Lactic acid bacteria are gut commensals that contribute to gut homeostasis. Research on symbiotic microorganisms in wild animals may help to understand both co-evolution and health beneficial characteristics of gut bacteria for the host. However, wild animals have rarely been studied in their natural habitats, due to the difficulty in culturing and the lack of access to appropriate microbiological tools in field conditions. Therefore, the isolation of microorganisms in the wild environment represents a challenge. The time between sampling and analysis as well as modalities for collection and storage of samples are important factors which negatively impact the viability of fecal microorganisms. In the present study, Indri indri the largest extant living lemur of Madagascar, a 'critically endangered' species, has been studied. Eighteen fecal samples were collected in MRS soft agar, stored at 4 - 25 °C for about 10 d and then maintained at -80°C. Samples were plated onto MRS agar with mupirocin, a selective medium for bifidobacteria. In only two subjects out of 18, bacterial growth has been observed. Fifteen isolates with bacillar morphology have been obtained (6 from Zafy, 6 years old and 7 from Dart, 6 months old) and grouped by BOX-PCR. All isolates showed the same profiles except D6. Isolates D1 and D6, from Dart, and Z3 from Zafy were identified by means of comparative 16S rRNA analysis and genome sequencing. The pairwise Average Nucleotide Identity to 100 genomes of Lp. plantarum confirmed the identification as Lp. plantarum, however, the isolates from lemurs clustered peripheral to other strains in the species. Genomic analyses further revealed that genes for metabolism of phenolic acids and more than 18 genes from the CARD database for antimicrobial resistance are part of the core genome of Lp. plantarum. Our findings expand the knowledge of occurrence of lactobacilli in wild primates.
Improved procedure for the isolation of beneficial bacteria in wild habitat: Lactiplantibacillus plantarum from Indri indri in Madagascar / Modesto Monica, Donatella Scarafile, Gautam Gaur, Nanzhen Qiao, Felicitas Pswarayi, Luigi Maria Borruso, Camillo Sandri, Caterina Spiezio, Cristina Giacoma, Paola Mattarelli, Michael Ganzle . - ELETTRONICO. - (2021). (Intervento presentato al convegno ECCO XXXIX - Virtual meeting tenutosi a London nel 22-24 September 2021).
Improved procedure for the isolation of beneficial bacteria in wild habitat: Lactiplantibacillus plantarum from Indri indri in Madagascar
Modesto Monica;Donatella Scarafile;Camillo Sandri;Paola Mattarelli;
2021
Abstract
Lactic acid bacteria are gut commensals that contribute to gut homeostasis. Research on symbiotic microorganisms in wild animals may help to understand both co-evolution and health beneficial characteristics of gut bacteria for the host. However, wild animals have rarely been studied in their natural habitats, due to the difficulty in culturing and the lack of access to appropriate microbiological tools in field conditions. Therefore, the isolation of microorganisms in the wild environment represents a challenge. The time between sampling and analysis as well as modalities for collection and storage of samples are important factors which negatively impact the viability of fecal microorganisms. In the present study, Indri indri the largest extant living lemur of Madagascar, a 'critically endangered' species, has been studied. Eighteen fecal samples were collected in MRS soft agar, stored at 4 - 25 °C for about 10 d and then maintained at -80°C. Samples were plated onto MRS agar with mupirocin, a selective medium for bifidobacteria. In only two subjects out of 18, bacterial growth has been observed. Fifteen isolates with bacillar morphology have been obtained (6 from Zafy, 6 years old and 7 from Dart, 6 months old) and grouped by BOX-PCR. All isolates showed the same profiles except D6. Isolates D1 and D6, from Dart, and Z3 from Zafy were identified by means of comparative 16S rRNA analysis and genome sequencing. The pairwise Average Nucleotide Identity to 100 genomes of Lp. plantarum confirmed the identification as Lp. plantarum, however, the isolates from lemurs clustered peripheral to other strains in the species. Genomic analyses further revealed that genes for metabolism of phenolic acids and more than 18 genes from the CARD database for antimicrobial resistance are part of the core genome of Lp. plantarum. Our findings expand the knowledge of occurrence of lactobacilli in wild primates.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
