Effects of crowding and temperature on Wolbachia infection density among life cycle stages of Aedes albopictus

Species of the genus Wolbachia are a group of Rickettsia-like, maternally-inherited bacteria (gram negative), which cause various reproductive alterations in their arthropod and nematode hosts including cytoplasmic incompatibility (CI), male-killing, parthenogenesis and feminization. They can be divided into supergroups such as A and B based on phylogenetic analysis of 16S rDNA sequences. In this study, we examined the relative infection densities of Wolbachia strains among life cycle stages in the mosquito, Aedes albopictus in terms of crowding effect and temperature effect. A. albopictus is known to be superinfected with both A- and B-supergroup Wolbachia which cause CI. The relative Wolbachia densities within each individual mosquito were determined and quantified by using real-time quantitative PCR assay based on the wsp gene. We found that B-supergroup Wolbachia strain densities in this host species were consistently and significantly higher than in the A-supergroup. Larval crowding also reduced adult size of mosquitoes. Our results show clearly that the higher densities of mosquito larvae cause lower densities of Wolbachia strains. Examination of the effect of temperature on Wolbachia density in each stage of the mosquito clearly revealed a significant decrease in bacterial density following exposure to elevated temperature (37 °C) in both males and females.     

Geographic distribution of wolbachial infections in mosquitoes from Thailand

Members of the genus Wolbachia are inherited intracellular bacterial endosymbionts that infect a diverse range of arthropods. Here I report the results of a survey of these endosymbionts in different mosquito species from six geographic regions of Northern, Northeastern, Western, Central, Eastern and Southern Thailand. Using gene amplification assays with wsp and groE gene primers, wolbachiae were detected in 999 mosquitoes representing 28 species of 1622 specimens collected representing 74 species of wild-caught mosquitoes from all regions of Thailand. Results using wsp primers were similar to those using groE primers in all cases. Wolbachiae had not been reported previously from five of the species tested, namely, Aedes lineatopennis, Aedes vexans, Aedes vittatus, Culex pallidothorax and Culex whitmorei. Infections were found in all major disease vector genera except Anopheles. These results indicate that wolbachial infections are distributed throughout many mosquito species in Thailand.     

Effects of temephos and temperature on Wolbachia load and life history traits of Aedes albopictus

Maternally inherited Wolbachia (gram-negative bacteria) often affect the reproductive fitness of their arthropod hosts and may cause cytoplasmic incompatibility (CI). Comparing Wolbachia-infected and uninfected strains of the mosquito Aedes albopictus (Skuse) (Diptera: Culicidae), we assessed the effects on fitness of two stressors: temperature elevation (25 degrees C vs. 37 degrees C) and exposure to temephos insecticide (concentration range 0.0017-0.0167 mg/L) during larval development. Fitness was measured in terms of life history traits: percentage survival, development time and wing size. Insecticide treatment was associated with reduction in survival rates and wing size in both sexes, but did not affect development time or Wolbachia load. Temperature elevation by 12 degrees C significantly reduced all four bionomic parameters observed in both sexes. Wolbachia density within individual adult mosquitoes was determined by using real-time quantitative polymerase chain reaction (PCR) based on the wsp gene. Both male and female adults had significantly lower densities of Wolbachia after larval rearing at the higher temperature.     

Distribution,diversity and density of wolbachial infections in cladocerans and copepods from Thailand

Species of the genus Wolbachia comprise a group of Rickettsia-like, maternally-inherited bacteria that cause several reproductive alterations in arthropod hosts. The best known are cytoplasmic incompatibility and feminization. Here, the first systematic surveys of wolbachial infections in cladocerans and copepods from six geographic regions of Thailand, including Northern, Northeastern, Western, Central, Eastern and Southern are reported. Using gene amplification assays with wsp and groE primers, wolbachiae were detected in 239 (4 spp.) of 1885 (57 spp.) copepods and cladocerans from all regions of Thailand surveyed. Screening results obtained with wsp primers or groE primers were similar in all cases. The presence of wolbachiae was only detected in copepods, not in cladocerans. Sex ratio analyses of the progeny of two species of copepods, Mesocyclops aspericornis and Mesocyclops thermocyclopoides, naturally or artificially infected with wolbachiae showed infection causes feminization (female-bias). The relative density if infection in naturally infected populations of three copepod species, M. thermocyclopoides, Heliodiaptomus elegans and Neodiaptomus blachei, were determined using real-time quantitative PCR assay based on the wsp gene. The density of wolbachiae in M. thermocyclopoides was significantly higher than in the other two species. These results suggest that wolbachial infections are distributed throughout Thailand, and that possibly the natural occurrence of these in copepods may be due to their predation on mosquito larvae. This apparent novel biology may have importance as a genetic drive system for control of vector borne diseases in the future.