Infected Ticks Research Articles

Sofosbuvir and its tri-phosphate metabolite inhibit the RNA-dependent RNA polymerase activity of non-structural protein 5 from the Kyasanur forest disease virus

Kyasanur forest disease is a neglected zoonotic disease caused by a single-stranded RNA-based flavivirus, the incidence of which was first recorded in 1957 in the Southern part of India. Kyasanur forest disease virus is transmitted to monkeys and humans through the infected tick bite of Haemophysalis spinigera. Kyasanur forest disease is a febrile illness, which in severe cases, results in neurological complications leading to mortality. The current treatment regimens are symptomatic and supportive, and no targeted therapies are available for this disease. In this study, we evaluated the ability of FDA-approved drugs sofosbuvir (and its active metabolite) and Dasabuvir to inhibit the RNA-dependent RNA polymerase activity of NS5 protein from the Kyasanur forest disease virus. NS5 protein containing the N-terminal methyl transferase domain and C-terminal RNA-dependent RNA polymerase domain was expressed in Escherichia coli, and RNA-dependent RNA polymerase activity was demonstrated with the purified protein. The RNA-dependent RNA polymerase assay conditions were optimized, followed by the determination of apparent Km,ATP to validate the enzyme preparation. Half maximal-inhibitory concentrations against RNA-dependent RNA polymerase activity were determined for Sofosbuvir and its active metabolite. Dasabuvir did not show detectable inhibition with the tested conditions. This is the first demonstration of the inhibition of RNA-dependent RNA polymerase activity of NS5 protein from the Kyasanur forest disease virus with small molecule inhibitors. These initial findings can potentially facilitate the discovery and development of targeted therapies for treating Kyasanur forest disease.

Low genetic diversity and population structuring of Amblyomma hebraeum and Rickettsia africae from coastal and inland regions in the Eastern Cape Province of South Africa.

Amblyomma hebraeum is the main vector of Rickettsia africae, the causative agent of African tick bite fever in southern Africa. Because pathogen dispersal is known to be influenced by tick adaptations to climate or host species, this study aimed to analyse the genetic diversity of A. hebraeum and R. africae infection of ticks collected from cattle in the Eastern Cape province of South Africa. DNA was extracted, amplified, and sequenced for the COI and ITS2 markers from A. hebraeum samples and the 17 kDa and ompA genes for rickettsial detection. Between six and ten haplotypes were identified from 40 COI and 31 ITS2 sequences; however, no population structuring was observed among sites (ΦST=0.22, p < 0.05). All A. hebraeum isolates clustered with southern Africa GenBank isolates. Rickettsia africae was detected in 46.92% (95% CI=41%-53%, n=260) of ticks. All R. africae isolates clustered with strain PELE and Chucks, which were reported previously from South Africa. These results confirm that A. hebraeum populations are undergoing a recent population expansion driven by cattle movement, facilitating local and long dispersal events across the Eastern Cape province.

Natural reservoir Rousettus aegyptiacus bat host model of orthonairovirus infection identifies potential zoonotic spillover mechanisms

The human-pathogenic Kasokero virus (KASV; genus Orthonairovirus) has been isolated from the sera of Egyptian rousette bats (ERBs; Rousettus aegyptiacus) captured in Uganda and unengorged Ornithodoros (Reticulinasus) faini ticks collected from the rock crevices of ERB colonies in South Africa and Uganda. Although evidence suggests that KASV is maintained in an enzootic transmission cycle between O. (R.) faini ticks and ERBs with potential for incidental virus spillover to humans through the bite of an infected tick, the vertebrate reservoir status of ERBs for KASV has never been experimentally evaluated. Furthermore, the potential for bat-to-bat and bat-to-human transmission of KASV is unknown. Herein, we inoculate two groups of ERBs with KASV; one group of bats is serially sampled to assess viremia, oral, fecal, and urinary shedding and the second group of bats is serially euthanized to assess virus-tissue tropism. Throughout the study, none of the bats exhibit overt signs of clinical disease. Following the detection of high KASV loads of long duration in blood, oral, fecal, and urine specimens collected from ERBs in the serial sampling group, all bats seroconvert to KASV. ERBs from the serial euthanasia group exhibit high KASV loads indicative of virus replication in the skin at the inoculation site, spleen, and inguinal lymph node tissue, and histopathology and in situ hybridization reveal virus replication in the liver and self-limiting, KASV-induced lymphohistiocytic hepatitis. The results of this study suggest that ERBs are competent, natural vertebrate reservoir hosts for KASV that can sustain viremias of appropriate magnitude and duration to support virus maintenance through bat-tick-bat transmission cycles. Viral shedding data suggests that KASV might also be transmitted bat-to-bat and highlights the potential for KASV spillover to humans through contact with infectious oral secretions, feces, or urine.

Ứng dụng kỹ thuật giải trình tự gen phát hiện Rickettsia gây bệnh sốt nổi mụn (Rickettsia SFG) từ mẫu ngoại ký sinh thu thập tại Hà Giang

Rickettsiosis is a zoonotic disease caused by Rickettsiae which are gram negative bacteria, obligate intracellular. These pathogens are transmitted to human through infected ticks, mites, fleas, lice, louse… In this study, The 17kDa and OmpA partial genes of Rickettsia were amplified to detect Rickettsial DNA in buffalo ticks and louse (Haematopinus tuberculatus) collected in Ha Giang province. The PCR products were sequenced, analyzed and build phylogenetic tree by Bioedit, Mega-X softwares; were compared and identified the reference strain sequences by BLAST- NCBI, USA. The results indicated that 14/25 tick samples and 2/14 louse samples were positive for Rickettsia SFG by PCR. Sequence analysis of OmpA gen showed that 14 clone isolated from the tick samples exhibites a 98% - 100% similarity to Rickettsia japonica, whereas 2 clone isolated from the louse samples were 98% similar to Rickettsia massiliae. Thus, our research showed that Rickettsia SFG including Rickettsia japonica and Rickettsia massiliae were detected in ticks and louse, respectively collected in Ha Giang province, Vietnam.

Northern piroplasmosis of domestic reindeer in Southern Yakutia

In South Yakutia, in the Aldansky district, at the end of July, more than 50 deer fell to the reindeer herd No. 7 of OAO KMNS "Khatystyr". Earlier in Central Yakutia, in 2008, an outbreak of pyroplasmosis was recorded for the first time in domestic reindeer of the farm of JSC NAOK "Taba" at the Tomtor site of the Gorny district and the YaGSHA Tabsylyn educational and production farm at the Kangkeme site of the Yakutsk city district. There was a mass death of the main deer population in the "Kangkeme" section up to 90% and "Tomtor" up to 30%. During laboratory studies, a parasitic disease, pyroplasmosis, was registered. Pyroplasmosis is one of the most severe and acute parasitic diseases of animals. Especially in the abnormal summer heat, blood parasites multiply very quickly, piroplasmids destroy red blood cells in the body of deer, the process of oxidation and intoxication occurs, bilirubin is formed from the released hemoglobin by complex transformations, 2-3 days after infection, in the absence of medical care, a lethal outcome is observed. Carriers of parasites can be ticks (Ixodidea, Ixodes ricinus) and other blood-sucking. Blood parasitic diseases (hemosporidiosis) of animals in Yakutia have not been previously recorded, a case of infection of domestic reindeer in 2008 in Central Yakutia is known. Isolated cases of tick attacks on people have been recorded in the Aldan, Neryungrinsky, Olekminsky and Suntarsky districts. A case of tick attack, infection and disease with tick-borne borreliosis of one person was registered. In the warm season, a distinctive feature of the temperature regime in most of the territory of Yakutia is a rapid increase in average daily temperatures in spring, and the maximum summer temperature can reach +38 ° C. At this time, there is a massive flight of blood-sucking insects and an attack of ticks on animals and people on the territory of Yakutia.

ИССЛЕДОВАНИЯ КЛЕЩЕЙ РОДА DERMACENTOR (ACARI; IXODIDAE) НА ЕСТЕСТВЕННУЮ ВСТРЕЧАЕМОСТЬ ВОЗБУДИТЕЛЯ ЛИХОРАДКИ КУ В УСЛОВИЯХ ЦЕНТРАЛЬНОГО ПРЕДКАВКАЗЬЯ

The purpose of the research: to assess the natural occurrence of Coxiella burnetii in ticks of the genus Dermacentor; clarify of the relationship between the physiological age of ticks and their infection with the Q fever patogen. Materials and methods: In the period from 2017 to 2019, we examined 4908 specimens of Dermacentor marginatus (616 pools) 4208 specimens of Dermacentor reticulatus (566 pools) and 43 specimens of Dermacentor niveus (18 pools) were examined for infection with the Q fever patogen. To assess the dependence of the detection of Coxiella burnetii in ticks depending on the physiological age, we examined 3048 specimens D. reticulatus (381 pools) and 2319 specimens D. marginatus (366 pools). The study of ixodid ticks for the presence of Coxiella burnetii DNA of the causative agent of Q fever was carried out by the molecular genetic method, namely the polymerase chain reaction (PCR), in accordance with the regulatory guidelines. Results and discussion: The results of the research showed that the infection of ixodid ticks with Coxiella in the Central Ciscaucasian natural populations on D. reticulatus and D. marginatus fluctuates over the years and does not exceed one percent. When considering the dependence of the detection of Coxiella burnetii in ticks on their physiological age, statistically significant results were obtained only for D. reticulatus. There is a significant correlation between the more frequent detection of infected ticks of physiological age II compared with ticks of physiological age III and IV. For D. marginatus ticks, there was no statistically significant relationship between the detection of Q fever pathogen and physiological age. Key words: ixodid ticks, Dermacentor reticulatus, Dermacentor marginatus, Dermacentor niveus, Q fever, physiological age.

Pathogen Detection in Ornithodoros sonrai Ticks and Invasive House Mice Mus musculus domesticus in Senegal.

Ornithodoros sonrai (O. sonrai) ticks are the only known vectors of Borrelia crocidurae, an agent of tick-borne relapsing fever (TBRF) borreliosis. Rodents serve as principal natural reservoirs for Borrelia. Our research objective was to detect TBRF Borrelia and other zoonotic bacterial infections in ticks and in house mice Mus musculus domesticus, an invasive species currently expanding in rural northern Senegal. Real-time and conventional PCR were utilized for detecting Borrelia and other bacterial taxa. The analyses were performed on 253 specimens of O. sonrai and 150 samples of brain and spleen tissue from rodents. Borrelia crocidurae was found in one O. sonrai tick and 18 Mus musculus domesticus samples, with prevalences of 0.39 percent and 12 percent, respectively, as well as Ehrlichia sp. in one Mus musculus domesticus. Further, we were able to detect the presence of a potentially infectious novel species belonging to the Anaplasmataceae family for the first time in O. sonrai ticks. More attention should be paid to the house mouse and O. sonrai ticks, as they can be potential hosts for novel species of pathogenic bacteria in humans.

Occurrence and genotyping of Theileria equi in dogs and associated ticks in Egypt.

Theileriosis is a tick-borne protozoal disease caused by a piroplasm of the genus Theileria. Hard ticks are obligate hematophagous ectoparasites that serve as vectors of Theileria spp. Studies of the presence of theileriosis in Egyptian dogs and associated ticks are scarce. This study was conducted to detect and identify Theileria spp. in dogs and Rhipicephalus sanguineus ticks and to monitor the epidemiological data of this disease. The prevalence rates of Theileria equi infection were 12.02%, 0.73%, 2.93%, and 1.83% by microscopic examination of dog blood, tick hemolymph, tick midgut, and tick salivary smears, respectively. Conversely, the T. equi prevalence in dog blood and associated ticks assessed by PCR was 25.81% and 10.42%, respectively. Epidemiological data about Theileria infection revealed a significant difference in the infection between different seasons and different dog breeds (p value <0.05), whereas host, sex, and age of dogs had no significant effect on the infection. Sequencing of PCR products showed that all PCR positive samples were infected with T. equi. Transmission electron microscopy (TEM) described the different stages of Theileria in the midgut and salivary gland of infected ticks. The current study confirmed that T. equi is not specific to equine hosts, and confirmed that dogs are a susceptible host to T. equi.

Anaplasma phagocytophilum Transmission Activates Immune Pathways While Repressing Wound Healing in the Skin.

Anaplasma phagocytophilum, the causative agent of human granulocytic anaplasmosis (HGA), is an obligate intracellular bacterium transmitted by the bite of black-legged ticks, Ixodes scapularis. The main host cells in vertebrates are neutrophils. However, the first site of entry is in the skin during tick feeding. Given that the initial responses within skin are a crucial determinant of disease outcome in vector-borne diseases, we used a non-biased approach to characterize the transcriptional changes that take place at the bite during I. scapularis feeding and A. phagocytophilum transmission. Experimentally infected ticks were allowed to feed for 3 days on C57BL/6J mice to allow bacterial transmission and establishment. Skin biopsies were taken from the attachment site of uninfected ticks and A. phagocytophilum-infected ticks. Skin without ticks (intact skin) was used as baseline. RNA was isolated and sequenced using next-generation sequencing (NGS). The differentially expressed genes were used to identify over-represented pathways by gene ontology (GO) and pathway enrichment (PE). Anaplasma phagocytophilum transmission resulted in the activation of interferon signaling and neutrophil chemotaxis pathways in the skin. Interestingly, it also led to the downregulation of genes encoding extracellular matrix (ECM) components, and upregulation of metalloproteinases, suggesting that A. phagocytophilum delays wound healing responses and may increase vascular permeability at the bite site.

Hematophagy and tick-borne Rickettsial pathogen shape the microbial community structure and predicted functions within the tick vector, Amblyomma maculatum.

Ticks are the primary vectors of emerging and resurging pathogens of public health significance worldwide. Analyzing tick bacterial composition, diversity, and functionality across developmental stages and tissues is crucial for designing new strategies to control ticks and prevent tick-borne diseases. Here, we explored the microbial communities across the developmental timeline and in different tissues of the Gulf-Coast ticks (Amblyomma maculatum). Using a high-throughput sequencing approach, the influence of blood meal and Rickettsia parkeri, a spotted fever group rickettsiae infection in driving changes in microbiome composition, diversity, and functionality was determined. This study shows that the core microbiome of Am. maculatum comprises ten core bacterial genera. The genus Rickettsia, Francisella, and Candidatus_Midichloria are the key players, with positive interactions within each developmental stage and adult tick organ tested. Blood meal and Rickettsia parkeri led to an increase in the bacterial abundance in the tissues. According to functional analysis, the increase in bacterial numbers is positively correlated to highly abundant energy metabolism orthologs with blood meal. Correlation analysis identified an increase in OTUs identified as Candidatus Midichloria and a subsequent decrease in Francisella OTUs in Rickettsia parkeri infected tick stages and tissues. Results demonstrate the abundance of Rickettsia and Francisella predominate in the core microbiome of Am. maculatum, whereas Candidatus_Midichloria and Cutibacterium prevalence increase with R. parkeri-infection. Network analysis and functional annotation suggest that R. parkeri interacts positively with Candidatus_Midichloria and negatively with Francisella. We conclude that tick-transmitted pathogens, such as R. parkeri establishes infection by interacting with the core microbiome of the tick vector.

Molecular detection of tick-borne rickettsial pathogens in ticks collected from domestic animals from Cauca, Colombia

Some hard ticks’ species can act as vectors of a wide variety of pathogens of human and animal importance such as Anaplasma, Ehrlichia and Rickettsia spp. In Colombia, a total of forty-six tick species have been described, and some of them have been implicated as vectors of some infectious agents. The department of Cauca is one of the thirty-two departments of Colombia. Most of its population lives in rural areas and depends on agriculture as the main economic activity, favoring exposure to ticks and tick-borne pathogens. Thus, the present study aimed to determine the tick species and tick-borne pathogens circulating in this region. From August to November 2017, ticks were collected from dogs, horses and cattle from eight rural areas of four municipalities in the department of Cauca. All collected ticks were classified according to taxonomic keys and organized in pools. DNA was extracted from all tick pools for molecular confirmation of tick species and detection of Anaplasma, Ehrlichia and Rickettsia spp. A total of 2809 ticks were collected which were grouped in 602 pools. Ticks were morphologically identified as Amblyomma cajennense sensu lato, Dermacentor nitens, Rhipicephalus microplus and Rhipicephalus sanguineus sensu lato. The molecular identity of A. cajennense s.l. was confirmed as Amblyomma patinoi. A total of 95% of the pools scored positive for members of the Anaplasmataceae family, of which, 7.8% and 7.3% were positive to Anaplasma and Ehrlichia spp., respectively, being identified as Anaplasma marginale, Ehrlichia minasensis and Ehrlichia canis; and 16.1% were positive for Rickettsia spp. with high identity for Rickettsia asembonensis, Rickettsia felis and Candidatus Rickettsia senegalensis. This is the first report describing the natural infection of ticks with rickettsial pathogens and the occurrence of A. patinoi ticks in Cauca department, Colombia.

Integrated human behavior and tick risk maps to prioritize Lyme disease interventions using a 'One Health' approach

Lyme disease (LD) risk is emerging rapidly in Canada due to range expansion of its tick vectors, accelerated by climate change. The risk of contracting LD varies geographically due to variability in ecological characteristics that determine the hazard (the densities of infected host-seeking ticks) and vulnerability of the human population determined by their knowledge and adoption of preventive behaviors. Risk maps are commonly used to support public health decision-making on Lyme disease, but the ability of the human public to adopt preventive behaviors is rarely taken into account in their development, which represents a critical gap. The objective of this work was to improve LD risk mapping using an integrated social-behavioral and ecological approach to: (i) compute enhanced integrated risk maps for prioritization of interventions and (ii) develop a spatially-explicit assessment tool to examine the relative contribution of different risk factors.The study was carried out in the Estrie region located in southern Québec. The blacklegged tick, Ixodes scapularis, infected with the agent of LD is widespread in Estrie and as a result, regional LD incidence is the highest in the province. LD knowledge and behaviors in the population were measured in a cross-sectional health survey conducted in 2018 reaching 10,790 respondents in Estrie. These data were used to create an index for the social-behavioral component of risk in 2018. Local Empirical Bayes estimator technique were used to better quantify the spatial variance in the levels of adoption of LD preventive activities. For the ecological risk analysis, a tick abundance model was developed by integrating data from ongoing long-term tick surveillance programs from 2007 up to 2018. Social-behavioral and ecological components of the risk measures were combined to create vulnerability index maps and, with the addition of human population densities, prioritization index maps. Map predictions were validated by testing the association of high-risk areas with the current spatial distribution of human cases of LD and reported tick exposure. Our results demonstrated that social-behavioral and ecological components of LD risk have markedly different distributions within Estrie. The occurrence of human LD cases or reported tick exposure in a municipality was positively associated with tick density and the prioritization risk index (p < 0.001).This research is a second step towards a more comprehensive integrated LD risk assessment approach, examining social-behavioral risk factors that interact with ecological risk factors to influence the management of emerging tick-borne diseases, an approach that could be applied more widely to vector-borne and zoonotic diseases.

Use of the Western blot technique to identify the immunogenic proteins of Borrelia burgdorferi for developing a Lyme disease vaccine

BackgroundLyme disease is a serious infectious disease having a restricted worldwide distribution for which there is no vaccine available for human use. ObjectiveThis study was designed to determine common reactive antigens involved in Borrelia burgdorferi (Bb) infection that are recognized in mammalian sera that may be useful for vaccine development. MethodsBlood samples were collected from patients with documented Lyme disease, and from rabbits and mice experimentally infected with either tick-transmitted or culture-grown Borrelia burgdorferi. All samples were then processed for sera. For performing the Western blots, sonicated Bb organisms (whole cell lysates) and protein ladders were separated by protein gel electrophoresis. Immune reactivities of the electrophoresed proteins with the serum samples were then probed with anti-HRP IgG reagent. ResultsRabbit, mouse and human sera consistently reacted with the 41 kDa band of Bb which corresponded to the flagellin protein – the major protein component of this organism’s periplasmic flagella, also known as axial filaments or fibrils. Various other Bb antigens of wide molecular weight ranges were also recognized by rabbit and human sera, and less frequently with mouse sera. ConclusionThe strong immune response to the 41 kDa flagellin protein by the different mammalian species suggests the utility of a possible vaccine targeting this protein, although other proteins may also be appropriate, for preventing Lyme disease following a bite from an infected tick.

Seroepidemiology of Borrelia burgdorferi s.l. among German National Cohort (NAKO) Participants, Hanover.

Lyme borreliosis is the leading tick-related illness in Europe, caused by Borrelia Burgdorferi s.l. Lower Saxony, Germany, including its capital, Hanover, has a higher proportion of infected ticks than central European countries, justifying a research focus on the potential human consequences. The current knowledge gap on human incident infections, particularly in Western Germany, demands serological insights, especially regarding a potentially changing climate-related tick abundance and activity. We determined the immunoglobulin G (IgG) and immunoglobulin M (IgM) serostatuses for 8009 German National Cohort (NAKO) participants from Hanover, examined in 2014-2018. We used an enzyme-linked immunosorbent assay (ELISA) as the screening and a line immunoblot as confirmation for the Borrelia Burgdorferi s.l. antibodies. We weighted the seropositivity proportions to estimate general population seropositivity and estimated the force of infection (FOI). Using logistic regression, we investigated risk factors for seropositivity. Seropositivity was 3.0% (IgG) and 2.1% (IgM). The FOI varied with age, sharply increasing in participants aged ≥40 years. We confirmed advancing age and male sex as risk factors. We reported reduced odds for seropositivity with increasing body mass index and depressive symptomatology, respectively, pointing to an impact of lifestyle-related behaviors. The local proportion of seropositive individuals is comparable to previous estimates for northern Germany, indicating a steady seroprevalence.

County-level surveillance for the lone star tick, Amblyomma americanum, and its associated pathogen, Ehrlichia chaffeensis, in Kentucky

Kentucky experiences some of the highest incidence rates for ehrlichiosis nationwide. Ehrlichiosis is a bacterial infection caused primarily by the pathogen Ehrlichia chaffeensis and can be transmitted to humans through the bite of an infected tick, notably Amblyomma americanum. Amblyomma americanum, the lone star tick, is common to Kentucky and much of the southeast, but has expanded farther north in recent years. As an abundant and aggressive nondiscriminatory biter, this species is of major public health concern for transmission of pathogens to humans. As this vector's range expands, surveillance remains a necessary tool providing data that allows researchers to track this expansion over time. The historical information on tick distribution in Kentucky is variable with very little data on a statewide scale. From January 2019 to December 2020, we conducted surveillance for A. americanum in Kentucky through field collections and the establishment of a statewide tick submission program with the help of the Kentucky Department for Public Health and screened for E. chaffeensis on a county-level throughout the state. We collected 5,726 A. americanum ticks in 77 counties and detected E. chaffeensis in 32 counties. The minimum infection rate was 1.8%. With the expansion of A. americanum and increasing cases of tick-borne diseases, future surveillance is needed to monitor this important tick vector over time.

Transmission Cycle of Tick-Borne Infections and Co-Infections, Animal Models and Diseases.

Tick-borne pathogens such as species of Borrelia, Babesia, Anaplasma, Rickettsia, and Ehrlichia are widespread in the United States and Europe among wildlife, in passerines as well as in domestic and farm animals. Transmission of these pathogens occurs by infected ticks during their blood meal, carnivorism, and through animal bites in wildlife, whereas humans can become infected either by an infected tick bite, through blood transfusion and in some cases, congenitally. The reservoir hosts play an important role in maintaining pathogens in nature and facilitate transmission of individual pathogens or of multiple pathogens simultaneously to humans through ticks. Tick-borne co-infections were first reported in the 1980s in white-footed mice, the most prominent reservoir host for causative organisms in the United States, and they are becoming a major concern for public health now. Various animal infection models have been used extensively to better understand pathogenesis of tick-borne pathogens and to reveal the interaction among pathogens co-existing in the same host. In this review, we focus on the prevalence of these pathogens in different reservoir hosts, animal models used to investigate their pathogenesis and host responses they trigger to understand diseases in humans. We also documented the prevalence of these pathogens as correlating with the infected ticks' surveillance studies. The association of tick-borne co-infections with other topics such as pathogens virulence factors, host immune responses as they relate to diseases severity, identification of vaccine candidates, and disease economic impact are also briefly addressed here.

Rickettsial infection in ticks from a natural area of Atlantic Forest biome in southern Brazil.

From June 2013 to January 2014, blood sera samples and ticks were collected from domestic dogs and wild small mammals, and ticks from the vegetation in a preservation area of the Atlantic Forest biome (Turvo State Park), and the rural area surrounding the Park in Derrubadas municipality, state of Rio Grande do Sul, southern Brazil. Dogs were infested by Amblyomma ovale and Amblyomma aureolatum adult ticks, whereas small mammals were infested by immature stages of A. ovale, Amblyomma yucumense, Amblyomma brasiliense, Ixodes loricatus, and adults of I. loricatus. Ticks collected on vegetation were A. brasiliense, A. ovale, A. yucumense, Amblyomma incisum, and Haemaphysalis juxtakochi. Three Rickettsia species were molecularly detected in ticks: Rickettsia bellii in I. loricatus (also isolated through cell culture inoculation), Rickettsia amblyommatis in A. brasiliense, and Rickettsia rhipicephali in A. yucumense. The latter two are tick-rickettsia associations reported for the first time. Seroreactivity to Rickettsia antigens were detected in 33.5% (55/164) small mammals and 8.3% (3/36) canine sera. The present study reveals a richness of ticks and associated-rickettsiae in the largest Atlantic Forest Reserve of the state of Rio Grande do Sul, which is characterized by a rich fauna of wild mammals, typical of more preserved areas of this biome. Noteworthy, none of the detected Rickettsia species have been associated to human or animal diseases. This result contrasts to other areas of this biome in Brazil, which are endemic for tick-borne spotted fever caused by Rickettsia rickettsii or Rickettsia parkeri.

Efficacy and Immune Correlates of OMP-1B and VirB2-4 Vaccines for Protection of Dogs from Tick Transmission of Ehrlichia chaffeensis.

Ehrlichia chaffeensis, an obligatory intracellular bacterium, causes human monocytic ehrlichiosis, an emerging disease transmitted by the Lone Star tick, Amblyomma americanum. Here, we investigated the vaccine potential of OMP-1B and VirB2-4. Among the highly expressed and immunodominant E. chaffeensis porin P28s/OMP-1s, OMP-1B is predominantly expressed by E. chaffeensis in A. americanum ticks, whereas VirB2-4 is a pilus protein of the type IV secretion system essential for E. chaffeensis infection of host cells. Immunization with recombinant OMP-1B (rOMP-1B) or recombinant VirB2-4 (rVirB2-4) protected mice from E. chaffeensis infection as effectively as Entry-triggering protein of Ehrlichia immunization. Dogs vaccinated with a nanoparticle vaccine composed of rOMP-1B or rVirB2-4 and an immunostimulating complex developed high antibody titers against the respective antigen. Upon challenge with E. chaffeensis-infected A. americanum ticks, E. chaffeensis was undetectable in the blood of rOMP-1B or rVirB2-4 immunized dogs on day 3 or 6 post-tick attachment and for the duration of the experiment, whereas dogs sham-vaccinated with the complex alone were persistently infected for the duration of the experiment. E. chaffeensis exponentially replicates in blood-feeding ticks to facilitate transmission. Previously infected ticks removed from OMP-1B-immunized dogs showed significantly lower bacterial load relative to ticks removed from sham-immunized dogs, suggesting in-tick neutralization. Peripheral blood leukocytes from rVirB2-4-vaccinated dogs secreted significantly elevated amounts of interferon-γ soon after tick attachment by ELISpot assay and reverse transcription-quantitative PCR, suggesting interferon-γ-mediated Ehrlichia inhibition. Thus, Ehrlichia surface-exposed proteins OMP-1B and VirB2-4 represent new potential vaccine candidates for blocking tick-borne ehrlichial transmission. IMPORTANCE Ehrlichia are tick-borne pathogens that cause a potentially fatal illness-ehrlichiosis-in animals and humans worldwide. Currently, no vaccine is available for ehrlichiosis, and treatment options are limited. Ticks are biological vectors of Ehrlichia, i.e., Ehrlichia exponentially replicates in blood-sucking ticks before infecting animals. Ticks also inoculate immunomodulatory substances into animals. Thus, it is important to study effects of candidate vaccines on Ehrlichia infection in both animals and ticks and the immune responses of animals shortly after infected tick challenge. Here, we investigated the efficacy of vaccination with functionality-defined two surface-exposed outer membrane proteins of Ehrlichia chaffeensis, OMP-1B and VirB2-4, in a mouse infection model and then in a dog-tick transmission model. Our results begin to fill gaps in our understanding of Ehrlichia-derived protective antigens against tick-transmission and immune correlates and mechanisms that could help future development of vaccines for immunization of humans and animals to counter tick-transmitted ehrlichiosis.

Minimal Rickettsial Infection Rates and Distribution of Ticks in Uganda: An Assessment of the Seasonal Effects and Relevance to Tick-Borne Disease Risk in East Africa.

Tick-borne diseases (TBDs) pose a significant risk to humans and represent one of the major factors influencing readiness within the United States' military worldwide. Additionally, ticks and TBDs constitute major animal health problems leading to economic losses at multiple levels affecting low- and middle-income countries the hardest. Tick control is frequently hampered by issues ranging from acaricide resistance to lack of data on tick distribution and infection rates. We conducted a cross-sectional study to assess tick species distribution, host use, and rickettsial pathogen infection rate of ticks in different areas of the Uganda Cattle Corridor. We identified 4,425 hard ticks (Ixodida: Ixodidae) comprised of seven species by morphological characters with 3,315 ticks collected from four locations during the dry season and 1,110 ticks from one location during the wet season. Rickettsial pathogen prevalence was assessed in ticks collected from two districts to determine the minimum infection rate compared across seasons, village location, and tick species. We found statistically significant differences in the abundance and distribution of tick species among districts in the dry season, host animal species, and the proportion of rickettsial positive pools between villages. Seasonality, village location, and tick species do not affect the minimum infection rate of rickettsial pathogens of ticks in Uganda, but village location affects the proportion of positive tick pools. These results indicate geographical and seasonal differences among pathogen-harboring ticks contributing to our understanding of the current distribution of ticks and TBDs in Uganda.

Tick-Borne Encephalitis (TBE) - Fundamentals

Tick-Borne Encephalitis (TBE) - Fundamentals Abstract. There is widespread endemicity of tick-borne encephalitis (TBE) in all German-speaking countries. In most regions there is a gradual increase in incidence and further territorial spread. As a proportion of the cases is not diagnosed and since the clinical course is getting worse with growing age, substantial underreporting occurs particularly in the pediatric and adolescent patient population. Infected ticks are transmitting the TBE virus, a flavivirus, within a few minutes after the bite. After an incubation period of 4 to 28 days uncharacteristic flu-like symptoms usually occur during a first viremic phase. This is followed by a brief asymptomatic interval before a minority of patients suffer of a variety of symptoms associated with damage of different parts of the central nervous system. This often results in permanent neurological injury and in about 1% the clinical course is fatal. To establish a diagnosis, it is essential to suspect the infection and to obtain a history of exposure. Depending on the phase of illness IgM and/or IgG antibodies can be detected in the serum and/or in the cerebrospinal fluid. In the early phase of the infection the TBE virus may be detected by PCR.