Host-Feeding Behavior of Mosquitoes in the Florida Everglades.

Background: West Nile virus (WNV), Everglades virus (EVEV), and five species of Orthobunyavirus were isolated from mosquitoes collected in the Everglades in 2016-2017. Prior studies of blood meals of mosquitoes in southern Florida have related findings to acquisition and transmission of EVEV, St. Louis encephalitis virus, and WNV, but not the Orthobunyavirus viruses associated with the subgenus Melanoconion of the genus Culex. Materials and Methods: In the present study, blood-fed mosquitoes were collected in the Everglades in 2016, 2017, 2021, and 2022, and from an industrial site in Naples, FL in 2017. Blood meals were identified to host species by PCR assays using mitochondrial cytochrome b gene. Results: Blood meals were identified from Anopheles crucians complex and 11 mosquito species captured in the Florida Everglades and from 3 species collected from an industrial site. The largest numbers of blood-fed specimens were from Culex nigripalpus, Culex erraticus, Culex cedecei, and Aedes taeniorhynchus. Cx. erraticus fed on mammals, birds, and reptiles, particularly American alligator. This mosquito species could transmit WNV to American alligator in the wild. Cx. nigripalpus acquired blood meals primarily from birds and mammals and frequently fed on medium-sized mammals and white-tailed deer. Water and wading birds were the primary avian hosts for Cx. nigripalpus and Cx. erraticus in the Everglades. Wading birds are susceptible to WNV and could serve as reservoir hosts. Cx. cedecei fed on five species of rodents, particularly black and hispid cotton rats. EVEV and three different species of Orthobunyavirus have been isolated from the hispid cotton rat and Cx. cedecei in the Everglades. Cx. cedecei is likely acquiring and transmitting these viruses among hispid cotton rats and other rodents. The marsh rabbit was a frequent host for An. crucians complex. An. crucians complex, and other species could acquire Tensaw virus from rabbits. Conclusions: Our study contributes to a better understanding of the host and viral associations of mosquito species in southwestern Florida.

Comparison of acarological risk metrics derived from active and passive surveillance and their concordance with tick-borne disease incidence.

Tick-borne diseases continue to threaten human health across the United States. Both active and passive tick surveillance can complement human case surveillance, providing spatio-temporal information on when and where humans are at risk for encounters with ticks and tick-borne pathogens. However, little work has been done to assess the concordance of the acarological risk metrics from each surveillance method. We used data on Ixodes scapularis and its associated human pathogens from Connecticut (2019-2021) collected through active collections (drag sampling) or passive submissions from the public to compare county estimates of tick and pathogen presence, infection prevalence, and tick abundance by life stage. Between the surveillance strategies, we found complete agreement in estimates of tick and pathogen presence, high concordance in infection prevalence estimates for Anaplasma phagocytophilum, Borrelia burgdorferi sensu stricto, and Babesia microti, but no consistent relationships between actively and passively derived estimates of tick abundance or abundance of infected ticks by life stage. We also compared nymphal metrics (i.e., pathogen prevalence in nymphs, nymphal abundance, and abundance of infected nymphs) with reported incidence of Lyme disease, anaplasmosis, and babesiosis, but did not find any consistent relationships with any of these metrics. The small spatial and temporal scale for which we had consistently collected active and passive data limited our ability to find significant relationships. Findings are likely to differ if examined across a broader spatial or temporal coverage with greater variation in acarological and epidemiological outcomes. Our results indicate similar outcomes between some actively and passively derived tick surveillance metrics (tick and pathogen presence, pathogen prevalence), but comparisons were variable for abundance estimates.

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.

Evidence of Uranotaenia sapphirina (Diptera: Culicidae) feeding on annelid worms in the Northeastern United States.

Mosquito host-feeding behavior is an important parameter for determining the vector potential of mosquito species in a given locale. Despite the recent discovery of Uranotaenia sapphirina Osten Sacken feeding on annelid hosts in Florida, host association studies for this mosquito species in the United States remain limited. To investigate the blood-feeding pattern of Ur. sapphirina in the northeastern United States, mosquitoes were collected from Massachusetts, Connecticut, and New Jersey using CDC miniature light traps, peat fiber resting boxes, gravid traps, and backpack aspirators. Vertebrate and invertebrate hosts of this mosquito species were identified through PCR amplification and nucleotide sequencing of portions of the mitochondrial cytochrome b gene and the 28S ribosomal RNA gene, respectively. Of 21 (24.7%) specimens successfully identified to host species, 47.6% contained solely annelid blood, 14.3% mammalian blood, 14.3% avian blood, and 23.8% with mixed blood of annelid and avian origin. The mud earthworm, Sparganophilus tennesseensis Reynolds (Haplotaxida: Sparganophilidae), was identified as the most common host (n = 14, including mixed bloods), followed by American robin, Turdus migratorius (n = 7, including mixed bloods). Testing of these blood engorged mosquitoes for West Nile virus and eastern equine encephalitis virus did not result in any positive specimens. This is the first report of Ur. sapphirina feeding on annelids and on both vertebrate and invertebrate hosts in mixed bloodmeals in the northeastern United States. Our findings support the recent report of Ur. sapphirina feeding on invertebrates and further emphasizes the inclination of some mosquito species to feed on a wider range of hosts spanning nontraditional taxonomic groups.

Anomalous morphologies in Ixodes scapularis feeding on human hosts.

Cases of anomalous morphologies in the blacklegged tick, Ixodes scapularis, have been reported in both field-collected and human-biting specimen in the Northeastern and Midwestern United States, complicating the identification of this medically important tick species. We herein describe four cases of morphological anomalies in I. scapularis females exhibiting nanism and abnormally small genital apertures. We also report a female I. scapularis displaying slight asymmetry in the lower abdomen oriented toward the right side and an abnormal anal groove completely enclosing the anus. The identity of each specimen was confirmed using taxonomic keys, high resolution light and scanning electron microscopy imaging, and DNA sequencing of the 18S rRNA gene. All specimens described in this study were found parasitizing human hosts and were submitted to the Connecticut Agricultural Experiment Station-Tick Testing Laboratory in 2021 for species identification and pathogen screening. Here, we also discuss recent reports of teratological abnormalities in I. scapularis as well as likely causes for such deformities and potential implications.

Host Associations of Culex pipiens: A Two-Year Analysis of Bloodmeal Sources and Implications for Arboviral Transmission in Southeastern Virginia.

Understanding vector-host interactions is crucial for evaluating the role of mosquito species in enzootic cycling and epidemic/epizootic transmission of arboviruses, as well as assessing vertebrate host contributions to maintenance and amplification in different virus foci. To investigate blood-feeding pattern of Culex pipiens, engorged mosquitoes were collected on a weekly basis at 50 sites throughout Suffolk, Virginia, using Centers for Disease Control and Prevention miniature light traps, BG-Sentinel traps, and modified Reiter gravid traps. Vertebrate hosts of mosquitoes were identified by amplifying and sequencing portions of the mitochondrial cytochrome b gene. Of 281 Cx. pipiens bloodmeals successfully identified to species, 255 (90.7%) contained solely avian blood, 13 (4.6%) mammalian, 1 (0.4%) reptilian, and 12 (4.3%) both avian and mammalian blood. Nineteen avian species were identified as hosts for Cx. pipiens with American robin (n = 141, 55.3% of avian hosts) and northern cardinal (n = 57, 22.4%) as the most common hosts. More American robin feedings took place in areas of higher development. Three mammalian species were also identified as hosts for Cx. pipiens with Virginia opossum and domestic cat as the most common hosts in this class (each n = 6, 46.2% of mammalian hosts). There was no significant seasonal difference in the proportion of bloodmeals obtained from avian hosts, but there was a decrease in the proportion of bloodmeals from mammalian hosts from spring to fall. One engorged specimen of Cx. pipiens with Virginia opossum-derived bloodmeal tested positive for West Nile virus (WNV), and another with black-and-white warbler-derived bloodmeal tested positive for eastern equine encephalitis virus. Our findings, in conjunction with the results of vector competence studies and virus isolation from field-collected mosquitoes, lend additional support that Cx. pipiens serves as the principal enzootic vector and potential epizootic/epidemic vector of WNV in southeastern Virginia.

Established Population of the Gulf Coast Tick, Amblyomma maculatum (Acari: Ixodidae), Infected with Rickettsia parkeri (Rickettsiales: Rickettsiaceae), in Connecticut.

We identified an established population of the Gulf Coast tick (Amblyomma maculatum Koch) infected with Rickettsia parkeri in Connecticut, representing the northernmost range limit of this medically relevant tick species. Our finding highlights the importance of tick surveillance and public health challenges posed by geographic expansion of tick vectors and their pathogens.