In Vitro Characterization and Safety Assessment of <i>Streptococcus salivarius</i>, <i>Levilactobacillus brevis</i> and <i>Pediococcus pentosaceus</i> Isolated from the Small Intestine of Broiler Breeders

In poultry production, antibiotics have been excessively used as growth promoters to support well-being and decrease mortality caused by pathogenic microorganisms. The overuse of antibiotics has led to the emergence of antibiotic-resistant bacteria and the presence of antibiotic residues in poultry...

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Main Authors: Nwabisa Happiness Kokwe, Freedom Tshabuse, Feroz Mahomed Swalaha
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Microorganisms
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Online Access:https://www.mdpi.com/2076-2607/13/6/1231
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author Nwabisa Happiness Kokwe
Freedom Tshabuse
Feroz Mahomed Swalaha
author_facet Nwabisa Happiness Kokwe
Freedom Tshabuse
Feroz Mahomed Swalaha
author_sort Nwabisa Happiness Kokwe
collection DOAJ
description In poultry production, antibiotics have been excessively used as growth promoters to support well-being and decrease mortality caused by pathogenic microorganisms. The overuse of antibiotics has led to the emergence of antibiotic-resistant bacteria and the presence of antibiotic residues in poultry products. To counteract this problem, probiotics could be used as adjuncts or as substitutes for preserving a diverse and balanced microflora to prevent the colonization and multiplication of pathogenic bacteria in the GI tract. This study aimed to isolate and characterize the potential probiotic properties of lactic acid bacteria from the small intestine of 23-week-old broiler breeders, with the goal of identifying potential probiotic candidates. Four phenotypically healthy broiler breeders were selected, and intestinal contents were aseptically collected and cultured on MRS agar. From the initial pool of 39 colonies, six isolates were identified based on Gram-positive and catalase-negative characteristics and further classified using 16S rRNA sequencing as <i>Levilactobacillus brevis</i> (<i>n</i> = 3), <i>Pediococcus pentosaceus</i> (<i>n</i> = 2), and <i>Streptococcus salivarius</i> (<i>n</i> = 1). These strains were further evaluated for probiotic properties such as transit resistance to simulated upper gastrointestinal conditions, antagonist activity, haemolytic activity, and cell surface properties such as autoaggregation, co-aggregation and hydrophobicity, <i>in vitro</i>. <i>L. brevis</i> NKFS8 showed good tolerance to pH 3, while <i>P. pentosaceus</i> NKSF10 exhibited good tolerance to pH 4 acidic conditions. All isolates demonstrated good survivability in bile salt concentration of 3% (<i>w</i>/<i>v</i>), with <i>P. pentosaceus</i> NKSF10 exhibiting the highest tolerance. The isolates showed a wide range of antagonistic activity against the test pathogens <i>Pseudomonas aeruginosa</i> (ATCC 27853), <i>Salmonella typhimurium</i>, <i>Salmonella enterica</i> (ATCC 13314), <i>Staphylococcus aureus</i> (ATCC 29213), and <i>Listeria monocytogenes</i> (ATCC 7644). Furthermore, these strains exhibited good auto-aggregation, co-aggregation, and hydrophobicity properties. In conclusion, lactic acid bacteria from the small intestine of broiler breeders present a valuable prospect for the development of effective probiotics. These probiotics can be utilized as a supplementary inclusion in poultry feed, obviating the need for antibiotics as growth promoters. Nevertheless, additional in vivo studies are required to closely monitor and assess the effects of probiotics on the gastrointestinal system of chickens.
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spelling doaj-art-b0f7b31868424c76bab75ea23311a2d12025-08-20T03:16:34ZengMDPI AGMicroorganisms2076-26072025-05-01136123110.3390/microorganisms13061231In Vitro Characterization and Safety Assessment of <i>Streptococcus salivarius</i>, <i>Levilactobacillus brevis</i> and <i>Pediococcus pentosaceus</i> Isolated from the Small Intestine of Broiler BreedersNwabisa Happiness Kokwe0Freedom Tshabuse1Feroz Mahomed Swalaha2Department of Biotechnology and Food Science, Durban University of Technology, Durban 4001, South AfricaDepartment of Biochemistry and Microbiology, University of Zululand, KwaDlangezwa 3886, South AfricaDepartment of Biotechnology and Food Science, Durban University of Technology, Durban 4001, South AfricaIn poultry production, antibiotics have been excessively used as growth promoters to support well-being and decrease mortality caused by pathogenic microorganisms. The overuse of antibiotics has led to the emergence of antibiotic-resistant bacteria and the presence of antibiotic residues in poultry products. To counteract this problem, probiotics could be used as adjuncts or as substitutes for preserving a diverse and balanced microflora to prevent the colonization and multiplication of pathogenic bacteria in the GI tract. This study aimed to isolate and characterize the potential probiotic properties of lactic acid bacteria from the small intestine of 23-week-old broiler breeders, with the goal of identifying potential probiotic candidates. Four phenotypically healthy broiler breeders were selected, and intestinal contents were aseptically collected and cultured on MRS agar. From the initial pool of 39 colonies, six isolates were identified based on Gram-positive and catalase-negative characteristics and further classified using 16S rRNA sequencing as <i>Levilactobacillus brevis</i> (<i>n</i> = 3), <i>Pediococcus pentosaceus</i> (<i>n</i> = 2), and <i>Streptococcus salivarius</i> (<i>n</i> = 1). These strains were further evaluated for probiotic properties such as transit resistance to simulated upper gastrointestinal conditions, antagonist activity, haemolytic activity, and cell surface properties such as autoaggregation, co-aggregation and hydrophobicity, <i>in vitro</i>. <i>L. brevis</i> NKFS8 showed good tolerance to pH 3, while <i>P. pentosaceus</i> NKSF10 exhibited good tolerance to pH 4 acidic conditions. All isolates demonstrated good survivability in bile salt concentration of 3% (<i>w</i>/<i>v</i>), with <i>P. pentosaceus</i> NKSF10 exhibiting the highest tolerance. The isolates showed a wide range of antagonistic activity against the test pathogens <i>Pseudomonas aeruginosa</i> (ATCC 27853), <i>Salmonella typhimurium</i>, <i>Salmonella enterica</i> (ATCC 13314), <i>Staphylococcus aureus</i> (ATCC 29213), and <i>Listeria monocytogenes</i> (ATCC 7644). Furthermore, these strains exhibited good auto-aggregation, co-aggregation, and hydrophobicity properties. In conclusion, lactic acid bacteria from the small intestine of broiler breeders present a valuable prospect for the development of effective probiotics. These probiotics can be utilized as a supplementary inclusion in poultry feed, obviating the need for antibiotics as growth promoters. Nevertheless, additional in vivo studies are required to closely monitor and assess the effects of probiotics on the gastrointestinal system of chickens.https://www.mdpi.com/2076-2607/13/6/1231gastrointestinal systemprobioticslactic acid bacteriapoultry
spellingShingle Nwabisa Happiness Kokwe
Freedom Tshabuse
Feroz Mahomed Swalaha
In Vitro Characterization and Safety Assessment of <i>Streptococcus salivarius</i>, <i>Levilactobacillus brevis</i> and <i>Pediococcus pentosaceus</i> Isolated from the Small Intestine of Broiler Breeders
Microorganisms
gastrointestinal system
probiotics
lactic acid bacteria
poultry
title In Vitro Characterization and Safety Assessment of <i>Streptococcus salivarius</i>, <i>Levilactobacillus brevis</i> and <i>Pediococcus pentosaceus</i> Isolated from the Small Intestine of Broiler Breeders
title_full In Vitro Characterization and Safety Assessment of <i>Streptococcus salivarius</i>, <i>Levilactobacillus brevis</i> and <i>Pediococcus pentosaceus</i> Isolated from the Small Intestine of Broiler Breeders
title_fullStr In Vitro Characterization and Safety Assessment of <i>Streptococcus salivarius</i>, <i>Levilactobacillus brevis</i> and <i>Pediococcus pentosaceus</i> Isolated from the Small Intestine of Broiler Breeders
title_full_unstemmed In Vitro Characterization and Safety Assessment of <i>Streptococcus salivarius</i>, <i>Levilactobacillus brevis</i> and <i>Pediococcus pentosaceus</i> Isolated from the Small Intestine of Broiler Breeders
title_short In Vitro Characterization and Safety Assessment of <i>Streptococcus salivarius</i>, <i>Levilactobacillus brevis</i> and <i>Pediococcus pentosaceus</i> Isolated from the Small Intestine of Broiler Breeders
title_sort in vitro characterization and safety assessment of i streptococcus salivarius i i levilactobacillus brevis i and i pediococcus pentosaceus i isolated from the small intestine of broiler breeders
topic gastrointestinal system
probiotics
lactic acid bacteria
poultry
url https://www.mdpi.com/2076-2607/13/6/1231
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