Targeted sequencing analysis of Mycoplasma gallisepticum isolates in chicken layer and breeder flocks in Thailand

Abstract

Mycoplasma gallisepticum (MG) is one of the most economically important pathogens worldwide. MG affects the respiratory system and impairs growth performance in poultry. In developing countries, the most widely used technique to identify MG is the conventional PCR assay. In this study, 24 MG isolates collected from Thailand farms with unvaccinated chickens during 2002-2020 were characterized by gene-targeted sequencing (GTS), followed by phylogenetic analysis using unweighted pair group method with arithmetic mean. These 24 Thai MG isolates differed from vaccine strains, including the F, ts-11 and 6/85 strains. One isolate showed 99.5-100% genetic similarity to the F strain with 4 partial gene analyses. This result may have been due to contamination from vaccinated flocks because the F strain is the most commonly used vaccine strain in Thailand. However, the GTS analysis using the partial MG genes in this study showed that the isolates could be grouped into different patterns based on individual gene sequences. The phylogenetic analysis of partial mgc2, gapA, pvpA and lp gene sequences classified the Thai MG isolates into 7, 11, 7 and 2 groups, respectively. In conclusion, at least 2 partial MG genes, especially partial gapA and mgc2 genes, are needed to differentiate MG isolates.

Figure 1

A phylogenetic tree of Thai MG and reference strains based on the alignment of the partial mgc2 gene was constructed with the unweighted pair group method with arithmetic mean (UPGMA) using Bionumeric version 7.6 software.

Figure 2

A phylogenetic tree of Thai MG and reference strains based on the alignment of the partial gapA gene was constructed with the unweighted pair group method with arithmetic mean (UPGMA) using Bionumeric version 7.6 software.

Figure 3

A phylogenetic tree of Thai MG and reference strains based on the alignment of the partial pvpA gene was constructed with the unweighted pair group method with arithmetic mean (UPGMA) using Bionumeric version 7.6 software.

Figure 4

A phylogenetic tree of Thai MG and reference strains based on the alignment of the partial pvpA gene was constructed with the unweighted pair group method with arithmetic mean (UPGMA) using Bionumeric version 7.6 software.