Balanced Th1/Th2 immune response induced by MSP1a functional motif coupled to multiwalled carbon nanotubes as anti-anaplasmosis vaccine in murine model.

Anaplasmosis is one of the most prevalent tick-borne diseases of cattle caused by Anaplasma marginale. MSP1a surface protein has been shown to be involved in eliciting immunity to infected cattle. Carbon nanotubes (CNTs) has been increasingly highlighted due to their needle like structure, which contain multiple attachment sites for biomolecules and may interact with or cross biological membranes, increasing antigen availability to immune system. Here, we have successfully designed a nanocomplex of a synthetic peptide noncovalently attached to multiwalled CNT (MWCNT). Peptide comprising the core motif of the MSP1a was efficiently adsorb onto the nanoparticle surface. The MWCNT-Am1 nanocomplex exhibited high stability and good dispersibility and in vivo immunization showed high levels of IgG1 and IgG2a, followed by increased expression of pro-inflammatory and anti-inflammatory cytokines. This is a proof-of-concept of a nanovaccine that was able to generate a strong immune response compared to the common antigen-adjuvant vaccine without the nanoparticles.

Innate Immune Response to Tick-Borne Pathogens: Cellular and Molecular Mechanisms Induced in the Hosts.

Many pathogens are transmitted by tick bites, including Anaplasma spp., Ehrlichia spp., Rickettsia spp., Babesia and Theileria sensu stricto species. These pathogens cause infectious diseases both in animals and humans. Different types of immune effector mechanisms could be induced in hosts by these microorganisms, triggered either directly by pathogen-derived antigens or indirectly by molecules released by host cells binding to these antigens. The components of innate immunity, such as natural killer cells, complement proteins, macrophages, dendritic cells and tumor necrosis factor alpha, cause a rapid and intense protection for the acute phase of infectious diseases. Moreover, the onset of a pro-inflammatory state occurs upon the activation of the inflammasome, a protein scaffold with a key-role in host defense mechanism, regulating the action of caspase-1 and the maturation of interleukin-1ß and IL-18 into bioactive molecules. During the infection caused by different microbial agents, very similar profiles of the human innate immune response are observed including secretion of IL-1α, IL-8, and IFN-α, and suppression of superoxide dismutase, IL-1Ra and IL-17A release. Innate immunity is activated immediately after the infection and inflammasome-mediated changes in the pro-inflammatory cytokines at systemic and intracellular levels can be detected as early as on days 2-5 after tick bite. The ongoing research field of "inflammasome biology" focuses on the interactions among molecules and cells of innate immune response that could be responsible for triggering a protective adaptive immunity. The knowledge of the innate immunity mechanisms, as well as the new targets of investigation arising by bioinformatics analysis, could lead to the development of new methods of emergency diagnosis and prevention of tick-borne infections.

Molecular epidemiology, associated risk factors, and phylogenetic analysis of anaplasmosis in camel.

Camel Anaplasmosis is caused by members of family Anaplasmatacae, a tick transmitted, obligate intracellular bacteria. The etiological bacteria are transmitted by ixodid tick species. The species have multi host range distribution that is why it is crucial to diagnose it timely. The aim of present study was to investigate the molecular epidemiology i.e. prevalence and risk factors analysis of camel anaplasmosis. Furthermore, variations in hematological standards were also evaluated. The study found an overall 13.33% prevalence in camels. The confirmation of PCR positive samples for Anaplasma spp. was made through sequencing, the study isolatesshowed high homology with Iranian, Chinese, Philippines and South African isolates of Anaplasmatacae (Accession numbers'; KX765882, KP062964, KY242456, LC007100 and U54806) on BLAST queries. The phylogenetic analysis revealedthree study isolates of present study clustered with each other and the cluster was found closer to Chinese isolate of A. phagocytophilum (KY242456), A. marginale (KU586048), and Mongolian isolates of A. ovis (LC194134). Two of the isolates resembled Iranian isolate of Candidatus Anaplasmacamelii (KX765882), while one isolate resembled with Chinese isolates of A. Platys (KX987336) and Croatian isolates of A. Platys (KY114935). The key risk factors odds ratio (OR>1) identified for occurrence of camel anaplasmosis using regression model found sex and age of animal, previous tick history, tick infestation and tick control status, housing type, cracks in walls, rearing system and other species in surrounding as the key risk factors. The hematological parameters like lymphocytes, monocytes, granulocytes and platelets count were significantly decreased (p < 0.05) in diseased camels than healthy. This is the first ever molecular data on camel anaplasmosis in Pakistan. The disease should be monitored unceasingly as the etiologies have multi host distribution. Prompt attention should be offered to animals because neutropenia, lymphopenia and thrombocytopenia can exacerbate the disease by making the animal predisposed to otherdiseases.

Prevalence of Tick-Borne Pathogens from Ticks Collected from Cattle and Wild Animals in Tanzania in 2012.

This study was aimed to disclose the prevalence rate of tick-borne pathogens from ticks collected from cattle and wild animals in Tanzania in 2012. Ticks were collected from slaughtered cattle and dead wild animals from November 5 to December 23, 2012 and identified. PCR for detecting Anaplasmataceae, Piroplamidae, Rickettsiaceae, Borrelia spp., and Coxiella spp. were done. Among those tested, Rickettsiaceae, Piroplasmidae, and Anaplasmataceae, were detected in ticks from the 2 regions. Rickettsiaceae represented the major tick-borne pathogens of the 2 regions. Ticks from animals in Maswa were associated with a higher pathogen detection rate compared to that in ticks from Iringa. In addition, a higher pathogen detection rate was observed in ticks infesting cattle than in ticks infesting wild animals. All examined ticks of the genus Amblyomma were infected with diverse pathogens. Ticks of the genera Rhipicephalus and Hyalomma were infected with 1 or 2 pathogens. Collectively, this study provides important information regarding differences in pathogen status among various regions, hosts, and tick species in Tanzania. Results in this study will affect the programs to prevent tick-borne diseases (TBD) of humans and livestock in Tanzania.

Genetic profiling for Anaplasma and Ehrlichia species in ticks collected in the Eastern Cape Province of South Africa.

BACKGROUND: Anaplasma and Ehrlichia are emerging tick-borne pathogens that cause anaplasmosis and ehrlichiosis in humans and other animals worldwide. Infections caused by these pathogens are deadly if left untreated. There has been relatively no systematic survey of these pathogens among ticks in South Africa, thus necessitating this study. The presence of Anaplasma and Ehrlichia species were demonstrated by PCR in ticks collected from domestic ruminants at some selected communities in the Eastern Cape of South Africa. The ticks were identified by morphological characteristics and thereafter processed to extract bacterial DNA, which was analyzed for the presence of genetic materials of Anaplasma and Ehrlichia. RESULTS: Three genera of ticks comprising five species were identified. The screening yielded 16 positive genetic materials that were phylogenetically related to Ehrlichia sequences obtained from GenBank, while no positive result was obtained for Anaplasma. The obtained Ehrlichia sequences were closely related to E. chaffeensis, E. canis, E. muris and the incompletely described Ehrlichia sp. UFMG-EV and Ehrlichia sp. UFMT. CONCLUSION: The findings showed that ticks in the studied areas were infected with Ehrlichia spp. and that the possibility of transmission to humans who might be tick infested is high.

DHR-ICMR Guidelines for diagnosis & management of Rickettsial diseases in India.

Rickettsial diseases, caused by a variety of obligate intracellular, gram-negative bacteria from the genera Rickettsia, Orientia, Ehrlichia, Neorickettsia, Neoehrlichia, and Anaplasma, belonging to the Alphaproteobacteria, are considered some of the most covert emerging and re-emerging diseases and are being increasingly recognized. Among the major groups of rickettsioses, commonly reported diseases in India are scrub typhus, murine flea-borne typhus, Indian tick typhus and Q fever. Rickettsial infections are generally incapacitating and difficult to diagnose; untreated cases have case fatality rates as high as 30-45 per cent with multiple organ dysfunction, if not promptly diagnosed and appropriately treated. The vast variability and non-specific presentation of this infection have often made it difficult to diagnose clinically. Prompt antibiotic therapy shortens the course of the disease, lowers the risk of complications and in turn reduces morbidity and mortality due to rickettsial diseases. There is a distinct need for physicians and health care workers at all levels of care in India to be aware of the clinical features, available diagnostic tests and their interpretation, and the therapy of these infections. Therefore, a Task Force was constituted by the Indian Council of Medical Research (ICMR) to formulate guidelines for diagnosis and management of rickettsial diseases. These guidelines include presenting manifestations, case definition, laboratory criteria (specific and supportive investigations) and treatment.

Anaplasma capra: a new emerging tick-borne zoonotic pathogen.

The genus Anaplasma includes A. marginale, A. centrale, A. bovis, A. ovis, A. platys, and A. phagocytophilum transmitted by ticks, some of which are zoonotic and cause anaplasmosis in humans and animals. In 2012, a new species was discovered in goats in China. In 2015, the same agent was detected in humans in China, and it was provisionally named Anaplasma capra, referring to 2012. The studies conducted to date have revealed the existence of A. capra in humans, domestic animals, wild animals, and ticks from three different continents (Asia, Europe, and Africa). Phylogenetic analyses based on gltA and groEL sequences show that A. capra clearly includes two different genotypes (A. capra genotype-1 and A. capra genotype-2). Although A. capra human isolates are in the genotype-2 group, goat, sheep, and cattle isolates are in both groups, making it difficult to establish a host genotype-relationship. According to current data, it can be thought that human isolates are genotype-2 and while only genotype-1 is found in Europe, both genotypes are found in Asia. Anaplasma capra causes clinical disease in humans, but the situation is not yet sufficient to understand the zoonotic importance and pathogenicity in animals. In the present review, the history, hosts (vertebrates and ticks), molecular prevalence, pathogenic properties, and genetic diversity of A. capra were evaluated from a broad perspective.

Detection of Anaplasma and Ehrlichia bacteria in humans, wildlife, and ticks in the Amazon rainforest.

Tick-borne bacteria of the genera Ehrlichia and Anaplasma cause several emerging human infectious diseases worldwide. In this study, we conduct an extensive survey for Ehrlichia and Anaplasma infections in the rainforests of the Amazon biome of French Guiana. Through molecular genetics and metagenomics reconstruction, we observe a high indigenous biodiversity of infections circulating among humans, wildlife, and ticks inhabiting these ecosystems. Molecular typing identifies these infections as highly endemic, with a majority of new strains and putative species specific to French Guiana. They are detected in unusual rainforest wild animals, suggesting they have distinctive sylvatic transmission cycles. They also present potential health hazards, as revealed by the detection of Candidatus Anaplasma sparouinense in human red blood cells and that of a new close relative of the human pathogen Ehrlichia ewingii, Candidatus Ehrlichia cajennense, in the tick species that most frequently bite humans in South America. The genome assembly of three new putative species obtained from human, sloth, and tick metagenomes further reveals the presence of major homologs of Ehrlichia and Anaplasma virulence factors. These observations converge to classify health hazards associated with Ehrlichia and Anaplasma infections in the Amazon biome as distinct from those in the Northern Hemisphere.

Epidemiological aspects on vector-borne infections in stray and pet dogs from Romania and Hungary with focus on Babesia spp.

Canine arthropod-borne infections are of major interest in small animal practice and have been widely investigated in Central and Western Europe. However, only limited epidemiological data are available from South-Eastern European countries, although diseases including babesiosis or dirofilariosis are widely recognised as important canine infections in these countries. A steadily increasing number of dogs imported from South-Eastern Europe into Germany require particular attention by small animal practitioners. In this study, a total of 216 dogs [29 local Romanian pet dogs presented at Salvavet Veterinary Clinic in Bucharest, Romania, and 187 imported stray dogs from Romania (n = 109) and Hungary (n = 78) into Germany] were screened by molecular biological, serological and haematological methods for canine arthropod-borne infections. Eleven different parasitic and bacterial vector-borne pathogens-Babesia canis canis, Babesia canis vogeli, Babesia gibsoni, Babesia felis-like, Hepatozoon canis, Leishmania spp., Dirofilaria immitis, Dirofilaria repens, Acanthocheilonema reconditum, Anaplasma phagocytophilum and Mycoplasma haemocanis-were detected. Fifty-six percent of the dogs were positive by direct methods. B. canis canis was the most prevalent pathogen in dogs imported to Germany (42.8%) and dogs submitted for clinical consultation in Bucharest (44.8%). Our data strongly suggest the introduction of an adjusted screening panel in dogs from South-East Europe in view of increasing importation of dogs into Germany.

Proportion and seasonality of blood parasites in animals in Mosul using the Veterinary Teaching Hospital Lab data.

Several local studies have examined evidence of blood parasites in different animals in Mosul; however, information about the most prevalent parasite and the seasonality of the infection remains limited. The objective of the study conducted here was to investigate the proportion and seasonality of blood parasites in animals in Mosul using the Veterinary Teaching Hospital Lab data. Laboratory records for a period of 25 months were used for data retrieval. In all included animals, Giemsa-stained blood smears were examined by an attending clinical pathologist for the presence of parasites. Seasons were assigned on a basis of examination date, and the seasonality was quantified by estimating season-to-season ratio. The results indicated that 61.77% of examined animals were tested positive for blood parasites. The most evident parasites were Trypanosoma spp., Theileria spp., Babesia spp., and then Anaplasma spp., with evidence of mixed infection. The odds of the infection did not significantly vary in different age groups. There was a marked linear pattern in the seasonality of the infection with Trypanosoma spp. and Anaplasma spp. An increase of the infection during spring and autumn with Theileria spp. and Babesia spp. was also evident. In conclusion, infection with blood parasites in different animals in Mosul is common with substantial burden, the effect of age-related infection is negligible, and the seasonality of the infection is evident.

Associations between coinfection prevalence of Borrelia lusitaniae, Anaplasma sp., and Rickettsia sp. in hard ticks feeding on reptile hosts.

An increasing number of studies reveal that ticks and their hosts are infected with multiple pathogens, suggesting that coinfection might be frequent for both vectors and wild reservoir hosts. Whereas the examination of associations between coinfecting pathogen agents in natural host-vector-pathogen systems is a prerequisite for a better understanding of disease maintenance and transmission, the associations between pathogens within vectors or hosts are seldom explicitly examined. We examined the prevalence of pathogen agents and the patterns of associations between them under natural conditions, using a previously unexamined host-vector-pathogen system--green lizards Lacerta viridis, hard ticks Ixodes ricinus, and Borrelia, Anaplasma, and Rickettsia pathogens. We found that immature ticks infesting a temperate lizard species in Central Europe were infected with multiple pathogens. Considering I. ricinus nymphs and larvae, the prevalence of Anaplasma, Borrelia, and Rickettsia was 13.1% and 8.7%, 12.8% and 1.3%, and 4.5% and 2.7%, respectively. The patterns of pathogen prevalence and observed coinfection rates suggest that the risk of tick infection with one pathogen is not independent of other pathogens. Our results indicate that Anaplasma can play a role in suppressing the transmission of Borrelia to tick vectors. Overall, however, positive effects of Borrelia on Anaplasma seem to prevail as judged by higher-than-expected Borrelia-Anaplasma coinfection rates.

Recombinase polymerase amplification (RPA) with lateral flow detection for three Anaplasma species of importance to livestock health.

Anaplasma marginale, A. ovis, and A. phagocytophilum are the causative agents of bovine anaplasmosis, ovine anaplasmosis, and granulocytic anaplasmosis, respectively. The gold standard for diagnosis of post-acute and long-term persistent infections is the serological cELISA, which does not discriminate between Anaplasma species and requires highly equipped laboratories and trained personnel. This study addresses the development of a rapid, isothermal, sensitive, species-specific RPA assays to detect three Anaplasma species in blood and cELISA A. marginale-positive serum samples. Three RPA primer and probe sets were designed targeting msp4 genes of each Anaplasma species and the internal control (GAPDH gene) for each assay. The limit of detection of gel-based or RPA-basic assays is 8.99 × 104 copies/µl = A. marginale, 5.04 × 106 copies/µl = A. ovis, and 4.58 × 103 copies/µl = A. phagocytophilum, and for each multiplex lateral flow or RPA-nfo assays is 8.99 × 103 copies/µl of A. marginale, 5.04 × 103 copies/µl of A. ovis, 4.58 × 103 copies/µl of A. phagocytophilum, and 5.51 × 103 copies/µl of internal control (GAPDH). Although none of the 80 blood samples collected from Oklahoma cattle were positive, the RPA-nfo assays detected all A. marginale cattle blood samples with varying prevalence rates of infection, 83% of the 24 cELISA A. marginale-positive serum samples, and all A. phagocytophilum cell culture samples. Overall, although early detection of three Anaplasma species was not specifically addressed, the described RPA technique represents an improvement for detection of three Anaplasma in regions where access to laboratory equipment is limited.

Frequency and factors associated of potential zoonotic pathogens (Borrelia spp., Rickettsia spp., Leishmania spp., and Anaplasma phagocytophilum) in equids in the state of Bahia, Brazil.

BACKGROUND: Currently, various zoonotic diseases are classified as emerging or reemerging. Because equids have a direct relationship with various vectors, they are possibly more frequently exposed to zoonotic agents than are humans. The undeniable importance of diseases such as human granulocytic anaplasmosis, spotted fever, and leishmaniasis for both public and animal health, as well as the possibility of equids acting as sources, reservoirs, or even sentinels for these pathogens, justifies the detection of their frequency and factors associated with infection in equids from northeastern Brazil. METHODS: Blood samples were collected from 569 equids (528 horses, 33 donkeys, and 8 mules), 516 from a rural area and 53 from an urban area. Pathogen detection was carried out as follows: Borrelia spp. and Rickettsia spp., serological analysis; Leishmania spp., serological analysis and polymerase chain reaction (PCR); Anaplasma phagocytophilum, PCR. Determination of associated factors was carried out through generalized linear models. RESULTS: The frequencies of positivity for the pathogens observed in equids were as follows: Borrelia spp., 13.9% (79/569); Leishmania spp., 3.5% (20/569); Rickettsia spp. 33.4% (190/569). Regarding factors associated with infection, male sex was associated with protection against Borrelia spp.; donkeys and mules were associated with protection against Rickettsia spp., while a younger age was a risk factor. The infection of A. phagocytophilum was not detected in the sampled population. Co-infection was detected in 5.1% (29/569) of the animals. CONCLUSIONS: Most of the studied pathogenic agents are present in the prospected area, indicating a possible risk for both human and animal health. This demonstrates that equids can be considered important sentinels in the assessment of pathogens with zoonotic potential in the region.

Ticks infesting humans and associated pathogens: a cross-sectional study in a 3-year period (2017-2019) in northwest Italy.

BACKGROUND: Tick-borne diseases are common throughout Europe. Ticks transmit pathogens to the host while feeding and together with mosquitoes, they are major vectors of infectious agents worldwide. In recent years, there has been a marked increase in the incidence of tick-bite events and tick-borne disease in northwest Italy, but information on the prevalence of tick-borne pathogens in ticks removed from humans remains scarce. To fill this gap, we report here the prevalence of tick bites and tick-borne pathogens documented for humans in Piedmont, northwest Italy, in the 3-year period 2017-2019. METHODS: Ticks attached to humans during 2017-2019 were collected from residents of urban and rural area by physicians and veterinarians working with local veterinary agencies. All ticks (n = 1290) were morphologically identified to the species level. A subset of ticks removed from children (age 0-18 years) and the elderly (> 70 years), both age groups considered to be at-risk populations, was screened by biomolecular analysis to detect pathogens (e.g. Rickettsia spp., Borrelia spp., Anaplasma spp.). Pathogen identity was confirmed by Sanger sequencing. RESULTS: Ticks were taxonomically assigned to ten species of six genera (Amblyomma, Dermacentor, Haemaphysalis, Hyalomma, Ixodes and Rhipicephalus). Most belonged to the genus Ixodes: 1009 ticks (78.22%) were classified as Ixodes ricinus. A subset of 500 ticks collected from the two at-risk populations were subjected to PCR assay to determine the presence of Rickettsia spp., Borrelia spp., and Anaplasma spp. The overall prevalence of infection was 22.8% (n = 114; 95% confidence interval [CI]: 19.19-26.73%), meaning that at least one pathogen was detected: Rickettsia spp. (prevalence 15%, n = 76; 95% CI 12.17-18.65%); Borrelia spp. (prevalence 6.4%, n = 32; 95% CI 4.42-8.92%); and Anaplasma spp. (prevalence 1.2%, n = 6; 95% CI 0.44-2.6%). CONCLUSIONS: Our data underline the importance of surveillance in the epidemiology of tick-borne diseases and the implementation of strategies to control tick infestation and associated pathogens.

Anaplasma platys-Like Infection in Goats, Beijing, China.

As one of the important tick-borne zoonotic pathogens, Anaplasma has both veterinary and public health significance. Here, we performed a survey of Anaplasma infection in the goats from a farm in Beijing, China, and found 44.6% (41/92) were infected with Anaplasma capra, and 22.8% (21/92) were infected with Anaplasma sp. This Anaplasma sp. bacterium was close to a recently emerging Anaplasma platys strain based on gltA and groEL gene phylogenetic analysis. As to further understand the characteristics of Anaplasma sp., we raised a couple of positive goats (n = 2) in the laboratory with tick-free settings. We observed inappetence, vomiting, high fever, and weakness of limbs in the goat's offspring (n = 3). In addition, the blood samples from all offspring were all positive of this Anaplasma spp. We did not see any intracellular morulae in neutrophils, monocytes, and erythrocytes, but we identified some in the platelets of the blood smears from the positive goats by light microscopy. We named it A. platys-like and suggested it may infect platelets and be transmitted vertically through the placenta of goats. These findings deserve further evaluation.

Seroprevalence of viral and vector-borne bacterial pathogens in domestic dogs (Canis familiaris) in northern Botswana.

BACKGROUND: Domestic dogs (Canis familiaris) have the potential to act as disease reservoirs for wildlife and are important sentinels for common circulating pathogens. Therefore, the infectious disease seroprevalence among domestic dogs in northern Botswana may be indicative of pathogen exposure of various wildlife species. The objective of this study was to assess the seroprevalence of Ehrlichia spp., Borrelia burgdorferi, Anaplasma spp., Dirofilaria immitis, canine adenovirus, canine parvovirus, and canine distemper virus in domestic dogs as proxies of disease prevalence in the local wildlife in the Okavango Delta region of Botswana. Statistical analysis assessed crude and factor-specific seroprevalence proportions in relation to age, sex, and geographical location as predictors of seropositivity. Logistic regression was used to identify adjusted predictors of seropositivity for each of the pathogens of interest. RESULTS: Samples from 233 dogs in a total of seven locations in Maun, Botswana, and surrounding villages were collected and serologically analyzed. No dogs were seropositive for B. burgdorferi, while low seroprevalence proportions were observed for Anaplasma spp. (2.2%) and D. immitis (0.9%). Higher seroprevalence proportions were observed for the tick-borne pathogen Ehrlichia spp. (21.0%), and 19.7% were seropositive for canine adenovirus (hepatitis). The highest seroprevalence proportions were for canine parvovirus (70.0%) and canine distemper virus (44.8%). The predictors of seropositivity revealed that adults were more likely to be seropositive for canine adenovirus, canine distemper virus, and canine parvovirus than juveniles, and location was a risk factor for canine adenovirus, canine distemper virus, canine parvovirus, and Ehrlichia spp. CONCLUSIONS: Results indicate that increasing tick control and vaccination campaigns for domestic dogs may improve the health of domestic animals, and potentially wildlife and humans in the Okavango Delta since viral and vector-borne bacterial pathogens can be transmitted between them.

Sir Arnold Theiler and the discovery of anaplasmosis: a centennial perspective.

Sir Arnold Theiler's research in 1908/09 led to the discovery of the first rickettsial pathogen, Anaplasma marginale, and set the stage for his development and implementation of an effective live vaccine based on a less virulent strain, A. marginale ss. centrale. His 1910 report, describing A. marginale, is among the classic monographs in infectious disease research, presenting not only observations in exacting detail but also highlighting the deductive reasoning leading to association of a new pathogen with a specific disease. With a centennial perspective and both conceptual frameworks and molecular tools unimaginable in Theiler's time, the significance of several observations in the original report--cyclic bacteremia, strain superinfection, and taxonomic position--is now clear and highlight the broad applicability of key principles of pathogen biology.

First detection and molecular identification of the zoonotic Anaplasma capra in deer in France.

Cervids are known to be reservoirs of zoonotic bacteria transmitted by ticks. This study aimed to identify the Anaplasma species carried by captive red deer and swamp deer in a wild fauna reserve in France. Blood from 59 red deer and 7 swamp deer was collected and analyzed over a period of two years. A semi-nested PCR targeting the 23S rRNA was performed to detect and characterize Anaplasma spp. and determine the presence of zoonotic species. Anaplasma phagocytophilum was identified in 14/59 red deer (23.7%) but it was not identified in any of the swamp deer (7 animals). Three sequences could not be assigned to any particular species based on the 23S rRNA sequences. Complementary nested PCR targeting 16S rRNA, gltA and groEL genes and sequencing analysis then identified these sequences as a recently reported zoonotic species, Anaplasma capra; this species was found in 2 red deer (Cervus elaphus) and 1 swamp deer (Rucervus duvaucelii). This is the first report of the tick-borne zoonotic bacterium A. capra in France, a species otherwise described only in China, Japan, Malaysia and South Korea in goats, sheep, deer, cattle and Japanese serows (Capricornis crispus). While this bacterium may have been introduced into the reserve by infected imported animals, its local epidemiological cycle via tick transmission seems possible as locally born deer were found infected. Diagnostic methods, especially molecular ones, should take into account the potential infection of animals and humans with this species.

Novel variants of the newly emerged Anaplasma capra from Korean water deer (Hydropotes inermis argyropus) in South Korea.

BACKGROUND: Anaplasma spp. are tick-borne Gram-negative obligate intracellular bacteria that infect humans and a wide range of animals. Anaplasma capra has emerged as a human pathogen; however, little is known about the occurrence and genetic identity of this agent in wildlife. The present study aimed to determine the infection rate and genetic profile of this pathogen in wild animals in the Republic of Korea. METHODS: A total of 253 blood samples [198 from Korean water deer (Hydropotes inermis argyropus), 53 from raccoon dogs (Nyctereutes procyonoides) and one sample each from a leopard cat (Prionailurus bengalensis) and a roe deer (Capreolus pygargus)] were collected at Chungbuk Wildlife Center during the period 2015-2018. Genomic DNA was extracted from the samples and screened for presence of Anaplasma species by PCR/sequence analysis of 429 bp of the 16S rRNA gene marker. Anaplasma capra-positive isolates were genetically profiled by amplification of a longer fragment of 16S rRNA (rrs) as well as partial sequences of citrate synthase (gltA), heat-shock protein (groEL), major surface protein 2 (msp2) and major surface protein 4 (msp4). Generated sequences of each gene marker were aligned with homologous sequences in the database and phylogenetically analyzed. RESULTS: Anaplasma capra was detected in blood samples derived from Korean water deer, whereas samples from other animal species were negative. The overall infection rate in tested samples was 13.8% (35/253) and in the water deer the rate was 17.8% (35/198), distributed along the study period from 2015 to 2018. Genetic profiling and a phylogenetic analysis based on analyzed gene markers revealed the occurrence of two distinct strains, clustered in a single clade with counterpart sequences of A. capra in the database. CONCLUSIONS: Anaplasma capra infection were detected in Korean water deer in the Republic of Korea, providing insight into the role of wildlife as a potential reservoir for animal and human anaplasmosis. However, further work is needed in order to evaluate the role of Korean water deer as a host/reservoir host of A. capra.

Molecular detection and characterization of Anaplasma platys and Ehrlichia canis in dogs from northern Colombia.

Ehrlichia canis and Anaplasma platys are intracellular tick-transmitted bacteria that infect dogs; there is evidence for limited zoonotic potential as well. The prevalence of E. canis in Colombia has been evaluated in different regions; however little is known about the prevalence or distribution of A. platys. Neither pathogen has been studied in the Magdalena region, thus the purpose of our study was to assess the prevalence of these pathogens in dogs attending veterinary clinics from the cities of Santa Marta and Ciénaga, and to assess possible associated risk factors for infection. A. platys and E. canis infections in blood were evaluated by Taqman PCR assays. E. canis was detected in 26/170 (15.3%, 95% CI 10.4%-21.8%) and A. platys in 34/168 (20.2%, 95% CI 14.6%-27.3%) of all dogs tested. Eleven dogs (6.5%, 95% CI 3.4-11.7%) were coinfected with both pathogens. Sequencing results showed low diversity within E. canis and within A. platys strains, however a strain of E. canis detected in our study area is genetically distinct from strains reported in another city of Colombia. Our results suggest that for A. platys, Santa Marta dogs were at greater risk than Ciénaga dogs, and that purebred dogs were at slightly lower risk in both areas. The confirmation of these pathogens in northern Colombia should cause concern for the possible co-transmission of these agents to humans or animals in the region.