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

Abstract

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.

Molecular detection and genetic characterization of Anaplasma marginale and Anaplasma platys-like (Rickettsiales: Anaplasmataceae) in water buffalo from eight provinces of Thailand

“…The 16S rRNA gene is a common target for pathogen detection and species identification and is also used to infer phylogenetic relationships. However, this gene is relatively highly conserved, as demonstrated by its relatively low level of polymorphism and genetic diversity compared to other gene targets, such as the outer membrane protein msp1α [34][35][36], and msp4 [14,34]. Indeed, although msp1α, msp4, and msp5 are relatively conserved genes, they were shown to be useful for phylogenetic analysis among different geographic isolates of A. marginale strains [35].…”

First report on the molecular detection and genetic diversity of Anaplasma marginale in healthy dairy cattle in Khon Kaen province, Thailand

“…The prevalence of anaplasmosis in dairy cattle in Khon Kaen province by molecular detection was 12.73% (49/385) with a 95% CI of 0.51–1.94 for A. marginale based on the msp4 gene. In addition to msp1a , msp1b , msp2 , and msp5 [ 23 , 31 , 32 ], the msp4 gene was used in this study as a good marker for providing sufficient variation to create a large-scale phylogeographic pattern [ 15 ]. It is useful for epidemiological analysis of anaplasmosis [ 33 , 34 ].…”

“…It is suitable for clinically ill cattle, usually during the acute phase of the disease, but parasites are rarely detected microscopically in chronic infection. Polymerase chain reaction (PCR) is a molecular technique that detects organism DNA with high sensitivity and specificity, particularly in persistently infected cattle [14][15][16].…”

Detection and study of genetic diversity of selected vector-borne protozoa and rickettsia in buffaloes in Thailand by biomolecular method

“…The present study was successful in detecting the prevalence of rickettsia and protozoa in buffalo bloods. Nevertheless, the 16S rRNA and 18S rRNA gene sequences had a relatively low level of polymorphism and genetic diversity when compared to other DNA targets such as outer membrane protein: msp1α (de Souza Ramos et al, 2019;Fedorina et al, 2019;Fernandes et al, 2019), msp4 (Ramos et al, 2019, msp5 and major piroplasm surface protein (MPSP)…”