First phylogenetic characterization of unusual bacterial taxa detected in ticks

Ticks are significant vectors of zoonotic pathogens and harbor diverse microbial communities. The ecological origin of unusual bacteria, whether derived from the host, environment, human-associated sources, or tick-associated microbiota, remains uncertain. This study aimed to phylogenetically characterize unusual bacteria present in ticks infesting cattle, goats, sheep, dogs, domestic fowls, and bats in Khyber Pakhtunkhwa (KP), Pakistan. Tick specimens were collected, morphologically identified, and subjected to genomic DNA isolation for bacterial screening using the groEL sequence. The obtained groEL sequences were analyzed using BLAST, phylogenetic analysis, and evolutionary distance estimation. A total of 726 ticks belonging to nine species were identified. Based on BLAST results, thirteen distinct taxa of unusual bacteria were detected, namely Acinetobacter pseudolwoffii , Aggregatibacter sp., Burkholderia contaminans , Enterobacter sp., Enterobacter bugandensis , Enterobacter hormaechei , Enterobacter ludwigii , Klebsiella sp., Neisseria sp., Pantoea agglomerans , Providencia manganoxydans, and two distinct isolates of Psychrobacter sp. (MO1 and MO17). These bacteria showed low genetic distance and clustered with the corresponding high-identity sequences. This study provides the first phylogenetic insights into the unusual bacterial species in ticks from KP, Pakistan. Their detection highlights the need for further studies to determine their epidemiological significance and implications for animals and public health.