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We investigated the species diversity of Mycobacteriaceae in surface water samples from six environments at the zoological park in São Paulo, Brazil. Three hundred and eighty isolates were cultivated and identified by phenotypic characteristics (growth rate and pigmentation) and sequencing of hsp65, rpoB and 16S rRNA genes. The results revealed that almost 48% of the isolates could be identified at the species level; about 50% were classified at the genus level, and only less than 2% of the isolates showed an inconclusive identification. The isolates classified at the genus level and not identified were then evaluated by phylogenetic analyses using the same three concatenated target genes. The results allowed us to identify at the genus level some isolates that previously had inconclusive identification, and they also suggested the presence of putative candidate species within the sample, demonstrating that this zoological park is an important source of diversity.
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31935265
???displayArticle.pmcLink???PMC6959594 ???displayArticle.link???PLoS One
Fig 1. Distribution of Mycobacteriaceae isolates according to origin and period of analysis.cS: crude sewage; tS: treated sewage; L70: Lake70; SS: Smaller Spring; BS: Bigger Spring and WTS: water treatment station.
Fig 2. Classification of the isolates into 78 groups by sequencing the hsp65 gene.Similarity dendrogram was based on the multiple alignment matrix of the hsp65 gene sequences using the UPGMA method. In the first column are the isolates randomly selected to be analyzed by sequencing the 16S rRNA and rpoB genes. Column 2: Number of isolates included in each group. Column 3: Result of the comparison of the hsp65 gene sequences of isolates with sequences deposited in the GenBank database with their percentages of identity. Column 4: GenBank accession number of each sequence.
Fig 3. Phylogeny of Mycobacteriaceae based on concatenated sequences of hsp65, rpoB and 16S rRNA genes (2,511 bp), according to maximum-likelihood method (Lnl = 36046.26).Values above branches are bootstrap proportions (≥70%) and those in parenthesis after the terminal names refer to number of isolates belonging to each group. * Groups with more than one isolate studied by phylogenetic analysis.
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