Anaplasma Capra Research Articles

Molecular identification and characterization of Anaplasma capra and Anaplasma platys-like in Rhipicephalus microplus in Ankang, Northwest China

BackgroundFour species within Anaplasma genus are emerging zoonotic pathogens, which are transmitted by ticks and generate veterinary and public health concerns. Here, we performed a molecular survey of Anaplasma in Ankang, Northwest China.MethodsHard ticks were collected and identified using morphological and molecular methods. Human-pathogenic Anaplasma species were tested using nested polymerase chain reaction. The nearly complete rrs, gltA, and groEL genes sequences from revealed Anaplasma species were amplified and sequenced to determine their molecular characteristics and their phylogeny.ResultsAll ticks collected in Ankang belonged to the Rhipicephalus microplus. Novel unclassified Anaplasma strains genetically related to A. platys and A. capra were detected in these ticks. Co-infection of these two organisms was also found. The novel unclassified Anaplasma strains identified in this study formed a distinct phylogenetic lineage based on the groEL gene and two lineages based on the gltA gene within A. platys and related strains group. The revealed A. capra strains identified in this study were most closely related to those detected in humans and other vertebrate animals.ConclusionWe revealed the presence of A. capra, a novel human pathogens in R. microplus ticks in previously unrecognized endemic regions. We also detected a novel unclassified Anaplasma species genetically related to A. platys. The epidemiology of anaplasmosis caused by these two Anaplasma species in humans should be assessed in future studies.

A novel genotype of “Anaplasma capra” in wildlife and its phylogenetic relationship with the human genotypes

Anaplasma spp. are gram-negative obligate intracellular bacteria of remarkable importance in both human and veterinary health. Six Anaplasma species have currently been classified in the genus Anap...

Differential identification of Anaplasma in cattle and potential of cattle to serve as reservoirs of Anaplasma capra, an emerging tick-borne zoonotic pathogen

Bovine anaplasmosis is a tick-borne, infectious, non-contagious disease caused by Anaplasma marginale, A. centrale, A. bovis, and zoonotic A. phagocytophilum. Recently, Anaplasma capra detected in goats was identified as a novel zoonotic pathogen. To determine whether A. capra can infect bovines, we used PCR to differentially diagnose Anaplasma spp. in 1219 South Korean cattle by performing multilocus gene typing and restriction fragment length polymorphism (RFLP). Nucleotide sequencing and phylogenetic analysis detected the 16S rRNA gene of A. bovis and four genes from A. capra in 12 (1.0%) and five (0.4%) cattle, respectively. Supplementary discrimination between A. bovis and A. capra was accomplished by RFLP. The 16S rRNA, msp4, groEL, and gltA genes of A. capra identified in this study had much lower degrees of identity to those in A. centrale and other Anaplasma spp. A. phagocytophilum was not detected in any of the tested cattle. Although the prevalence was low, this study suggests the potential of cattle to serve as reservoirs of A. capra. Thus, further studies are needed to clarify the pathogenesis of A. capra in cattle and its possible involvement in transmission to humans.

Detection and Phylogenetic Characterization of Anaplasma capra: An Emerging Pathogen in Sheep and Goats in China.

Anaplasma capra is an emerging pathogen, which can infect ruminants and humans. This study was conducted to determine the occurrence of A. capra in the blood samples of sheep and goats in China. Using nested polymerase chain reaction (nested-PCR) targeting the gltA gene and conventional PCR targeting the heat shock protein (groEL) gene and the major surface protein4 gene (msp4), A. capra was detected in 129 (8.9%) of 1453 sheep and goat blood samples. The positive rate was higher in goats (9.4%, 89/943) than in sheep (7.8%, 40/510) (χ2 = 1.04, p > 0.05, df = 1). For sheep, A. capra was found in 17 sites from 2 provinces. The prevalence was 28.6% in sheep from Liaoning province, which was higher than in Henan Province (7.3%). For goats, A. capra was detected in 35 sites from 7 provinces. The prevalence varied from 0 to 19.4% in the goat sites examined. The prevalence rates were 19.4, 19.3, 10, 8.8, 6.8, 1.8, and 0% in goats from Guizhou province, Henan Province, Inner Mongolia Autonomous Region, Shanxi Province, Xinjiang Uygur Autonomous Region, Yunnan province, and Gansu province, respectively. Based on the analysis of the A. capra citrate synthase gene (gltA), two variants were identified. Variant I showed a high sequence similarity to the A. capra, which were previously reported in sheep, goats, Ixodes persulcatus, Haemaphysalis longicornis, Haemaphysalis qinghaiensis, and humans. Variant II was only found in Luoyang, Anyang, and Sanmengxia, of Henan province. To our knowledge, this is the first detection of this variant of A. capra in sheep and goat blood in China. Phylogenetic analysis based on groEL and msp4 genes showed that the Anaplasma sp. sequences clustered independently from A. capra and other Anaplasma species with high bootstrap values. We found A. capra DNA in sheep and goats in China, providing evidence that sheep and goats can be infected by A. capra. We also found that this zoonotic pathogen is widely distributed in China. This study provides information for assessing the public health risks for human anaplasmosis.

Molecular investigation of Anaplasma spp. in domestic and wildlife animals in Peninsular Malaysia.

Anaplasma spp. are Gram-negative obligate intracellular, tick-borne bacteria which are of medical and veterinary importance. Little information is available on Anaplasma infection affecting domestic and wildlife animals in Malaysia. This study investigated the presence of Anaplasma spp. in the blood samples of domestic and wildlife animals in Peninsular Malaysia, using polymerase chain reaction (EHR-PCR) assays targeting the 16S rRNA gene of Anaplasmataceae. High detection rates (60.7% and 59.0%, respectively) of Anaplasma DNA were noted in 224 cattle (Bos taurus) and 78 deer (77 Rusa timorensis and one Rusa unicolor) investigated in this study. Of the 60 amplified fragments obtained for sequence analysis, Anaplasma marginale was exclusively detected in cattle while Anaplasma platys/Anaplasma phagocytophilum was predominantly detected in the deer. Based on sequence analyses of the longer fragment of the 16S rRNA gene (approximately 1000 bp), the occurrence of A. marginale, Anaplasma capra and Candidatus Anaplasma camelii in cattle, Candidatus A. camelii in deer and Anaplasma bovis in a goat was identified in this study. To assess whether animals were infected with more than one species of Anaplasma, nested amplification of A. phagocytophilum, A. bovis and Ehrlichia chaffeensis DNA was performed for 33 animal samples initially screened positive for Anaplasmataceae. No amplification of E. chaffeensis DNA was obtained from animals investigated. BLAST analyses of the 16S rDNA sequences from three deer (R. timorensis), a buffalo (Bubalus bubalis) and a cow (B. taurus) reveal similarity with that of Candidatus Anaplasma boleense strain (GenBank accession no.: KX987335). Sequence analyses of the partial gene fragments of major surface protein (msp4) gene from two deer (R. timorensis) and a monitor lizard (Varanus salvator) show the detection of a strain highly similar (99%) to that of A. phagocytophilum strain ZJ-China (EU008082). The findings in this study show the occurrence of various Anaplasma species including those newly reported species in Malaysian domestic and wildlife animals. The role of these animals as reservoirs/maintenance hosts for Anaplasma infection are yet to be determined.

A novel zoonotic Anaplasma species is prevalent in small ruminants: potential public health implications

BackgroundTick-borne diseases currently represent an important issue for global health. A number of emerging tick-transmitted microbes continue to be discovered, and some of these are already identified as the cause of human infections. Over the past two decades, Anaplasma phagocytophilum is considered to be mainly responsible for human anaplasmosis. However, a novel zoonotic pathogen provisionally named “Anaplasma capra” has recently been identified in China. In this study, we did an active surveillance of A. capra in goats and sheep in different geographical regions of China.MethodsThe presence of A. capra was determined by nested PCR in 547 blood samples collected from goats and sheep from 24 counties distributed in 12 provinces in China. The molecular characterization of A. capra isolates in sheep and goats was achieved based on four conventional genetic markers (16S rRNA, gltA, groEL and msp4 genes).ResultsAnaplasma capra was identified in 75 of 547 animals, with an overall prevalence of 13.7%. The infection rates in the survey sites ranged from 0 to 78.6%, and were significantly different (P < 0.01). Phylogenetic analysis revealed that the isolates obtained from goats, sheep, Ixodes persulcatus ticks and humans create a separate clade within the genus Anaplasma and distinct from other recognized Anaplasma species. These findings indicated that these A. capra isolates possess the same molecular characteristics, suggesting that this organism could be a substantial health threat to both animals and humans.ConclusionsAnaplasma capra is an emerging tick-transmitted zoonotic pathogen. This novel Anaplasna species is widespread across China with an overall prevalence of 13.7% in goats and sheep with isolates indistinguishable from those found in humans. These findings warrant increased public health awareness for human anaplasmosis.

Prevalence of Severe Febrile and Thrombocytopenic Syndrome Virus, Anaplasma spp. and Babesia microti in Hard Ticks (Acari: Ixodidae) from Jiaodong Peninsula, Shandong Province.

The presence of tick-borne pathogens and their possible coinfections were evaluated among host-seeking ticks in seven cities from Jiaodong peninsula, Shandong Province, with specific PCR or reverse transcription-PCR tests. Among 2107 ticks collected, four species of three genera were identified with Haemaphysalis longicornis as predominant species, and total of 63 H. longicornis and 10 Rhicephalus microplus were confirmed infected with tick-borne pathogens. These pathogens were consequently identified as severe febrile and thrombocytopenic syndrome virus (SFTSV), Anaplasma capra, Anaplasma phygocytophilum, and Babesia microti, respectively, with high phylogenetic scores on some fragments of species-specific genes. The infection rates of the pathogens in H. longicornis were presented as 1.03%, 0.84%, 0.58%, and 1.66%, respectively, close related to its field density and clump distribution pattern. Furthermore, coinfection of A. capra and SFTSV was also discovered from two female H. longicornis in Pingdu city. These results indicated that the potential human pathogens other than severe febrile and thrombocytopenic syndrome might be transmitted by hard ticks separately or in combination, and more reliable differential diagnosis, proper administrations, rational prevention, and control measures should be developed with the support of precision laboratory tests.

Molecular survey and characterization of a novel Anaplasma species closely related to Anaplasma capra in ticks, northwestern China.

Background Anaplasma spp. are tick-transmitted bacteria that infect a wide variety of wild and domestic animals. These pathogens exhibit a high degree of biological diversity, broad geographical distribution, and represent a serious threat to veterinary and public health worldwide.ResultsA novel Anaplasma species was identified in Haemaphysalis qinghaiensis (Ixodidae) in northwestern China and was molecularly characterized by comparison of 16S rRNA, gltA, and groEL gene sequences. Of the 414 samples tested, 24 (5.8%) were positive for this Anaplasma species. On the basis of the 16S rRNA gene, this organism has been found to be closely related to and exhibit the highest sequence similarity with A. capra (99.8–99.9%) that was identified in goats and humans in northern China, but was distinct from other known Anaplasma species. Sequence analysis of the gltA and groEL genes revealed that this Anaplasma species was distinct from A. capra considering the lower sequence identity (88.6–88.7% for gltA and 90.6–91.0% for groEL) and a divergent phylogenetic position. Therefore, we described this Anaplasma species as A. capra-like bacteria.ConclusionsThe present study reports a potential novel Anaplasma species closely related to A. capra in H. qinghaiensis in northwestern China. The zoonotic potential of A. capra-like bacteria needs to be further determined.

Human infection with a novel tick-borne Anaplasma species in China: a surveillance study

Anaplasma phagocytophilum and Anaplasma ovis cause human infections. We investigated the potential for human pathogenicity of a newly discovered Anaplasma species infecting goats in China. We collected blood samples from patients with a history of tick bite in the preceding 2 months at Mudanjiang Forestry Central Hospital of Heilongjiang Province from May 1, to June 10, 2014, to detect the novel Anaplasma species by PCR. We inoculated positive samples into cell cultures. We characterised the isolated pathogen by morphological and phylogenetic analyses. We tested serum antibodies by indirect immunofluorescence assay. 28 (6%) of 477 patients assessed were infected with the novel Anaplasma species according to PCR and sequencing. We isolated the pathogen in vitro from three patients. Phylogenetic analyses of rrs, gltA, groEL, msp2, and msp4 showed that the pathogen was distinct from all known Anaplasma species. We provisionally nominate it "Anaplasma capra". 22 (92%) of 24 patients with data available had seroconversion or a four-fold increase in antibody titres. All 28 patients developed non-specific febrile manifestations, including fever in 23 (82%), headache in 14 (50%), malaise in 13 (46%), dizziness in nine (32%), myalgia in four (14%), and chills in four (14%). Additionally, ten (36%) of 28 patients had rash or eschar, eight (29%) had lymphadenopathy, eight (29%) had gastrointestinal symptoms, and three (11%) had stiff neck. Five patients were admitted to hospital because of severe disease. Six (35%) of 17 patients with data available had high hepatic aminotransferase concentrations. The emergence of "A capra" as a cause of human disease suggests that individuals living in or travelling to endemic regions in northern China should take precautions to reduce their risk of exposure to this novel tick-borne pathogen. Natural Science Foundation of China and the US National Institutes of Health.

Of goats and men: rethinking anaplasmoses as zoonotic infections

Ticks transmit the widest variety of pathogens among arthropod vectors, leading to substantial public health threats and economic loss in livestock industries.1 Novel tick-transmitted microbes with links to human disease continue to be discovered. In The Lancet Infectious Diseases, Hao Li and colleagues2 describe a new tick-transmitted bacterium from patients in northern China. PCR and sequencing showed nucleotide variations signifying a phylogenetically distinct Anaplasma species. The provisionally nominated “Anaplasma capra”—so named for its discovery in goats (Capra aegagrus hircus)—joins the growing list of human anaplasmosis pathogens with pastoral origins. For decades, Anaplasma phagocytophilum has been recognised as a veterinary pathogen causing tick-borne fever among ruminants. But only recently has it been associated with human infection,3 with seroepidemiological evidence suggesting incidence as high as 36% in endemic regions of the USA and 28% in populations across 15 European countries.4,5 In 2010, a variant of Anaplasma ovis, which infects goats and sheep worldwide, was detected in a Cypriot patient following a tick bite,6 although no additional human cases have been reported. Anaplasma species organisms have been detected in several ruminant species worldwide. Such reports could lead one to think that anaplasmoses are associated only with agricultural communities, but that is far from the case. Surveys of European green spaces suggest that tick-borne pathongens are common in urban areas.7,8 At present, 54% of the world’s population lives in urban areas and that proportion is expected to reach two-thirds by 2050.9 With greater urbanisation, human beings increasingly share environments with wild and domestic mammals that maintain tick populations and also serve as reservoirs for tick-borne pathogens. Aside from ruminants, common animals in cities, including rodents, birds, cats, and dogs, can host Anaplasma species.10 Considering the wide geographical spread of more than 19 tick species that can harbour anaplasmosis pathogens and the broad range of mammalian reservoirs,10,11 these pathogens are virtually ubiquitous in nature. Li and colleagues sought “A capra” in patients who lived in areas where it had been identified in ticks and goats, rather than beginning with patients and seeking the cause of their tick-related febrile disease. In doing so, they discovered a new causative agent of human anaplasmosis. They noted several features of “A capra” illness that differ from Anaplasma phagocytophilum infection: unreliable visualisation of morulae (intracytoplasmic vacuolar inclusions of anaplasma) in peripheral blood samples; lower incidence of leucopenia, thrombocytopenia, and increased hepatic aminotransferase concentrations; low-to-undetectable seroreactivity to A phagocytophilum antigen; and more common rash and eschar. “A capra” infection presented with symptoms that could make diagnosis difficult. A phagocytophilum infection is not often lethal in immunocompetent individuals, but delaying antibiotic treatment can be fatal.12 Morbidity and mortality associated with “A capra” is not yet known. Five of 28 patients with “A capra” infection were admitted to hospital, one of whom had CNS complications, suggesting that “A capra” could pose a substantial threat to public health.2 The big question remains: how many Anaplasma species that contribute to human disease remain undiscovered? The study by Li and colleagues provides us with a 2-month snapshot from a single region. With over 200 anaplasmosis and related ehrlichiosis cases of undetermined cause reported by the US Centers for Disease Control and Prevention in 2013,13 undiscovered anaplasma pathogens might already be causing human disease in the USA and elsewhere. Because human cases are generally under-reported,12 the global burden of anaplasmoses on human health is probably underestimated. With increasing urbanisation and interactions between people and ticks, additional zoonotic pathogens, including Anaplasma species, are likely to be discovered.