Insights in the development and uses of alternatives to antibiotic growth promoters in poultry and swine production

The overuse and misuse of antibiotics has contributed to the rise and spread of multidrugresistant bacteria. To address this global public health threat, many countries have restricted the use of antibiotics as growth promoters and promoted the development of alternatives to antibiotics in human and veterinary medicine and animal farming. In food-animal production, acidifiers, bacteriophages, enzymes, phytochemicals, probiotics, prebiotics, and antimicrobial peptides have shown hallmarks as alternatives to antibiotics. This review reports the current state of these alternatives as growth-promoting factors for poultry and swine production and describes their mode of action. Recent findings on their usefulness and the factors that presently hinder their broader use in animal food production are identified by SWOT (strength, weakness, opportunity, and threat) analysis. The potential for resistance development as well as co- and cross-resistance with currently used antibiotics is also discussed. Using predetermined keywords, we searched specialized databases including Scopus, Web of Science, and Google Scholar. Antibiotic resistance cannot be stopped, but its spreading can certainly be hindered or delayed with the development of more alternatives with innovative modes of action and a wise and careful use of antimicrobials in a One Health approach

Comparative effects of exopolysaccharides from lactic acid bacteria and fructo-oligosaccharides on infant gut microbiota tested in an in vitro colonic model with immobilized cells

The aim of this study was to compare the effects of purified exopolysaccharides from Lactobacillus rhamnosus RW-9595M with those of a well-known prebiotic (short-chain fructo-oligosaccharides) on infant colonic microbiota using a new three-stage chemostat model with immobilized infant faecal microbiota. Two continuous cultures with different faecal inocula were tested with different compositions of carbohydrate media. During the first fermentation (F1), fructo-oligosaccharides tested at a concentration of 9.8 g L−1 increased the number of lactobacilli and decreased coliforms both in gel beads and in effluent from all three reactors, in agreement with data from the literature. During the second fermentation (F2), the effect of fructo-oligosaccharides tested at a lower concentration (7.5 g L−1) was reduced compared with F1. Fructo-oligosaccharides also increased total organic acid concentration and decreased ammonia production. Results obtained for exopolysaccharide tested at 1.5 g L−1 indicate that exopolysaccharides from L. rhamnosus RW-9595M was not metabolized by infant microbiota and lacked any prebiotic effec

New three-stage in vitro model for infant colonic fermentation with immobilized fecal microbiota

The development and validation of a new three-stage culture system with immobilized fecal microbiota to simulate infant colonic ecosystem is described. Two continuous cultures with different fecal inocula were used to assess the validity and stability of the intestinal model. The total anaerobe populations measured in beads and effluent fermentations reached high concentrations similar to infant feces. Fluorescence in situ hybridization analyses and denaturing gradient gel electrophoresis profiles of effluent samples from the three reactors revealed complex patterns similar to that observed in the inoculum, indicating that fecal bacterial diversity was well-preserved and that dominant bacterial populations showed good stability among reactors. For both experiments, the bacterial populations and fermentation product concentrations were in the range of published data for infant feces. These results demonstrate that this new three-stage continuous culture with immobilized cells provides a useful tool for studying the infant colon ecosyste

Bacteriocin-based synergetic consortia : a promising strategy to enhance antimicrobial activity and broaden the spectrum of inhibition

Bacteria-derived natural antimicrobial compounds such as bacteriocins, reruterin, and organic acids have recently received substantial attention as food preservatives or therapeutic alternatives in human or animal sectors. This study aimed to evaluate the antimicrobial activity of different bacteria-derived antimicrobials, alone or in combination, against a large panel of Gram-negative and Grampositive bacteria. Bacteriocins, including microcin J25, pediocin PA-1, nisin Z, and reuterin, were investigated alone or in combination with lactic acid and citric acid, using a checkerboard assay. Concentrations were selected based on predetermined MICs against Salmonella enterica subsp. enterica serovar Newport ATCC 6962 and Listeria ivanovii HPB28 as Gram-negative and Gram-positive indicator strains, respectively. The results demonstrated that the combination of microcin J25 1 citric acid 1 lactic acid; microcin J25 1 reuterin 1 citric acid; and microcin J25 1 reuterin 1 lactic acid tested against S. Newport ATCC 6962 showed synergistic effects (FIC index = 0.5). Moreover, a combination of pediocin PA-1 1 citric acid 1 lactic acid; and reuterin 1 citric acid 1 lactic acid against L. ivanovii HPB28 showed a partially synergistic interactions (FIC index = 0.75). Nisin Z exerted a partially synergistic effect in combination with acids (FIC index = 0.625 -0.75), whereas when it was combined with reuterin or pediocin PA-1, it showed additive effects (FIC index = 1) against L. ivanovii HPB28. The inhibitory activity of synergetic consortia were tested against a large panel of Gram-positive and Gram-negative bacteria. According to our results, combining different antimicrobials with different mechanisms of action led to higher potency and a broad spectrum of inhibition, including multidrug-resistance pathogens

In vitro assessment of skin sensitization, irritability and toxicity of bacteriocins and reuterin for possible topical applications

Bacteriocins and reuterin are promising antimicrobials for application in food, veterinary, and medical sectors. In the light of their high potential for application in hand sanitizer, we investigated the skin toxicity of reuterin, microcin J25, pediocin PA-1, bactofencin A, and nisin Z in vitro using neutral red and LDH release assays on NHEK cells. We determined their skin sensitization potential using the human cell line activation test (h-CLAT). Their skin irritation potential was measured on human epidermal model EpiDerm™. We showed that the viability and membrane integrity of NHEK cells remained unaltered after exposure to bacteriocins and reuterin at concentrations up to 400 µg/mL and 80 mg/mL, respectively. Furthermore, microcin J25 and reuterin showed no skin sensitization at concentrations up to 100 µg/mL and 40 mg/mL, respectively, while pediocin PA-1, bactofencin A, and nisin Z caused sensitization at concentrations higher than 100 µg/mL. Tissue viability was unafected in presence of bacteriocins and reuterin at concentrations up to 200 µg/mL and 40 mg/ mL, respectively, which was confrmed by measuring cytokine IL-1α and IL-8 levels and by histological analysis. In conclusion, the current study provides scientifc evidence that some bacteriocins and reuterin, could be safely applied topically as sanitizers at recommended concentration

Classification of a moderately oxygen-tolerant isolate from baby faeces as Bifidobacterium thermophilum

<p>Abstract</p> <p>Background</p> <p>Bifidobacteria are found at varying prevalence in human microbiota and seem to play an important role in the human gastrointestinal tract (GIT). Bifidobacteria are highly adapted to the human GIT which is reflected in the genome sequence of a <it>Bifidobacterim longum </it>isolate. The competitiveness against other bacteria is not fully understood yet but may be related to the production of antimicrobial compounds such as bacteriocins. In a previous study, 34 <it>Bifidobacterium </it>isolates have been isolated from baby faeces among which six showed proteinaceous antilisterial activity against <it>Listeria monocytogenes</it>. In this study, one of these isolates, RBL67, was further identified and characterized.</p> <p>Results</p> <p><it>Bifidobacterium </it>isolate RBL67 was classified and characterized using a polyphasic approach. RBL67 was classified as <it>Bifidobacterium thermophilum </it>based on phenotypic and DNA-DNA hybridization characteristics, although 16S rDNA analyses and partial <it>gro</it>EL sequences showed higher homology with <it>B. thermacidophilum </it>subsp. <it>porcinum </it>and <it>B. thermacidophilum </it>subsp. <it>thermacidophilum</it>, respectively. RBL67 was moderately oxygen-tolerant and was able to grow at pH 4 and at a temperature of 47°C.</p> <p>Conclusion</p> <p>In order to assign RBL67 to a species, a polyphasic approach was used. This resulted in the classification of RBL67 as a <it>Bifidobacterium thermophilum </it>strain. To our knowledge, this is the first report about <it>B. thermophilum </it>isolated from baby faeces since the <it>B. thermophilum </it>strains were related to ruminants and swine faeces before. <it>B. thermophilum </it>was previously only isolated from animal sources and was therefore suggested to be used as differential species between animal and human contamination. Our findings may disapprove this suggestion and further studies are now conducted to determine whether <it>B. thermophilum </it>is distributed broader in human faeces. Furthermore, the postulated differentiation between human and animal strains by growth above 45°C is no longer valid since <it>B. thermophilum </it>is able to grow at 47°C. In our study, 16S rDNA and partial <it>gro</it>EL sequence analysis were not able to clearly assign RBL67 to a species and were contradictory. Our study suggests that partial <it>gro</it>EL sequences may not be reliable as a single tool for species differentiation.</p

Bacteriocins as a new generation of antimicrobials : toxicity aspects and regulations

In recent decades, bacteriocins have received substantial attention as antimicrobial compounds. Although bacteriocins have been predominantly exploited as food preservatives, they are now receiving increased attention as potential clinical antimicrobials and as possible immune-modulating agents. Infections caused by antibiotic-resistant bacteria have been declared as a global threat to public health. Bacteriocins represent a potential solution to this worldwide threat due to their broad- or narrow-spectrum activity against antibiotic-resistant bacteria. Notably, despite their role in food safety as natural alternatives to chemical preservatives, nisin remains the only bacteriocin legally approved by regulatory agencies as a food preservative. Moreover, insufficient data on the safety and toxicity of bacteriocins represent a barrier against the more widespread use of bacteriocins by the food and medical industry. Here, we focus on the most recent trends relating to the application of bacteriocins, their toxicity and impacts

Gastrointestinal stability and cytotoxicity of bacteriocins from gram-positive and gram-negative bacteria : a comparative in vitro study

Bacteriocins are receiving increased attention as potent candidates in food preservation and medicine. Although the inhibitory activity of bacteriocins has been studied widely, little is known about their gastrointestinal stability and toxicity toward normal human cell lines. The aim of this study was to evaluate the gastrointestinal stability and activity of microcin J25, pediocin PA-1, bactofencin A and nisin using in vitro models. In addition cytotoxicity and hemolytic activity of these bacteriocins were investigated on human epithelial colorectal adenocarcinoma cells (Caco-2) and rat erythrocytes, respectively. Pediocin PA-1, bactofencin A, and nisin were observed to lose their stability while passing through the gastrointestinal tract, while microcin J25 is only partially degraded. Besides, selected bacteriocins were not toxic to Caco-2 cells, and integrity of cell membrane was observed to remain unaffected in presence of these bacteriocins at concentrations up to 400 µg/mL. In hemolysis study, pediocin PA-1, bactofencin A, and nisin were observed to lyse rat erythrocytes at concentrations higher than 50 µg/mL, while microcin J25 showed no effect on these cells. According to data indicating gastrointestinal degradation and the absence of toxicity of pediocin PA-1, bactofencin A, and microcin J25 they could potentially be used in food or clinical applications

Structure-activity studies of the bacteriocin bactofencin A and its interaction with the bacterial membrane

The antimicrobial peptide bactofencin A is an unmodified non-pediocin-like bacteriocin that inhibits several clinically relevant pathogens, including Listeria monocytogenes and Staphylococcus aureus. Here we report the synthesis and structure–activity relationship studies of bactofencin A and novel analogues thereof. Synthetic bactofencin A was a potent inhibitor of L. monocytogenes (MIC = 8.0 μM) and S. aureus (MIC = 4.0 μM), similar to the bacteriocin produced naturally by Lactobacillus salivarius. Of particular interest is the fact that linear analogues lacking the disulfide bond found in bactofencin A were as potent and also active against several strains of methicillin-resistant S. aureus (MRSA) and one strain of vancomycin-resistant S. aureus (VRSA). Supported by the structure–activity relationship study, investigation of the interaction of bactofencin A with bacterial membrane by molecular dynamics simulations showed the importance of the positively charged N-terminal tail for peptide–membrane interaction. These results suggest that the C-terminal macrocycle is involved in target protein binding and bacterial growth inhibition