Detection of single nucleotide polymorphism in the para-sodium channel gene of Rhipicephalus annulatus populations from Egypt resistant to deltamethrin.

Rhipicephalus annulatus field populations collected from small cattle farms in Beni-Suef province in Egypt were evaluated for deltamethrin resistance by toxicological in vitro bioassays (adult immersion test-AIT and larval packet test-LPT). Moreover, a quantitative PCR high resolution melting (PCR-HRM) technique was used to detect nucleotide substitutions in the voltage-gated sodium channel (Na-channel) gene. By the in vitro bioassays, the examined ticks were phenotypically categorized as deltamethrin susceptible (populations El-Wasta - A, and El-Hakamna - C) or resistant (populations El-Wasta - B, El-Hakamna - D, EL-Halabia - E, and Kom-abokhalad - F). Based on LPT findings, the phenotypic resistant populations were found to have a resistance ratio between 6.5 - 10.8. The PCR-HRM genotyping of the ticks showed variable melting curves among the populations in domain II of the Na-channel gene. Analysis of the curves showed the presence of wild type, mutant homozygous, and mutant heterozygous tick individuals. By sequencing the PCR amplified fragments, the C190A mutation was the only detected nucleotide polymorphism of domain II among the phenotypically resistant populations, which was present in 39.5 % (34/86) of the ticks tested. On the other hand, the phenotypically susceptible populations A and C did not show C190A mutant homozygous (RR) individuals. Meanwhile, in domain III all of the examined populations revealed melting curves like the wild type. Furthermore, the sequence analysis of these populations confirmed the absence of SNPs in domain III. The C190A single point mutation was detected for the first time in domain II of the Na-channel gene of deltamethrin-resistant R. annulatus in Egypt using PCR-HRM. Screening for efficacy of chemical compounds used by farmers to control ticks on cattle should be considered as part of animal health programs to manage the emerging resistance to acaricides in R. annulatus populations.

Comparison of Some Molecular Markers for Tick Species Identification.

BACKGROUND: Ticks are obligate blood-sucking ectoparasites of vertebrates. Since many tick identification studies are based on the analysis of 16S rDNA, 12S rDNA and ITS-1, 2 rDNA genes, we aimed to compare the performance of these molecular markers of common use for the identification of ticks, under a diagnostic laboratory environment. METHODS: Overall, 192 tick specimens were collected through the state of Texas from January 2014 to August 2015 and the species was determined by both morphology and molecular amplification using the 16S rDNA, 12S rDNA, ITS1 and ITS2. RESULTS: The species collected were identified by molecular techniques as Dermacentor albipictus, D. variabilis, Amblyomma americanum, Ixodes scapularis, A. cajennense, Rhipicephalus sanguineus and Carios capensis. ITS1 and ITS2 were not able to prove consistent amplification and therefore have been considered as potential markers for tick identification. CONCLUSION: The use of mitochondrial genes in tick identification showed to provide more consistent results in the diagnostic environment.

Test for Borrelia spp. in bats in an urban area in the South of Brazil.

INTRODUCTION: We investigated the occurrence of relapsing fever (RF) causing Borrelia genus spirochetes in synanthropic bats from the municipality of Maringá, Paraná, South of Brazil. METHODS: Tissue samples from the wings of bats were collected monthly from April 2013 to February 2014 and extracted DNA was used to evaluate the presence of RF causing Borrelia spp. RESULTS: All bat tissues tested negative for RF causing Borrelia spp. CONCLUSIONS: Borrelia spp. do not occur in chiropterans from Maringá.

Searching for the Immature Stages of Ixodes scapularis (Acari: Ixodidae) in Leaf Litter and Soil in Texas.

The standard tick collection methods of flagging and dragging are successful for collecting all stages of the blacklegged tick, Ixodes scapularis (Say) (Acari: Ixodidae), in the northern United States. However, for unknown reasons, these methods are unsuccessful for collecting the immature stages of I. scapularis in the southern United States. Thus, a different collection strategy was employed to search for the immature stages of I. scapularis in the southern state of Texas. Monthly sampling of three types of microhabitats potentially harboring ticks was conducted for 17 mo at the Big Thicket National Preserve. Samples of leaf litter, topsoil, and subsoil were placed within Berlese funnels to determine if the immature stages of I. scapularis are residing in these layers. No ticks were found in any of the 600 substrate samples examined. Along nearby trail edges in the same area, 656 adult I. scapularis (an average of 22.6 per 1,000 m2), as well as 268 immatures of other species (i.e., Amblyomma americanum (Linnaeaus) (Acari: Ixodidae) and Dermacentor variabilis (Say) (Acari: Ixodidae)) were collected using flagging and dragging. These results suggest that unlike speculations from previous studies in the southern United States, the immature stages of I. scapularis may not be residing in the leaf litter and soil layers in Texas. We hypothesize that they may be residing in their host's nests and burrows. Perhaps I. scapularis in the south is exhibiting a stage specific mixed host-seeking strategy by residing in nests and burrows as immatures, contributing to the geographical difference in Lyme disease prevalence between the northern and southern United States.

Genetic structure analysis of Amblyomma mixtum populations in Veracruz State, Mexico.

Amblyomma mixtum Koch, 1844 parasitizes livestock, humans, and wildlife in Mexico. However, information on population genetics for this tick species in the country is missing. Tick samples were collected from livestock in ten regions across the state of Veracruz (22°28'N, 17°09'S, 93°36'E, 98°39'W) to analyze the genetic structure of A. mixtum populations. Ticks were morphologically identified using taxonomic keys. In order to test the intra-specific variability of A. mixtum fragments of the mitochondrial gene 16S-rRNA and cytochrome oxidase subunit 1 (COI) were amplified. Ninety-six sequences were amplified from the 50 specimens' analyzed (96% amplification success). Eleven haplotypes were detected in 16S-rRNA gene and 10 more for COI. Neutrality tests showed negative results in most of the locations analyzed, which is indicative of an excess of recently derived haplotypes. However, these results were not statistically significant. Minimal union network analysis revealed that there is no separation of populations by geography, and that there is an overlap of several haplotypes among diverse populations. Significant genetic differentiation was not detected in the A. mixtum populations sampled in the state of Veracruz, Mexico, this may be due to the frequent movement of livestock hosts. This is the first report on the genetic structure of A. mixtum populations in Mexico.

Test for Borrelia spp. in bats in an urban area in the South of Brazil

Abstract INTRODUCTION: We investigated the occurrence of relapsing fever (RF) causing Borrelia genus spirochetes in synanthropic bats from the municipality of Maringá, Paraná, South of Brazil. METHODS: Tissue samples from the wings of bats were collected monthly from April 2013 to February 2014 and extracted DNA was used to evaluate the presence of RF causing Borrelia spp. RESULTS: All bat tissues tested negative for RF causing Borrelia spp. CONCLUSIONS: Borrelia spp. do not occur in chiropterans from Maringá.

Free-living ticks in a cerrado fragment, state of Mato Grosso do Sul, Brazil / Carrapatos de vida livre em fragmento de cerrado, no Estado de Mato Grosso do Sul, Brasil

The fatal cases of spotted fever reported in some Brazilian municipalities are generally associated with the presence of ticks and capybaras (Hydrochoerus hydrochaeris). The urban forest fragments where this rodent is present are of great concern. We investigated the diversity of free ticks as well as the occurrence of ticks infected by Rickettsia in one of these fragments, in Campo Grande city, Mato Grosso do Sul state, Brazil. From winter 2012 until the autumn of 2015, using CO2 traps, we collect 1219 ticks, of which 1045 belonged to the species Amblyomma sculptum and 174 belonged to A. dubitatum. Both species were more abundant in spring (September to December) and summer (December to March). Rickettsial DNA was not detected in any of the specimens collected; however, one of the species found, A. sculptum is considered the main vector of Brazilian Spotted Fever, thus deserving constant vigilance by the public health organs.

Casos fatais de febre maculosa que foram relatados em alguns municípios brasileiros geralmente estão associados à presença de carrapatos e capivaras (Hydrochoerus hydrochaeris). Os fragmentos florestais urbanos onde este roedor está presente são de grande preocupação. Investigou-se aqui a diversidade de carrapatos livres e a ocorrência de carrapatos infectados por riquétsias em um desses fragmentos em Campo Grande, Mato Grosso do Sul. Desde o inverno de 2012 até o outono de 2015, usando armadilhas de CO2, foram coletados 1219 carrapatos, dos quais 1045 pertenciam à espécie Amblyomma sculptum e 174 pertenciam a A. dubitatum. Ambas as espécies foram mais abundantes na primavera (Setembro a Dezembro) e no verão (Dezembro a Março) . O DNA riquetsial não foi detectado em nenhum dos espécimes coletados; entretanto, uma das espécies encontradas, o A. sculptum é considerado o principal vetor da Febre Maculosa Brasileira, merecendo assim vigilância constante pelos Orgãos de Saúde Pública.

Species Identity Supersedes the Dilution Effect Concerning Hantavirus Prevalence at Sites across Texas and México.

Recent models suggest a relationship exists between community diversity and pathogen prevalence, the proportion of individuals in a population that are infected by a pathogen, with most inferences tied to assemblage structure. Two contrasting outcomes of this relationship have been proposed: the "dilution effect" and the "amplification effect." Small mammal assemblage structure in disturbed habitats often differs from assemblages in sylvan environments, and hantavirus prevalence is often negatively correlated with habitats containing high species diversity via dilution effect dynamics. As species richness increases, prevalence of infection often is decreased. However, anthropogenic changes to sylvan landscapes have been shown to decrease species richness and/or increase phylogenetic similarities within assemblages. Between January 2011 and January 2016, we captured and tested 2406 individual small mammals for hantavirus antibodies at 20 sites across Texas and México and compared differences in hantavirus seroprevalence, species composition, and assemblage structure between sylvan and disturbed habitats. We found 313 small mammals positive for antibodies against hantaviruses, evincing an overall prevalence of 9.7% across all sites. In total, 40 species of small mammals were identified comprising 2 taxonomic orders (Rodentia and Eulipotyphla). By sampling both habitat types concurrently, we were able to make real-world inferences into the efficacy of dilution effect theory in terms of hantavirus ecology. Our hypothesis predicting greater species richness higher in sylvan habitats compared to disturbed areas was not supported, suggesting the characteristics of assemblage structure do not adhere to current conceptions of species richness negatively influencing prevalence via a dilution effect.

Prevalencia de Borrelia burgdorferi sensu lato en roedores sinantrópicos de dos comunidades rurales de Yucatán, México / Prevalence of Borrelia burgdorferi sensu lato in synanthropic rodents in two rural communities of Yucatán, México

Introducción. La enfermedad de Lyme es una zoonosis multisistémica causada por Borrelia burgdorferi sensu lato. Esta espiroqueta circula en un ciclo enzoótico entre un reservorio vertebrado primario y las garrapatas. Se ha encontrado que varias especies de roedores son eficientes reservorios naturales de B. burgdorferi s.l. Objetivo. Estimar la prevalencia de B. burgdorferi s.l. en roedores sinantrópicos en dos comunidades rurales de Yucatán, México. Materiales y métodos. Se capturaron 123 roedores (94 Mus musculus y 29 Rattus rattus ) para obtener muestras de tejidos de oreja y vejiga. Para la detección de B. burgdorferi s.l. en las muestras, se amplificaron los genes de la flagelina B ( fla B ) y las lipoproteínas de membrana externa, ospC y p66 , mediante reacción en cadena de la polimerasa, y se secuenciaron los amplicones obtenidos. Resultados. La frecuencia de infección por B. burgdorferi s.l. en roedores fue de 36,5 % para flaB (45/123), de 10,5 % (13/123) para p66 y de 3,2 % (4/123) para ospC . En R. rattus la frecuencia de infección fue de 17,2 % y en M. musculus fue de 42,5 %. La frecuencia de infección de B. burgdorferi s.l. en los tejidos estudiados fue de 11,3 % (14/123) en muestras de tejido de vejiga y de 17,0 % (21/123) en las de oreja. No se encontraron diferencias estadísticas (p>0,05) en la frecuencia de infección entre los dos tipos de muestras de tejido utilizadas para el diagnóstico. El gen ospC presentó 98 % de homología con la especie Borrelia garinii , una de las especies heterogéneas del complejo B. burgdorferi s.l. Conclusiones. Los roedores presentaron una alta prevalencia de infección con B. burgdorferi s.l.; las especies M. musculus y R. rattus podrían jugar un papel importante en la continuidad de la presencia de esta bacteria en comunidades rurales de Yucatán, México.

Introduction: Lyme disease is a multisystemic zoonotic disease caused by Borrelia burgdorferi sensu lato. This spirochete circulates in an enzootic cycle between the primary vertebrate reservoir and its tick vectors. Different species of rodents are known to be efficient natural reservoirs for B. burgdorferi s.l. Objective: To estimate the prevalence of B. burgdorferi s.l. in synanthropic rodents from two rural communities of Yucatán, México. Materials and methods: A total of 123 rodents (94 Mus musculus and 29 Rattus rattus ) were trapped, and ear and bladder samples were collected. Flagelin B ( flaB ) genes and outer membrane lipoproteins ospC y p66 were amplified in order to detect B. burgdorferi s.l. presence in the samples. The obtained amplicons were sequenced. Results: The overall infection rates in rodents were 36.5% for flaB (45/123), 10.5% (13/123) for p66, and 3.2% (4/123) for ospC . Rattus rattus had 17.2% of infection and M. musculus , 42.5%. From all examined tissue, 11.3% (14/123) of bladders, and 17.0% (21/123) of ears were infected with the spirochete Borrelia burgdorferi s.l. No statistical differences (p>0.05) were found between the two tissue samples used for diagnosis. The ospC gen was 98% homologous to Borrelia garinii , one species of the B. burgdorferi s.l. complex. Conclusions: We concluded that rodents have a high prevalence of B. burgdorferi s.l. infection, and both species of rodents, M. musculus and R. rattus, might be playing an important role in the maintenance of this bacterium in rural communities of Yucatán, México.

Prevalence of Borrelia burgdorferi sensu lato in synanthropic rodents in two rural communities of Yucatán, México.

INTRODUCTION: Lyme disease is a multisystemic zoonotic disease caused by Borrelia burgdorferi sensu lato. This spirochete circulates in an enzootic cycle between the primary vertebrate reservoir and its tick vectors. Different species of rodents are known to be efficient natural reservoirs for B. burgdorferi s.l.  OBJECTIVE: To estimate the prevalence of B. burgdorferi s.l. in synanthropic rodents from two rural communities of Yucatán, México.  MATERIALS AND METHODS: A total of 123 rodents (94 Mus musculus and 29 Rattus rattus) were trapped, and ear and bladder samples were collected. Flagelin B (flaB) genes and outer membrane lipoproteins ospC y p66 were amplified in order to detect B. burgdorferi s.l. presence in the samples. The obtained amplicons were sequenced.  RESULTS: The overall infection rates in rodents were 36.5% for flaB (45/123), 10.5% (13/123) for p66, and 3.2% (4/123) for ospC. Rattus rattus had 17.2% of infection and M. musculus, 42.5%. From all examined tissue, 11.3% (14/123) of bladders, and 17.0% (21/123) of ears were infected with the spirochete Borrelia burgdorferi s.l. No statistical differences (p>0.05) were found between the two tissue samples used for diagnosis. The ospC gen was 98% homologous to Borrelia garinii, one species of the B. burgdorferi s.l. complex.  CONCLUSIONS: We concluded that rodents have a high prevalence of B. burgdorferi s.l. infection, and both species of rodents, M. musculus and R. rattus, might be playing an important role in the maintenance of this bacterium in rural communities of Yucatán, México.

Seroprevalence of Borrelia burgdorferi antibodies in white-tailed deer from Texas.

Lyme Disease is caused by the bacterial pathogen Borrelia burgdorferi, and is transmitted by the tick-vector Ixodes scapularis. It is the most prevalent arthropod-borne disease in the United States. To determine the seroprevalence of B. burgdorferi antibodies in white-tailed deer (Odocoileus virginianus) from Texas, we analyzed serum samples (n = 1493) collected during the 2001-2015 hunting seasons, using indirect ELISA. Samples with higher sero-reactivity (0.803 and above) than the negative control group (0.662) were further tested using a more specific standardized western immunoblot assay to rule out false positives. Using ELISA, 4.7% of the samples were sero-reactive against B. burgdorferi, and these originated in two eco-regions in Texas (Edwards Plateau and South Texas Plains). However, only 0.5% of the total samples were sero-reactive by standardized western immunoblot assay. Additionally, both ELISA and standardized western immunoblot assay results correlated with an increased incidence in human Lyme Disease cases reported in Texas. This is the first longitudinal study to demonstrate fluctuation in sero-reactivity of white-tailed deer to B. burgdorferi sensu stricto antigens in southern United States. Future ecological and geographical studies are needed to assess the environmental factors governing the prevalence of Lyme Disease in non-endemic areas of the southern United States.

Anti-Lyme Subunit Vaccines: Design and Development of Peptide-Based Vaccine Candidates.

Vaccinology today has been presented with several avenues to improve protection against infectious disease. The recent employment of the reverse vaccinology technique has changed the face of vaccine development against many pathogens, including Borrelia burgdorferi, the causative agent of Lyme disease. Using this technique, genomics and in silico analyses come together to identify potentially antigenic epitopes in a high-throughput fashion. The forward methodology of vaccine development was used previously to generate the only licensed human vaccine for Lyme disease, which is no longer on the market. Using reverse vaccinology to identify new antigens and isolate specific epitopes to protect against B. burgdorferi, subunit vaccines will be generated that lack reactogenic and nonspecific epitopes, yielding more effective vaccine candidates. Additionally, novel epitopes are being utilized and are presently in the commercialization pipeline both for B. burgdorferi and other spirochaetal pathogens. The versatility and methodology of the subunit protein vaccine are described as it pertains to Lyme disease from conception to performance evaluation.

TRANSLATING ECOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND POPULATION GENETICS RESEARCH TO MEET THE CHALLENGE OF TICK AND TICK-BORNE DISEASES IN NORTH AMERICA.

Emerging and re-emerging tick-borne diseases threaten public health and the wellbeing of domestic animals and wildlife globally. The adoption of an evolutionary ecology framework aimed to diminish the impact of tick-borne diseases needs to be part of strategies to protect human and animal populations. We present a review of current knowledge on the adaptation of ticks to their environment, and the impact that global change could have on their geographic distribution in North America. Environmental pressures will affect tick population genetics by selecting genotypes able to withstand new and changing environments and by altering the connectivity and isolation of several tick populations. Research in these areas is particularly lacking in the southern United States and most of Mexico with knowledge gaps on the ecology of these diseases, including a void in the identity of reservoir hosts for several tick-borne pathogens. Additionally, the way in which anthropogenic changes to landscapes may influence tick-borne disease ecology remains to be fully understood. Enhanced knowledge in these areas is needed in order to implement effective and sustainable integrated tick management strategies. We propose to refocus ecology studies with emphasis on metacommunity-based approaches to enable a holistic perspective addressing whole pathogen and host assemblages. Network analyses could be used to develop mechanistic models involving multihost-pathogen communities. An increase in our understanding of the ecology of tick-borne diseases across their geographic distribution will aid in the design of effective area-wide tick control strategies aimed to diminish the burden of pathogens transmitted by ticks.

Exploratory spatial analysis of Lyme disease in Texas -what can we learn from the reported cases?

BACKGROUND: Lyme disease (LD) is a tick-borne zoonotic illness caused by the bacterium Borrelia burgdorferi. Texas is considered a non-endemic state for LD and the spatial distribution of the state's reported LD cases is unknown. METHODS: We analyzed human LD cases reported to the Texas Department of State Health Services (TX-DSHS) between 2000 and 2011 using exploratory spatial analysis with the objective to investigate the spatial patterns of LD in Texas. Case data were aggregated at the county level, and census data were used as the population at risk. Empirical Bayesian smoothing was performed to stabilize the variance. Global Moran's I was calculated to assess the presence and type of spatial autocorrelation. Local Indicator of Spatial Association (LISA) was used to determine the location of spatial clusters and outliers. RESULTS AND DISCUSSION: There was significant positive spatial autocorrelation of LD incidence in Texas with Moran's I of 0.41 (p = 0.001). LISA revealed significant variation in the spatial distribution of human LD in Texas. First, we identified a high-risk cluster in Central Texas, in a region that is thought to be beyond the geographical range of the main vector, Ixodes scapularis. Second, the eastern part of Texas, which is thought to provide the most suitable habitat for I. scapularis, did not appear to be a high-risk area. Third, LD cases were reported from several counties in western Texas, a region considered unsuitable for the survival of Ixodes ticks. CONCLUSIONS: These results emphasize the need for follow-up investigations to determine whether the identified spatial pattern is due to: clustering of misdiagnosed cases, clustering of patients with an out-of state travel history, or presence of a clustered unknown enzootic cycle of B. burgdorferi in Texas. This would enable an improved surveillance and reporting of LD in Texas.

Absence of sodA Increases the Levels of Oxidation of Key Metabolic Determinants of Borrelia burgdorferi.

Borrelia burgdorferi, the causative agent of Lyme disease, alters its gene expression in response to environmental signals unique to its tick vector or vertebrate hosts. B. burgdorferi carries one superoxide dismutase gene (sodA) capable of controlling intracellular superoxide levels. Previously, sodA was shown to be essential for infection of B. burgdorferi in the C3H/HeN model of Lyme disease. We employed two-dimensional electrophoresis (2-DE) and immunoblot analysis with antibodies specific to carbonylated proteins to identify targets that were differentially oxidized in the soluble fractions of the sodA mutant compared to its isogenic parental control strain following treatment with an endogenous superoxide generator, methyl viologen (MV, paraquat). HPLC-ESI-MS/MS analysis of oxidized proteins revealed that several proteins of the glycolytic pathway (BB0057, BB0020, BB0348) exhibited increased carbonylation in the sodA mutant treated with MV. Levels of ATP and NAD/NADH were reduced in the sodA mutant compared with the parental strain following treatment with MV and could be attributed to increased levels of oxidation of proteins of the glycolytic pathway. In addition, a chaperone, HtpG (BB0560), and outer surface protein A (OspA, BBA15) were also observed to be oxidized in the sodA mutant. Immunoblot analysis revealed reduced levels of Outer surface protein C (OspC), Decorin binding protein A (DbpA), fibronectin binding protein (BBK32), RpoS and BosR in the sodA mutant compared to the control strains. Viable sodA mutant spirochetes could not be recovered from both gp91/phox-/- and iNOS deficient mice while borrelial DNA was detected in multiple tissues samples from infected mice at significantly lower levels compared to the parental strain. Taken together, these observations indicate that the increased oxidation of select borrelial determinants and reduced levels of critical pathogenesis-associated lipoproteins contribute to the in vivo deficit of the sodA mutant in the mouse model of Lyme disease. This study, utilizing the sodA mutant, has provided insights into adaptive capabilities critical for survival of B. burgdorferi in its hosts.

Prevalence of Borrelia burgdorferi-infected ticks from wildlife hosts, a response to Norris et al.

In a recent Letter to the Editor, Norris et al. questioned the validity of some of our data reported by Feria-Arroyo et al. The main issue investigated by us was the potential impact of climate change on the probable distribution of the tick vector Ixodes scapularis in the Texas-Mexico transboundary region. As an ancillary issue, an analysis of sequence data for the intergenic spacer of Borrelia burgdorferi was conducted. In the present letter, we provide further evidence supporting our original results, and advocate that extensive study of the population genetics of B. burgdorferi is needed in the Texas-Mexico transboundary region.

Pathogenic Landscape of Transboundary Zoonotic Diseases in the Mexico-US Border Along the Rio Grande.

Transboundary zoonotic diseases, several of which are vector borne, can maintain a dynamic focus and have pathogens circulating in geographic regions encircling multiple geopolitical boundaries. Global change is intensifying transboundary problems, including the spatial variation of the risk and incidence of zoonotic diseases. The complexity of these challenges can be greater in areas where rivers delineate international boundaries and encompass transitions between ecozones. The Rio Grande serves as a natural border between the US State of Texas and the Mexican States of Chihuahua, Coahuila, Nuevo León, and Tamaulipas. Not only do millions of people live in this transboundary region, but also a substantial amount of goods and people pass through it everyday. Moreover, it occurs over a region that functions as a corridor for animal migrations, and thus links the Neotropic and Nearctic biogeographic zones, with the latter being a known foci of zoonotic diseases. However, the pathogenic landscape of important zoonotic diseases in the south Texas-Mexico transboundary region remains to be fully understood. An international perspective on the interplay between disease systems, ecosystem processes, land use, and human behaviors is applied here to analyze landscape and spatial features of Venezuelan equine encephalitis, Hantavirus disease, Lyme Borreliosis, Leptospirosis, Bartonellosis, Chagas disease, human Babesiosis, and Leishmaniasis. Surveillance systems following the One Health approach with a regional perspective will help identifying opportunities to mitigate the health burden of those diseases on human and animal populations. It is proposed that the Mexico-US border along the Rio Grande region be viewed as a continuum landscape where zoonotic pathogens circulate regardless of national borders.

Implications of climate change on the distribution of the tick vector Ixodes scapularis and risk for Lyme disease in the Texas-Mexico transboundary region.

BACKGROUND: Disease risk maps are important tools that help ascertain the likelihood of exposure to specific infectious agents. Understanding how climate change may affect the suitability of habitats for ticks will improve the accuracy of risk maps of tick-borne pathogen transmission in humans and domestic animal populations. Lyme disease (LD) is the most prevalent arthropod borne disease in the US and Europe. The bacterium Borrelia burgdorferi causes LD and it is transmitted to humans and other mammalian hosts through the bite of infected Ixodes ticks. LD risk maps in the transboundary region between the U.S. and Mexico are lacking. Moreover, none of the published studies that evaluated the effect of climate change in the spatial and temporal distribution of I. scapularis have focused on this region. METHODS: The area of study included Texas and a portion of northeast Mexico. This area is referred herein as the Texas-Mexico transboundary region. Tick samples were obtained from various vertebrate hosts in the region under study. Ticks identified as I. scapularis were processed to obtain DNA and to determine if they were infected with B. burgdorferi using PCR. A maximum entropy approach (MAXENT) was used to forecast the present and future (2050) distribution of B. burgdorferi-infected I. scapularis in the Texas-Mexico transboundary region by correlating geographic data with climatic variables. RESULTS: Of the 1235 tick samples collected, 109 were identified as I. scapularis. Infection with B. burgdorferi was detected in 45% of the I. scapularis ticks collected. The model presented here indicates a wide distribution for I. scapularis, with higher probability of occurrence along the Gulf of Mexico coast. Results of the modeling approach applied predict that habitat suitable for the distribution of I. scapularis in the Texas-Mexico transboundary region will remain relatively stable until 2050. CONCLUSIONS: The Texas-Mexico transboundary region appears to be part of a continuum in the pathogenic landscape of LD. Forecasting based on climate trends provides a tool to adapt strategies in the near future to mitigate the impact of LD related to its distribution and risk for transmission to human populations in the Mexico-US transboundary region.

Immunization with a Borrelia burgdorferi BB0172-derived peptide protects mice against lyme disease.

Lyme disease is the most prevalent arthropod borne disease in the US and it is caused by the bacterial spirochete Borrelia burgdorferi (Bb), which is acquired through the bite of an infected Ixodes tick. Vaccine development efforts focused on the von Willebrand factor A domain of the borrelial protein BB0172 from which four peptides (A, B, C and D) were synthesized and conjugated to Keyhole Limpet Hemocyanin, formulated in Titer Max® adjuvant and used to immunize C3H/HeN mice subcutaneously at days 0, 14 and 21. Sera were collected to evaluate antibody responses and some mice were sacrificed for histopathology to evaluate vaccine safety. Twenty-eight days post-priming, protection was evaluated by needle inoculation of half the mice in each group with 10³ Bb/mouse, whereas the rest were challenged with 105Bb/mouse. Eight weeks post-priming, another four groups of similarly immunized mice were challenged using infected ticks. In both experiments, twenty-one days post-challenge, the mice were sacrificed to determine antibody responses, bacterial burdens and conduct histopathology. Results showed that only mice immunized with peptide B were protected against challenge with Bb. In addition, compared to the other the treatment groups, peptide B-immunized mice showed very limited inflammation in the heart and joint tissues. Peptide B-specific antibody titers peaked at 8 weeks post-priming and surprisingly, the anti-peptide B antibodies did not cross-react with Bb lysates. These findings strongly suggest that peptide B is a promising candidate for the development of a new DIVA vaccine (Differentiate between Infected and Vaccinated Animals) for protection against Lyme disease.

BB0172, a Borrelia burgdorferi outer membrane protein that binds integrin α3ß1.

Lyme disease is a multisystemic disorder caused by Borrelia burgdorferi infection. Upon infection, some B. burgdorferi genes are upregulated, including members of the microbial surface components recognizing adhesive matrix molecule (MSCRAMM) protein family, which facilitate B. burgdorferi adherence to extracellular matrix components of the host. Comparative genome analysis has revealed a new family of B. burgdorferi proteins containing the von Willebrand factor A (vWFA) domain. In the present study, we characterized the expression and membrane association of the vWFA domain-containing protein BB0172 by using in vitro transcription/translation systems in the presence of microsomal membranes and with detergent phase separation assays. Our results showed evidence of BB0172 localization in the outer membrane, the orientation of the vWFA domain to the extracellular environment, and its function as a metal ion-dependent integrin-binding protein. This is the first report of a borrelial adhesin with a metal ion-dependent adhesion site (MIDAS) motif that is similar to those observed in eukaryotic integrins and has a similar function.