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Background  Tsetse flies harbor at least three bacterial symbionts:  Wigglesworthia glossinidia ,  Wolbachia pipientis  and  Sodalis glossinidius. Wigglesworthia  and  Sodalis  reside in the gut in close association with trypanosomes and may influence establishment and development of midgut parasite infections.  Wolbachia  has been shown to induce reproductive effects in infected tsetse. This study was conducted to determine the prevalence of these endosymbionts in natural populations of  G. austeni  and  G. pallidipes  and to assess the degree of concurrent infections with trypanosomes.    Methods  Fly samples analyzed originated from Kenyan coastal forests (trapped in 2009–2011) and South African  G. austeni  collected in 2008. The age structure was estimated by standard methods.  G. austeni  (n=298) and  G. pallidipes  (n= 302) were analyzed for infection with  Wolbachia  and  Sodalis  using PCR. Trypanosome infection was determined either by microscopic examination of dissected organs or by PCR amplification.    Results  Overall we observed that  G. pallidipes  females had a longer lifespan (70 d) than  G. austeni  (54 d) in natural populations.  Wolbachia  infections were present in all  G. austeni  flies analysed, while in contrast, this symbiont was absent from  G. pallidipes . The density of  Wolbachia  infections in the Kenyan  G. austeni  population was higher than that observed in South African flies. The infection prevalence of  Sodalis  ranged from 3.7% in  G. austeni  to about 16% in  G. pallidipes . Microscopic examination of midguts revealed an overall trypanosome infection prevalence of 6% (n = 235) and 5% (n = 552), while evaluation with ITS1 primers indicated a prevalence of about 13% (n = 296) and 10% (n = 302) in  G. austeni  and  G. pallidipes,  respectively. The majority of infections (46%) were with  T. congolense.  Co-infection with all three organisms was observed at 1% and 3.3% in  G. austeni  and  G. pallidipes,  respectively .  Eleven out of the thirteen (85%) co-infected flies harboured  T. congolense  and  T. simiae  parasites. While the association between trypanosomes and  Sodalis  infection was statistically significant in  G. pallidipes  (P = 0.0127), the number of co-infected flies was too few for a definite conclusion.    Conclusions  The tsetse populations analyzed differed in the prevalence of symbionts, despite being sympatric and therefore exposed to identical environmental factors. The density of infections with  Wolbachia  also differed between  G. austeni  populations. There were too few natural co-infections detected with the  Sodalis  and trypanosomes to suggest extensive inter-relations between these infections in natural populations. We discuss these findings in the context of potential symbiont-mediated control interventions to reduce parasite infections and/or fly populations.

Wolbachia, Sodalis and trypanosome co-infections in natural populations of Glossina austeni and Glossina pallidipes