Zoonotic Tick-borne Pathogens Research Articles

Seromolecular survey and risk factor analysis of Crimean-Congo haemorrhagic fever orthonairovirus in occupationally exposed herdsmen and unexposed febrile patients in Kwara State, Nigeria.

Crimean-Congo haemorrhagic fever virus (CCHFV) is a globally significant tick-borne zoonotic pathogen that causes fatal haemorrhagic disease in humans. Despite constituting an ongoing public health threat, limited research exists on the presence of CCHFV among herdsmen, an occupationally exposed population that has prolonged contact with ruminants and ticks. This cross-sectional study, conducted between October 2018 and February 2020 in Kwara State, Nigeria, was aimed at assessing CCHFV seroprevalence among herdsmen and non-herdsmen febrile patients, and identifying the associated risk factors. Blood samples from herdsmen (n = 91) and febrile patients in hospitals (n = 646) were analyzed for anti-CCHFV IgG antibodies and CCHFV S-segment RNA using ELISA and RT-PCR, respectively. Results revealed a remarkably high CCHFV seroprevalence of 92.3% (84/91) among herdsmen compared to 7.1% (46/646) in febrile patients. Occupational risk factors like animal and tick contact, tick bites, and hand crushing of ticks significantly contributed to higher seroprevalence in the herdsmen (p<0.0001). Herdsmen were 156.5 times more likely (p<0.0001) to be exposed to CCHFV than febrile patients. Notably, the odds of exposure were significantly higher (OR = 191.3; p<0.0001) in herdsmen with a history of tick bites. Although CCHFV genome was not detectable in the tested sera, our findings reveal that the virus is endemic among herdsmen in Kwara State, Nigeria. CCHFV should be considered as a probable cause of febrile illness among humans in the study area. Given the nomadic lifestyle of herdsmen, further investigations into CCHF epidemiology in this neglected population are crucial. This study enhances our understanding of CCHFV dynamics and emphasizes the need for targeted interventions in at-risk communities.

Frequency of occurrence of zoonotic tick-borne pathogens in ticks – Latvian 2023 year experience

Frequency of occurrence of zoonotic tick-borne pathogens in ticks – Latvian 2023 year experience

Mapping the risk of exposure to Crimean-Congo haemorrhagic fever virus in the Iberian Peninsula using Eurasian wild boar (Sus scrofa) as a model

Crimean-Congo haemorrhagic fever (CCHF) virus (CCHFV) is a tick-borne zoonotic pathogen that can cause a lethal haemorrhagic disease in humans. Although the virus appears to be endemically established in the Iberian Peninsula, CCHF is an emerging disease in Spain. Clinical signs of CCHFV infection are mainly manifested in humans, but the virus replicates in several animal species. Understanding the determinants of CCHFV exposure risk from animal models is essential to predicting high-risk exposure hotspots for public health action. With this objective in mind, we designed a cross-sectional study of Eurasian wild boar (Sus scrofa) in Spain and Portugal. The study analysed 5,291 sera collected between 2006 and 2022 from 90 wild boar populations with a specific double-antigen ELISA to estimate CCHFV serum prevalence and identify the main determinants of exposure probability. To do so, we statistically modelled exposure risk with host- and environment-related predictors and spatially projected it at a 10 × 10 km square resolution at the scale of the Iberian Peninsula to map foci of infection risk. Fifty-seven (63.3 %) of the 90 populations had at least one seropositive animal, with seroprevalence ranging from 0.0 to 88.2 %. Anti-CCHFV antibodies were found in 1,026 of 5,291 wild boar (19.4 %; 95 % confidence interval: 18.3–20.5 %), with highest exposure rates in southwestern Iberia. The most relevant predictors of virus exposure risk were wild boar abundance, local rainfall regime, shrub cover, winter air temperature and soil temperature variation. The spatial projection of the best-fit model identified high-risk foci as occurring in most of western and southwestern Iberia and identified recently confirmed risk foci in eastern Spain. The results of the study demonstrate that serological surveys of CCHFV vector hosts are a powerful, robust and highly informative tool for public health authorities to take action to prevent human cases of CCHF in enzootic and emergency settings.

Ticks and Tick-Borne Zoonotic Pathogens from Wild Birds in Northwestern Coastal Spain

Migratory and local birds may disperse ticks and their associated pathogens. The aim of this study was to provide information regarding ticks infesting birds in Asturias, a region that accounts for most of the Lyme disease hospitalizations in Spain. From September 2021 to April 2023, trained and experienced bird-banders collected ticks from birds in two estuary and four forest locations. A total of 1698 birds (52 species, 38 genera, and 26 families) were captured. A total of 51 ticks (28 larvae, 20 nymphs, and 3 females) were collected from 43 birds, belonging to three species: Ixodes ricinus (31), Ixodes frontalis (18), and Haemaphysalis concinna (1). The average prevalence of tick infestation was 2.5% and the average tick burden was 1.2 ticks per infested host. The bird species Turdus merula, Parus major, Luscinia svecica, and Anthus pratensis were among the most infested. Anaplasma phagocytophilum was detected in one I. ricinus nymph collected from Phylloscopus collybita. We have not detected Borrelia burgdorferi s.l., Rickettsia spp., Coxiella burnetii, or piroplasmids in any of the 51 analyzed ticks. These results suggest low infestation rate in migratory/estuary birds and a higher rate in forest/sedentary ones. Despite this, the detection of pathogens, although with low prevalence, can pose a risk to public health.

Wild ungulates as sentinels of flaviviruses and tick-borne zoonotic pathogen circulation: an Italian perspective

BackgroundVector-borne zoonotic diseases are a concerning issue in Europe. Lyme disease and tick-borne encephalitis virus (TBEV) have been reported in several countries with a large impact on public health; other emerging pathogens, such as Rickettsiales, and mosquito-borne flaviviruses have been increasingly reported. All these pathogens are linked to wild ungulates playing roles as tick feeders, spreaders, and sentinels for pathogen circulation. This study evaluated the prevalence of TBEV, Borrelia burgdorferi sensu lato, Rickettsia spp., Ehrlichia spp., and Coxiella spp. by biomolecular screening of blood samples and ticks collected from wild ungulates. Ungulates were also screened by ELISA and virus neutralization tests for flaviviral antibody detection.ResultsA total of 274 blood samples were collected from several wild ungulate species, as well as 406 Ixodes ricinus, which were feeding on them. Blood samples tested positive for B. burgdorferi s.l. (1.1%; 0-2.3%) and Rickettsia spp. (1.1%; 0-2.3%) and showed an overall flaviviral seroprevalence of 30.6% (22.1–39.2%): 26.1% (17.9–34.3%) for TBEV, 3.6% (0.1–7.1%) for Usutu virus and 0.9% (0-2.7%) for West Nile virus. Ticks were pooled when possible and yielded 331 tick samples that tested positive for B. burgdorferi s.l. (8.8%; 5.8–11.8%), Rickettsia spp. (26.6%; 21.8–31.2%) and Neoehrlichia mikurensis (1.2%; 0-2.4%). TBEV and Coxiella spp. were not detected in either blood or tick samples.ConclusionsThis research highlighted a high prevalence of several tick-borne zoonotic pathogens and high seroprevalence for flaviviruses in both hilly and alpine areas. For the first time, an alpine chamois tested positive for anti-TBEV antibodies. Ungulate species are of particular interest due to their sentinel role in flavivirus circulation and their indirect role in tick-borne diseases and maintenance as Ixodes feeders and spreaders.

Spotted Fever Group Rickettsiae in Ticks and Small Mammals from Grassland and Forest Habitats in Central Germany.

Rickettsiae of the spotted fever group (SFG) are zoonotic tick-borne pathogens. Small mammals are important hosts for the immature life stages of two of the most common tick species in Europe, Ixodes ricinus and Dermacentor reticulatus. These hosts and vectors can be found in diverse habitats with different vegetation types like grasslands and forests. To investigate the influence of environmental and individual factors on Rickettsia prevalence, this study aimed to analyse the prevalence of SFG rickettsiae in ticks and small mammals in different small-scale habitats in central Germany for the first time. Small mammals of ten species and ticks of two species were collected from grasslands and forests in the Hainich-Dün region, central Germany. After species identification, DNA samples from 1098 ticks and ear snips of 1167 small mammals were screened for Rickettsia DNA by qPCR targeting the gltA gene. Positive samples were retested by conventional PCR targeting the ompB gene and sequencing. Rickettsia DNA was detected in eight out of ten small mammal species. Small mammal hosts from forests (14.0%) were significantly more often infected than those from grasslands (4.4%) (p < 0.001). The highest prevalence was found in the mostly forest-inhabiting genus Apodemus (14.8%) and the lowest in Microtus (6.6%), which inhabits grasslands. The prevalence was higher in D. reticulatus (46.3%) than in the I. ricinus complex (8.6%). Adult ticks were more often infected than nymphs (p = 0.0199). All sequenced rickettsiae in I. ricinus complex ticks were R. helvetica, and the ones in D. reticulatus were R. raoultii. Unlike adults, questing nymphs have had only one blood meal, which explains the higher prevalence in I. ricinus adults. Interestingly, habitat type did influence infection probability in small mammals, but did not in ticks. A possible explanation may be the high prevalence in Apodemus flavicollis and A. sylvaticus which were more abundant in the forest.

­­Primary Detection of the Establishment of Blacklegged Ticks, Ixodes scapularis, in British Columbia, Canada

Ticks transport and transmit microbial pathogens that inflict malevolent diseases on domestic and wildlife animals, and humans. We reveal the first record of the blacklegged tick, Ixodes scapularis, in British Columbia (BC) and, concurrently, far western North America. We unveil the primary tick-host record of I. scapularis parasitizing a Mallard duck, Anas platyrhynchos. In our study, the most pronounced Ixodes species was I. scapularis (61%) followed by the western blacklegged tick, Ixodes pacificus (34%). The most frequently occurring mammalian host parasitized by I. scapularis was the eastern cottontail, Sylvilagus floridanus, a lagomorph of grassland habitats. Healthcare professionals must be aware that both I. pacificus, and I. scapularis bite humans in BC, and transmit at least six tick-borne human zoonotic pathogens that cause insidious diseases.

Study of tick-borne zoonotic pathogens in questing and feeding ticks in Tenerife, Canary Islands, Spain.

Ticks are vectors of many zoonotic pathogens of clinical relevance, including Anaplasma and Rickettsia species. Since few data about these tick-borne pathogens are available in the Canary Islands, the aim of the present study was to screen their presence in questing and feeding ticks on the island of Tenerife. A total of 81 ticks was removed from six hedgehogs, and eight ticks were collected from the grass. PCR assays for tick species identification based on the LSU rRNA gene, and detection of Anaplasma spp. and Rickettsia spp. by targeting the 16s rRNA and gltA gene, respectively, were carried out. Rhipicephalus sanguineus and Haemaphysalis formosensis tick species were identified, with Rickettsia hoogstraalii detected in H. formosensis. In addition, Rickettsia helvetica and one unidentified Rickettsia species were detected. The overall prevalence of Rickettsia in ticks was 12.2%, and none harbored Anaplasma sp. This work constitutes the first identification in the Canary Islands of pathogenic R. helvetica and the species of unknown pathogenicity R. hoogstraalii in questing ticks. The clinical relevance of the pathogens identified highlights the need for studies with increased sample size and locations, including potential hosts, as well as warning the population about the relevance of ticks as vectors.

Clinical usefulness of metagenomic next-generation sequencing for Rickettsia and Coxiella burnetii diagnosis

Rickettsia and Coxiella burnetii are zoonotic tick-borne pathogens that cause febrile illnesses in humans. Metagenomic next-generation sequencing (mNGS) is a new technology used to diagnose infectious diseases. However, clinical experience with applying the test to rickettsioses and Q fever is relatively limited. Therefore, this study aimed to explore the diagnostic performance of mNGS in detecting Rickettsia and C. burnetii. We retrospectively studied patients with rickettsioses or Q fever between August 2021 and July 2022. Peripheral blood mNGS and polymerase chain reaction (PCR) were performed for all patients. Clinical data were retrieved for analysis. Thirteen patients were included in this study (eleven confirmed cases and two suspected cases). Signs and symptoms included fever (13, 100%), rash (7, 53.8%), muscle soreness (5, 38.5%), headache (4, 30.8%), skin eschar (3, 23.1%), and disturbance of consciousness (2, 15.4%). In addition, eight patients (61.6%) had thrombocytopenia, ten (76.9%) had liver function impairment, and two (15.4%) had renal function impairment. The results of mNGS revealed seven patients with R. japonica (53.8%), five with C. burneti (38.5%), two with R. heilongjiangensis (15.4%), and one with R. honei (7.7%). PCR results were positive in 11 patients (84.6%). After receiving doxycycline-based treatment, 12 (92.3%) patients returned to a normal temperature within 72 h. All patients were discharged in better health. Therefore, mNGS can help diagnose Rickettsia and C. burnetii and shorten the diagnosis time, especially for patients with atypical clinical manifestations and unclear epidemiologic evidence of a tick bite or exposure.

Migratory Songbirds Transport Amblyomma longirostre and Amblyomma maculatum Ticks to Canada

Birds transport ticks into Canada during northward spring migration, and some of these ticks are infected with tick-borne zoonotic pathogens. Some Amblyomma species harbour pathogens that cause debilitating diseases that can be fatal to humans, and domestic and wildlife animals. At least 65 Amblyomma spp. are indigenous in the Western Hemisphere, and approximately half bite humans. Amblyomma longirostre carries Rickettsia amblyommatis which causes spotted fever group rickettsiosis, a febrile disease in humans. Additionally, Amblyomma maculatum harbors and transmits Rickettsia parkeri, a spotted fever group rickettsiosis, and this tick bites humans. In the present study, we use two technologies to identify ticks. To confirm identification, we took microphotographs followed by DNA barcoding of the cytochrome c oxidase I gene. Based on molecular analysis, we confirmed that the two Amblyomma spp. were Amblyomma longirostre, a neotropical tick and Amblyomma maculatum, the Gulf Coast tick. Based on our tick-bird findings, we confirm that migratory songbirds transport Amblyomma ticks into Canada, and have the potential, either directly or indirectly, to transmit tick-borne zoonotic pathogens to humans.

Occurrence of Rickettsia spp., Hantaviridae, Bartonella spp. and Leptospira spp. in European Moles (Talpa europaea) from the Netherlands.

The European mole (Talpa europaea) has a widespread distribution throughout Europe. However, little is known about the presence of zoonotic pathogens in European moles. We therefore tested 180 moles from the middle and the south of the Netherlands by (q)PCR for the presence of multiple (tick-borne) zoonotic pathogens. Spotted fever Rickettsia was found in one (0.6%), Leptospira spp. in three (1.7%), Bartonella spp. in 69 (38.3%) and Hantaviridae in 89 (49.4%) of the 180 moles. Infections with Anaplasma phagocytophilum, Babesia spp., Neoehrlichia mikurensis, Borrelia spp., Spiroplasma spp. and Francisella tularensis were not found. In addition, in a subset of 35 moles no antibodies against Tick-borne encephalitis virus were found. The obtained sequences of Bartonella spp. were closely related to Bartonella spp. sequences from moles in Spain and Hungary. The Hantaviridae were identified as the mole-borne Nova virus, with high sequence similarity to sequences from other European countries, and Bruges virus. Though the zoonotic risk from moles appears limited, our results indicate that these animals do play a role in multiple host-pathogen cycles.

Molecular Detection of Anaplasma phagocytophilum, Babesia odocoilei, Babesia species and Borrelia burgdorferi Sensu Lato in Songbirds

The blacklegged tick, Ixodes scapularis, is known to carry various tick-borne zoonotic pathogens with the potential to cause debilitating human and animal diseases. Juvenile I. scapularis parasitize songbirds and, perhaps, these avifauna are competent hosts of common microbial pathogens. We extracted brachial venous blood from 18 ground-foraging passerine birds that were parasitized by I. scapularis larvae and nymphs. Using molecular identification, namely PCR, DNA sequencing, and Basic Local Alignment Search Tool (BLAST), we targeted Anaplasma phagocytophilum, Babesia spp. and Borrelia burgdorferi sensu lato. Overall, 15 (83%) of 18 passerine birds were positive for 3 microbial zoonotic pathogens that comprised of A. phagocytophilum (n = 8), Babesia odocoilei (n = 6), Babesia spp. 20-5A74 (n = 1), and B. burgdorferi sensu lato (n = 9). The pathogen load consisted of 8 singles, 5 doubles, and 2 triples. One novel Babesia sp. (Babesia spp. 20-5A74) was found, and the remaining Babesia infections were B. odocoilei. Our findings reveal that ground-foraging, passerine birds are avian hosts of zoonotic pathogens. We provide the first-ever documentation that songbirds are hosts of B. odocoilei. Based on our data, B. odocoilei outnumbered other Babesia spp., and elucidated the authentic fact that B. odocoilei is the predominant Babesia sp. in North America. As avian hosts, passerine birds play a significant role in the enzootic transmission cycle of B. burgdorferi sensu lato, A. phagocytophilum, and Babesia species.

Patterns of adult tick parasitization of coexisting European (Erinaceus europaeus) and Algerian (Atelerix algirus) hedgehog populations in eastern Iberia

The availability of data regarding population abundance and dynamics of ticks in wild vertebrate populations is crucial to understand the host-tick relationship and to assess the risk for domestic animals and humans from possible zoonotic diseases. In this study we analyse several host-intrinsic and environmental factors affecting the tick population of two sympatric hedgehog species (the European and the Algerian hedgehog) in two differently anthropized habitats in eastern Spain. We captured 215 hedgehogs and sampled 356 adult ticks of four different species (Rhipicephalus sanguineus, R. turanicus, R. bursa and R. pusillus). We analysed by General Linear Mixed Models how the hosts’ species, sex, capture month and site and their interactions affected tick presence and tick load. Further, we carried out Spearman correlation tests to analyse how environmental temperature and precipitation affect tick presence and load on hedgehogs. We found that, in general, male hedgehogs were more infested than females. However, the effects of the different factors depended on the tick species, especially related to their endophilic or exophilic character. High values of general tick infestation can be explained by the coincidence of tick activity peaks and higher male host activity levels, especially within areas with higher habitat diversity and species richness. We also discuss how the potential immunosuppressive effect of testosterone could be affecting our results. Our results show that in highly anthropized environments hedgehogs potentially act as important mixing vessels for tick-borne zoonotic pathogens and that monitoring ticks in periurban wildlife is important to assess potential health risks for pets and humans.

Detection of Endosymbiont Candidatus Midichloria mitochondrii and Tickborne Pathogens in Humans Exposed to Tick Bites, Italy.

During 2021, we collected blood and serum samples from 135 persons exposed to tick bites in southern Italy. We serologically and molecularly screened for zoonotic tickborne pathogens and only molecularly screened for Candidatus Midichloria mitochondrii. Overall, 62 (45.9%) persons tested positive for tickborne pathogens. Coxiella burnetii was detected most frequently (27.4%), along with Rickettsia spp. (21.5%) and Borrelia spp. (10.4%). We detected Candidatus M. mitochondrii DNA in 46 (34.1%) participants who had statistically significant associations to tickborne pathogens (p<0.0001). Phylogenetic analysis of Candidatus M. mitochondrii sequences revealed 5 clades and 8 human sequence types that correlated with vertebrates, Ixodes spp. ticks, and countries in Europe. These data demonstrated a high circulation of tickborne pathogens and Candidatus M. mitochondrii DNA in persons participating in outdoor activities in southern Italy. Our study shows how coordinated surveillance among patients, clinicians, and veterinarians could inform a One Health approach for monitoring and controlling the circulation of tickborne pathogens.

Toxoplasma gondii and Rickettsia spp. in ticks collected from migratory birds in the Republic of Korea

Migratory birds disperse ticks and associated tick-borne pathogens along their migratory routes. Four selected pathogens of medical importance (Coxiella burnetii, Rickettsia spp., Francisella tularensis, and Toxoplasma gondii) were targeted for detection in 804 ticks (365 pools) collected from migratory birds at Hong and Heuksan Islands in the Republic of Korea (ROK) from 2010 to 2011 and 2016. Toxoplasma gondii and Rickettsia spp., were detected in 1/365 (0.27%) and 34/365 (9.32%) pools of ticks, respectively. T. gondii and five rickettsial species were recorded in ticks collected from migratory birds for the first time in ROK. The five rickettsial species (R. monacensis, Candidatus Rickettsia longicornii, R. japonica, R. raoultii, and R. tamurae) were identified using sequence and phylogenetic analysis using ompA and gltA gene fragments. Rickettsia spp. are important pathogens that cause rickettsiosis in humans, with cases recorded in the ROK. These results provide important evidence for the potential role of migratory birds in the introduction and dispersal of T. gondii and Rickettsia spp. along their migratory routes and raise awareness of potential transmission of zoonotic tick-borne pathogens associated with migratory birds in the ROK.

Molecular survey on the presence of arthropod-borne bacteria and protozoans in roe deer (Capreolus capreolus) and ticks from Central Italy

Environmental changes, due to climatic emergency and to anthropogenic activities severely impact on the epidemiology of vector borne diseases, mostly when transmitted by ticks. The data about the distribution of microorganisms responsible for them in roe deer (Capreolus capreolus) population living in Italy are scanty and completely lacking in Tuscany, so a molecular survey was carried out to estimate the prevalence of some zoonotic tick-borne pathogens in roe deer, and ticks removed from them, living in areas of Central Italy with high risk of arthropod exposure. Spleen samples from 72 roe deer were tested by PCR for Anaplasma phagocytophilum, Borrelia burgdorferi s.l., Francisella tularensis and piroplasms. Moreover, 345 ticks were removed from 65 roe deer, morphologically or molecularly identified and grouped into 162 pools that were submitted to PCR for detecting the same pathogens. Forty-six (63.88%) roe deer were positive for at least one investigated pathogen: 43 (59.72%) for A. phagocytophilum, 2 (2.78%) for Babesia capreoli, 1 (1.39%) for B. burgdorferi, and 1 (1.39%) for Babesia sp. No animals were PCR positive for F. tularensis. All ticks were identified as Ixodes ricinus. Seventy-six (46.91%) tick pools showed DNA of one or more pathogens: 66 (40.74%) were positive for A. phagocytophilum, 22 (13.58%) for B. burgodorferi s.l., 6 (3.70%) for B. venatorum and 3 (1.85%) for B. capreoli. No pools were positive for F. tularensis. Two or three pathogens were detected in 23 (14.19%) pools.

Tick-borne pathogens in neotropical animals in Trinidad, West Indies

BackgroundTicks are important vectors of many pathogens that have contributed to the morbidity and mortality of humans and domestic animals worldwide. Wildlife species have also been implicated as reservoir hosts of a variety of tick-borne pathogens. The objective of this study was to determine which tick-transmitted pathogens were present in the animals harvested from the forest in Trinidad for human consumption.MethodsThin blood smears from 43 neotropical animals were examined microscopically for tick-borne pathogens. Additionally, DNA extraction and PCR amplification of the 16S rRNA gene were used for amplification of Anaplasma and Ehrlichia while the gltA gene was used for Bartonella, and Rickettsia spp. and the 18S rRNA gene for Babesia, Hepatozoon and Theileria species.ResultsPathogen DNA was amplified from four samples (a deer, collared peccary and two agoutis). Sequencing of the amplified products from the deer and collared peccary revealed 99.8% homology to Anaplasma bovis and 98.8% homology to Ehrlichia canis, respectively. Sequences from two agoutis revealed 90.4% homology to Theileria spp. DNA of Hepatozoon spp., Bartonella spp. Babesia spp. and Rickettsia spp. was not detected in any of the screened samples. An incidental finding in this study was the presence of bacteria in the blood of animals.ConclusionsThe results indicate that the DNA of tick-transmitted pathogens is present at a frequency of about 10% in the study population and suggests that neotropical mammals may serve as a source for the potential transmission of tick-borne pathogens to domestic animals and humans. In addition, physicians and hunters should be aware of the symptoms associated with zoonotic tick-borne pathogens so that these infections can be recognised, diagnosed and treated promptly. Bacteria present in carcasses can pose a food safety hazard and hunters should be trained in proper harvesting and handling of carcasses.Graphical

Current knowledge of vector-borne zoonotic pathogens in Zambia: A clarion call to scaling-up "One Health" research in the wake of emerging and re-emerging infectious diseases.

Although vector-borne zoonotic diseases are a major public health threat globally, they are usually neglected, especially among resource-constrained countries, including those in sub-Saharan Africa. This scoping review examined the current knowledge and identified research gaps of vector-borne zoonotic pathogens in Zambia. Major scientific databases (Web of Science, PubMed, Scopus, Google Scholar, CABI, Scientific Information Database (SID)) were searched for articles describing vector-borne (mosquitoes, ticks, fleas and tsetse flies) zoonotic pathogens in Zambia. Several mosquito-borne arboviruses have been reported including Yellow fever, Ntaya, Mayaro, Dengue, Zika, West Nile, Chikungunya, Sindbis, and Rift Valley fever viruses. Flea-borne zoonotic pathogens reported include Yersinia pestis and Rickettsia felis. Trypanosoma sp. was the only tsetse fly-borne pathogen identified. Further, tick-borne zoonotic pathogens reported included Crimean-Congo Haemorrhagic fever virus, Rickettsia sp., Anaplasma sp., Ehrlichia sp., Borrelia sp., and Coxiella burnetii. This study revealed the presence of many vector-borne zoonotic pathogens circulating in vectors and animals in Zambia. Though reports of human clinical cases were limited, several serological studies provided considerable evidence of zoonotic transmission of vector-borne pathogens in humans. However, the disease burden in humans attributable to vector-borne zoonotic infections could not be ascertained from the available reports and this precludes the formulation of national policies that could help in the control and mitigation of the impact of these diseases in Zambia. Therefore, there is an urgent need to scale-up "One Health" research in emerging and re-emerging infectious diseases to enable the country to prepare for future epidemics, including pandemics.

Ehrlichia spp. disülfid oksidoredüktaz genini hedefleyen polimeraz zincir reaksiyonunun analitik duyarlılığının belirlenmesi: Klinik olarak leishmaniasis'ten şüphelenilen bir köpekte ehrlichiosis'in moleküler tanısı

Ehrlichia spp. is tick-borne zoonotic pathogen that can infect humans and animals. Nowadays, among the tests used in the diagnosis of ehrlichiosis, the importance of molecular methods is increasing steadily due to their high sensitivity and specificity. The aim of this study was to determine the analytical sensitivity of a conventional polymerase chain reaction (PCR) targeting Ehrlichia spp. disulfide oxidoreductase (DSB) gene. Ehrlichia spp. DSB gene was cloned into the TOPO vector. After TOPO plasmid containing DSB gene were serially diluted, PCR targeting the Ehrlichia spp. DSB gene was performed. While working on this research, blood and skin scraping samples of a stray dog clinically suspected with leishmaniasis as well as treated for leishmaniasis arrived to our laboratory. Thereafter, PCRs targeting Ehrlichia spp. DSB and 16S rRNA and Leishmania kinetoplast DNA (kDNA) genes were performed to identify the pathogen in blood and skin scraping samples of the stray dog. The analytical sensitivity of the PCR assay targeting Ehrlichia spp. DSB gene was 1 ≥ copy plasmid/reaction using serially diluted TOPO plasmid containing DSB gene. PCR targeting the Ehrlichia spp. DSB gene was positive and PCR targeting Leishmania spp. kDNA was negative in blood and skin samples of the stray dog clinically suspected with leishmaniasis. Using nested PCR targeting Ehrlichia spp. 16S rRNA, E. canis was identified in blood and skin scraping samples of the stray dog. In this study, PCR targeting Ehrlichia spp. DSB gene has been shown to have high sensitivity. Also it was shown molecular methods can help clinicians in differential diagnosis of ehrlichiosis and leishmaniasis to prevent inappropriate treatment.

Bacteriome in Ticks Collected from Domestic Livestock in Kenya

Background: Metagenomics approaches are increasingly being utilized as “dipstick” for microbial carriage. In this study, 16S rRNA metagenomics was used to probe for microbial community that resides in the ticks, those they pick from the environment, wildlife and livestock and to identify potential tick borne zoonoses. Methods: Tick DNA from 463 tick pools collected from domestic animals between 2007 and 2008 were amplified with primers that target the 16S rRNA V3-V4 domain and then sequenced on Illumina Miseq platform using 300 cycles version 3 kits. Ticks were pooled according to species and animal from which they were collected. A non-target control was used to track laboratory contaminants. Sequence data were analyzed using Mothur v1.3 pipeline and R v3.3.1 software and taxonomy determined using SILVA rRNA database. Shannon diversity index was used to compute bacterial diversity in each tick species before computing the means. Results: A total of 645 bacteria genera grouped into 27 phyla were identified. Four phyla contributed 97.4% of the 36,973,934 total sequences. Proteobacteria contributed 61.2% of these sequences that tarried to 33.8% genera, compared to 15.9% (23.4% genera) for Firmicutes, 15.6% (20% genera) for Actinobacteria and 4.7% (11.6% genera) for Bacteroidetes. The remaining 23 phyla only contributed 2.6% of the sequence reads (11.2% genera). Amongst the 645 genera, three groups were discernible, with the biggest group comprised commensals/symbionts that contributed 93.6% of the genera, but their individual sequence contribution was very low. Group two comprised genera that are known to contain pathogenic species, with Coxiella contributing 15,445,204 (41.8%) sequences, Corynebacterium (13.6%), Acinetobacter (4.3%), Staphylococcus (3.9%), Bacillus (2.7%) and Porphyromonas (1.6%), Ralstonia (1.5%), Streptococcus (1.3%), Moraxella (1.3%), amongst others. Group three comprised genera known to contain tick borne zoonotic pathogens (TBZ): Rickettsiae, Anaplasma, Francisella, Ehrlichia, Bartonella and Borrelia. Individually the TBZ contributed Amblyomma variegatum carried the least diverse bacteria (mean Shannon diversity index of 2.69 ± 0.92) compared to 3.79 ± 1.10 for A. gemma, 3.71 ± 1.32 for A. hebraeum, 4.15 ± 1.08 for other Amblyomma spp, 3.79 ± 1.37 for Hyalomma truncatum, 3.67 ± 1.38 for other Hyalomma spp, 3.86 ± 1.27 for Rhipicephalus annulatus, 3.56 ± 1.21 for Rh. appendiculatus, and 3.65 ± 1.30 for Rh. Pulchellus, but the difference was not significant (p = 0.443). Conclusion: This study illustrates the utility of 16S rRNA metagenomics in revealing the complexity of bacteria communities that reside and/or transit through the tick having been picked from the environment, livestock and/or wild animals, some with potential to cause zoonoses.