title: Zebrafish gut colonization by mCherry-labelled lactic acid bacteria.
abstract: A critical feature of probiotic microorganisms is their ability to colonize the intestine of the host. Although the microbial potential to adhere to the human gut lumen has been investigated in in vitro models, there is still much to discover about their in vivo behaviour. Zebrafish is a vertebrate model that is being widely used to investigate various biological processes shared with humans. In this work, we report on the use of the zebrafish model to investigate the in vivo colonization ability of previously characterized probiotic lactic acid bacteria. Lactobacillus plantarum Lp90, L. plantarum B2 and Lactobacillus fermentum PBCC11.5 were fluorescently tagged by transfer of the pRCR12 plasmid, which encodes the mCherry protein and which was constructed in this work. The recombinant bacteria were used to infect germ-free zebrafish larvae. After removal of bacteria, the colonization ability of the strains was monitored until 3 days post-infection by using a fluorescence stereomicroscope. The results indicated differential adhesion capabilities among the strains. Interestingly, a displacement of bacteria from the medium to the posterior intestinal tract was observed as a function of time that suggested a transient colonization by probiotics. Based on fluorescence observation, L. plantarum strains exhibited a more robust adhesion capability. In conclusion, the use of pRCR12 plasmid for labelling Lactobacillus strains provides a powerful and very efficient tool to monitor the in vivo colonization in zebrafish larvae and to investigate the adhesion ability of probiotic microorganisms.
authors.affiliation: Promis Biotech srl, via Napoli 25, 71122, Foggia, Italy.
authors.forename: Pasquale
authors.initials: P
authors.lastname: Russo
authors.forename: Iñaki
authors.initials: I
authors.lastname: Iturria
authors.forename: Maria Luz
authors.initials: ML
authors.lastname: Mohedano
authors.forename: Graziano
authors.initials: G
authors.lastname: Caggianiello
authors.forename: Sandra
authors.initials: S
authors.lastname: Rainieri
authors.forename: Daniela
authors.initials: D
authors.lastname: Fiocco
authors.forename: Miguel
authors.initials: M
authors.lastname: Angel Pardo
authors.forename: Paloma
authors.initials: P
authors.lastname: López
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Spano
mesh_terms: D000818:Animals; D041981:Gastrointestinal Tract / Q000382:microbiology*; D017930:Genes, Reporter; D052197:Limosilactobacillus fermentum / Q000235:genetics / Q000254:growth & development*; D048191:Lactiplantibacillus plantarum / Q000235:genetics / Q000254:growth & development*; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010957:Plasmids; D017422:Sequence Analysis, DNA; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors; D015027:Zebrafish / Q000382:microbiology*; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein