Phylogeography of the blacklegged tick (Ixodes scapularis) throughout the USA identifies candidate loci for differences in vectorial capacity.

The blacklegged tick (Ixodes scapularis (Journal of the Academy of Natural Sciences of Philadelphia, 1821, 2, 59)) is a vector of Borrelia burgdorferi sensu stricto (s.s.) (International Journal of Systematic Bacteriology, 1984, 34, 496), the causative bacterial agent of Lyme disease, part of a slow-moving epidemic of Lyme borreliosis spreading across the northern hemisphere. Well-known geographical differences in the vectorial capacity of these ticks are associated with genetic variation. Despite the need for detailed genetic information in this disease system, previous phylogeographical studies of these ticks have been restricted to relatively few populations or few genetic loci. Here we present the most comprehensive phylogeographical study of genome-wide markers in I. scapularis, conducted by using 3RAD (triple-enzyme restriction-site associated sequencing) and surveying 353 ticks from 33 counties throughout the species' range. We found limited genetic variation among populations from the Northeast and Upper Midwest, where Lyme disease is most common, and higher genetic variation among populations from the South. We identify five spatially associated genetic clusters of I. scapularis. In regions where Lyme disease is increasing in frequency, the I. scapularis populations genetically group with ticks from historically highly Lyme-endemic regions. Finally, we identify 10 variable DNA sites that contribute the most to population differentiation. These variable sites cluster on one of the chromosome-scale scaffolds for I. scapularis and are within identified genes. Our findings illuminate the need for additional research to identify loci causing variation in the vectorial capacity of I. scapularis and where additional tick sampling would be most valuable to further understand disease trends caused by pathogens transmitted by I. scapularis.

Environmental Drivers of Gulf Coast Tick (Acari: Ixodidae) Range Expansion in the United States.

In the United States, the Gulf Coast tick (Amblyomma maculatum Koch) is a species of growing medical and veterinary significance, serving as the primary vector of the pathogenic bacterium, Rickettsia parkeri (Rickettsiales: Rickettsiaceae), in humans and the apicomplexan parasite, Hepatozoon americanum, in canines. Ongoing reports of A. maculatum from locations outside its historically reported distribution in the southeastern United States suggest the possibility of current and continuing range expansion. Using an ecological niche modeling approach, we combined new occurrence records with high-resolution climate and land cover data to investigate environmental drivers of the current distribution of A. maculatum in the United States. We found that environmental suitability for A. maculatum varied regionally and was primarily driven by climatic factors such as annual temperature variation and seasonality of precipitation. We also found that presence of A. maculatum was associated with open habitat with minimal canopy cover. Our model predicts large areas beyond the current distribution of A. maculatum to be environmentally suitable, suggesting the possibility of future northward and westward range expansion. These predictions of environmental suitability may be used to identify areas at potential risk for establishment and to guide future surveillance of A. maculatum in the United States.

Recently Evolved Francisella-Like Endosymbiont Outcompetes an Ancient and Evolutionarily Associated Coxiella-Like Endosymbiont in the Lone Star Tick (Amblyomma americanum) Linked to the Alpha-Gal Syndrome.

Background: Ticks are hematophagous arthropods that transmit various bacterial, viral, and protozoan pathogens of public health significance. The lone star tick (Amblyomma americanum) is an aggressive human-biting tick that transmits bacterial and viral pathogens, and its bites are suspected of eliciting the alpha-gal syndrome, a newly emerged delayed hypersensitivity following consumption of red meat in the United States. While ongoing studies have attempted to investigate the contribution of different tick-inherent factors to the induction of alpha-gal syndrome, an otherwise understudied aspect is the contribution of the tick microbiome and specifically obligate endosymbionts to the establishment of the alpha-gal syndrome in humans. Materials and Methods: Here we utilized a high-throughput metagenomic sequencing approach to cataloging the entire microbial communities residing within different developmental stages and tissues of unfed and blood-fed ticks from laboratory-maintained ticks and three new geographical locations in the United States. The Quantitative Insights Into Microbial Ecology (QIIME2) pipeline was used to perform data analysis and taxonomic classification. Moreover, using a SparCC (Sparse Correlations for Compositional data) network construction model, we investigated potential interactions between members of the microbial communities from laboratory-maintained and field-collected ticks. Results: Overall, Francisellaceae was the most dominant bacteria identified in the microbiome of both laboratory-raised and field-collected Am. americanum across all tissues and developmental stages. Likewise, microbial diversity was seen to be significantly higher in field-collected ticks compared with laboratory-maintained ticks as seen with a higher number of both Operational Taxonomic Units and measures of species richness. Several potential positive and negative correlations were identified from our network analysis. We observed a strong positive correlation between Francisellaceae, Rickettsiaceae, and Midichloriaceae in both developmental stages and tissues from laboratory-maintained ticks, whereas ovarian tissues had a strong positive correlation of bacteria in the family Xanthobacteraceae and Rhizobiaceae. A negative interaction was observed between Coxiellaceae and Francisellaceae in Illinois, and all the bacteria detected from ticks from Delaware were negatively correlated. Conclusion: This study is the first to catalog the microbiome of Am. americanum throughout its developmental stages and different tissue niches and report the potential replacement of Coxiellaceae by Francisellaceae across developmental stages and tissues tested except in ovarian tissues. These unique and significant findings advance our knowledge and open a new avenue of research to further understand the role of tick microbiome in tick-borne diseases and develop a holistic strategy to control alpha-gal syndrome.

Comparing Contributions of Passive and Active Tick Collection Methods to Determine Establishment of Ticks of Public Health Concern Within Illinois.

In Illinois, between 1990 and 2017, tick-borne diseases in humans increased 10-fold, yet we have insufficient information on when and where people are exposed to vector ticks (Ixodida: Ixodidae). The aims of our research were to compare contributions of passive and active tick collection methods in determining establishment of ticks of public health concern and obtain information on tick distributions within Illinois. We used three surveillance strategies within the Illinois Tick Inventory Collaboration Network to gather information about the ticks of public health concern: 1) passive collection (voluntary submission by the public); 2) systematic collection (biweekly active surveillance); and 3) special collections (active collections in locations of special interest). Of collected adult and nymphal ticks, 436 were from passive collections, 142 from systematic collections, and 1,270 from special collections. Tick species distribution status changed in 36 counties. Our data provide noteworthy updates to distribution maps for use by public health agencies to develop prevention and control strategies. Additionally, the program built a network of collaborations and partnerships to support future tick surveillance efforts within Illinois and highlighted how the combination of the three surveillance strategies can be used to determine geographic spread of ticks, pinpoint locations in need of more surveillance, and help with long-term efforts that support phenology studies.

Heartland Virus in Humans and Ticks, Illinois, USA, 2018-2019.

In 2018, Heartland disease virus infected 2 persons in Illinois, USA. In 2019, ticks were collected at potential tick bite exposure locations and tested for Heartland and Bourbon viruses. A Heartland virus-positive pool of adult male Amblyomma americanum ticks was found at 2 locations, 439 km apart, suggesting widespread distribution in Illinois.

Documentation of the Expansion of the Gulf Coast Tick (Amblyomma maculatum) and Rickettsia parkeri: First Report in Illinois.

The Gulf Coast tick, Amblyomma maculatum, is of public and veterinary health concern, as it is the primary vector of Rickettsia parkeri and Hepatozoon americanum, causative agents of Rickettsiosis and American canine hepatozoonosis. The Gulf Coast tick's range has expanded over the last 50 yr into the mid-Atlantic states, and its expansion is expected to continue northward. We are reporting the presence of A. maculatum for the first time in Illinois, including a total of 18 specimens collected at 6 different sites during surveys in 2013 and 2019. Fourteen of these specimens were screened for Rickettsia parkeri, which resulted in the detection of this bacteria in 8 samples from 4 counties. By depositing these specimens in scientific collections, we provide materialistic evidence of their establishment in 2 counties. We urge health officials to rely on and use scientific collections to document the expansion of these and other vectors across the country. Additionally, we recommend that health practitioners become aware of the clinical similarities between Rocky Mountain Spotted Fever (caused by Rickettsia rickettsii) and "tidewater" fever (caused by R. parkeri).

Impact of Unexplored Data Sources on the Historical Distribution of Three Vector Tick Species in Illinois.

We updated the Illinois historical (1905-December 2017) distribution and status (not reported, reported or established) maps for Amblyomma americanum (L.) (Acari: Ixodidae), Dermacentor variabilis (Say) (Acari: Ixodidae), and Ixodes scapularis (Say) (Acari: Ixodidae) by compiling publicly available, previously unexplored or newly identified published and unpublished data (untapped data). Primary data sources offered specific tick-level information, followed by secondary and tertiary data sources. For A. americanum, D. variabilis, and I. scapularis, primary data contributed to 90% (4,045/4,482), 80% (2,124/2,640), and 32% (3,490/10,898) tick records vs 10%, 20%, and 68%, respectively from secondary data; primary data updated status in 95% (62/65), 94% (51/54) and in 90% (9/10) of the updated counties for each of these tick species; by 1985 there were tick records in 6%, 68%, and 0% of the counties, compared to 20%, 72%, and 58% by 2004, and 77%, 96%, and 75% of the counties by 2017, respectively for A. americanum, D. variabilis, and I. scapularis. We document the loss of tick records due to unidentified, not cataloged tick collections, unidentified ticks in tick collections, unpublished data or manuscripts without specific county location, and tick-level information, to determine distribution and status. In light of the increase in tick-borne illnesses, updates in historical distributions and status maps help researchers and health officials to identify risk areas for a tick encounter and suggest targeted areas for public outreach and surveillance efforts for ticks and tick-borne diseases. There is a need for a systematic, national vector surveillance program to support research and public health responses to tick expansions and tick-borne diseases.

Effects of Pyriproxifen on Aedes japonicus Development and Its Auto-Dissemination by Gravid Females in Laboratory Trials.

In a series of laboratory experiments, we investigated the effects of the juvenile-hormone analog pyriproxyfen on the inhibition of Aedes japonicus adult emergence after exposure of late-stage (3rd/4th) larvae (either field-collected or lab-reared from field-collected eggs) to pyriproxyfen. Emergence inhibition was 74%, 83%, 86%, and 92% at 0.01, 0.5, 2.5, and 5 µg/liter, respectively. Additionally, following a 1-day exposure of larvae to pyriproxyfen-containing water (5 µg/liter), and for a separate cohort exposed to that same water 10 days later, significantly more pupae died resulting in fewer adults emerged in the treatment versus control group. In 4 tent trials, gravid adult females were able to auto-disseminate a 5% pyriproxyfen powder to larval development habitats at high enough concentrations to cause a significant increase in inhibition of adult emergence. Therefore, we conclude that pyriproxyfen has a great potential for use in controlling this invasive cryptic breeder.

Identification of phlebotomine sand flies using one MALDI-TOF MS reference database and two mass spectrometer systems.

BACKGROUND: Rapid, accurate and high-throughput identification of vector arthropods is of paramount importance in surveillance programmes that are becoming more common due to the changing geographic occurrence and extent of many arthropod-borne diseases. Protein profiling by MALDI-TOF mass spectrometry fulfils these requirements for identification, and reference databases have recently been established for several vector taxa, mostly with specimens from laboratory colonies. METHODS: We established and validated a reference database containing 20 phlebotomine sand fly (Diptera: Psychodidae, Phlebotominae) species by using specimens from colonies or field-collections that had been stored for various periods of time. RESULTS: Identical biomarker mass patterns ('superspectra') were obtained with colony- or field-derived specimens of the same species. In the validation study, high quality spectra (i.e. more than 30 evaluable masses) were obtained with all fresh insects from colonies, and with 55/59 insects deep-frozen (liquid nitrogen/-80 °C) for up to 25 years. In contrast, only 36/52 specimens stored in ethanol could be identified. This resulted in an overall sensitivity of 87 % (140/161); specificity was 100 %. Duration of storage impaired data counts in the high mass range, and thus cluster analyses of closely related specimens might reflect their storage conditions rather than phenotypic distinctness. A major drawback of MALDI-TOF MS is the restricted availability of in-house databases and the fact that mass spectrometers from 2 companies (Bruker, Shimadzu) are widely being used. We have analysed fingerprints of phlebotomine sand flies obtained by automatic routine procedure on a Bruker instrument by using our database and the software established on a Shimadzu system. The sensitivity with 312 specimens from 8 sand fly species from laboratory colonies when evaluating only high quality spectra was 98.3 %; the specificity was 100 %. The corresponding diagnostic values with 55 field-collected specimens from 4 species were 94.7 % and 97.4 %, respectively. CONCLUSIONS: A centralized high-quality database (created by expert taxonomists and experienced users of mass spectrometers) that is easily amenable to customer-oriented identification services is a highly desirable resource. As shown in the present work, spectra obtained from different specimens with different instruments can be analysed using a centralized database, which should be available in the near future via an online platform in a cost-efficient manner.

Habitat characteristics of larval mosquitoes in zoos of South Carolina, USA.

To investigate whether the unique assemblage of habitats in zoos could affect mosquito oviposition behavior and to provide zoos with suggestions for mosquito control, larvae were sampled and associated habitat variables were measured in 2 zoos in South Carolina, U.S.A. Fifty-nine sites were sampled from March 2008 to January 2009. A total of 1630 larvae representing 16 species was collected and identified. The dominant species was Aedes albopictus (46.0%), followed by Ae. triseriatus (23.6%), Culex restuans (12.4%), and Cx. pipiens complex (9.7%). Principal components and multiple logistic regression analyses showed that across both zoos the distribution of Ae. albopictus larvae was predicted by ambient and site temperature, precipitation, dissolved oxygen, and container habitats. The distribution of Ae. triseriatus larvae was predicted by natural containers and shade height < or =2 m. Overall larval mosquito presence (regardless of species) was predicted by ambient and site temperature, precipitation, dissolved oxygen, presence of natural habitats, and absence of aquatic vegetation. Additionally, C8 values of pairwise species associations indicated significant habitat-based relationships between Ae. albopictus and Ae. triseriatus, and Cx. pipiens complex and Cx. restuans. In general, species-habitat associations conformed to previously published studies. Recommendations to zoo personnel include elimination of artificial container habitats, reduction of shade sources < or =2 m over aquatic habitats, use of approved mosquito larvicides, and training in recognizing and mitigating larval mosquito habitats.

Arthropods of medicoveterinary importance in zoos.

Zoos present a unique assemblage of arthropods, captive vertebrates, free-roaming wildlife, humans, and plants, each with its own biota of symbiotic organisms. Arthropods of medicoveterinary importance are well represented in zoos, and an ample literature documents their influence in these animal-rich environments. Mosquitoes are of greatest significance because of the animal and human pathogens they transmit, followed by ectoparasites, many of which are exotic and present health risks to captive and native animals. Biting flies, cockroaches, filth flies, and triatomid bugs represent additional concerns. Integrated management programs for arthropods in zoos are commonplace. Zoos can play a role in biosurveillance, serving as an advanced guard for detecting exotic arthropods and vector-borne diseases. We provide the first review of arthropods of medicoveterinary importance in zoos. A case is made for the value of collaborations between entomologists and zoo personnel as a means of enhancing research and public education while safeguarding the health of captive animals and the public.

Cuterebrid myiasis (Diptera: Oestridae) in captive ring-tailed lemurs (Lemur catta) at a South Carolina zoo.

In September 2008, two ring-tailed lemurs (Lemur catta), comprising a mother-daughter pair, at the Greenville Zoo, Greenville, South Carolina, USA, were diagnosed with cuterebrid myiasis (Diptera: Oestridae) subsequent to sudden death of the adult lemur. On necropsy, a single bot warble was discovered in the subcutis of the axillary region. Histopathology revealed a severe eosinophilic bronchopneumonia. The juvenile lemur was inspected and found to have warbles on three separate dates in late September. One representative bot fly larva was identified as a Cuterebra sp. that normally infests lagomorphs in the southeastern United States. Cuterebrid myiasis is rarely reported in lemurs and has not been previously associated with pneumonia or death in these animals.