Investigating the influence of blood meal sources on the composition of culturable haemolytic gut bacteria of a wild-caught BTV vector Culicoides oxystoma Kieffer (Diptera: Ceratopogonidae).

Culicoides oxystoma Kieffer (Diptera: Ceratopogonidae) transmits many pathogens, including seven viruses, four protozoa and one nematode. This species has a wide distribution range across northern Afro-tropical, Palearctic, Australian, Indo-Malayan realms with a broad host spectrum, including cattle, buffaloes, sheep, pigs, dogs, horses and even humans. The heterogeneous nature of Culicoides' blood-feeding patterns is well documented, but the influence of various host blood meal sources on gut bacterial composition remains scant. Adult midges were collected during April (2023) by operating UV light traps in cattle, buffalo sheds and poultry farm in Purulia (India). Besides C. oxystoma, eleven Culicoides species were collected across the sheds and farm, seven of which are vectors. Culicoides liui Wirth and Hubert and C. thurmanae Wirth and Hubert are reported from India for the first time. In all the sheds, engorged females of C. oxystoma were ubiquitous. Identification of culturable gut bacteria and the host blood meal of C. oxystoma were done through the polymerase chain reaction (PCR)-based method. Blood meal analysis confirmed the following hosts: cattle, buffaloes and humans. Identification of blood meal of engorged C. oxystoma caught from poultry farm showed positive results for humans but not for birds. Among bacteria, Bacillus cereus was abundant in all of the engorged females. Bacillus paramycoides and Enterococcus faecium were identified from females feeding on cattle and buffaloes' blood, while Alcaligenes faecalis was found in the gut contents of females that fed on cattle and human blood. The gut bacteria Alcaligenes faecalis exhibited alpha haemolytic activity. In contrast, Bacillus sp., B. cereus, B. flexus, B. licheniformis, B. thuringiensis, B. paramycoides, E. faecium, Paenibacillus sp. and Pseudomonas sp. exhibited beta haemolysis. This is the first report on the composition of gut bacteria, with particular emphasis on the haemolytic bacteria of C. oxystoma with different host blood meals. The pathogenic bacteria B. cereus, B. licheniformis and A. faecalis within the females could potentially impact pathogen acquisition and increase the probability of their zoonotic transmissibility.