A Two Bacteriocinogenic Ligilactobacillus Strain Association Inhibits Growth, Adhesion, and Invasion of Salmonella in a Simulated Chicken Gut Environment

Abstract

In this study, we aimed to develop a protective probiotic coculture to inhibit the growth of Salmonella enterica serovar Typhimurium in the simulated chicken gut environment. Bacterial strains were isolated from the digestive mucosa of broilers and screened in vitro against Salmonella Typhimurium ATCC 14028. A biocompatibility coculture test was performed, which identified two biocompatible strains, Ligilactobacillus salivarius UO.C109 and Ligilactobacillus saerimneri UO.C121 with high inhibitory activity against Salmonella. The cell-free supernatant (CFS) of the selected isolates exhibited dose-dependent effects, and the inhibitory agents were confirmed to be proteinaceous by enzymatic and thermal treatments. Proteome and genome analyses revealed the presence of known bacteriocins in the CFS of L. salivarius UO.C109, but unknown for L. saerimneri UO.C121. The addition of these selected probiotic candidates altered the bacterial community structure, increased the diversity of the chicken gut microbiota challenged with Salmonella, and significantly reduced the abundances of Enterobacteriaceae, Parasutterlla, Phascolarctobacterium, Enterococcus, and Megamonas. It also modulated microbiome production of short-chain fatty acids (SCFAs) with increased levels of acetic and propionic acids after 12 and 24 h of incubation compared to the microbiome challenged with S. Typhimurium. Furthermore, the selected probiotic candidates reduced the adhesion and invasion of Salmonella to Caco-2 cells by 37–39% and 51%, respectively, after 3 h of incubation, compared to the control. These results suggest that the developed coculture probiotic strains has protective activity and could be an effective strategy to control Salmonella infections in poultry.

Full text

Vol.:(0123456789)
1 3
Probiotics and Antimicrobial Proteins (2024) 16:2021–2038
https://doi.org/10.1007/s12602-023-10148-5
RESEARCH
A Two Bacteriocinogenic Ligilactobacillus Strain Association Inhibits
Growth, Adhesion, andInvasion ofSalmonella inaSimulated Chicken
Gut Environment
SabaMiri1· HebatoallahHassan1· GalalAliEsmail1· EmmanuelN.Njoku1· MariemChiba1· BasitYousuf1·
TamerA.E.Ahmed1· MaxwellHincke2,3· WalidMottawea1,4· RiadhHammami1,5
Accepted: 22 August 2023 / Published online: 30 August 2023
© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2023
Abstract
In this study, we aimed to develop a protective probiotic coculture to inhibit the growth of Salmonella enterica serovar Typh-
imurium in the simulated chicken gut environment. Bacterial strains were isolated from the digestive mucosa of broilers and
screened invitro against Salmonella Typhimurium ATCC 14028. A biocompatibility coculture test was performed, which
identified two biocompatible strains, Ligilactobacillus salivarius UO.C109 and Ligilactobacillus saerimneri UO.C121 with
high inhibitory activity against Salmonella. The cell-free supernatant (CFS) of the selected isolates exhibited dose-dependent
effects, and the inhibitory agents were confirmed to be proteinaceous by enzymatic and thermal treatments. Proteome and
genome analyses revealed the presence of known bacteriocins in the CFS of L. salivarius UO.C109, but unknown for L.
saerimneri UO.C121. The addition of these selected probiotic candidates altered the bacterial community structure, increased
the diversity of the chicken gut microbiota challenged with Salmonella, and significantly reduced the abundances of Entero-
bacteriaceae, Parasutterlla, Phascolarctobacterium, Enterococcus, and Megamonas. It also modulated microbiome produc-
tion of short-chain fatty acids (SCFAs) with increased levels of acetic and propionic acids after 12 and 24h of incubation
compared to the microbiome challenged with S. Typhimurium. Furthermore, the selected probiotic candidates reduced the
adhesion and invasion of Salmonella to Caco-2 cells by 37–39% and 51%, respectively, after 3h of incubation, compared
to the control. These results suggest that the developed coculture probiotic strains has protective activity and could be an
effective strategy to control Salmonella infections in poultry.
Keywords Salmonellatyphimurium· Bacteriocinogenic probiotic· Protective coculture· Ligilactobacillus salivarius·
Ligilactobacillus saerimneri· Gut mucosa· Chicken
Introduction
The increasing prevalence of antibiotic resistance among
common pathogenic microbes is a growing public health
concern, with the potential to reduce the effectiveness of
future antimicrobial treatment. The overuse of antibiotics
in poultry production is a leading cause of bacterial anti-
biotic resistance, and pathogenic bacteria, such as Salmo-
nella, which are present in the intestinal tract of animals,
including poultry, can be transmitted to humans via the
food chain or fecal contamination, posing a threat to public
health [1]. Recent studies have revealed a link between
salmonellosis outbreaks and exposure to poultry products,
making it a risk factor for sporadic infections [2]. In recent
years, antibiotic-resistant Salmonella strains have devel-
oped, complicating the treatment of infections caused by
* Riadh Hammami
riadh.hammami@uottawa.ca
1 NuGut Research Platform, School ofNutrition Sciences,
Faculty ofHealth Sciences, University ofOttawa,
K1N6N5Ottawa, ON, Canada
2 Department ofInnovation inMedical Education, Faculty
ofMedicine, University ofOttawa, K1H8M5Ottawa, ON,
Canada
3 Department ofCellular andMolecular Medicine, Faculty
ofMedicine, University ofOttawa, K1H8M5Ottawa, ON,
Canada
4 Department ofMicrobiology andImmunology, Faculty
ofPharmacy, Mansoura University, Mansoura, Egypt
5 Department ofBiochemistry, Microbiology andImmunology,
Faculty ofMedicine, University ofOttawa, Ottawa, ON,
Canada
Content courtesy of Springer Nature, terms of use apply. Rights reserved.