Description of microsporidia in simulids: molecular and morphological characterization of microsporidia in the larvae of Simulium pertinax Kollar (Diptera: Simuliidae)

Introduction Microsporidia constitute the most common black fly pathogens, although the species' diversity, seasonal occurrence and transmission mechanisms remain poorly understood. Infections by this agent are often chronic and non-lethal, but they can cause reduced fecundity and decreased longevity. The objective of this study was to identify microsporidia infecting Simulium (Chirostilbia) pertinax (Kollar, 1832) larvae from Caraguatatuba, State of São Paulo, Brazil, by molecular and morphological characterization. Methods Larvae were collected at a single point in a stream in a rural area of the city and were kept under artificial aeration until analysis. Polydispyrenia spp. infection was characterized by the presence of at least 32 mononuclear spores measuring 6.9 ± 1.0 × 5.0 ± 0.7µm in persistent sporophorous vesicles. Similarly, Amblyospora spp. were characterized by the presence of eight uninucleate spores measuring 4.5 × 3.5µm in sporophorous vesicles. Results The molecular analysis confirmed the presence of microsporidian DNA in the 8 samples (prevalence of 0.51%). Six samples (Brazilian larvae) were related to Polydispyrenia simulii and Caudospora palustris reference sequences but in separate clusters. One sample was clustered with Amblyospora spp. Edhazardia aedis was the positive control taxon. Conclusions Samples identified as Polydispyrenia spp. and Amblyospora spp. were grouped with P. simulii and Amblyospora spp., respectively, corroborating previous results. However, the 16S gene tree showed a considerable distance between the black fly-infecting Amblyospora spp. and the mosquito-infecting spp. This distance suggests that these two groups are not congeneric. Additional genomic region evaluation is necessary to obtain a coherent phylogeny for this group. .

Influence of temperature on microsporidia infections in a natural population of Simulium pertinax Kollar, 1832 (Diptera; Simuliidae)

During the studies involving the correlation between the water temperature of the breeding site of Simulium pertinax larvae and the infection prevalence by microsporidia, developed in the Andorinhas river, Magé, RJ, weekly samples of blackfly larvae were taken within a two-year period (2001-2002 and 2003-2004), and it was noticed that the infections by Amblyospora sp. were more prevalent when compared to infections by Polydispyrenia sp. in larvae. It was also observed that the infections do not follow the same pattern, since the genus Amblyospora was recorded almost every month during the study with the exception of December, 2001. In the results of correlation between the environmental water temperature and the microsporidia infection rates, it was observed that for the first period studied, there was a high negative correlation, while during the second period there was absence correlation. On the other hand, the Amblyospora sp. infection rates prove that the correlation was high and significant in the first period, but was not significant in the second sampling period and Polydispyrenia sp. showed absence correlation in both periods.

Este estudo envolvendo a correlação entre a temperatura da água do criadouro de larvas de Simulium pertinax e a prevalência de infecção por microsporídeos, foi desenvolvido no rio Andorinhas, Magé, RJ, onde foram realizadas coletas semanais de larvas de simulídeos no período de dois anos (2001-2002 e 2003-2004), e foi observado que as infecções por Amblyospora sp. apresentaram maior prevalência quando comparadas com as infecções por Polydispyrenia sp. Com isso, verificou-se que as infecções não seguem o mesmo padrão, onde o gênero Amblyospora foi relatado em quase todos os meses de desenvolvimento do estudo, com exceção apenas de dezembro de 2001. Nos resultados de correlação entre a temperatura ambiente e a taxa de infecção por microsporídeos no primeiro período de estudo, foi observada uma forte correlação negativa, porém foi verificada ausência de correlação no segundo período. Embora tenha sido evidenciada uma forte e significante correlação com a taxa de infecção por Amblyospora sp. no primeiro período, esta correlação mostrou-se não significativa no segundo período de amostragem e Polydispyrenia sp. apresentou ausência de correlação em ambos os períodos de estudo.