Continued Susceptibility of the wMel Wolbachia Infection in Aedes aegypti to Heat Stress Following Field Deployment and Selection

Aedes aegypti mosquitoes infected with the wMel strain of Wolbachia are being deployed to control the spread of arboviruses around the world through blockage of viral transmission. Blockage by Wolbachia in some scenarios may be affected by the susceptibility of wMel to cyclical heat stress during mosquito larval development. We therefore evaluated the potential to generate a heat-resistant strain of wMel in Ae. aegypti through artificial laboratory selection and through exposure to field temperatures across multiple generations. To generate an artificially selected strain, wMel-infected females reared under cyclical heat stress were crossed to wMel-infected males reared at 26 °C. The low proportion of larvae that hatched founded the next generation, and this process was repeated for eight generations. The wMel heat-selected strain (wMel-HS) was similar to wMel (unselected) in its ability to induce cytoplasmic incompatibility and restore compatibility when larvae were reared under cyclical heat stress, but wMel-HS adults exhibited reduced Wolbachia densities at 26 °C. To investigate the effects of field exposure, we compared the response of wMel-infected Ae. aegypti collected from Cairns, Australia where the infection has been established for seven years, to a wMel-infected population maintained in the laboratory for approximately 60 generations. Field and laboratory strains of wMel did not differ in their response to cyclical heat stress or in their phenotypic effects at 26 °C. The capacity for the wMel infection in Ae. aegypti to adapt to high temperatures therefore appears limited, and alternative strains may need to be considered for deployment in environments where high temperatures are regularly experienced in mosquito breeding sites.

Effect of age and cyclical heat stress on the serum biochemical profile of broiler chickens

This study aimed to evaluate the influence of age and cyclical heat stress for 1 hour per day on the levels of serum proteins, metabolites, and enzymes in broiler chickens of 21-42 days of age. We used 420 male broiler chickens, Cobb Avian48TM breed, in a completely randomized experimental design, composed of two treatments and six replicates. The treatments consisted of two thermal environments: one in which broiler chickens were raised under natural conditions of temperature and humidity for the first 42 days of age (control) and another in which chickens were subjected to heat stress at 36°C, from days 16 to 42, for 1 hour per day (1200-1300 hours). At 21, 28, 35, and 42 days of age, approximately 5 mL of blood was collected by cardiac puncture in two birds per replicate. In each serum sample, levels of total protein, albumin, globulin, albumin:globulin ratio, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and gamma-glutamyltransferase (GGT) activity and uric acid, creatinine, total cholesterol, triglycerides, high-density lipoproteins (HDL), very low density lipoproteins (VLDL), low-density lipoproteins (LDL), and glucose levels were determined. Concerning the environment and the age, no significant differences in the levels of glucose, uric acid, ALT, triglycerides, and very low density lipoproteins were observed. There was a significant interaction between the environment and the age for cholesterol and LDL values. At 21 days of age, cholesterol and LDL-cholesterol (LDL-C) were higher in birds under stress than in the control treatment birds, which did not occur in other age groups. Aspartate aminotransferase activity was higher in chickens under cyclical heat stress than in chickens subjected to control treatment and displayed a cubic response regarding the environment. The age of the birds influenced the values of total protein, albumin, globulin, albumin:globulin (A:G) ratio, creatinine, GGT, and HDL. In conclusion, cyclical heat stress, at 36ºC, for one hour, from days 16 to 42 of age increases serum AST, whereas cholesterol and LDL-C levels increase only at day 21, indicating that birds may have adapted to heat stress. Age influenced the activity of AST and GGT and levels of creatinine, total protein, albumin, globulin, A:G ratio, cholesterol, HDL-C, and LDL-C.

Wolbachia infections in Aedes aegypti differ markedly in their response to cyclical heat stress

Aedes aegypti mosquitoes infected with Wolbachia bacteria are currently being released for arbovirus suppression around the world. Their potential to invade populations and persist will depend on interactions with environmental conditions, particularly as larvae are often exposed to fluctuating and extreme temperatures in the field. We reared Ae. aegypti larvae infected with different types of Wolbachia (wMel, wAlbB and wMelPop) under diurnal cyclical temperatures. Rearing wMel and wMelPop-infected larvae at 26-37°C reduced the expression of cytoplasmic incompatibility, a reproductive manipulation induced by Wolbachia. We also observed a sharp reduction in the density of Wolbachia in adults. Furthermore, exposure to 26-37°C over two generations eliminated both the wMel and wMelPop infections. In contrast, the wAlbB infection was maintained at a high density, exhibited complete cytoplasmic incompatibility, and was transmitted from mother to offspring with a high fidelity under this temperature cycle. These findings have implications for the success of Wolbachia interventions across different environments and highlight the importance of temperature control in rearing.