Isolation and comparative genomic analysis of reuterin-producingLactobacillus reuterifrom poultry gastrointestinal tract

Abstract

Lactobacillus reuteri is a natural inhabitant of selected animal and human gastrointestinal tract (GIT). Certain strains have the capacity to transform glycerol to 3-hydroxypropionaldehyde (3-HPA), further excreted to form reuterin, a potent antimicrobial system. Reuterin-producing strains may be applied as a natural antimicrobial in feed to prevent pathogen colonization of animals, such as in poultry, and replace added antimicrobials. To date, only seven L. reuteri strains isolated from poultry have been characterized which limits phylogenetic studies and host-microbes interactions characterization. This study aimed to isolate L. reuteri strains from poultry GIT and to characterize their reuterin production and antimicrobial resistance (AMR) profiles using phenotypic and genetic methods. Seventy reuterin-producing strains were isolated from poultry crop, faeces and caeca and twenty-five selected for further characterization. Draft genomes were generated for the new 25 isolates and integrated in a phylogenetic tree of 40 strains from different hosts. Phylogenetic analysis based on gene content as well as on core genomes showed grouping of the selected 25 L.reuteri poultry isolates within the poultry/human lineage VI. Strains harbouring pducob-cbi-hem genes (23/25) produced between 156 mM ± 11 and 330 mM ± 14 3-HPA, from 600 mM of glycerol, in the conditions of the test. All 25 poultry strains were sensitive to cefotaxime (MIC between 0.016 and 1 µg/mL) and penicillin (MIC between 0.02 and 4 µg/mL). Akin to the reference strains DSM20016 and SD2112, the novel isolates were resistant to penicillin, possibly associated with identified point mutations in ponA, pbpX, pbpF and pbpB. All strains resistant to erythromycin (4/27) carried the ermB gene, and it was only present in poultry strains. All strains resistant to tetracycline (5/27) harbored tetW gene. This study confirms the evolutionary history of poultry/human lineage VI and identifies pdu-cob-cbi-hem as a frequent trait but not always present in this lineage. L. reuteri poultry strains producing high 3-HPA yield may have potential to prevent enteropathogen colonization of poultry.

Isolation and Comparative Genomic Analysis of Reuterin-Producing Lactobacillus reuteri From the Chicken Gastrointestinal Tract

“…This manuscript has been released as a Pre-Print at BioRxiv ( Greppi et al, 2019 ). The authors are grateful to Alfonso Die for the HPLC measurements.…”

Reuterin Demonstrates Potent Antimicrobial Activity Against a Broad Panel of Human and Poultry Meat Campylobacter spp. Isolates

“…These strains phylogenetically cluster together in the poultry/human VI clade of L. reuteri, previously described [18]. Details on the isolation, characterisation and efficiency of reuterin production of these strains were reported by Greppi et al [17].…”

“…The production of reuterin by L. reuteri is a strain-specific characteristic [37]. The ability of chicken-derived reuterin-producing L. reuteri PTA5_F13 to synthesize reuterin, and inactivate C.jejuni in co-culture was evaluated and compared with L. reuteri PTA8_1, which does not produce reuterin [17]. The phenotypic similarity of L. reuteri PTA8_1 and L. reuteri PTA5_F13 was confirmed during mono-cultures without glycerol where there were no significant difference in cell growth and main metabolite (lactate and acetate) production.…”

“…reuteri DSM20016 was obtained from the DSM strain collection (Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany). L. reuteri PTA8_11 (non-reuterin producer) and L. reuteri PTA5_F13, PTA6_C2 and PTA4_C2 (all three reuterin-producers) were isolated from poultry gut and shown to share average nucleotide identity (ANI) of 95% with reference strain L. reuteri DSM20016T [17]. They were obtained from our culture collection (Laboratory of Food Biotechnology, ETH-Zürich, Switzerland).…”

Hybridization alters the gut microbial and metabolic profile concurrent with modifying intestinal functions in Tunchang pigs

“…Here, we gave several conjectures to learn more about the alteration of the colonization of Lactobacillus. KEGG pathway of the metagenome showed that 'tetracycline biosynthesis' was enriched in TC pigs, such antibiotics affected microbial colonization, including Prevotella and Lactobacillus (Takahashi et al, 2006;Campedelli et al, 2019;Li et al, 2019;Greppi et al, 2020). On the other hand, several steroidrelated metabolites were negatively correlated with Lactobacillus.…”