Statewide surveillance of Ixodes scapularis (Acari: Ixodidae) for the presence of the human pathogen Borrelia miyamotoi (Spirochaetales: Spirochaetaceae), a relapsing fever spirochete in Pennsylvania, USA, 2019-2020.

Borrelia miyamotoi disease is an emerging tick-borne human illness in the United States caused by Borrelia miyamotoi (Spirochaetales: Spirochaetaceae) bacterium. With Pennsylvania reporting thousands of tick-borne disease cases annually, determining the minimum infection rate (MIR) of B. miyamotoi in Ixodes scapularis (Say, Acari: Ixodidae) adults within Pennsylvania is of utmost importance. Active surveillance was performed from October 2019 to April 2020 to collect a minimum of 50 I. scapularis ticks from every county within Pennsylvania and then screened for B. miyamotoi via qPCR. Ticks were collected from all 67 counties with the majority of those being adult I. scapularis. Additional ticks collected were Dermacentor albipictus (Packard, Acari: Ixodidae), Haemaphysalis longicornis (Neumann, Acari: Ixodidae), and immature I. scapularis. Adult I. scapularis were pooled and tested for B. miyamotoi. MIR for positive B. miyamotoi pools and density of infected adult I. scapularis varied by county, with positive pools from 38 Pennsylvania counties. This is the first statewide evaluation of B. miyamotoi in Pennsylvania in questing adult I. scapularis. These prevalence and distribution data will aid health care practitioners within the state of Pennsylvania and the northeast United States to understand potential risk and bring awareness to the lesser known human Borrelia illness, Borrelia miyamotoi disease.

EVIDENCE OF PROTOZOAN AND BACTERIAL INFECTION AND CO-INFECTION AND PARTIAL BLOOD FEEDING IN THE INVASIVE TICK HAEMAPHYSALIS LONGICORNIS IN PENNSYLVANIA.

The Asian longhorned tick, Haemaphysalis longicornis, an invasive tick species in the United States, has been found actively host-seeking while infected with several human pathogens. Recent work has recovered large numbers of partially engorged, host-seeking H. longicornis, which together with infection findings raises the question of whether such ticks can reattach to a host and transmit pathogens while taking additional bloodmeals. Here we conducted molecular blood meal analysis in tandem with pathogen screening of partially engorged, host-seeking H. longicornis to identify feeding sources and more inclusively characterize acarological risk. Active, statewide surveillance in Pennsylvania from 2020 to 2021 resulted in the recovery of 22/1,425 (1.5%) partially engorged, host-seeking nymphal and 5/163 (3.1%) female H. longicornis. Pathogen testing of engorged nymphs detected 2 specimens positive for Borrelia burgdorferi sensu lato, 2 for Babesia microti, and 1 co-infected with Bo. burgdorferi s.l. and Ba. microti. No female specimens tested positive for pathogens. Conventional PCR blood meal analysis of H. longicornis nymphs detected avian and mammalian hosts in 3 and 18 specimens, respectively. Mammalian blood was detected in all H. longicornis female specimens. Only 2 H. longicornis nymphs produced viable sequencing results and were determined to have fed on black-crowned night heron, Nycticorax nycticorax. These data are the first to molecularly confirm H. longicornis partial blood meals from vertebrate hosts and Ba. microti infection and co-infection with Bo. burgdorferi s.l. in host-seeking specimens in the United States, and the data help characterize important determinants indirectly affecting vectorial capacity. Repeated blood meals within a life stage by pathogen-infected ticks suggest that an understanding of the vector potential of invasive H. longicornis populations may be incomplete without data on their natural host-seeking behaviors and blood-feeding patterns in nature.

Semi-field and surveillance data define the natural diapause timeline for Culex pipiens across the United States.

Reproductive diapause serves as biological mechanism for many insects, including the mosquito Culex pipiens, to overwinter in temperate climates. While Cx. pipiens diapause has been well-studied in the laboratory, the timing and environmental signals that promote diapause under natural conditions are less understood. In this study, we examine laboratory, semi-field, and mosquito surveillance data to define the approximate timeline and seasonal conditions that contribute to Cx. pipiens diapause across the United States. While confirming integral roles of temperature and photoperiod in diapause induction, we also demonstrate the influence of latitude, elevation, and mosquito population genetics in shaping Cx. pipiens diapause incidence across the country. Coinciding with the cessation of WNV activity, these data can have important implications for mosquito control, where targeted efforts prior to diapause induction can decrease mosquito populations and WNV overwintering to reduce mosquito-borne disease incidence the following season.

Recovery of Partially Engorged Haemaphysalis longicornis (Acari: Ixodidae) Ticks from Active Surveillance.

The invasive Asian longhorned tick, Haemaphysalis longicornis, has rapidly spread across the northeastern United States and is associated with pathogens of public health and veterinary concern. Despite its importance in pathogen dynamics, H. longicornis blood-feeding behavior in nature, specifically the likelihood of interrupted feeding, remains poorly documented. Here, we report the recovery of partially engorged, questing H. longicornis from active tick surveillance in Pennsylvania. Significantly more engorged H. longicornis nymphs (1.54%) and adults (3.07%) were recovered compared to Ixodes scapularis nymphs (0.22%) and adults (zero). Mean Scutal Index difference between unengorged and engorged nymph specimens was 0.65 and 0.42 for I. scapularis and H. longicornis, respectively, suggesting the questing, engorged H. longicornis also engorged to a comparatively lesser extent. These data are among the first to document recovery of engorged, host-seeking H. longicornis ticks and provide initial evidence for interrupted feeding and repeated successful questing events bearing implications for pathogen transmission and warranting consideration in vector dynamics models.

First detection of human pathogenic variant of Anaplasma phagocytophilum in field-collected Haemaphysalis longicornis, Pennsylvania, USA.

The Asian longhorned tick, Haemaphysalis longicornis, an invasive species associated with human pathogens, has spread rapidly across the eastern USA. Questing H. longicornis ticks recovered from active surveillance conducted from 1 May to 6 September, 2019 throughout Pennsylvania were tested for rickettsial pathogens. Of 265 ticks tested by PCR for pathogens, 4 (1.5%) were positive for Anaplasma phagocytophilum. Sequence analysis of the 16S rRNA gene confirmed two positives as A. phagocytophilum-human agent variant. This is the first reported detection of A. phagocytophilum-human pathogenic strain DNA in exotic H. longicornis collected in the USA.

Borrelia burgdorferi Sensu Stricto DNA in Field-Collected Haemaphysalis longicornis Ticks, Pennsylvania, United States.

We collected questing Haemaphysalis longicornis ticks from southeastern counties of Pennsylvania, USA. Of 263 ticks tested by PCR for pathogens, 1 adult female was positive for Borrelia burgdorferi sensu stricto, yielding a 0.4% infection rate. Continued monitoring of this invasive tick is essential to determine its public health role.

Distribution and Density of Haemaphysalis longicornis (Acari: Ixodidae) on Public Lands in Pennsylvania, United States.

Since the recent introduction of the Asian longhorned tick (Haemaphysalis longicornis Neumann) in the United States, quantitative surveillance information remains lacking, which hinders accurate estimates of population structure and entomological risk. We conducted statewide, active tick surveillance from May to August 2019 and report data on H. longicornis geographical distribution and population density in Pennsylvania. In total, 615 H. longicornis were collected from four counties. Across samples recovering H. longicornis, mean density of H. longicornis was 9.2/100 m2, comparably greater than Ixodes scapularis Say (8.5/100 m2). Density of H. longicornis was also significantly greater in August, largely driven by larvae, and greater in recreational habitat types (12.6/100 m2) and in Bucks County (11.7/100 m2), situated adjacent to the location of the first U.S. discovery of intense infestations. These data are among the first to document H. longicornis from statewide tick surveillance and provide initial measures of population density enabling more quantitative characterizations of distributional patterns.