Molecular detection of hemoplasmas in domestic cats from different Brazilian regions

Abstract

Domestic cats can be parasitized by Mycoplasma haemofelis, 'Candidatus Mycoplasma haemominutum', and 'Candidatus Mycoplasma turicensis'. Although the molecular occurrence of hemoplasmas in domestic cats has been investigated in the five geographical regions of Brazil, no studies have been conducted in the states of Rondônia and Minas Gerais to date. The present work aimed to investigate the molecular occurrence of hemoplasmas in cats from four Brazilian states in the Northern, Central-Western and Southeastern regions of the country. Among 486 blood samples - 80 from Rondônia (RO), 100 from Goiás (GO), 155 from Minas Gerais (MG), and 151 from São Paulo (SP) - submitted to the endogenous gapdh gene-based PCR, 94.44% (459/486) were positive, of which 1.96% (9/459; 1 from RO, 2 from GO, 5 from MG, and 1 from SP) were positive in the qPCR assay for hemoplasmas based on the 16S rRNA gene. In the phylogenetic analyses, the obtained 16S rRNA (two genotypes) and 23S rRNA (three genotypes) sequences were positioned together with 'Candidatus Mycoplasma haemominutum'. The present study showed the first molecular evidence of infection by hemoplasmas in cats from MG and RO, contributing to a better understanding of the epidemiology of feline hemoplasmosis agents in Brazil.

Figure 1. Map generated by QGIS 2.18.2.8, showing the states of Brazil (in different colors) and the location of cities by state where cats were sampled in the present study.

Figure 2. Phylogenetic tree based on an alignment of 989 bp length of Mycoplasma spp. 16S rRNA sequences, created using the Bayesian method, and GTR+I+G as the evolutionary model. The sequences obtained in this study are highlighted in red. Mycoplasmoides pneumoniae, Bacillus subtilis and Bacillus mycoides were used as an outgroup.

Figure 3. Phylogenetic tree based on an alignment of 759 bp length of Mycoplasma spp. 23S rRNA sequences, created using the Bayesian method and GTR+G as the evolutionary model. The sequences obtained in this study are highlighted in red. Bacillus cereus and Bacillus subtilis were used as an outgroup.

Figure 4. (A) ‘Candidatus Mycoplasma haemominutum’ split-network distance analysis of the 16S rRNA sequences. The analysis was performed applying the NeighborNet method based on a 469 bp alignment. The sequences obtained in the present study are highlighted in red; (B) Genotype network obtained from 16S rRNA sequences (469 bp) of 'Candidatus Mycoplasma haemominutum' detected in domestic cats, wild felids and a dog worldwide. The size of the circles varies according to the number of sequences comprising each genotype. Each color represents the location where each sequence was sampled. The black vertical lines represent mutational events between each genotype. The 16S rRNA genotypes obtained in the present study are represented by an illustration of a cat (), followed by the accession number on GenBank.

Figure 5. (A) ‘Candidatus Mycoplasma haemominutum’ split-network distance analysis of the 23S rRNA sequences. The analysis was performed applying the NeighborNet method based on a 671 bp alignment. The sequences obtained in the present study are highlighted in red; (B) Genotype network obtained from 23S rRNA sequences (671 bp) of 'Candidatus Mycoplasma haemominutum' detected in domestic cats and humans worldwide. The size of the circles varies according to the number of sequences from each genotype. Each color represents the location where each sequence was sampled. The black vertical lines represent mutational events between each genotype. The 23S rRNA genotypes obtained in the present study are represented by an illustration of a cat (), followed by the accession number on GenBank.