Molecular characterization of Anaplasma phagocytophilum infection in the cervids and feeding ticks from Lithuania

Anaplasma phagocytophilum is a bacterial pathogen, which is a major cause of zoonotic disease, anaplasmosis. The main vectors of A. phagocytophilum are ticks of the Ixodes ricinus complex. A. phagocytophilum has a broad geographic distribution and a high degree of biological and clinical diversity. Epidemiological studies in multiple countries have shown that the prevalence of A. phagocytophilum highly depends on the density of ticks and their potential hosts such as the cervids, which are one of the main sources of nutrition for Ixodes ticks. In Lithuania, the cervids are important game animals but their contribution as reservoirs for A. phagocytophilum remains unknown. The objectives of the study were to investigate the prevalence of A. phagocytophilum infections in the cervids and feeding ticks and to characterize the A. phagocytophilum strains obtained from the cervids and ticks based on sequence analysis of msp4 gene. A total of 187 ticks were collected from 44 cervids (roe deer, red deer, and moose) harvested by professional hunters during the hunting seasons of 2010–2013 and 2016–2017 in Lithuania. Blood and spleen samples were collected from 29 animals (27 roe deer and two red deer). A. phagocytophilum DNA was identified in ten (37.04%) of the 27 roe deer. The overall prevalence of A. phagocytophilum in I. ricinus and D. reticulatus ticks was 39.3% (70/178) and 22.2% (2/9) respectively. The sequence analysis of the msp4 gene of A. phagocytophilum revealed nine different sequence types: five msp4 sequence types were detected in ticks and seven in roe deer.

Phylogeography of msp4 genotypes of Anaplasma marginale in beef cattle from the Brazilian Pantanal

The msp4 gene of A. marginale is unicodon, stable and mostly homogeneous, being considered as a useful marker for phylogeographic characterization of this bacterium. The objective of this work was to analyze the phylogeography of A. marginale based on the msp4 gene in beef cattle from the Brazilian Pantanal, compared to those found in other regions worldwide. The blood samples investigated were collected from 400 animals (200 cows and 200 calves) reared in five extensive breeding farms in this region. The results indicated that of the evaluated samples, 56.75% (227/400) were positive for A. marginale based on the msp1β gene by quantitatitve PCR (qPCR), while 8.37% (19/227) were positive for the msp4 gene in the conventional PCR. In the Network distance analysis, 14 sequences from the Brazilian Pantanal were grouped into a single group with those from Thailand, India, Spain, Colombia, Parana (Brazil), Mexico, Portugal, Argentina, China, Venezuela, Australia, Italy and Minas Gerais (Brazil). Among 68 sequences from Brazil and the world, 15 genotypes were present while genotype number one (#1) was the most distributed worldwide. Both Splitstree and network analyses showed that the A. marginale msp4 sequences detected in beef cattle from the Brazilian Pantanal showed low polymorphism, with the formation of one genogroup phylogenetically related to those found in ruminants from South and Central America, Europe, and Asia.

Anaplasma ovis as the suspected cause of mortality in a neonatal elk calf

Anaplasma ovis infection is known to occur in elk experimentally, but without clinical signs or significant clinicopathologic changes. An elk farm in southern Indiana experienced the death of 3 neonates. Gross findings suggested hemolytic anemia as the cause of death. Splenic impression smears revealed numerous intra-erythrocytic parasites compatible with Anaplasma spp. Products of a semi-nested PCR targeting the msp4 gene of A. ovis were sequenced and had 100% identity with published A. ovis sequences. Given the clinical presentation, vertical transmission of A. ovis was suspected. Pathologic and molecular findings confirmed that natural A. ovis infection occurred in an elk calf.

Occurrence of Anaplasmosis (Anaplasma Marginale) in cattle in Sulaimani province, Kurdistan region of Iraq

The goal of this study was to determine both the incidence of anaplasmosis (Anaplasma Marginale) and phylogenetic relationship between A. marginale isolates from cattle in Sulaimani province, Kurdistan Region- Iraq during (March 10th to April 10th 2021) and those from other Anaplasma spp. A total of two isolates were tested for the major surface protein (msp4) gene for this purpose. Eighty blood samples of cattle (51 males and 29 females) were examined using both microscopic examination and PCR tests. Overall results were 23/80 (28.7 5%) and 8/80 (10 %) using microscopic examination and PCR assay, respectively. Age and sex were not significant factors in the appearance of infection, since no statistically significant difference in infection rate has been observed among sex and age group of cattle (P value >0.05). The results also revealed that the accuracies of traditional method and PCR assays in the diagnosis of the disease were 81 %, and 100 respectively. There was moderate correlation (0.43) between both techniques by the Kappa (k) test. However, The PCR technique recorded the highest sensitivity (100%) and specify (100%) for A. marginale detection. In conclusion, by the findings of the present study, it has been confirmed for the first time that A. marginale is the causative agent of anaplasmosis of cattle in the study areas and the best technique for the detection of either acute or chronic cases in cattle was the PCR assay.

Important frequency of Anaplasma phagocytophilum infection in a population of domiciled dogs in an urbanized area in south-eastern Brazil

ABSTRACT: Anaplasma phagocytophilum is responsible for granulocytic anaplasmosis in humans and various animal species. The aim of the present study was to determine the prevalence of A. phagocytophilum-infected dogs in a residential area of Belo Horizonte, Minas Gerais state, Brazil. A total of 62 dogs were submitted to serological (indirect fluorescent-antibody -IFI) and molecular (PCR) tests. Anti-A. phagocytophilum antibodies were detected in 43.8% of the dogs. Seven dogs (10.9%) were PCR-positive for the msp4 gene, six and four of these were positive for the for the msp2/p44 gene of A. phagocytophilum and 16S rRNA region of granulocytic Anaplasmataceae respectively. This study confirms a relatively high frequency of A. phagocytophilum infection in a population of domiciled dogs in an urbanized area in south-eastern Brazil and highlights the need for further studies on the role of Rhipicephalus sanguineus sensu lato ticks in the transmission of this bacterium to dogs in urban Brazilian areas.