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Despite a goat population of approximately 80 million in Pakistan during 2020−2021, the prevalence of vector-borne pathogens in goats remains largely underexplored. This study aimed to assess the molecular prevalence and phylogenetic characteristics of Anaplasma ovis, Anaplasma marginale and Anaplasma phagocytophilum in goat blood samples (N = 239) collected from three districts (Muzaffargarh, Rajanpur, and Dera Ghazi Khan) in Punjab between September 2023 and October 2024. Blood samples were first screened with generic and then with species specific primers. Molecular analyses revealed a prevalence of 39% for Anaplasma spp. and 14% for A. ovis . A. marginale and A. phagocytophilum were not detected. DNA sequencing, by targeting 16S rRNA and msp4 genes, and BLAST analysis confirmed the presence of Anaplasma spp. and A. ovis, respectively. For both screening, bacterial prevalence rates varied significantly across sampling sites (P = 0.01 for Anaplasma spp. and P = 0.04 for A. ovis ). Additionally, the prevalence of Anaplasma spp. significantly differed among goat breeds (P = 0.004), while no association was found between goat sex and bacterial infections (P > 0.05 for both screening). Notably, Anaplasma spp. infection was associated with a significant decrease in red blood cell count and hemoglobin concentration, while A. ovis infection did not affect the complete blood count profile. Phylogenetic analysis revealed that our Anaplasma spp. isolates clustered with those from Iran, Cyprus and China while our A. ovis isolates clustered with those from Pakistan, China, and Sudan. In conclusion, this study reports the presence of Anaplasma spp. and A. ovis in Pakistani goats and recommends large-scale studies across diverse geo-climatic regions to further investigate the epidemiology, genetic diversity and host-parasite interactions for effective control of these infections in local goat populations.

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 × 10 4 copies/µl =  A. marginale , 5.04 × 10 6 copies/µl =  A. ovis , and 4.58 × 10 3 copies/µl =  A. phagocytophilum , and for each multiplex lateral flow or RPA- nfo assays is 8.99 × 10 3 copies/µl of A. marginale, 5.04 × 10 3 copies/µl of A. ovis , 4.58 × 10 3 copies/µl of A. phagocytophilum, and 5.51 × 10 3 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.

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.

Canine ehrlichiosis and anaplasmosis are important tick-borne diseases with a worldwide distribution. Information has been continuously collected on these infections in Europe, and publications have increased in recent years. Prevalence rates are high for Ehrlichia and Anaplasma spp. infections in dogs from different European countries. The goal of this article was to provide a practical guideline for veterinary practitioners on the diagnosis, treatment, and prevention of ehrlichiosis and anaplasmosis in dogs from Europe. This guideline is intended to answer the most common questions on these diseases from a practical point of view.

Anaplasmosis is a tick-borne disease (TBDs) caused by Anaplasma spp. In areas where TBDs are endemic, it is crucial to consider the animals’ immunological status in relation to these diseases. The true prevalence of bovine anaplasmosis, the percentage of animals with protective antibodies against this TBD, and the diagnostic characteristics of three tests (multiplex polymerase chain reaction (mPCR), competitive-inhibition enzyme-linked immunosorbent assay (cELISA), and blood smear (BS)) were estimated using a Bayesian approach. A total of 620 samples were collected in two subtropical areas of Ecuador. A significant finding of this study is that approximately 70% of cattle in those endemic areas harbored protective antibodies against Anaplasma marginale . This elevated percentage may stem from persistent exposure with a high pathogen prevalence in ticks. The decline in cELISA specificity must be attributed to cross-reactivity with protective antibodies against Anaplasma spp. It is crucial to interpret this test outcome alongside exposure history and clinical manifestations. The elevated apparent prevalence detected by cELISA and BS should be contextualized with mPCR results. The high seroprevalence and infrequent clinical outbreaks suggest that the pathogen has achieved endemic stability. This study provides valuable insights into the dynamics of anaplasmosis in endemic areas and may serve as a foundation for devising TBDs control strategies in these areas.

Anaplasma phagocytophilum is the agent of tick-borne fever, equine, canine and human granulocytic anaplasmosis. The common route of A. phagocytophilum transmission is through a tick bite, the main vector in Europe being Ixodes ricinus . Despite the apparently ubiquitous presence of the pathogen A. phagocytophilum in ticks and various wild and domestic animals from Europe, up to date published clinical cases of human granulocytic anaplasmosis (HGA) remain rare compared to the worldwide status. It is unclear if this reflects the epidemiological dynamics of the human infection in Europe or if the disease is underdiagnosed or underreported. Epidemiologic studies in Europe have suggested an increased occupational risk of infection for forestry workers, hunters, veterinarians, and farmers with a tick-bite history and living in endemic areas. Although the overall genetic diversity of A. phagocytophilum in Europe is higher than in the USA, the strains responsible for the human infections are related on both continents. However, the study of the genetic variability and assessment of the difference of pathogenicity and infectivity between strains to various hosts has been insufficiently explored to date. Most of the European HGA cases presented as a mild infection, common clinical signs being pyrexia, headache, myalgia and arthralgia. The diagnosis of HGA in the USA was recommended to be based on clinical signs and the patient’s history and later confirmed using specialized laboratory tests. However, in Europe since the majority of cases are presenting as mild infection, laboratory tests may be performed before the treatment in order to avoid antibiotic overuse. The drug of choice for HGA is doxycycline and because of potential for serious complication the treatment should be instituted on clinical suspicion alone.

Anaplasma phagocytophilum , an obligate intracellular bacterial pathogen that lives in a host cell-derived vacuole, causes human and veterinary diseases of global importance. In the pathogen-occupied vacuole, A. phagocytophilum transitions from a replicative, non-infectious morphotype to a non-replicative, infectious morphotype that is released to spread infection. We established that distinct modes of bacterial cell division drive not only A. phagocytophilum replication but also its differentiation to the infectious form and dissemination to naïve cells. How pleomorphism is regulated in most vacuole-adapted bacterial pathogens is poorly understood. Therefore, this study advances fundamental knowledge of vacuole-adapted pleomorphic bacteria pathobiology and could ultimately identify common novel antibiotic targets for treating the diseases they cause.

Background Anaplasma marginale is an obligate intracellular bacterium and the main cause of bovine anaplasmosis in tropical and subtropical regions. In Egypt, data regarding the prevalence of A. marginale in ruminant hosts and of the circulating genotypes is lacking. This study therefore aimed to (i) investigate the presence, epidemiology and genotypes of A. marginale in cattle and buffaloes in Egypt, (ii) to evaluate suitable diagnostic tools and (iii) to identify co-infections of A. marginale with other selected tick-borne pathogens. Methods Blood samples were collected from 394 animals (309 cattle and 85 buffaloes) from three different areas in Egypt. For the detection of A. marginale infection, several tests were compared for their sensitivity and specificity: blood smear analysis, enzyme-linked immunosorbent assay (ELISA), PCR, real-time PCR and reverse line blot (RLB) assay. Co-infections with A. marginale, piroplasms and other Anaplasmataceae were surveyed by RLB while A. marginale genotypes were identified by amplifying and sequencing the partial msp1α gene. Results Anaplasma marginale DNA was amplified by qPCR in 68.3% of cattle and 29.4% of buffaloes. RLB showed infection with A. marginale in 50.2% of cattle and 42.5% of buffaloes. Blood smear analysis detected this agent in 16.2% of cattle and 2.4% of buffaloes. ELISA showed specific antibodies against A. marginale in 54.9% of cattle. Anaplasma marginale was associated, in cattle and buffaloes, with several tick-borne pathogens ( Theileria annulata , Babesia bovis , Babesia bigemina , Babesia occultans and Anaplasma platys ). A significant difference of A. marginale infection level was noticed in cattle, where animals between 3–5-years-old had a higher prevalence (79.2%) compared to those older than 5 years (36.4%) and younger than 3 years (59.7%) and one year (64.5%), respectively ( P  = 0.002281). Microsatellite analysis identified 15 different genotypes. Conclusions The epidemiological findings revealed high prevalence of A. marginale in cattle and buffaloes in all the investigated areas. The circulation of diverse genotypes was observed, most of these A. marginale genotypes being specific for Egypt. The qPCR assay was confirmed to be the most sensitive tool for detection of A. marginale in cattle and buffaloes even in the carrier state, highlighting the importance of using suitable diagnostic tests.

The bacterium Anaplasma phagocytophilum has for decades been known to cause the disease tick-borne fever (TBF) in domestic ruminants in Ixodes ricinus-infested areas in northern Europe. In recent years, the bacterium has been found associated with Ixodes-tick species more or less worldwide on the northern hemisphere. A. phagocytophilum has a broad host range and may cause severe disease in several mammalian species, including humans. However, the clinical symptoms vary from subclinical to fatal conditions, and considerable underreporting of clinical incidents is suspected in both human and veterinary medicine. Several variants of A. phagocytophilum have been genetically characterized. Identification and stratification into phylogenetic subfamilies has been based on cell culturing, experimental infections, PCR, and sequencing techniques. However, few genome sequences have been completed so far, thus observations on biological, ecological, and pathological differences between genotypes of the bacterium, have yet to be elucidated by molecular and experimental infection studies. The natural transmission cycles of various A. phagocytophilum variants, the involvement of their respective hosts and vectors involved, in particular the zoonotic potential, have to be unraveled. A. phagocytophilum is able to persist between seasons of tick activity in several mammalian species and movement of hosts and infected ticks on migrating animals or birds may spread the bacterium. In the present review, we focus on the ecology and epidemiology of A. phagocytophilum, especially the role of wildlife in contribution to the spread and sustainability of the infection in domestic livestock and humans.

Background Anaplasmosis, an animal disease caused by rickettsial bacteria in the genus Anaplasma , is of considerable economic importance in livestock animals in many countries worldwide. The objectives of this study were to determine the identity, prevalence, and geographic distribution of Ehrlichia and Anaplasma in naturally infected water buffalo in Thailand using PCR amplification and sequencing of the 16S ribosomal RNA and heat shock protein groEL genes. A total of 456 buffalo blood samples from Thailand were investigated. Species identification and genetic differentiation of intra-population and inter-population with the global isolates were conducted based on nucleotide sequences. Interplay between the infection and host factors was also assessed. Results Overall, 41% of water buffalo were found to be infected with rickettsial organisms in the family Anaplasmataceae , but Ehrlichia spp. , Neorickettsia spp., and Wolbachia spp. were not found in any of the sequenced samples in this study. Female buffalo were more frequently infected with bacteria in the family Anaplasmataceae than males [71 out of 176 females (40.3%) versus 11 out of 47 males (23.4%)]. The Odds Ratio value indicated that the risk of infection for female buffalo was 2.2-fold higher than that for males ( p  < 0.05). We detected three haplotypes of A. marginale 16S rRNA gene and they were placed in a clade that was closely related to the A. marginale in buffalo in China; and cattle in Thailand, Uganda, and China. Homology searching of groEL sequences against the GenBank™ database using the BLASTn algorithm revealed that the obtained sequences had a high percentage similarity (98.36–99.62%) to A. platys sequences. The groEL sequences of three A. platys -like isolates were clustered in the same clade as the A. platys from the tick Rhipicephalus microplus in China . Conclusions Our data showed that the apparently healthy buffalo were naturally infected by bacteria in the family Anaplasmataceae at a relatively high prevalence. We also report the finding of A. platys -like infections in water buffalo in Thailand for the first time. Water buffalo serving as the reservoir host of anaplasmosis is of concern for managing the disease control and prevention in ruminants.