Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Apr;87(2):127-145.

Identification and characterization of potential probiotic lactic acid bacteria isolated from pig feces at various production stages

José D Uezen  1 , Cecilia Aristimuño Ficoseco  1 , María E Fátima Nader-Macías  1 , Graciela M Vignolo  1
Affiliations

Identification and characterization of potential probiotic lactic acid bacteria isolated from pig feces at various production stages

José D Uezen et al. Can J Vet Res. 2023 Apr.

Abstract
in English, French

Lactic acid bacteria (LAB) were isolated, identified, and characterized from pig feces at various growth stages and feed rations in order to be used as probiotic feed additives. Lactic acid bacteria numbers ranged from 7.10 ± 1.50 to 9.40 log CFUs/g for growing and lactating pigs, respectively. Isolates (n = 230) were identified by (GTG)5-polymerase chain reaction and partial sequence analysis of 16S rRNA. Major LAB populations were Limosilactobacillus reuteri (49.2%), Pediococcus pentosaceus (20%), Lactobacillus amylovorus (11.4%), and L. johnsonii (8.7%). In-vitro assays were performed, including surface characterization and tolerance to acid and bile salts. Several lactobacilli exhibited hydrophobic and aggregative characteristics and were able to withstand gastrointestinal tract conditions. In addition, lactobacilli showed starch- and phytate-degrading ability, as well as antagonistic activity against Gram-negative pathogens and the production of bacteriocin-like inhibitory substances. When resistance or susceptibility to antibiotics was evaluated, high phenotypic resistance to ampicillin, gentamicin, kanamycin, streptomycin, and tetracycline and susceptibility towards clindamycin and chloramphenicol was observed in the assayed LAB. Genotypic characterization showed that 5 out of 15 resistance genes were identified in lactobacilli; their presence did not correlate with phenotypic traits. Genes erm(B), strA, strB, and aadE conferring resistance to erythromycin and streptomycin were reported among all lactobacilli, whereas tet(M) gene was harbored by L. reuteri and L. amylovorus strains. Based on these results, 6 probiotic LAB strains (L. reuteri F207R/G9R/B66R, L. amylovorus G636T/S244T, and L. johnsonii S92R) can be selected to explore their potential as direct feed additives to promote swine health and replace antibiotics.

Des bactéries lactiques (LAB) ont été isolées, identifiées et caractérisées à partir de matières fécales de porc à différents stades de croissance et de rations alimentaires afin d’être utilisées comme additifs alimentaires probiotiques. Le nombre de bactéries lactiques variait de 7,10 ± 1,50 à 9,40 log UFC/g pour les porcs en croissance et en lactation, respectivement. Les isolats (n = 230) ont été identifiés par réaction d’amplification en chaîne par la (GTG)5-polymérase et analyse partielle de la séquence de l’ARNr 16S. Les principales populations de LAB étaient Limosilactobacillus reuteri (49,2 %), Pediococcus pentosaceus (20 %), Lactobacillus amylovorus (11,4 %) et L. johnsonii (8,7 %). Des tests in vitro ont été effectués, y compris la caractérisation de surface et la tolérance aux acides et aux sels biliaires. Plusieurs lactobacilles présentaient des caractéristiques hydrophobes et agrégatives et étaient capables de résister aux conditions du tractus gastro-intestinal. De plus, les lactobacilles ont montré une capacité de dégradation de l’amidon et des phytates, ainsi qu’une activité antagoniste contre les agents pathogènes à Gram négatif et la production de substances inhibitrices de type bactériocine. Lorsque la résistance ou la sensibilité aux antibiotiques a été évaluée, une résistance phénotypique élevée à l’ampicilline, à la gentamicine, à la kanamycine, à la streptomycine et à la tétracycline et une sensibilité à la clindamycine et au chloramphénicol ont été observées dans les LAB testés. La caractérisation génotypique a montré que cinq gènes de résistance sur 15 ont été identifiés dans les lactobacilles; leur présence n’était pas corrélée aux traits phénotypiques. Les gènes erm(B), strA, strB et aadE conférant une résistance à l’érythromycine et à la streptomycine ont été signalés parmi tous les lactobacilles, tandis que le gène tet(M) était hébergé par les souches L. reuteri et L. amylovorus. Sur la base de ces résultats, six souches probiotiques LAB (L. reuteri F207R/G9R/B66R, L. amylovorus G636T/S244T et L. johnsonii S92R) peuvent être sélectionnées pour explorer leur potentiel en tant qu’additifs alimentaires directs pour promouvoir la santé des porcs et remplacer les antibiotiques.(Traduit par Docteur Serge Messier).

PubMed Disclaimer

Figures

Figure 1
Figure 1
Polymerase chain reaction amplification of repetitive bacterial DNA element fingerprinting using the (GTG)5 primer of lactic acid bacteria isolated from pigs at various stages. (1 kb DNA ladder, Invitrogen). a — (GTG)5-RAPD profiles including the following biotypes: Bt1-Bt9 (L. reuteri); Bt10 (L. brevis); Bt11-Bt16 (L. amylovorus) and Bt17 (L. crispatus). b — (GTG)5-RAPD profiles including Bt18-Bt22 (L. johnsonii); Bt23 (L. curvatus); Bt24 (E. faecalis); Bt25 (E. hirae); Bt26 (P. pentosaceus); Bt27 (B. subtilis); Bt28 (S. alactolyticus), and Bt29 (W. cibaria). MW — Molecular weight marker.
Figure 2
Figure 2
Surface characterization of lactic acid bacteria isolated from pig feces at various stages. Auto-aggregation and hydrophobicity (toluene and xylene) indexes for pig feces (■) and feed rations (●) * Pearson correlation values.
Figure 3
Figure 3
Biofilm production by lactic acid bacteria from pig feces. a — Lactobacilli and b — Pediococcus, Streptococcus, Weissella, Enterococcus, and Bacillus strains in MRS (red) and MRS-T (MRS with Tween) (blue). The bars indicate the standard deviation obtained from experiments done in triplicate. Different letters indicate significant differences (P < 0.05) between strains under the same culture media conditions.
Figure 4
Figure 4
Resistance to gastrointestinal tract conditions of lactic acid bacteria from pig feces. a — Different pH; b — Different bile salt concentrations.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Centro de Información de Actividades Porcinas [homepage on the Internet] [Last accessed February 11, 2023]. Available from: http://www.ciap.org.ar/Sitio/Sipu/Noticias/Busqueda-Noticias.jsp.
    1. Cao G, Tao F, Hu Y, et al. Positive effects of a Clostridium butyricum-based compound probiotic on growth performance, immune responses, intestinal morphology, hypothalamic neurotransmitters, and colonic microbiota in weaned piglets. Food Funct. 2019;10:2926–2934. - PubMed
    1. Jayaraman B, Nyachoti CM. Husbandry practices and gut health outcomes in weaned piglets: A review. Anim Nutr. 2017;3:205–211. - PMC - PubMed
    1. Guevarra RB, Lee JH, Lee SH, et al. Piglet gut microbial shifts early in life: Causes and effects. J Anim Sci Biotechnol. 2019;10:1. - PMC - PubMed
    1. Rhouma M, Fairbrother JM, Beaudry F, Letellier A. Post weaning diarrhea in pigs: Risk factors and non-colistin-based control strategies. Acta Vet Scand. 2017;59:31. - PMC - PubMed

LinkOut - more resources