Targeted next-generation sequencing reveals hemotropic mycoplasmas, Bartonella spp., and Babesia in shelter dogs from Texas, USA.

The screening of vector-borne pathogens (VBPs) in shelter dog populations is crucial for early detection and informs proper management and control efforts. In this study, we used a targeted next-generation sequencing (tNGS) assay to detect VBPs in shelter dogs from Texas. Genomic DNA from whole blood (n = 153) was extracted and subjected to the tNGS assay to search for 15 pathogens. A total of 26.1 % (n = 40/153) of dogs were positive for at least one pathogen, with 80 % (32/40) and 20 % (8/40) samples positive for single and co-infections, respectively. Considering single infections, Mycoplasma haemocanis (13.0 %; n = 20/153) was the most detected pathogen, followed by Bartonella vinsonii subsp. berkhoffii (5.8 %; n = 9/153), and Candidatus Mycoplasma haematoparvum (1.9 %; n = 3/153). Among co-infections, Cand. M. haematoparvum + M. haemocanis (2.6 %; n = 4/153), followed by B. v. berkhoffii + M. haemocanis (1.3 %; n = 2/153), B. v. berkhoffii + Cand. M. haematoparvum (0.7 %; n = 1/153) and B. vulpes + M. haemocanis (0.7 %; n = 1/153). We did not find any association between demographic variables (e.g., age, sex, spayed or neutered status, and breed) and pathogen detection. This study underscores the importance of hemotropic mycoplasma and co-infecting pathogen surveillance in shelter dogs and emphasizes the need for robust and comprehensive assays that allow the simultaneous detection of multiple canine VBPs. Additionally, based on the detection of some pathogens with zoonotic potential, prevention strategies, including comprehensive screenings and targeted biosecurity measures, are recommended for minimizing the risk of exposure to their future owners.