The prevalence of selected vector-borne diseases in dromedary camels (Camelus dromedarius) in the United Arab Emirates.

Vector-borne diseases (VBDs) affecting dromedary camels (Camelus dromedarius) have considerable importance in the United Arab Emirates (UAE) because of the consequences associated with production decline and economic losses. Our study aimed to determine the prevalence of selected VBDs in camels in the UAE and identify risk factors. This research is currently affected by the low number of epidemiological molecular surveys addressing this issue. Blood samples were obtained from 425 dromedary camels from different locations across the UAE. Whole genomic DNA was isolated, and PCR screening was done to detect piroplasmids (Babesia/Theileria spp.), Trypanosoma spp., and Anaplasmataceae spp. (Anaplasma, Ehrlichia, Neorickettsia and Wolbachia spp.). Amplicons were sequenced, and phylogenetic trees were constructed. Trypanosoma sequences were identified as T. brucei evansi, whereas Anaplasmataceae sequences were identified as A. platys-like. All camels were negative for Babesia/Theileria spp. (0%); however, 18 camels were positive for T. b. evansi (4%) and 52 were positive for A. platys-like (12%). Mixed infection with T. b. evansi and A. platys-like was found in one camel. Statistical analyses revealed that camels with a brown coat colour were significantly more prone to acquire the A. platys-like strain compared with those having a clearer coat. A similar finding was observed when comparing urban moving camels with desert indoor and urban indoor camels. Continuous disease surveillance is required to ensure and maintain the good health status of the camels in the UAE. Nonetheless, the risk of disease outbreak remains if the misuse of drugs continues.

A community approach of pathogens and their arthropod vectors (ticks and fleas) in dogs of African Sub-Sahara.

BACKGROUND: Arthropod-borne pathogens and their vectors are present throughout Africa. They have been well-studied in livestock of sub-Saharan Africa, but poorly in companion animals. Given the socio-economic importance of companion animals, the African Small Companion Animal Network (AFSCAN), as part of the WSAVA Foundation, initiated a standardized multi-country surveillance study. METHODS: Macro-geographic variation in ectoparasite (ticks and fleas) and pathogen communities in dogs was assessed through molecular screening of approximately 100 infested dogs in each of six countries (Ghana, Kenya, Nigeria, Tanzania, Uganda and Namibia), both in rural and urban settings. The most important intrinsic and extrinsic risk factors within the subpopulation of infested dogs were evaluated. RESULTS: Despite the large macro-geographic variation in the dogs screened, there was no consistent difference between East and West Africa in terms of the diversity and numbers of ticks. The highest and lowest numbers of ticks were found in Nigeria and Namibia, respectively. Most often, there was a higher diversity of ticks in rural habitats than in urban habitats, although the highest diversity was observed in an urban Uganda setting. With the exception of Namibia, more fleas were collected in rural areas. We identified tick species (including Haemaphysalis spinulosa) as well as zoonotic pathogens (Coxiella burnetti, Trypanosoma spp.) that are not classically associated with companion animals. Rhipicephalus sanguineus was the most abundant tick, with a preference for urban areas. Exophilic ticks, such as Haemaphysalis spp., were more often found in rural areas. Several multi-host ticks occurred in urban areas. For R. sanguineus, housing conditions and additional pets were relevant factors in terms of infestation, while for a rural tick species (Haemaphysalis elliptica), free-roaming dogs were more often infested. Tick occurrence was associated to the use of endoparasiticide, but not to the use of ectoparasiticide. The most prevalent tick-borne pathogen was Hepatozoon canis followed by Ehrlichia canis. High levels of co-parasitism were observed in all countries and habitats. CONCLUSIONS: As dogs share a common environment with people, they have the potential to extend the network of pathogen transmission to humans. Our study will help epidemiologists to provide recommendations for surveillance and prevention of pathogens in dogs and humans.

Competency of Neural Networks for the Numerical Treatment of Nonlinear Host-Vector-Predator Model.

The aim of this work is to introduce a stochastic solver based on the Levenberg-Marquardt backpropagation neural networks (LMBNNs) for the nonlinear host-vector-predator model. The nonlinear host-vector-predator model is dependent upon five classes, susceptible/infected populations of host plant, susceptible/infected vectors population, and population of predator. The numerical performances through the LMBNN solver are observed for three different types of the nonlinear host-vector-predator model using the authentication, testing, sample data, and training. The proportions of these data are chosen as a larger part, i.e., 80% for training and 10% for validation and testing, respectively. The nonlinear host-vector-predator model is numerically treated through the LMBNNs, and comparative investigations have been performed using the reference solutions. The obtained results of the model are presented using the LMBNNs to reduce the mean square error (MSE). For the competence, exactness, consistency, and efficacy of the LMBNNs, the numerical results using the proportional measures through the MSE, error histograms (EHs), and regression/correlation are performed.

Data analysis workflow for the detection of canine vector-borne pathogens using 16 S rRNA Next-Generation Sequencing.

BACKGROUND: Vector-borne diseases (VBDs) impact both human and veterinary medicine and pose special public health challenges. The main bacterial vector-borne pathogens (VBPs) of importance in veterinary medicine include Anaplasma spp., Bartonella spp., Ehrlichia spp., and Spotted Fever Group Rickettsia. Taxon-targeted PCR assays are the current gold standard for VBP diagnostics but limitations on the detection of genetically diverse organisms support a novel approach for broader detection of VBPs. We present a methodology for genetic characterization of VBPs using Next-Generation Sequencing (NGS) and computational approaches. A major advantage of NGS is the ability to detect multiple organisms present in the same clinical sample in an unsupervised (i.e. non-targeted) and semi-quantitative way. The Standard Operating Procedure (SOP) presented here combines industry-standard microbiome analysis tools with our ad-hoc bioinformatic scripts to form a complete analysis pipeline accessible to veterinary scientists and freely available for download and use at https://github.com/eltonjrv/microbiome.westernu/tree/SOP . RESULTS: We tested and validated our SOP by mimicking single, double, and triple infections in genomic canine DNA using serial dilutions of plasmids containing the entire 16 S rRNA gene sequence of (A) phagocytophilum, (B) v. berkhoffii, and E. canis. NGS with broad-range 16 S rRNA primers followed by our bioinformatics SOP was capable of detecting these pathogens in biological replicates of different dilutions. These results illustrate the ability of NGS to detect and genetically characterize multi-infections with different amounts of pathogens in a single sample. CONCLUSIONS: Bloodborne microbiomics & metagenomics approaches may help expand the molecular diagnostic toolbox in veterinary and human medicine. In this paper, we present both in vitro and in silico detailed protocols that can be combined into a single workflow that may provide a significant improvement in VBP diagnostics and also facilitate future applications of microbiome research in veterinary medicine.

Molecular detection of Anaplasma phagocytophilum in hair and spleen of cats revealed a possible underestimation of feline vector-borne pathogens.

Feline Vector-Borne Diseases show increased global prevalence and some Anaplasma and Ehrlichia species may pose a risk to human health. The diagnosis of Anaplasma and Ehrlichia species infection in cats is achieved by the combined use of different methods as cytologic examination evidencing intracytoplasmic morulae, serologic tests and molecular assays. The peripheral whole blood is considered the sample of choice for Anaplasma and Ehrlichia species DNA detection in cats, but false negative results are reported leading to underestimation of infection prevalence. In order to have a more accurate assessment of the spread of feline vector-borne pathogens, the presence of Anaplasma spp. and Ehrlichia spp. DNA in 37 owner and shelter cats subjected to necropsy were prospectively investigated by testing in end-point PCR spleen, bone marrow, blood clot and hair samples. The bacteria identified were genetically characterised. Three shelter cats tested positive for A. phagocytophilum DNA in spleen (one cat) or in hair samples (two cats). None of the cats tested positive in bone marrow and blood samples. From the results obtained, it can be assumed that the use of spleen or hair samples could allow a more reliable detection of A. phagocytophilum DNA in cats with blood tested negative. In the phylogeny constructed with a fragment of the heat shock (groEL) gene nucleotide sequences, all the identified A. phagocytophilum clustered with bacteria infecting a wide range of hosts, including humans, showing a potential zoonotic role.

Convergence Research for Emerging Zoonoses.

Global emergence of vector-borne and zoonotic diseases presents a rapidly growing 'wicked' problem. We outline the need for a transdisciplinary research program that is grounded in ecological and evolutionary theory but integrates fundamentally with research perspectives spanning the health, social, and natural sciences.

Ticks, mites, fleas, and vector-borne pathogens in free-ranging neotropical wild felids from southern Brazil.

The study of vectors and vector-borne diseases associated with ectoparasites from free-living Neotropical small wild felids is scarce, and the few existing studies on this theme focused on the genus Panthera, Paleartic species or in captive animals. For this reason, the aim of this study was to identify the diversity of ectoparasites and potentially associated Rickettsia and Bartonella species in free-ranging neotropical wild cats collected in the State of Rio Grande do Sul, southern Brazil. A total of 82 ticks, 10 fleas and 22 mites were collected from 18 Geoffroy's cats (Leopardus geoffroyi), 11 margays (Leopardus wiedii), two southern tiger cats (Leopardus guttulus), two jaguarundis (Herpailurus yagouaroundi), three ocelots (Leopardus pardalis) and two pumas (Puma concolor). We identified four tick species: Rhipicephalus microplus and three species from genus Amblyomma, the most frequent being Amblyomma aureolatum; three flea species corresponding to Ctenocephalides felis, Xenopsylla cheopis, and Pulex irritans; and one mite of genus Eutrombicula. In ectoparasites we found DNA of Rickettsia parkeri and Rickettsia asembonensis in ticks and DNA of Bartonella clarridgeiae, Bartonella sp. and R. asembonensis in fleas. Our results highlight the evidences of vectors and vector-borne agents in wild animals, and their potential wide distribution in the Pampa biome and the southernmost portion of the Atlantic Rainforest in Brazil.

Molecular survey of vector-borne diseases in two groups of domestic dogs from Lisbon, Portugal.

BACKGROUND: Canine vector-borne diseases (CVBDs) are caused by a wide range of pathogens transmitted by arthropods. They have been an issue of growing importance in recent years; however, there is limited information about the vector-borne pathogens circulating in Portugal. The aim of the present study was to detect canine vector-borne bacteria and protozoa of veterinary and zoonotic importance using molecular methods. METHODS: One hundred and forty-two dogs from Lisbon, southern Portugal, were tested: 48 dogs from a veterinary hospital clinically suspected of vector-borne diseases and 94 apparently healthy dogs from shelters. Anaplasma spp./Ehrlichia spp., Babesia/Theileria spp., Hepatozoon spp., and Mycoplasma spp. infections were detected by PCR from blood samples and examined under light microscopy. Other information including clinical status and diagnostic test results were collected for each animal. RESULTS: Infections were detected by PCR in 48 (33.80%) dogs. Single infections were found in 35 dogs (24.64%), and co-infections were found in 13 (9.15%) dogs. Twenty-nine (20.42%) dogs were positive for Hepatozoon spp., 15 (10.56%) for Mycoplasma spp., 11 (7.75%) for Anaplasma spp./Ehrlichia spp., and six (4.21%) for Babesia spp. DNA sequencing was used to identify Babesia vogeli (2.81%), Babesia canis (1.40%), Hepatozoon canis (20.42%), Mycoplasma haematoparvum (2.11%), Mycoplasma haemocanis (8.45%), Anaplasma platys (7.04%), and Ehrlichia canis (0.70%). CONCLUSIONS: This is the first molecular identification of B. canis and M. haematoparvum in dogs from southern Portugal. This study highlights the importance of molecular methods to identify CVBD pathogens in endemic areas and helps to guide the clinical approach of veterinarians in practice.

Evaluation of a commercial microbial enrichment kit used prior DNA extraction to improve the molecular detection of vector-borne pathogens from naturally infected dogs.

Accurate detection of vector-borne pathogens (VBPs) is extremely important as the number of reported cases in humans and animals continues to rise in the US and abroad. Validated PCR assays are currently the cornerstone of molecular diagnostics and can achieve excellent analytical sensitivity and specificity. However, the detection of pathogens at low parasitemia still presents a challenge for VBP diagnosis, especially given the very low volume of specimens tested by molecular methods. The objective of this study is to determine if a commercially available microbial enrichment kit, used prior DNA extraction, is capable of expanding the overall microbial community and increasing detectable levels of VBPs in canine blood samples through host DNA depletion. This study used EDTA-whole blood samples from dogs naturally infected with varying parasitemia levels of either Anaplasma phagocytophilum, Babesia gibsoni, or Ehrlichia ewingii. For two VBPs, EDTA-blood samples were diluted to determine the effect of microbial concentration at low parasitemia. Paired EDTA-blood samples from each dog were subjected to traditional, automated DNA extraction with or without the microbial concentrating kit (MolYsis®) prior DNA extraction. Relative amounts of pathogen DNA in paired samples were determined by real-time PCR and Next-Generation Sequencing targeting conserved regions of 16S rRNA (for bacteria) and 18S rRNA (for protozoa). Results from the three molecular methods suggest that the microbial concentrating kit did not improve the detection of VBPs, although significantly reduced the presence of host DNA. Alternative methods for VBP enrichment in clinical samples prior to molecular testing should continue to be investigated, as it may significantly improve clinical sensitivity and reduce the number of false-negative results.

Descriptive epidemiology of rickettsial infections in Japan: Scrub typhus and Japanese spotted fever, 2007-2016.

OBJECTIVE: This study aimed to describe the epidemiological and clinical characteristics of endemics of two rickettsial diseases, scrub typhus (ST) and Japanese spotted fever (JSF), in Japan. METHODS: We conducted a retrospective, descriptive epidemiological assessment of cases notified via national surveillance from 2007-2016. RESULTS: Over the 10-year period, 4185 ST and 1765 JSF cases were notified; of these, 20 (0.48%) cases of ST and 16 (0.91%) cases of JSF were fatal at the time of reporting. The elderly had higher notification rates and fatalities. While the annual number of ST notifications was stable and cases were reported from a broad geographic range, the number of JSF reports increased three-fold, expanding from the southwest to the east. The seasonality of ST varied by region and was more common during spring/summer in the north and autumn/winter in the south; 78% of cases occurred during autumn/winter, mainly in the southern region. Most of the fatal ST cases occurred in the spring/summer and occurred in the northern region. CONCLUSION: Our analysis identified seasonal and regional variations in the distribution of rickettsiosis. These variations were most likely to be related to the ecology of the vectors and etiological agents. Knowing the recent epidemiological and clinical features of ST and JSF can support clinical diagnosis and guide preventative activities against these vector-borne diseases.

Canine infection with Dirofilaria immitis, Borrelia burgdorferi, Anaplasma spp., and Ehrlichia spp. in the United States, 2013-2019.

BACKGROUND: Dogs in the US are commonly infected with vector-borne pathogens, including heartworm and tick-borne disease agents. The geographic distribution of both arthropod vectors and the pathogens they transmit continues to expand. METHODS: To describe the current geographic distribution and prevalence of antigen of Dirofilaria immitis and antibody to Borrelia burgdorferi, Ehrlichia spp., and Anaplasma spp. in dogs, we summarized over 144 million test results from 2013 to 2019, inclusive, by county, state, and region. Canine seroprevalence by state was compared to population-adjusted human reports of tick-borne diseases. RESULTS: Results varied regionally, with D. immitis antigen and Ehrlichia spp. antibodies more frequently detected in the Southeast (2.6% and 5.2%, respectively) and antibody to B. burgdorferi and Anaplasma spp. most common in the Northeast (12.1% and 7.3%, respectively). Overall, percent positive test results to D. immitis decreased in the Southeast by 33.3% when compared to earlier summaries using the same strategy (from 3.9 to 2.6%). Geographic expansion of areas where dogs commonly test positive for Ehrlichia spp. was evident, likely because of a change in the test made in 2012 to allow detection of antibodies to E. ewingii concomitant with expansion of vector tick populations. Percent positive test results to Ehrlichia spp. increased in every region; this shift was particularly pronounced in the Southeast, where percent positive test results increased fourfold (from 1.3 to 5.2%). Continued geographic expansion of B. burgdorferi and A. phagocytophilum was apparent in the Northeast, Midwest, and Upper South, although canine seroprevalence of antibody to B. burgdorferi was much lower than prior surveys in many Lyme-endemic areas. Annual reports of human cases of Lyme disease, ehrlichiosis, and anaplasmosis were associated with percent positive canine results by state for the three tick-borne disease agents (R2 = 0.812, 0.521, and 0.546, respectively). Within endemic areas, percent positive test results for all three tick-borne agents demonstrated evidence of geographic expansion. CONCLUSIONS: Large scale analysis of results from screening dogs in practice for evidence of vector-borne infections, including those with zoonotic importance, continues to be a valuable strategy for understanding geographic trends in infection risk over time.

Vector-borne diseases in Iran: epidemiology and key challenges.

Vector-borne diseases have become a global health concern in recent decades as a result of global warming, globalization, growth in international trade and travel, use of insecticide and drug resistance. This review study addressed the key vector-borne diseases and their current status in Iran to emphasize the requirements for further research on vector-borne diseases. The dispersion patterns of these diseases differ in various regions. Some of them such as Crimean-Congo hemorrhagic fever, and Q fever are distributed all across Iran, whereas some others such as plague, leishmaniasis, tularemia, and malaria are restricted to specific areas. The high prevalence of vectors throughout the country necessitates enhancing the monitoring and surveillance of emerging and reemerging vector-borne diseases and their potential vectors.

Fleas (Siphonaptera) Parasitizing Peridomestic and Indigenous Mammals in Panamá and Screening of Selected Fleas for Vector-Borne Bacterial Pathogens.

In total, 341 fleas belonging to 16 species were collected from 78 host mammals belonging to 10 species in Panamá from 2010 to 2016. The cat flea, Ctenocephalides felis (Bouché) predominated on domestic dogs and was also recorded from domestic cats, the raccoon, Procyon lotor (Linnaeus) and the common opossum, Didelphis marsupialis Linnaeus. The largest number of flea species (7) was recorded from D. marsupialis and the most common flea on that host was the ctenophthalmid, Adoratopsylla intermedia copha Jordan. One Oriental rat flea, Xenopsylla cheopis (Rothschild), was collected from D. marsupialis. Native rodents were parasitized by indigenous ceratophyllid, rhopalopsyllid, and stephanocircid fleas. The Mexican deermouse, Peromyscus mexicanus (Saussure), was parasitized by six species of ceratophyllids belonging to the mostly Central American genera, Baculomeris, Jellisonia, Kohlsia and Plusaetis. The long-tailed singing mouse, Scotinomys xerampelinus (Bangs), was parasitized by Plocopsylla scotinomi Tipton and Méndez, the only species of stephanocircid flea known from Central America. Twenty-six pools of extracted flea DNA representing 5 flea species (C. felis, Pulex echidnophagoides (Wagner), Pulex simulans Baker, A. intermedia copha, and P. scotinomi) and 79 individual fleas were all real-time polymerase chain reaction negative for Rickettsia felis, Rickettsia typhi, and Bartonella henselae.

Vector-borne pathogens in dogs from Guatemala, Central America.

Vector-borne pathogens (VBPs) of dogs have been extensively studied worldwide, though scientific information for some countries, as is the case for Guatemala, is almost nonexistent. From 2012 to 2015, 975 dogs residing in different departments of Guatemala were sampled and screened using a rapid ELISA for detecting antigen of Dirofilaria immitis and antibodies against Anaplasma spp., Ehrlichia spp. and Borrelia burgdorferi. Out of 975 samples analyzed, 46.4% (n = 452) scored positive for at least one pathogen, with D. immitis being the most prevalent (29.7%), followed by Ehrlichia spp. (11.5%) and Anaplasma spp. (5.1%). None of the dogs were positive to B. burgdorferi. Co-infections were observed in 18.1% (n = 176) of dogs, with the simultaneous detection of D. immitis and Ehrlichia spp. being most common. The frequency of VBPs was higher in the department of Santa Rosa (southeast coast of Guatemala), which has a tropical savanna climate. In this region, 59.7% of the dogs surveyed were infected with D. immitis. Our results suggest that the prevalence of VBPs in dogs in Guatemala may be affected by the climate, with dogs living in the southeast coast being at higher risk, as compared to other regions studied. Increased awareness regarding the risk of VBPs in dogs in Guatemala is advocated and the adoption of preventive strategies should be encouraged.

An outbreak of relapsing fever unmasked by microbial paleoserology, 16th century, France.

OBJECTIVES: Depicting past epidemics currently relies on DNA-based detection of pathogens, an approach limited to pathogens with well-preserved DNA sequences. We used paleoserology as a complementary approach detecting specific antibodies under a mini line-blot format including positive and negative control antigens. METHODS: Mini line blot assay incorporated skim milk as negative control, Staphylococcus aureus as positive control, and antigens prepared from lice-borne pathogens Rickettsia prowazekii, Borrelia recurrentis, Bartonella quintana, and Yersinia pestis. Paleoserums were extracted from rehydrated dental pulp recovered from buried individuals. Mini line blots observed with the naked eye, were quantified using a scanner and appropriate software. Paleoserology was applied to the indirect detection of lice-borne pathogens in seven skeletons exhumed from a 16th-17th century suspected military burial site (Auxi-le-Château); and 14 civils exhumed from a 5th-13th century burial site (Saint-Mont). Direct detection of pathogens was performed using quantitative real-time PCR. RESULTS: In Auxi-le-Château, paleoserology yielded 7/7 interpretable paleoserums including 7/7 positives for B. recurrentis including one also positive for B. quintana. In Saint-Mont, paleoserology yielded 8/14 interpretable paleoserums and none reacted against any of the four pathogens. Antibodies against R. prowazekii and Y. pestis were not detected. The seroprevalence was significantly higher in the military burial site of Auxi-le-Château than in the civil burial site of Saint-Mont. Real-time PCR detection of B. quintana yielded 5/21 positive (3 at Saint-Mont and 2 at Auxi-le-Château) whereas B. recurrentis was not detected. CONCLUSIONS: Paleoserology unmasked an outbreak of relapsing B. recurrentis fever in one 16th - 17th century military garrison, missed by real-time PCR. Paleoserology offers a new tool for investigating past epidemics, in complement to DNA sequence-based approaches.

Seven years of evaluation of ectoparasites and vector-borne pathogens among ring-tailed coatis in an urban park in southeastern Brazil.

Wild animals have been recognized as potential reservoirs of vector-borne pathogens. Proximity between these animals and urban areas increases the need to know which pathogens these are and whether they can infect domestic animals and humans. In Mangabeiras Municipal Park in Belo Horizonte, Brazil, coatis live near the urban area, which is mainly occupied by human residents and their domestic animals. Therefore, the objective of this study was to detect, through molecular and direct methods, the presence of ectoparasites and hemoparasites in coatis. A total of 216 samples were collected, of which 209 samples were from first-captures and seven were from recaptures. The following parasites were found: ticks of the genus Amblyomma, lice of the species Neotrichodectes pallidus and fleas of the species Rhopalopsyllus lutzi lutzi and Ctenocephalides felis felis. All the samples were negative for the family Anaplasmataceae and the species Leishmania sp. and Trypanosoma cruzi. The hemoparasites Trypanosoma evansi, Hepatozoon procyonis, Babesia sp. and Sarcocystis neurona were found. The area of the present study is not endemic for T. evansi, which therefore suggests that these coatis may be acting as reservoirs or sentinels of this parasite. This finding is of great epidemiological importance and should be investigated more closely. Thus, this study showed that there is a great variety of pathogens in the park that transit among coatis and, probably, among other animals that inhabit or live close to the park.

Bacterial Vector-Borne Plant Diseases: Unanswered Questions and Future Directions.

Vector-borne plant diseases have significant ecological and economic impacts, affecting farm profitability and forest composition throughout the world. Bacterial vector-borne pathogens have evolved sophisticated strategies to interact with their hemipteran insect vectors and plant hosts. These pathogens reside in plant vascular tissue, and their study represents an excellent opportunity to uncover novel biological mechanisms regulating intracellular pathogenesis and to contribute to the control of some of the world's most invasive emerging diseases. In this perspective, we highlight recent advances and major unanswered questions in the realm of bacterial vector-borne disease, focusing on liberibacters, phytoplasmas, spiroplasmas, and Xylella fastidiosa.

Arthropod exosomes as bubbles with message(s) to transmit vector-borne diseases.

Ticks and mosquitoes are medically important vectors that transmit several pathogens, including arboviruses, to humans. Understanding how these blood-feeding arthropods transmit pathogens to humans requires knowledge on the molecular and cellular interplay at vector-host interface. Recent studies have highlighted the role of tick and mosquito small extracellular vesicles (EVs), including exosomes, facilitating arbovirus transmission within arthropod cells and from arthropod to mammalian cells. In this review, we summarize this emerging line of investigation in understanding the role of tick and mosquito exosomes in vector-pathogen-host tripartite interactions. Understanding the role of arthropod exosomes in pathogen interactions could lead to the discovery of novel therapeutic targets to interfere with the life cycle of several pathogens transmitted by vectors.

MOLECULAR INVESTIGATION OF VECTOR-BORNE PATHOGENS IN RED FOXES (VULPES VULPES) FROM SOUTHERN FRANCE.

Because of their free-ranging nature, the probability of wild animals being exposed to vector-borne pathogens is likely higher than that of humans and pets. Recently, the red fox (Vulpes vulpes) has been suspected as being a reservoir or host of several pathogens of veterinary and public health importance. We conducted a molecular survey on 93 red foxes hunted in 2008-18, in the departments of Bouches-du-Rhône and Var, in southeastern France, for pathogens including Leishmania infantum, Piroplasmida, Hepatozoon spp., nematodes, Coxiella burnetii, Borrelia spp., Rickettsia spp., and Anaplasmataceae. Spleen samples were screened for the presence of vector-borne pathogens by PCR followed by sequencing. Pathogens were detected in 94% (87/93) of red foxes, and coinfections were identified in 24% (22/93) of foxes. We identified DNA from Hepatozoon canis, L. infantum, and Babesia vogeli in 92% (86/93), 15% (14/93), and 3% (3/93) of red foxes, respectively. We also found DNA of nematodes in 3% (3/93) of foxes; Spirocerca vulpis was identified in one fox and Dirofilaria immitis in the two others. Interestingly, C. burnetii genotype 3, previously described in humans from the same region, was identified in 3% (3/93) of foxes and Anaplasma platys in 2% (2/93) of foxes. We did not detect DNA of Borrelia spp., Bartonella spp., or Rickettsia spp. In our study, the prevalence of pathogens did not vary by fox origin, sex, or tick carriage. Molecular evidence of B. vogeli, H. canis, S. vulpis, D. immitis, C. burnetii, and A. platys in red foxes has not previously, to our knowledge, been reported from southern France. We propose that red foxes are potential reservoirs for several pathogens, including major zoonotic agents such as L. infantum. They could be incidental hosts for pathogens, such C. burnetii. The high prevalence for H. canis suggests an important role of foxes in domestic dog (Canis lupus familiaris) infection. These animals may pose a threat to human and animal health.

Circulation of Rickettsia species and rickettsial endosymbionts among small mammals and their ectoparasites in Eastern Slovakia.

Bacteria belonging to the genus Rickettsia are known as causative agents of vector-borne zoonotic diseases, such as spotted fevers, epidemic typhus and endemic typhus. Different species of ticks, mites and fleas could act as reservoirs and arthropod vectors of different pathogenic Rickettsia species. The aim of this work was to establish active surveillance of Rickettsia spp. in mites, ticks and fleas collected from small mammals (rodents and shrews) in Eastern Slovakia. A total of 964 animal ear biopsies, 871 mites, 667 ticks and 743 fleas were collected from small mammals in the Kosice region, Eastern Slovakia. All specimens were identified using specialized taxonomic keys, and were conserved in ethanol until DNA extraction was performed. After DNA extraction, identification of Rickettsia species was performed by PCR-based methods. The total prevalence of rickettsiae from ear biopsies was 4.6% (95% CI, 3.2-5.9), in tested mites 9.3% (95% CI, 7.4-11.2), 17.2% (95% CI, 14.3-20.1) in I. ricinus ticks and 3.5% (95% CI, 2.2-4.8) in fleas. Sequence analysis of the partial gltA gene and Rickettsia helvetica-, Rickettsia slovaca-, Rickettsia raoultii- species specific real-time PCR tests revealed the presence of R. helvetica, R. slovaca, unidentified Rickettsia and rickettsial endosymbionts. These pathogenic and symbiotic species were confirmed in the following ectoparasite species-Laelaps jettmari, Haemogamasus nidi, Laelaps agilis and Eulaelaps stabularis mites, Ixodes ricinus ticks, Ctenophthalmus solutus, C. assimilis and Megabothris turbidus fleas infesting host-Apodemus agrarius, A. flavicollis, Microtus arvalis and Myodes glareolus small mammals. These results confirm the circulation of R. helvetica, R. slovaca, unidentified Rickettsia and rickettsial endosymbionts in mites, ticks and fleas collected on small mammals in the Kosice region, Eastern Slovakia.