Study of general practitioner consultations for tick bites at high,medium and low incidence areas for Lyme borreliosis in England and Wales

Lyme borreliosis (LB) is a tick‐borne disease caused by Borrelia burgdorferi sensu lato complex. In Europe, it is predominately transmitted by the sheep tick, Ixodes ricinus. Compared with other European countries, the United Kingdom (UK) is considered to have a low incidence of LB, although this varies regionally. To determine whether an association exists between tick bite consultations and LB incidence in the UK, retrospective questionnaires were sent to general practitioners (GPs) in high (Wiltshire), medium (Cumbria) and low (Wales) incidence areas. During 2011, the greatest incidence of consultations for tick bites was reported by GPs in Cumbria (204 consultations per 100,000 inhabitants), followed by Wiltshire (160 per 100,000 population) and Wales (54 per 100,000 population). In Wiltshire and Cumbria, GPs predominantly provided advice on tick removal, whilst Welsh GPs mostly advised patients on tick bite prevention. Focusing on Cumbria during 2011–2013, 72.5% of GPs removed ticks from patients (incidence of 101 consultations per 100,000 population), and more GPs diagnosed LB based on clinical features than laboratory‐confirmed diagnoses. To date, this is the first study to investigate the incidence of tick bite consultations and LB in England and Wales.     

Detection of Invasive Borrelia burgdorferi Strains in North‐Eastern Piedmont,Italy

Following reports of human cases of Lyme borreliosis from the Ossola Valley, a mountainous area of Piemonte, north‐western Italy, the abundance and altitudinal distribution of ticks, and infection of these vectors with Borrelia burgdorferi sensu lato were evaluated. A total of 1662 host‐seeking Ixodes ricinus were collected by dragging from April to September 2011 at locations between 400 and 1450 m above sea level. Additional 104 I. ricinus were collected from 35 hunted wild animals (4 chamois, 8 roe deer, 23 red deer). Tick density, expressed as the number of ticks per 100 m², resulted highly variable among different areas, ranging from 0 to 105 larvae and from 0 to 22 nymphs. A sample of 352 ticks (327 from dragging and 25 from wild animals) was screened by a PCR assay targeting a fragment of the 16S rRNA gene of B. burgdorferi s.l. Positive samples were confirmed with a PCR assay specific for the 5S‐23S rRNA intergenic spacer region and sequenced. Four genospecies were found: B. afzelii (prevalence 4.0%), B. lusitaniae (4.0%), B. garinii (1.5%) and B. valaisiana (0.3%). Phylogenetic analysis based on the ospC gene showed that most of the Borrelia strains from pathogenic genospecies had the potential for human infection and for invasion of secondary body sites.     

A morphological and molecular study of Anaplasma phagocytophilum transmission events at the time of Ixodes ricinus tick bite

Background

Anaplasma phagocytophilum is the causative agent of human granulocytic anaplasmosis (HGA) in humans and tick-borne fever (TBF) in ruminants. The bacterium invades and replicates in phagocytes, especially in polymorphonuclear granulocytes.

Methods

In the present study, skin biopsies and ticks (Ixodes ricinus) were collected from tick feeding lesions on 38 grazing lambs between two and three weeks after access to pastures. The histopathological changes associated with tick bites and A. phagocytophilum infection, were described. In addition the skin biopsies were examined by immunohistochemistry. Furthermore, samples from blood, skin biopsies and ticks were examined by serology, PCR amplification of msp2 (p44), genotyping of rrs (16S rRNA) variants, and compared with the results obtained from histological and immunohistochemical investigations.

Results

Tick bites were associated with chronic and hyperplastic inflammatory skin lesions in this study. A. phagocytophilum present in skin lesions were mainly associated with neutrophils and macrophages. Bacteria were occasionally observed in the Tunica media and Tunica adventitia of small vessels, but were rarely found in association with endothelial cells. PCR and genotyping of organisms present in blood, ticks and skin biopsies suggested a haematogenous and a local spread of organisms at the tick attachment sites.

Conclusions

The present study describes different aspects of A. phagocytophilum infection at the site of tick bite, and indicates that A. phagocytophilum rarely associates with endothelium during the early pathogenesis of infection.     

Distribution of Neoehrlichia mikurensis in Ixodes ricinus ticks along the coast of Norway: The western seaboard is a low‐prevalence region

Neoehrlichia mikurensis is a tick‐borne pathogen widespread among ticks and rodents in Europe and Asia. A previous study on Ixodes ricinus ticks in Norway suggested that N. mikurensis was scarce or absent on the south‐west coast of Norway, but abundant elsewhere. The aim of this study was to further investigate the prevalence and distribution of N. mikurensis along the western seaboard of Norway in comparison with more eastern and northern areas. The second aim of the study was to examine seasonal variation of the bacterium in one specific location in the south‐eastern part of Norway. Questing I. ricinus were collected from 13 locations along the coast of Norway, from Brønnøysund in Nordland County to Spjærøy in Østfold County. In total, 11,113 nymphs in 1,113 pools and 718 individual adult ticks were analysed for N. mikurensis by real‐time PCR. The mean prevalence of N. mikurensis in adult ticks was 7.9% while the estimated pooled prevalence in nymphs was 3.5%. The prevalence ranged from 0% to 25.5%, with the highest prevalence in the southernmost and the northernmost locations. The pathogen was absent, or present only at low prevalence (<5%), at eight locations, all located in the west, from 58.9°N to 64.9°N. The prevalence of N. mikurensis was significantly different between counties (p < .0001). No significant seasonal variation of N. mikurensis prevalence was observed in the period May to October 2015. Our results confirm earlier findings of a low prevalence of N. mikurensis in the western seaboard of Norway.     

Genetic parameters for tick counts across months for different tick species and anatomical locations in South African Nguni cattle

The objective of the study was to characterise genetic parameters across months for different tick species and anatomical locations in South African Nguni cattle. Tick counts were conducted monthly, over a 2-year period, on 586 Nguni cattle under natural infestation, from four herds located in different provinces of South Africa. The counts were recorded for six species of ticks (Amblyomma hebraeum, Rhipicephalus evertsi evertsi, Rhipicephalus decoleratus and microplus (Boofilids), Rhipicephalus appendiculatus, Rhipicephalus simus and Hyalomma marginatum) attached on eight anatomical locations on the animals and were summed by species and anatomical location. Heritability estimates, phenotypic and genetic correlations were estimated on a monthly basis using mixed linear models, fitting univariate and bivariate sire models. Fixed effects considered were location, sex, year and age as a covariate. Tick counts were higher in the hot months, and A. hebraeum was the most dominant tick species. Heritability estimates for tick count varied by month and trait and ranged from 0 to 0.89. Genetic correlations were mostly positive, and low to high, with some negative correlations with high standard error. Phenotypic correlations were low to moderate. In general, high genetic correlations were observed between whole body count and the anatomical location counts, suggesting that it may not be necessary to conduct whole body counts. Counts from the belly and perineum appeared to be the most suitable surrogate traits for whole body count. These findings provide useful information for developing strategies for the practical implementation of genetic selection, as a supplement to the traditional tick control measures.     

Discrepancies between self‐reported tick bites and evidence of tick‐borne disease exposure among nomadic Mongolian herders

Twenty‐six per cent of Mongolians live pastoral lifestyles, increasing their likelihood of exposure to ticks and placing them at a higher risk for contracting tick‐borne diseases (TBDs). Anaplasma spp. and Rickettsia spp. have been identified in ticks, livestock and humans in Mongolia, but no known qualitative research has been conducted investigating the association between nomadic herder characteristics, tick bite history and exposure to TBDs. To better understand the association between self‐reported tick bites and symptoms versus actual exposure to TBDs, this study paired serological data with 335 surveys administered to Mongolian herders, ages 12–69, from 2014 to 2015. Logistic regression results identified no significant associations between reported tick bites or symptoms with serological evidence of Anaplasma spp. and Rickettsia spp. controlling for age, gender and aimag. Among the 335 respondents who were seropositive to either Anaplasma spp. or Rickettsia spp., 32.9% self‐reported experiencing abnormal symptoms such as redness, inflammation, headache, arthritis or fever after being bitten. Alternatively, 17.3% (58/335) of individuals reported experiencing symptoms following a tick bite in instances where serological results indicated no exposure to Anaplasma spp. or Rickettsia spp. Results also identified inconsistencies in reporting and seroprevalence among different age groups, with children having the highest reporting and treatment seeking rates but low levels of exposure in comparison with other groups. While survey results showed that individuals were aware of peak tick seasons and tick species that inhabit specific areas, 58% of heads of households (49/84) were unaware that ticks can cause disease in livestock or dogs. This study suggests that herders are an at‐risk population in Mongolia with gaps in awareness of TBD risk. Increased surveillance paired with focused outreach to prevent TBDs targeted to the herder population is encouraged.     

Study on ticks and tick-borne zoonoses in public parks in Italy

A survey on tick density and on tick‐borne zoonoses was carried out in four public parks in the outskirts of Imola (northern Italy) from June to October 2006. All stages of Ixodes ricinus and only larvae of Riphicephalus sanguineus were recovered by dragging, performed on 100‐m transects. Almost all ticks (99%) were harvested in one park. I. ricinus density (nymphs/100 m²) ranged from 0 in park L to 6.3 in park F. Nymphs and adults of I. ricinus were subjected to PCR for Anaplasma phagocytophilum, Bartonella spp., Borrelia burgdorferi s. l. and Rickettsia spp. The observed prevalences were 38.3% for Bartonella henselae, 5.2% for Bartonella clarridgeiae, 10.4% for B. burgdorferi s. l., 2.6% for Rickettsia helvetica and 13% for Rickettsia monacensis, respectively. No DNA of A. phagocytophilum was found. Acarological risks (AR) were calculated as probabilities of collecting at least one infected nymph per transect. The AR values calculated for the various zoonotic agents were 11.4% for R. helvetica, 27.7% for B. clarridgeiae, 49.7% for B. burgdorferi s. l., 57.2% for R. monacensis and 90.4% for B. henselae, respectively. In this study, B. clarridgeiae was for the first time identified in I. ricinus ticks.     

First detection of Ixodes ricinus on beef cattle in Israel

This is the first report of the presence of Ixodes ricinus on beef cattle in Israel. Up to now, in the Middle East this tick was considered to be confined to Turkey and northern Iran. In the present study, tick samples collected from field-grazing beef cattle in western Galilee (northern Israel) were first examined morphologically for species-specific taxonomical features and then by molecular characterization. Ticks identified morphologically as I. ricinus were then examined by PCR with four different molecular markers: 12S rRNA, 16S rRNA, COX1 and cytochrome B. The PCR products were sequenced and compared with annotated I. ricinus sequences in GenBank? and the analyzed sequences from the collected samples shared 98–99% identity with reported I. ricinus sequences. In contrast, sequences from the collected ticks shared identity of 91% or less with annotated sequences from other Ixodes species. Multiple alignments and neighbor-joining analyses performed for each of the four markers reinforced the results obtained from pairwise alignments. These findings demonstrated for the first time the presence in Israel of the tick species I. ricinus – with results confirmed by a combination of morphological examination and molecular analyses.     

Knowledge,beliefs, and practices regarding tick bites in the Turkish population in a rural area of the Middle Anatolian Region

The aim of this study is to determine the knowledge, beliefs, and practices of a population living in a rural area in regards to tick bites and Crimean-Congo hemorrhagic fever (CCHF). The study was conducted in a rural area located in Central Anatolia in the region of Eskişehir. A total of 1,500 individuals aged 20 years and older chosen by a stratified random sample were enrolled. A questionnaire was administered in person. In 264 (17.4%) participants, there was a history of being bitten by a tick. This rate was higher in older persons, males, married persons, and farmers. The most commonly reported protective behavior was wearing long sleeves and long pants when wandering in rural areas (65.1% of participants). The least commonly reported behavior was using insect repellent on skin or clothes (3.3% of participants). Only 799 participants (54%) had heard about CCHF as a disease associated with ticks. Females, those with primary school education, housewives, and male farmers had a high frequency of having heard about CCHF. Tick bites and CCHF are important public health problems, yet the current knowledge of these problems is not sufficient in populations living in rural areas of the Middle Anatolian Region of Turkey.     

Anti-chemokine activities of ixodid ticks depend on tick species,developmental stage,and duration of feeding

Ixodid ticks require comparatively large bloodmeals for their development and survival. Blood-feeding elicits signaling events in the host leading to wound healing responses (hemostasis, inflammation, and tissue repair) and immunity. Bioactive molecules present in tick saliva sabotage these host responses at several levels. One of them is neutralization of cellular communication by binding of specific saliva molecules to cytokines that have important roles in innate and adaptive immunity. Chemokines are a subset of cytokines having chemotactic activities. We show anti-chemokine activities in salivary gland extracts (SGE) of adult Rhipicephalus appendiculatus ticks against human chemokines CXCL8, CCL2, CCL3, CCL5, and CCL11. At comparable protein concentrations, male Ixodes ricinus SGE showed activity against all the chemokines; SGE of female I. ricinus had comparatively lower levels of activity against all the chemokines but no detectable activity against CCL5 and CCL11. However, when the equivalent of a single pair of salivary glands was tested, male I. ricinus showed little or no activity against CCL3 and CCL5. No fundamental differences in activity were observed against mouse compared with human chemokines. A comparison with previously published data for Dermacentor reticulatus and Amblyomma variegatum indicates that the level of anti-cytokine activity depends on the species, developmental stage (adult or nymph), and amount of SGE used, as well as on the number of days the tick has been feeding.     

Evidence of Powassan/deer tick virus in adult black‐legged ticks (Ixodes scapularis) recovered from hunter‐harvested white‐tailed deer (Odocoileus virginianus) in Pennsylvania: A public health perspective

Studies reporting tick infection rates for Powassan virus (POWV), an emerging zoonotic arthropod‐borne pathogen responsible for POWV disease in the Commonwealth of Pennsylvania, are limited. To determine the presence and ascertain a statewide prevalence of POWV, ticks were collected from 9,912 hunter‐harvested white‐tailed deer (Odocoileus virginianus) heads presented to six regional Pennsylvania Game Commission Chronic Wasting Disease sampling stations in early December of 2013, 2014 and 2015. Of the 2,973 ticks recovered, 1,990 (66.9%) were identified as adult Ixodes scapularis (black‐legged tick). The 1,990 I. scapularis ticks were PCR‐tested for the presence of POWV. The ticks had a statewide Powassan/deer tick virus infection rate of 0.05%, providing evidence of this pathogen in Pennsylvania's adult I. scapularis ticks and supporting the need for more comprehensive pathogen prevalence assessment strategies, as well as increased public health awareness for this emerging zoonotic arthropod‐borne pathogen of public health concern.     

Modulation of cutaneous inflammation induced by ticks in contrasting phenotypes of infestation in bovines

Tick saliva contains molecules that are inoculated at the site of attachment on their hosts in order to modulate local immune responses and facilitate a successful blood meal. Bovines express heritable, contrasting phenotypes of infestations with the cattle tick, Rhipicephalus (Boophilus) microplus: breeds of Bos taurus indicus are significantly more resistant than those of Bos taurus taurus. Tick saliva may contain molecules that interfere with adhesion of leukocytes to endothelium and resistant hosts may mount an inflammatory profile that is more efficient to hamper the tick's blood meal. We show in vitro that adhesion of peripheral blood mononuclear cells to monolayers of cytokine-activated bovine umbilical endothelial cells was significantly inhibited by tick saliva. The inflammatory response to bites of adults of R. microplus mounted by genetically resistant and susceptible bovine hosts managed in the same pasture was investigated in vivo. The inflammatory infiltrates and levels of message coding for adhesion molecules were measured in biopsies of tick-bitten and control skin taken when animals of both breeds were exposed to low and high tick infestations. Histological studies reveal that cutaneous reactions of resistant hosts to bites of adult ticks contained significantly more basophils and eosinophils compared with reactions of the susceptible breed. Expression of the adhesion molecules – intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1) and P-selectin – was higher in adult-infested skin of susceptible hosts undergoing low infestations compared to resistant hosts; when host was exposed to high infestations expression of these adhesion molecules was down-regulated in both phenotypes of infestations. Expression of leukocyte adhesion glycoprotein-1 (LFA-1) was higher in skin from susceptible hosts undergoing low or high infestations compared to resistant hosts. Conversely, higher levels of E-selectin, which promotes adhesion of memory T cells, were expressed in skin of resistant animals. This finding may explain the resistant host's ability to mount more rapid and efficient secondary responses that limit hematophagy and infestations. The expression profiles observed for adhesion molecules indicate that there are differences in the kinetics of the inflammatory reactions mounted by resistant and susceptible hosts and the balance between tick and host is affected by the number of tick bites a host receives. We show that the contrasting phenotypes of infestations seen in bovines infested with R. microplus are correlated with differences in the cellular and molecular composition of inflammatory infiltrates elicited by bites with adult ticks.     

Anchoring tick salivary anti-complement proteins IRAC I and IRAC II to membrane increases their immunogenicity

Tick salivary proteins are promising targets for the development of anti-tick vaccines. Recently, we described two paralogous anti-complement proteins, called Ixodes ricinus anti-complement (IRAC) proteins I and II, that are co-expressed in tick I. ricinus salivary glands. However, our previous attempts to immunize rabbits against IRAC via infection with recombinant Bovine herpesvirus 4 (BoHV-4) vectors invariably failed although both recombinants expressed high levels of functional IRAC proteins in vitro. As IRAC are soluble monovalent antigens, one of the possible explanations is that monovalent ligation of the B-cell receptor induces receptor activation but fails to promote antigen presentation, a phenomenon that is thought to induce a state of B-cell tolerance. In the present study, we tried to increase IRAC immunogenicity by expressing them as oligovalent antigens. To this end, IRAC were fused to membrane anchors and BoHV-4 vectors expressing these recombinant forms were produced. The immunization potentials of recombinant viruses expressing either secreted or transmembrane IRAC proteins were then compared. While the former did not induce a detectable immune response against IRAC, the latter led to high titres of anti-IRAC antibodies that only marginally affected tick blood feeding. All together, the data presented in this study demonstrate that the immunogenicity of a soluble antigen can be greatly improved by anchoring it in membrane.     

Allochronic seasonal peak activities of Dermacentor and Haemaphysalis spp. under continental climate in Hungary

Hard ticks (Acari: Ixodidae) were collected from the vegetation at monthly intervals in 2008 on six places of three different biotopes in Hungary. Except for Haemaphysalis concinna and H. punctata, predominance of females was observed in the questing population. From among the species with biphasic activity period, metastriate ticks (Dermacentor spp. and H. inermis) had higher prevalence of host-seeking males in the autumn than in the spring, as opposed to prostriate Ixodes ricinus. In comparison with the permanently mild weather experienced at the beginning of 2007, sharply rising temperatures during consecutive winter days in January and February of 2008 appeared to be more efficient in triggering a 1–2 month earlier spring peak activity of hard ticks, except for I. ricinus. Regarding seasonality, D. marginatus was most numerous in February and March, whereas D. reticulatus in September and October; H. punctata showed peak activity in March–April, H. concinna in May, and H. inermis in November–December. Within these genera such a temporal difference (allochrony) between seasonal peak activities of sympatric species under continental climate is described for the first time.     

Geographical distribution and prevalence of tick‐borne encephalitis virus in questing Ixodes ricinus ticks and phylogeographic structure of the Ixodes ricinus vector in Norway

The tick‐borne encephalitis virus (TBEV), a zoonotic flaviviral infection, is endemic in large parts of Norway and Eurasia. Humans are mainly infected with TBEV via bites from infected ticks. In Norway, the main geographical distribution of ticks is along the Norwegian coastline from southeast (~59°N) and up to the southern parts of Nordland County (~65°N). In this study, we collected ticks by flagging along the coast from Østfold County to Nordland County. By whole‐genome sequencing of the mitochondrial genome of Ixodes ricinus, the phylogenetic tree suggests that there is limited phylogeographic structure both in Norway and in Europe. The overall TBEV prevalence is 0.3% for nymphs and 4.3% for adults. The highest estimated TBEV prevalence in adult ticks was detected in Rogaland and Vestfold County, while for nymphs it is highest in Vestfold, Vest‐Agder and Rogaland. The present work is one of the largest studies on distribution and prevalence of TBEV in ticks in Scandinavia, showing that the virus is wider distributed in Norway than previously anticipated.     

Reduction of <Emphasis Type="Italic">Theileria annulata</Emphasis> Infection in Ticks Fed on Calves Immunized with Purified Larval Antigens of <Emphasis Type="Italic">Hyalomma anatolicum anatolicum</Emphasis>

Larval antigen of Hyalomma anatolicum anatolicum, the vector of Theileria annulata, was purified by two-step affinity chromatography using anti-tick gut-specific rabbit IgG and IgG from immunized cattle. The purified antigen showed the presence of a single polypeptide of 37 kDa (GHLAgP) on SDS-PAGE. Two groups (I and II) of naive crossbred calves (Bos taurus × B. indicus) were immunized with 1 mg of GHLAgP in three divided doses. Immunized calves of group I were also infected with a sublethal dose of T. annulata along with a group of non-immunized calves (group III). Animals in groups I, II, III as well a control group (group IV) were challenged with live nymphs of H. a. anatolicum on the 10th day of immunization. There was a significant reduction in the number of emerging adults of 56.9% ± 1.67% in calves of group I (p < 0.01) and 63.09% ± 1.26% in calves of group II (p < 0.001) compared to the controls. The calves of groups I and II showed antibody responses to tick antigen up to day 70 post immunization. Infection with T. annulata was determined in the salivary glands of adult ticks that developed from the nymphs used for challenge infection. In ticks taken from group I calves, there was a 75.0% ± 0.00% infection compared with only 85.0% ± 2.88% infection in ticks taken from calves of group III. Using PCR, a lower infection (83.33% ± 3.33%) was detected in ticks that developed from calves of group I compared with calves from group III (90.00% ± 2.88%). The ground-up tick supernatants (GUTS) of the ticks taken from calves of group III yielded higher infection rate and exhibited higher infectivity titre in in vitro infection assay of bovine mononuclear cells than the GUTS of the ticks taken from calves of group I. The results suggest a partial reduction in growth rate of T. annulata in ticks feeding on calves immunized with GHLAgP.     

Active surveillance of pathogens from ticks collected in New York State suburban parks and schoolyards

Schoolyards and suburban parks are two environments where active tick surveillance may inform local management approaches. Even in a state such as New York with a robust active tick surveillance programme operated by the state Department of Health, these settings are not routinely covered. The goal of this study was to highlight the importance of active surveillance for tick‐borne pathogens by describing their prevalence in ticks collected from schoolyards and suburban parks and to guide the use of integrated pest management in these settings. Tick dragging was performed in three regions of New York State: Long Island, the Lower Hudson Valley and the Capital Region. A total of 19 schoolyards and 32 parks were sampled. The location, habitat and weather at the time of tick collection were recorded. Ticks were speciated and tested for the presence of 17 pathogens with a novel application of nanoscale real‐time PCR. The causative agents of Lyme disease, anaplasmosis, babesiosis and Powassan virus disease were all detected from Ixodes scapularis in various sites throughout the capital region and south‐eastern counties of New York state. The most common agent detected was Borrelia burgdorferi, and coinfection rates were as high as 36%. This surveillance study also captured the first of the invasive Asian longhorned tick species, Haemaphysalis longicornis, in New York state (collected 2 June 2017). Results from this study highlight the importance of collaborative efforts and data sharing for improvement of surveillance for tick‐borne disease agents.     

Knowledge and prevention of tickborne diseases among Hispanic and non‐Hispanic residents of Maryland and Virginia

Tickborne diseases (TBDs) such as Lyme disease (LD), babesiosis, ehrlichiosis and Rocky Mountain spotted fever cause substantial morbidity and even mortality in the USA. Data indicate that Hispanic populations may be at greater risk for occupational exposure to ticks and disseminated LD; however, information on knowledge and practices of Hispanic populations regarding TBDs is limited. We surveyed 153 Hispanic and 153 non‐Hispanic residents of Maryland and Virginia to assess awareness of TBDs, prevention practices and risk of tick encounters. Hispanic respondents were less likely than non‐Hispanics to report familiarity with LD symptoms (21% vs. 53%, p < 0.001) and correctly identify ticks as vectors of LD (40% vs. 85%, p < 0.001). Although there was no significant difference in overall proportion of respondents who routinely take one or more preventive measures to prevent tick bites (59% vs. 61%, p = 0.65), Hispanics were more likely to report showering after coming indoors (36% vs. 25%, p = 0.04) but less likely to conduct daily tick checks compared with non‐Hispanics (17% vs. 35%, p < 0.001). History of tick bite or finding a tick crawling on oneself or a household member in the past year did not significantly differ between Hispanics and non‐Hispanics (19% vs. 24%, p = 0.26). Notably, after controlling for Hispanic/non‐Hispanic ethnicity, primary language (English vs. Spanish) was a significant predictor of whether an individual had knowledge of LD symptoms, correctly identified ticks as vectors for LD and performed daily tick checks. These results provide guidance for future development of more targeted and effective TBD prevention education for both Hispanic and non‐Hispanic communities.     

Stable Transmission of Borrelia burgdorferi Sensu Stricto on the Outer Banks of North Carolina

The spirochaete (Borrelia burgdorferi) associated with Lyme disease was detected in questing ticks and rodents during a period of 18 years, 1991–2009, at five locations on the Outer Banks of North Carolina. The black‐legged tick (Ixodes scapularis) was collected at varied intervals between 1991 and 2009 and examined for B. burgdorferi. The white‐footed mouse (Peromyscus leucopus), house mouse (Mus musculus) marsh rice rat (Oryzomys palustris), marsh rabbit (Sylvilagus palustris), eastern cottontail (Sylvilagus floridanus) and six‐lined racerunner (Cnemidophorus sexlineatus) were live‐trapped, and their tissues cultured to isolate spirochaetes. Borrelia burgdorferi isolates were obtained from questing adult I. scapularis and engorged I. scapularis removed from P. leucopus, O. palustris and S. floridanus. The prevalence of B. burgdorferi infection was variable at different times and sites ranging from 7 to 14% of examined questing I. scapularis. Mitochondrial (16S) rRNA gene phylogenetic analysis from 65 adult I. scapularis identified 12 haplotypes in two major clades. Nine haplotypes were associated with northern/Midwestern I. scapularis populations and three with southern I. scapularis populations. Sixteen isolates obtained from tick hosts in 2005 were confirmed to be B. burgdorferi by amplifying and sequencing of 16S rRNA and 5S‐23S intergenic spacer fragments. The sequences had 98–99% identity to B. burgdorferi sensu stricto strains B31, JD1 and M11p. Taken together, these studies indicate that B. burgdorferi sensu stricto is endemic in questing I. scapularis and mammalian tick hosts on the Outer Banks of North Carolina.