Waterbirds are ubiquitous and globally distributed. Yet, studies on wild waterbirds’ gut microbiota are still rare. Our aim was to explore and compare the gut microbial community composition of wild waterbird species. Four wild waterbird species that are either wintering or all-year residents in Israel were studied: great cormorants, little egrets, black-crowned night herons and black-headed gulls. For each bird, three intestinal sections were sampled; anterior, middle and posterior. No significant differences were found among the microbiota compositions in the three intestine sections of each individual bird. Each waterbird species had a unique microbial composition. The gut microbiota of the black-headed gulls’ fundamentally deviated from that of the other bird species, probably due to a very high abundance (58.8%) of the genus Catellicoccus ( Firmicutes ). Our results suggest a correlation between the waterbird species’ phylogeny and their intestine microbial community hierarchical tree, which evinced phylosymbiosis. This recent coinage stands for eco-evolutionary patterns between the host phylogeny and its microbiota composition. We conclude that eco-evolutionary processes termed phylosymbiosis may occur between wild waterbird species and their gut microbial community composition.

Vibrio cholerae is the cause of cholera, a devastating epidemic and pandemic disease. Despite its importance, the way of its global dissemination is unknown. V. cholerae is abundant in aquatic habitats and is known to be borne by copepods, chironomids and fishes. Our aim was to determine if fish-eating birds act as vectors in the spread of V. cholerae by consuming infected fish. We determined the existence of V. cholerae in the microbiome of 5/7 wild cormorants’ intestine. In three of these V. cholerae-positive wild cormorants, the presence of a gene for cholera toxin (ctxA) was detected. We subsequently tested eight captive, hand-reared cormorants, divided into two equal groups. Prior to the experiment, the feces of the cormorants were V. cholerae-negative. One group was fed exclusively on tilapias, which are naturally infected with V. cholerae, and the other was fed exclusively on goldfish or on koi that were V. cholerae-negative. We detected V. cholerae in the feces of the tilapia-fed, but not in the goldfish/koi-fed, cormorants. Hence, we demonstrate that fish-eating birds can be infected with V. cholerae from their fish prey. The large-scale movements of many fish-eating birds provide a potential mechanism for the global distribution of V. cholerae.

Wild waterfowl species may carry pathogenic V. cholerae O1 and non-O1 serogroups and Aeromonas species in their intestine. The migration of waterfowl is a potential mechanism for global distribution of V. cholerae and Aeromonas.

Chironomids (Diptera; Chironomidae), also known as non-biting midges, are one of the most abundant insects in freshwater habitats. Our aim was to understand whether the metamorphosis developmental stages affect the endogenous microbiota composition of Chironomus transvaalensis. Toward our objective, we analyzed the endogenous microbiota composition of C. transvaalensis' four life stages: egg masses, larvae, pupae, and adults. Significant differences were found between the microbiota compositions of the different developmental stages of this Chironomus species. We observed a decline in bacterial diversity as the insect evolved from egg mass to adult, while the highest richness was observed in the pupal stage. Although there were significant differences between the microbiota compositions of each life stage, a bacterial core, which included 27 Amplicon Sequence Variants (ASVs), was found in all the developmental life stages (in ≥75% of samples). Chironomids are natural reservoirs of Vibrio cholerae and Aeromonas species, and the Vibrio and Aeromonas ASVs were part of the core bacteria. The presence of the ompW gene, which is specific to V. cholerae, confirmed the presence of this species in all the chironomid's life stages. Thus, the results provide important insights about the host-microbe interactions in chironomids with a specific understanding of chironomids-Vibrio-Aeromonas-microbiota interactions.

Infections due to Vibrio cholerae are rarely documented in Israel. Here we report a case of recurrent otitis media in a young male, caused by V. cholerae non-O1/O139. This extra-intestinal infection was caused by V. cholerae O100 and has been associated with freshwater exposure and travel. Symptoms of chronic periodic earaches along with purulent exudate began about one week after the patient suffered a water skiing accident on a river in Australia. The condition lasted for three years, until his ear exudate was examined in a clinical laboratory, diagnosed and treated. Five bacterial isolates were identified as V. cholerae O100. The isolates were screened for genetic characteristics and were found positive for the presence of hapA, hlyA, and ompU virulence genes. All isolates were negative for the presence of ctxA. Based on antibiogram susceptibility testing, ciprofloxacin ear drops were used until the patient’s symptoms disappeared. This case demonstrates that exposure to freshwater can cause otitis media by V. cholerae non-O1/O139 in young and otherwise healthy humans.