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• No pathogens were recovered from any life stage, questing or host-derived ticks. • Virus enrichment and shotgun bacterial metagenomics approaches were utilized and analyzed. • H. longicornis is potentially not a vector of tick-borne pathogens in NYC. Haemaphysalis longicornis , the Asian longhorned tick, is an invasive tick species that has spread rapidly across the northeastern and southeastern regions of the United States in recent years. This invasive pest species, known to transmit several tick-borne pathogens in its native range, is a potential threat to wildlife, livestock, domestic animals, and humans. Questing larval (n = 25), nymph (n = 10), and adult (n = 123), along with host-derived adult (n = 25) H. longicornis ticks were collected from various locations on Staten Island, NY. The pathobiome of each specimen was examined using two different high throughput sequencing approaches, virus enrichment and shotgun metagenomics. An average of 45,828,061 total reads per sample were recovered from the virus enriched samples and an average of 11,381,144 total reads per sample were obtained using shotgun metagenomics. Aside from endogenous viral sequences, no viruses were identified through either approach. Through shotgun metagenomics, Coxiella -like bacteria , Legionella , Sphingomonas , and other bacterial species were recovered. The Coxiella -like agent was ubiquitous and present at high abundances in all samples, suggesting it may be an endosymbiont. The other bacterial agents are not known to be transmitted by ticks. From these analyses, H. longicornis do not appear to host any endemic human tick-borne pathogens in the New York City region. [ABSTRACT FROM AUTHOR]

• Haemaphysalis longicornis were sampled from seven genera of large and medium-sized mammals. • Raccoons, opossums, and white-tailed deer fed a large proportion of H. longicornis. • Haemaphysalis longicornis did not acquire pathogens through co-feeding with native tick vectors. • Host species were infected with a range of pathogens of human and veterinary concern. • Host-derived H. longicornis engorged larvae were not infected with any pathogens. Haemaphysalis longicornis , the Asian longhorned tick, is an invasive ixodid tick that has rapidly spread across the northeastern and southeastern regions of the United States since first reported in 2017. The emergence of H. longicornis presents a potential threat for livestock, wildlife, and human health as the host associations and vector potential of this invasive pest in the United States are poorly understood. Previous field data from the United States has shown that H. longicornis was not associated with natural populations of small mammals or birds, but they show a preference for medium sized mammals in laboratory experiments. Therefore, medium and large sized mammals were sampled on Staten Island, New York, United States, to determine H. longicornis host associations and vector potential for a range of human and veterinary pathogens. A total of 97 hosts were sampled and five species of tick (Amblyomma americanum , Dermacentor variabilis , H. longicornis , Ixodes scapularis , Ixodes cookei) were found feeding concurrently on these hosts. Haemaphysalis longicornis was found in the highest proportions compared with other native tick species on raccoons (55.4%), Virginia opossums (28.9%), and white-tailed deer (11.5%). Tissue, blood, and engorged larvae were tested for 17 different pathogens using a nanoscale PCR platform. Infection with five pathogens (Borrelia burgdorferi , Anaplasma phagocytophilum , Rickettsia spp., Mycoplasma haemocanis , and Bartonella spp.) was detected in host samples, but no pathogens were found in any larval samples. These results suggest that although large and medium sized mammals feed large numbers of H. longicornis ticks in the environment, there is presently a low potential for H. longicornis to acquire pathogens from these wildlife hosts. [ABSTRACT FROM AUTHOR]