases in the early irradiation group (26 ± 2-h-old pupae). Wolbachia density also decreased as adults aged. This trend was confirmed in ovaries but not in extra-ovarian tissues. FISH analysis showed a strongly reduced Wolbachia-specific fluorescence in the ovaries of 13 ± 1-day-old females. CONCLUSIONS: These results suggest that, under sterile insect technique (SIT) programs, the vector capacity of a target population could increase with the frequency of the irradiated females co-released with the sterile males due to an increased biting rate. In the context of successful suppression, the related safety issues are expected to be generally negligible, but they should be conservatively evaluated when large-scale programs relying on imperfect sexing and high overflooding release ratios are run for long periods in areas endemic for arboviral diseases. Also, the effects of irradiation on the vector competence deserve further investigation.

A bacterium against the tiger: preliminary evidence of fertility reduction after release of Aedes albopictus males with manipulated Wolbachia infection in an Italian urban area.

BACKGROUND: Novel tools are needed to reduce the nuisance and risk of exotic arbovirus transmission associated with the colonization of temperate regions by Aedes albopictus. The incompatible insect technique (IIT) is a population suppression approach based on cytoplasmic incompatibility between males with manipulated endosymbionts and wild females. Here, we present the results of the first field experiment in Europe to assess the capacity of an Ae. albopictus line (ARwP) deprived of its natural endosymbiont Wolbachia and transinfected with a Wolbachia strain from the mosquito Culex pipiens, to sterilize wild females. RESULTS: We released ∼ 4500 ARwP males weekly for 6 weeks in a green area within urban Rome (Italy) and carried out egg (N = 13 442), female (N = 128) and male (N = 352) collections. Egg (N = 13 783) and female (N = 48) collections were also carried out at two untreated control sites. The percentage of viable eggs during release was, on average, significantly lower in treated sites than in control sites, with the greatest difference (16%) seen after the fourth release. The ARwP to wild male ratio in the release spots between day 3 after the first ARwP male release and day 7 after the last release was, on average, 7:10. Released males survived up to 2 weeks. Approximately 30% of females collected in the release spots showed 100% sterility and 20% showed strongly reduced fertility compared with control sites. CONCLUSIONS: Results support the potential of IIT as a tool contributing to Ae. albopictus control in the urban context, and stress the need for larger field trials to evaluate the cost-efficacy of the approach in suppressing wild populations. © 2019 Society of Chemical Industry.

Cytoplasmic incompatibility management to support Incompatible Insect Technique against Aedes albopictus.

BACKGROUND: The transinfection of the endosymbiotic bacterium Wolbachia provides a method to produce functionally sterile males to be used to suppress mosquito vectors. ARwP is a wPip Wolbachia infected Aedes albopictus which exhibits a bidirectional incompatibility pattern with wild-types. We coupled a modelistic approach with laboratory experiments to test ARwP as a control tool and evaluate the possible occurrence of population replacement following the release of ARwP females in a wild-type (SANG) population. Repeated male-only releases were simulated and tested in the laboratory in comparison with releases contaminated with 1% ARwP females. Model simulations also investigated how migration affects the outcome of contaminated releases. Finally, the mean level of egg fertility and the long-term evolution of populations constituted by two Wolbachia infection types were studied by testing SANG and ARwP Ae. albopictus and performing more general model simulations. RESULTS: The model was parametrized with laboratory data and simulations were compared with results of biological trials. Small populations of ARwP males and females were theoretically and experimentally demonstrated to rapidly become extinct when released in larger SANG populations. Male-only releases at a 5:1 ratio with respect to the wild-type males led to a complete suppression of the SANG population in a few generations. Contaminated releases were efficient as well but led to population replacement in the long term, when the wild-type population approached eradication. Migration significantly contrasted this trend as a 5% population turnover was sufficient to avoid any risk of population replacement. At equal frequencies between ARwP and SANG individuals, the mean egg fertility of the overall population was more than halved and suppression was self-sustaining until one of the two infection types extinguished. CONCLUSIONS: In the case of bidirectional incompatibility patterns, the repeated release of incompatible males with small percentages of contaminant females could lead to population replacement in confined environments while it could be managed to target high efficiency and sustainability in wild-type suppression when systems are open to migration. This possibility is discussed based on various contexts and taking into consideration the possibility of integration with other control methods such as SIT and the use of larvicides.

Combining Wolbachia-induced sterility and virus protection to fight Aedes albopictus-borne viruses.

Among the strategies targeting vector control, the exploitation of the endosymbiont Wolbachia to produce sterile males and/or invasive females with reduced vector competence seems to be promising. A new Aedes albopictus transinfection (ARwP-M) was generated by introducing wMel Wolbachia in the ARwP line which had been established previously by replacing wAlbA and wAlbB Wolbachia with the wPip strain. Various infection and fitness parameters were studied by comparing ARwP-M, ARwP and wild-type (SANG population) Ae. albopictus sharing the same genetic background. Moreover, the vector competence of ARwP-M related to chikungunya, dengue and zika viruses was evaluated in comparison with ARwP. ARwP-M showed a 100% rate of maternal inheritance of wMel and wPip Wolbachia. Survival, female fecundity and egg fertility did not show to differ between the three Ae. albopictus lines. Crosses between ARwP-M males and SANG females were fully unfertile regardless of male age while egg hatch in reverse crosses increased from 0 to about 17% with SANG males aging from 3 to 17 days. When competing with SANG males for SANG females, ARwP-M males induced a level of sterility significantly higher than that expected for an equal mating competitiveness (mean Fried index of 1.71 instead of 1). The overall Wolbachia density in ARwP-M females was about 15 fold higher than in ARwP, mostly due to the wMel infection. This feature corresponded to a strongly reduced vector competence for chikungunya and dengue viruses (in both cases, 5 and 0% rates of transmission at 14 and 21 days post infection) with respect to ARwP females. Results regarding Zika virus did not highlight significant differences between ARwP-M and ARwP. However, none of the tested ARwP-M females was capable at transmitting ZIKV. These findings are expected to promote the exploitation of Wolbachia to suppress the wild-type Ae. albopictus populations.

Analysis of the Mediterranean fruit fly [Ceratitis capitata (Wiedemann)] spatio-temporal distribution in relation to sex and female mating status for precision IPM.

The Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann), is a key pest of fruit crops in many tropical, subtropical and mild temperate areas worldwide. The economic importance of this fruit fly is increasing due to its invasion of new geographical areas. Efficient control and eradication efforts require adequate information regarding C. capitata adults in relation to environmental and physiological cues. This would allow effective characterisation of the population spatio-temporal dynamic of the C. capitata population at both the orchard level and the area-wide landscape. The aim of this study was to analyse population patterns of adult medflies caught using two trapping systems in a peach orchard located in central Italy. They were differentiated by adult sex (males or females) and mating status of females (unmated or mated females) to determine the spatio-temporal dynamic and evaluate the effect of cultivar and chemical treatments on trap catches. Female mating status was assessed by spermathecal dissection and a blind test was carried out to evaluate the reliability of the technique. Geostatistical methods, variogram and kriging, were used to produce distributional maps. Results showed a strong correlation between the distribution of males and unmated females, whereas males versus mated females and unmated females versus mated females showed a lower correlation. Both cultivar and chemical treatments had significant effects on trap catches, showing associations with sex and female mating status. Medfly adults showed aggregated distributions in the experimental field, but hot spots locations varied. The spatial pattern of unmated females reflected that of males, whereas mated females were largely distributed around ripening or ripe fruit. The results give relevant insights into pest management. Mated females may be distributed differently to unmated females and the identification of male hot spots through monitoring would allow localisation of virgin female populations. Based on our results, a more precise IPM strategy, coupled with effective sanitation practices, could represent a more effective approach to medfly control.

Characterization of a new Aedes albopictus (Diptera: Culicidae)-Wolbachia pipientis (Rickettsiales: Rickettsiaceae) symbiotic association generated by artificial transfer of the wPip strain from Culex pipiens (Diptera: Culicidae).

Wolbachia is a maternally inherited endosymbiont inducing various effects in insects and other invertebrate hosts that facilitate the invasion of naive host populations. One of the effects is a form of sterility known as cytoplasmic incompatibility (CI) through which females are effectively sterilized when they mate with males harboring a different Wolbachia strain. The repeated mass release of cytoplasmically incompatible males can be a tool to suppress insect populations. Here, we attempt to infect an Aedes albopictus (Skuse) (Diptera: Culicidae) strain, artificially deprived of the natural Wolbachia infection, with a new Wolbachia strain from Culex pipiens (L.) (Diptera: Culicidae). Further experiments were designed to study the effects of the new infection on Ae. albopictus fitness and evaluate key parameters that affect infection dynamics, including CI level and maternal inheritance. Using embryonic microinjection, the new Wolbachia strain was successfully established in Ae. albopictus. Crosses demonstrated a pattern of bidirectional CI between naturally infected and transinfected individuals. Specifically, egg hatch was essentially absent (i.e., CI was very high) in all crosses between the transinfected males and females with a different infection status. Furthermore, naturally infected Ae. albopictus males were incompatible with the transinfected females. Maternal inheritance was close to 100%. Moreover, the new infection did not affect immature and adult survivorship, but it significantly reduced female fecundity and egg hatch rate. The results are discussed in relation to the potential use of the new Ae. albopictus-Wolbachia symbiotic association as a suitable system for the study and development of CI-based strategies for suppressing populations of this important pest and disease vector.