Benefits and Inputs From Lactic Acid Bacteria and Their Bacteriocins as Alternatives to Antibiotic Growth Promoters During Food-Animal Production

Resistance to antibiotics is escalating and threatening humans and animals worldwide. Different countries have legislated or promoted the ban of antibiotics as growth promoters in livestock and aquaculture to reduce this phenomenon. Therefore, to improve animal growth and reproduction performance and to control multiple bacterial infections, there is a potential to use probiotics as non-antibiotic growth promoters. Lactic acid bacteria (LAB) offer various advantages as potential probiotics and can be considered as alternatives to antibiotics during food-animal production. LAB are safe microorganisms with abilities to produce different inhibitory compounds such as bacteriocins, organic acids as lactic acid, hydrogen peroxide, diacetyl, and carbon dioxide. LAB can inhibit harmful microorganisms with their arsenal, or through competitive exclusion mechanism based on competition for binding sites and nutrients. LAB endowed with specific enzymatic functions (amylase, protease…) can improve nutrients acquisition as well as animal immune system stimulation. This review aimed at underlining the benefits and inputs from LAB as potential alternatives to antibiotics in poultry, pigs, ruminants, and aquaculture production

Purification et caractérisations du mode d'action et de l'innocuité de l'entérocine S37, une bactériocine produite par Enterococcus faecalis S37

La sécurité et l hygiène des aliments est une préoccupation majeure des sociétés modernes. Le développement de bactéries pathogènes des aliments à l instar de Camplyobacter jejuni et Listeria monocytogenes exige la recherche de nouvelles méthodes de lutte contre ces microorganismes. Les bactériocines produites par les bactéries lactiques constituent une alternative intéressante qu il convient de continuer d explorer et valoriser. Dans cette thèse, nous nous sommes intéressés à l écosystème digestif du poulet en vue d isoler de nouvelles bactéries lactiques bactériocinogènes. Il résulte de notre étude une prédominance du genre Lactobacillus. Parmi les bactéries lactiques bactériocinogènes identifiées, il y a Enterococcus faecalis S37 qui présente une activité anti-Listeria forte et anti-Campylobacter variable. Cette activité est attribuée à la production d une bactériocine nommée entérocine S37. Nous avons purifié l entérocine S37 par des méthodes chromatographiques et caractérisé son mode d action. L entérocine S37 a un poids moléculaire compris entre 4 et 6 kDa, elle résiste aux traitements thermiques et pH extrêmes. Elle est active principalement contre les souches de Listeria monocytogenes et Enterococcus faecalis. Le mode d action de l entérocine S37, étudié grâce à l utilisation de modulateurs de canaux potassiques a montré une interaction avec les canaux potassiques ATP dépendants provoquant la fuite d ions K+ et des molécules d ATP intracellulaires. Par ailleurs, l innocuité de l entérocine S37 a été démontrée sur un modèle cellulaire entérocitaire Caco- 2/TC7 différenciées, faisant de cette bactériocine un candidat potentiel pour une utilisation en prévention de ces microorganismes. Nous avons également montré que la combinaison de la nisine A et/ou pédiocine PA-1 avec la polymyxine E permet un effet additif et/ou synergique sur des souches résistantes de Listeria monocytogenes.Development of foodborne pathogens such as Campylobacter jejuni and Listeria monocytogenes encourages the search of novel technologies in order to fight against the aforementioned bacteria. Bacteriocins produced by lactic acid bacteria stand as an interesting issue. In this project we studied and established the lactic acid bacteria present in chicken digestive tract. Consequently, the genus Lactobacillus was the predominant one, and Enterococcus faecalis S37 exhibited antagonism against Listeria monocytogenes while an unstable anti-Campylobacter activity was observed. This strain produces a bacteriocin, named enterocin S37, which was purified by a set of chromatographical methods. Enterocin S37 has a molecular weight comprised between 4 and 6 kDa, and a good resistance to elevated temperatures and pH extremes. Enterocin S37 was active against listerial and enterococcal strains. The mode of action of enterocin S37 was studied by using different potassium channels modulators. Such investigations concluded to ATP-dependent potassium channels interaction of enterocin S37, leading to leakage of K+ and intracellular ATP. Furthermore we established that enterocin S37 did not affect differentiated Caco-2/TC7 cells arguing on the capacities of enterocin S37 to be used in prevention of the aforementioned bacteria and certain infection. Additionally, we demonstrated the synergistic effect of nisin A and pediocin PA-1, when combined with polymyxin E (antibiotic) against resistant and sensitive listerial strains.NANTES-BU Sciences (441092104) / SudocSudocFranceF

Broadening and enhancing bacteriocins activities by association with bioactive substances

International audienceBacteriocins are antimicrobial peptides some of which are endowed with antiviral, anticancer and antibiofilm properties. These properties could be improved through synergistic interactions of these bacteriocins with other bioactive molecules such as antibiotics, phages, nanoparticles and essential oils. A number of studies are steadily reporting the effects of these combinations as new and potential therapeutic strategies in the future, as they may offer many incentives over existing therapies. In particular, bacteriocins can benefit from combination with nanoparticles which can improve their stability and solubility, and protect them from enzymatic degradation, reduce their interactions with other molecules and improve their bioavailability. Furthermore, the combination of bacteriocins with other antimicrobials is foreseen as a way to reduce the development of antibiotic resistance due to the involvement of several modes of action. Another relevant advantage of these synergistic combinations is that it decreases the concentration of each antimicrobial component, thereby reducing their side effects such as their toxicity. In addition, combination can extend the utility of bacteriocins as antiviral or anticancer agents. Thus, in this review, we report and discuss the synergistic effects of bacteriocin combinations as medicines, and also for other diverse applications including, antiviral, antispoilage, anticancer and antibiofilms

Clusters of Lactobacillus Strains from Vegetal Origins Are Associated with Beneficial Functions: Experimental Data and Statistical Interpretations

Nine strains of Lactiplantibacillus plantarum and one strain of Lacticaseibacillus paracasei that were recently isolated from prickly pears, fresh figs and blackberries, which are traditionally and largely consumed fruits in Kabylia (north of Algeria), were studied here for their antagonism and antioxidant properties as well as for production of exopolysaccharides. With respect to their inhibitory properties, these strains were tested against three food representative pathogens including Escherichia coli ATCC 8739, Staphylococcus aureus 2S6 and Listeria monocytogenes 162. The antagonism of these pathogens was attributable to lactic acid production, present in the cell free supernatant, at concentrations ranging from 9 to 16.74 g/L. The anti-adhesive properties observed on polystyrene or eukaryotic Caco-2 cells were exerted in a strain dependent-manner. Indeed, the scores obtained ranged from 27% to 75% for S. aureus 2S6, 54% to 95% for L. monocytogenes 162, and 50% to 97% for E. coli ATCC 8739. The co-aggregation of these Lactobacillus strains with the aforementioned target bacteria appeared to be exerted in a strain-dependent manner, with noticeably the upmost rate for Lb. paracasei FB1 on S. aureus 2S6. Interestingly, these novel Lactobacillus strains were able to produce a large amount (315.55 to 483.22 mg/L) of exopolysaccharides, and showed a significant scavenging activity on the 2,2-di-phényl-2-picrylhydrazyle (DPPH) synthetic free radical with rates of 51% to 56%. Of note, the highest antioxidant activity was observed for Lb. paracasei FB1 using the culture supernatants, intact cells or the intracellular extract. The statistical analysis of these data using the principal component analysis (ACP) enabled us to establish three distinct clusters with potential applications as bioprotective and/or probiotic agents, following further evaluation

Abundance of Lactobacillus plantarum Strains with Beneficial Attributes in Blackberries (Rubus sp.), Fresh Figs (Ficus carica), and Prickly Pears (Opuntia ficus-indica) Grown and Harvested in Algeria

This first study performed on traditional fruits consumed in North Africa reveals their richness in microorganisms with beneficial attributes like cholesterol lowering capabilities. Blackberries (Rubus sp.), fresh figs (Ficus carica), and prickly pears (Opuntia ficus-indica) are fruits largely and traditionally consumed in Kabylia, a beautiful northern Algerian region. Here, 85 lactic acid bacteria (LAB)-isolates were isolated and identified by MALDI-TOF mass spectrometry. The identified species belong to Lactobacillus and Leuconostoc genera. These 85 LAB-isolates were then assessed for their capabilities to grow under conditions mimicking the gastrointestinal tract, and the resulting data were statistically treated with principal component analysis (PCA). After which, only 26 LAB-isolates were selected and characterized for their genetic relatedness using random amplified polymorphic DNA (RAPD) method. Following the genetic relatedness assessment, only 10 LAB-strains, among which nine Lactobacillus plantarum and one Lactobacillus paracasei were studied for their pathoproperties and some probiotic features. Interestingly, all of these 10 LAB-strains were devoid of adverse effects, but capable to adhere to human epithelial colorectal adenocarcinoma Caco-2 cells. Of note, these 10 LAB-strains exhibited an important in vitro hypocholesteromia effect, in strain-dependent manner. Moreover, the Lactobacillus strains exhibited a high bile salt hydrolase (BSH) activity which was correlated with expression of bsh2, bsh3 and bsh4 genes

Lacticaseicin 30 and Colistin as a Promising Antibiotic Formulation against Gram-Negative β-Lactamase-Producing Strains and Colistin-Resistant Strains

Antimicrobial resistance is a global health concern across the world and it is foreseen to swell if no actions are taken now. To help curbing this well announced crisis different strategies are announced, and these include the use of antimicrobial peptides (AMP), which are remarkable molecules known for their killing activities towards pathogenic bacteria. Bacteriocins are ribosomally synthesized AMP produced by almost all prokaryotic lineages. Bacteriocins, unlike antibiotics, offer a set of advantages in terms of cytotoxicity towards eukaryotic cells, their mode of action, cross-resistance and impact of microbiota content. Most known bacteriocins are produced by Gram-positive bacteria, and specifically by lactic acid bacteria (LAB). LAB-bacteriocins were steadily reported and characterized for their activity against genetically related Gram-positive bacteria, and seldom against Gram-negative bacteria. The aim of this study is to show that lacticaseicin 30, which is one of the bacteriocins produced by Lacticaseibacillus paracasei CNCM I-5369, is active against Gram-negative clinical strains (Salmonella enterica Enteritidis H10, S. enterica Typhimurium H97, Enterobacter cloacae H51, Escherichia coli H45, E. coli H51, E. coli H66, Klebsiella oxytoca H40, K. pneumoniae H71, K. variicola H77, K. pneumoniae H79, K. pneumoniae H79), whereas antibiotics failed. In addition, lacticaseicin 30 and colistin enabled synergistic interactions towards the aforementioned target Gram-negative clinical strains. Further, the combinations of lacticaseicin 30 and colistin prompted a drastic downregulation of mcr-1 and mcr-9 genes, which are associated with the colistin resistance phenotypes of these clinical strains. This report shows that lacticaseicin 30 is active against Gram-negative clinical strains carrying a rainbow of mcr genes, and the combination of these antimicrobials constitutes a promising therapeutic option that needs to be further exploited

Heterologous biosynthesis of five new class II bacteriocins from Lactobacillus paracasei CNCM I-5369 with antagonistic activity against pathogenic Escherichia coli strains

International audienceLactobacillus paracasei CNCM I-5369 isolated from a traditional Algerian dairy product produces extracellular inhibitory substances, namely, bacteriocins, which are active against a panel of pathogenic Escherichia coli strains. This activity was observed only at a narrow pH 4.5-5, and resulted to be heat stable and sensitive to the action of proteolytic enzymes, which indicate a proteinaceous nature. This new strain has a genome of 2,752,975 bp, with a 46.6% G + C ratio and contains at least 2664 coding sequences. The Bagel software analysis identified five open reading frames (ORFs) that are translated to new class II bacteriocin. Each ORF was cloned in frame with a His-tag tail and expressed in E. coli BL21 (DE3) (pLysS) strain. Of note, each fusion protein carrying any of these ORFs at the C- or N-terminal position resulted to be active against E. coli 184 strain used as target organism. This manuscript reports the first multi-bacteriocinogenic strain producing five new class II bacteriocins with activity against Gram-negative bacilli (GNB), namely, E. coli. Heterologous expression and activity of each new class II bacteriocin were demonstrated

A Review on Enterocin DD14, the Leaderless Two-Peptide Bacteriocin with Multiple Biological Functions and Unusual Transport Pathway

Enterocin DD14 (EntDD14) is a two-peptide leaderless bacteriocin (LLB) produced by Enterococcus faecalis 14, a human strain isolated from meconium. Studies performed on EntDD14 enabled it to show its activity against Gram-positive bacteria such as Listeria monocytogenes, Clostridium perfringens, Enterococcus faecalis, and Staphylococcus aureus. EntDD14 was also shown to potentiate the activity of different antibiotics such as erythromycin, kanamycin, and methicillin when assessed against methicillin-resistant Staphylococcus aureus (MRSA) in vitro and in vivo in the NMRI-F holoxenic mouse model. Additionally, EntDD14 has an antiviral activity and decreased the secretion of pro-inflammatory IL-6 and IL-8 in inflamed human intestinal Caco-2 cells. The genome of E. faecalis 14 was sequenced and annotated. Molecular tools such as Bagel4 software enabled us to locate a 6.7kb-EntDD14 cluster. Transport of EntDD14 outside of the cytoplasm was shown to be performed synergistically by a channel composed of two pleckstrin-homology-domain-containing proteins, namely DdE/DdF and the ABC transporter DdGHIJ. This latter could also protect the bacteriocinogenic strain against extracellular EntDD14. Here, we focus on academic data and potential therapeutic issues of EntDD14, as a model of two-peptide LLB

In silico analyses of the genomes of three new bacteriocin-producing bacteria isolated from animal’s faeces

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New Bacteriocins from Lacticaseibacillus paracasei CNCM I-5369 Adsorbed on Alginate Nanoparticles Are Very Active against Escherichia coli

International audienceLacticaseibacillus paracasei CNCM I-5369, formerly Lactobacillus paracasei CNCM I-5369, produces bacteriocins that are remarkably active against Gram-negative bacteria, among which is the Escherichia coli-carrying mcr-1 gene that is involved in resistance to colistin. These bacteriocins present in the culture supernatant of the producing strain were extracted and semi-purified. The fraction containing these active bacteriocins was designated as E20. Further, E20 was loaded onto alginate nanoparticles (Alg NPs), leading to a highly active nano-antibiotics formulation named hereafter Alg NPs/E20. The amount of E20 adsorbed on the alginate nanoparticles was 12 wt.%, according to high-performance liquid chromatography (HPLC) analysis. The minimal inhibitory concentration (MIC) values obtained with E20 ranged from 250 to 2000 μg/mL, whilst those recorded for Alg NPs/E20 were comprised between 2 and 4 μg/mL, which allowed them to gain up to 500-fold in the anti-E. coli activity. The damages caused by E20 and/or Alg NPs/E20 on the cytology of the target bacteria were characterized by transmission electron microscopy (TEM) imaging and the quantification of intracellular proteins released following treatment of the target bacteria with these antimicrobials. Thus, loading these bacteriocins on Alg NPs appeared to improve their activity, and the resulting nano-antibiotics stand as a promising drug delivery system