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Growth increasement of LAB after incubation with monophasic S. Typhimurium in MRS medium compared with the control L. reuteri Protectis®. The significantly higher increase in LAB’s growth seen in LAB 8 (MZ602127) is marked by an asterisk (p < 0.05)
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Background
Salmonellosis is one of the most important food-borne zoonotic disease affecting both animals and humans. The objective of the present study was to identify gastrointestinal (GI) lactic acid bacteria (LAB) of canine-origin from Salmonella -negative dogs’ faeces able to inhibit monophasic Salmonella Typhimurium previously isolated from do...
Citations
... However, alternative therapies can help reduce antibiotic use and limit the transmission of antibiotic resistance (Cabassi et al., 2021;Jimenez-Trigos et al., 2022). Probiotics, particularly LAB microorganisms, offer a promising alternative to antibiotics for reducing bacterial infections such as Salmonella spp. in pets (Abraham et al., 2016;Czaplewski et al., 2016;Kumar et al., 2017). ...
... Probiotics, particularly LAB microorganisms, offer a promising alternative to antibiotics for reducing bacterial infections such as Salmonella spp. in pets (Abraham et al., 2016;Czaplewski et al., 2016;Kumar et al., 2017). Jimenez-Trigos et al. (2022) identified a potential antimicrobial LAB strain against S. typhimurium that was isolated from Salmonella-negative dogs and not found in Salmonella-positive dogs (Jimenez Trigos et al., 2022). As a preliminary approach, the researchers compared LAB in samples obtained from both Salmonella spp.-positive and Salmonella spp.-negative animals. ...
Background
Although salmonellosis is considered to be a foodborne zoonotic disease, pets can play a significant role in the dissemination of antimicrobial‐resistant Salmonella organisms to humans because of close contact with their owners.
Objectives
To determine the prevalence, risk factors, virulence factors, serotypes, and antimicrobial resistance profile of Salmonella in pet dogs and cats in Turkey and to assess the public health risk. Furthermore, to perform macroscopic comparison of lactic acid bacteria (LAB) in Salmonella‐positive and Salmonella‐negative animals.
Methods
International Standards Organization (ISO) 6579‐1:2017 and Food and Drug Administration (FDA) methods were used to compare the effectiveness of culture methods in the identification of Salmonella in 348 rectal swabs. Positive isolates were serotyped using the slide agglutination method according to the White‐Kauffmann‐Le Minor scheme and the presence of virulence genes (invA and stn) were evaluated by polymerase chain reaction (PCR). Antimicrobial activity was tested by Kirby–Bauer disk diffusion method according to Clinical and Laboratory Standards Institute (CLSI) guidelines.
Results
Salmonella prevalence was 5.73% (9/157) in dogs and 0.0% (0/191) in cats. Eight (8/9) isolates were cultured with the ISO method and 5 (5/9) isolates were cultured with the FDA method. Macroscopic results revealed that Salmonella agents had no effect on LAB. Three different serotypes were detected and all isolates were positive for virulence genes. Antibiotic resistance profiling indicated that 11.1% of the isolates were MDR and the highest resistance was found for ciprofloxacin. MDR‐resistant S. Virchow and carbapenem‐resistant S. Enteritidis were detected from dog isolates. There was a significant difference between raw meat consumption and Salmonella carriage (p < 0.01).
Conclusions
Dogs could be potential carriers of Salmonella infection. The isolation of Salmonella in healthy dogs instead of dogs suffering from diarrhoea indicates that attention should be paid to asymptomatic carriage. The emergence of resistance among zoonotic Salmonella isolates poses a significant threat to public health.
... They are safe microorganisms able to produce different compounds such as bacteriocins, organic acids as lactic acid, hydrogen peroxide, diacetyl, and carbon dioxide that favor the inhibition of pathogenic microorganisms. Lactic acid bacteria are Gram-positive bacteria, they can be found in the microbiota of various anatomical locations such as the oral cavity, the skin, the gastrointestinal tract, and the reproductive tract (19)(20)(21)(22)(23). Their presence in the raw milk of small ruminants is well known (24), and some strains have been tested in vitro for their potential probiotic characteristics (25,26). ...
... They were processed for genomic DNA extraction and identified based on PCR amplification and sequencing of 16S rRNA gene using bacterial universal primers (27F 5′-AGAGTTTGATCC TGGCTCAG and 1492R 5′-GGTT ACCTTGTTA CGACTT). The PCR was performed following the methodology previously described (21). The PCR products were purified, and sequenced and analyzed for sequence homology by BLAST. 1 The sequences were corrected and aligned by ClustalW with Molecular Evolutionary Genetics Analysis (MEGA) 7. Bacterial identification was carried out by comparing the problem sequence with the GenBank database through the Blast application. ...
... Indeed, these LAB populations can be found in the microbiota of the epithelium of the respiratory, mammary and reproductive tracts (23,24,60), which are anatomical locations that are colonized by mycoplasma associated with CA and linked to excretion route (34). In this sense, in a previous study involving Salmonella sp., the isolation of LAB was less important in dogs that were positive to this pathogen (21). Based on this hypothesis, the use of antibiotics could harm the natural barrier, that LAB with antimicrobial capacity represent, in locations such as the mammary gland of small ruminants. ...
Introduction
The complexity of fighting contagious agalactia (CA) has raised the necessity of alternative antimicrobial therapies, such as probiotics. Lactic acid bacteria (LAB) are present in the mammary gland of small ruminants and their antimicrobial effect have been previously described against species like Mycoplasma bovis but never against Mycoplasma agalactiae (Ma). This in vitro study aims to evaluate the antimicrobial activity against Ma of ovine and caprine LAB strains and a human commercial probiotic (L2) of Lactobacillus spp.
Methods
A total of 63 possible LAB strains were isolated from nine ovine and caprine farms in Spain, three isolates (33B, 248D, and 120B) from the 63 strains were selected, based on their capacity to grow in a specific medium in vitro, for an in vitro experiment to assess their antimicrobial activity against Ma in Ultra High Temperature (UHT) processed goat milk (GM). A women commercial vaginal probiotic was also included in the study. The inoculum of L2 was prepared at a concentration of 3.24 × 10⁸ CFU/mL and the average concentration of the inoculum of the wild LAB varied from 7.9 × 10⁷ to 8.4 × 10⁸ CFU/mL.
Results
The commercial probiotic L2 significantly reduced the concentration of Ma to 0.000 log CFU/mL (p < 0.001), strain 33B reduced it from 7.185 to 1.279 log CFU/mL (p < 0.001), and 120B from 6.825 to 6.466 log CFU/mL (p < 0.05). Strain 248D presented a bacteriostatic effect in GM. Moreover, the three wild strains and the commercial probiotic produced a significative reduction of the pH (p < 0.001).
Discussion
This is the first in vivo report of the antimicrobial potential of LAB strains against Ma and its interaction. Our results support possible future alternative strategies to antibiotic therapy, previously not contemplated, to fight CA in small ruminants. Further studies are necessary to elucidate the action mechanisms through which these LAB are able to inhibit Ma and to assess the safety of using these strains in possible in vivo studies.
... Molecular identification allowed the detection of a high proportion of L. murinus and one L. animalis. Specifically, L. murinus strains have been isolated from different niches, such as rodent gastrointestinal tract [39], dog feces [40], fermented dough [41], and marine sediments [19], while L. animalis has been found in the gastrointestinal tract of several animal species [42]. Furthermore, the identity matrix and the phylogenetic analysis determined the similarity between the sequences using the V2 variable region of the 16S rRNA gene from the 16 L. murinus strains and confirmed its close relationship with L. animalis, which has been described by several authors, identifying these two species at the same phylogenetic level with similarity percentages of up to 99% between 16S rRNA gene regions [41,43]. ...
Considering the objectives of “One Health” and the Sustainable development Goals “Good health and well-being” for the development of effective strategies to apply against bacterial resistance, food safety dangers, and zoonosis risks, this project explored the isolation and identification of Lactobacillus strains from the intestinal tract of recently weaned mice; as well as the assessment of antibacterial activity against clinical and zoonotic pathogens. For molecular identification, 16S rRNA gene-specific primers were used and, via BLAST-NCBI, 16 Ligilactobacillus murinus, one Ligilactobacillus animalis, and one Streptococcus salivarius strains were identified and registered in GenBank after the confirmation of their identity percentage and the phylogenetic analysis of the 16 Ligilactobacillus murinus strains and their association with Ligilactobacillus animalis. The 18 isolated strains showed antibacterial activity during agar diffusion tests against Listeria monocytogenes ATCC 15313, enteropathogenic Escherichia coli O103, and Campylobacter jejuni ATCC 49943. Electrophoretic and zymographic techniques confirmed the presence of bacteriolytic bands with a relative molecular mass of 107 kDa and another of 24 kDa in Ligilactobacillus murinus strains. UPLC-MS analysis allowed the identification of a 107 kDa lytic protein as an N-acetylmuramoyl-L-amidase involved in cytolysis and considered a bacteriolytic enzyme with antimicrobial activity. The 24 kDa band displayed similarity with a portion of protein with aminopeptidase function. It is expected that these findings will impact the search for new strains and their metabolites with antibacterial activity as an alternative strategy to inhibit pathogens associated with major health risks that help your solution.
... The antibacterial effect of probiotics has gained interest due to its potential to be used as safe bio-preservatives, which are easily degraded into the gastrointestinal tract (Botthoulath, Upaichit, & Thumarat, 2018). Furthermore, it has been reported that the antimicrobial activity of Lactobacillus could be an alternative for antibiotic treatments and, thus, avoid antibiotic resistances (Jimenez-Trigos et al., 2022). In this regard, LAB could have a bactericidal effect producing several postbiotic metabolites such as organic acids, peptides or bacteriocins (Liu, Zhang, Yang, & Huang, 2015;Sharma et al., 2017). ...
Lactiplantibacillus plantarum has been widely studied due to its beneficial effects on health such as protect against pathogens, enhance the immune system, or produce metabolites like γ-aminobutyric acid (GABA). The objective of this study was the evaluation of the GABA-producer L. plantarum K16 isolated from kimchi. The safety and probiotic characterisation of this strain was performed by analysing carbohydrates fermentation, enzymatic activity, antibiotics susceptibility, and haemolytic and antimicrobial activity. Likewise, GABA production was optimised following a one-factor-at-a-time procedure by changing relevant fermentation parameters like incubation temperature, yeast extract concentration and fermentation time. The results indicated that L. plantarum K16 has the potential to stimulate the digestion and absorption of several nutrients and it could have an inhibitory effect against pathogenic bacteria. The best results for GABA production by this strain was around 1000 mg/L, using 12 g/L of yeast extract, 34 °C of incubation temperature and 96 h of fermentation time.
... They are non-pathogenic organisms that may eliminate unfavorable microflora by several mechanisms such as the production of antimicrobial substances (lactic acid, bacteriocins, hydrogen peroxide), inhibition of bacterial adhesion to the mucosa, competition for nutrients, and stimulation of immunity [11]. There are several studies investigating the antimicrobial activity and metabolic potentials of Lactobacillus species isolated from the intestinal tract of dogs [12][13][14]. Currently, most of the probiotic microorganisms widely used in application studies on dogs are mainly of human origin [15,16]. ...
Background
Campylobacteriosis is currently the most frequently reported zoonosis. Dogs, especially puppies or those with diarrhea, are considered a possible source of human infection. Probiotic bacteria, such as Lactobacillus species, seem to be a valuable tool in controlling of intestinal pathogenic microorganisms in dogs. The main purpose of this study was to assess the anti-Campylobacter activity and some probiotic properties, like ability to produce H2O2, bile salt and low pH tolerance of Lactobacillus strains isolated from gastrointestinal tract of healthy dogs.
Results
A total of 39 rectal swabs derived from healthy dogs and 19 from dogs with diarrhea were examined to detect Lactobacillus and Campylobacter bacteria respectively. In total, 30 strains of Lactobacillus genus and four strains of Campylobacter genus were isolated and identified. Of the 30 strains of Lactobacillus, 22 showed an inhibitory effect towards Campylobacter. Four strains with the strongest antagonism towards Campylobacter bacteria (L. salivarius 25 K/L/1, L. rhamnosus 42 K/L/2, L. sakei 50 K/L/1 and L. agilis 55 K/L/1) were selected to assess their potential probiotic traits. Three out of four analyzed strains produced extracellular H2O2. All displayed very good or moderate survival at pH 3.0 and 2.0 and showed high tolerance to 0.5% and 1% bile salts.
Conclusions
Among selected Lactobacillus strains, all may have a potential probiotic application in reducing Campylobacter spp. in dogs and thus prevent transmission of infection to humans, although the best candidate for probiotic seems to be L. sakei 50 K/L/1. Further in vitro and in vivo studies are needed.
The spot on the law technique is of great importance for the study of probiotic bacteria. Therefore, the main objective of this research was to develop an adaptation of this technique with the prerogative of making its application more practical and executable on a large scale in addition, validating the adaptation of this methodology based on the quantitative and qualitative analysis of the data obtained. For statistical analysis, we used the Mann-Whitney Rank Sum Test, analogous to the T Test for non-parametric data. From the results obtained, it was possible to verify that the adaptation is equivalent to the classic technique, as there was no statistical difference between the two, and that the application of the adapted methodology facilitates the execution of the spot because it presents a lower degree of difficulty.
Background
This study aimed to characterize potential probiotic strains for use in dogs to prevent infectious enteropathies. Lactic acid bacteria (LAB) isolated from canine milk and colostrum were characterized according to their functional properties, including their resistance to gastrointestinal conditions, inhibitory effect against pathogens, and intestinal adhesion.
Methods
The immunomodulatory effects of the strains were also analyzed in in vitro and in vivo studies. Among the strains evaluated, two LAB strains (TUCO-16 and TUCO-17) showed remarkable resistance to pH 3.0, bile salts, and pancreatin, as well as inhibitory effects against pathogenic Escherichia coli, Salmonella sp., and Clostridium perfringens.
Results
The TUCO-16 and TUCO-17 strains induced a significant increase in the expression of TNF-α, IL-8, and TLR2 in canine macrophages. The oral administration of TUCO-16 and TUCO-17 strains to mice significantly augmented their resistance to pathogenic E. coli or Salmonella intestinal infections. Both canine strains reduced intestinal damage and pathogen counts in the liver and spleen and avoided their dissemination into the bloodstream. These protective effects were related to the ability of TUCO-16 and TUCO-17 strains to differentially modulate the production of IFN-γ, IFN-β, TNF-α, IL-6, KC, MCP-1, and IL-10 in the intestinal mucosa.
Conclusion
Both strains, TUCO-16 and TUCO-17, are potential probiotic candidates for improving intestinal health in dogs, particularly for their ability to inhibit the growth of Gram-negative pathogens common in gastrointestinal infections and modulate the animal's immune response. Further studies are required to effectively demonstrate the beneficial effects of TUCO-16 and TUCO-17 strains in dogs.
This study was conducted to determine the effect of animal protein inclusion rate and grain-free or grain-inclusive diets on macronutrient digestibility, fecal characteristics, metabolites, and microbiota in mixed-breed hounds and Beagles. Four experimental extruded kibble diets were made with varying amounts of animal protein and carbohydrates: 1) high animal protein, grain-inclusive (HA-GI), 2) low animal protein, grain-free (LA-GF), 3) low animal protein, grain-inclusive (LA-GI), and 4) high animal protein, grain-free (HA-GF). Thirty-two Beagles and 33 mixed-breed hounds were assigned to 1 of the 4 treatment groups in a completely randomized design that lasted 180 days. All diets were similar in chemical composition and well-digested by the animals. In general, for fecal metabolites, mixed-breed hounds had a greater concentration of total short-chain fatty acid (SCFA) and ammonia and lower indole concentration than Beagles (P < 0.05). In mixed-breed hounds, LA-GF had a greater (P < 0.05) total SCFA concentration than HA-GI and LA-GI; however, this was not observed in Beagles. There were greater concentrations of ammonia, phenol, and indole in HA-GI than in LA-GF (P < 0.05). Breed affected fecal primary bile acid concentration, as mixed-breed hounds had a greater concentration of cholic acid (CA) than Beagles (P < 0.05). Mixed-breed hounds fed LA-GF resulted in greater CA concentrations than HA-GI and LA-GI (P < 0.05). Dogs who consumed LA-GF had lower fecal secondary bile acid content than the other groups (P < 0.05). The distribution of the fecal microbiota community differed in LA-GF compared with the other groups, with lower α-diversity. However, dogs fed LA-GF had the largest difference in composition with greater Selenomonadaceae, Veillonellaceae, Lactobacillaceae, Streptococcus, Ligilactobacillus, Megamonas, Collinsella aerofaciens, and Bifidobacterium sp. than the other groups. A significant breed effect was noted on nutrient digestibility, fecal metabolites, and microbiota. A treatment effect was observed in LA-GF as it resulted in greater fecal SCFA, lower protein fermentative end-products, greater fecal primary bile acids, lower fecal secondary bile acid concentrations, and shifts in fecal microbiota.
The purpose of this research was the genotypic identification of lactic acid bacteria (LAB), isolated from the gastrointestinal tract (GIT) of healthy adult birds, and the study of their safety regarding antibiotic resistance, physiological and functional properties involved in the colonization of the GIT of poultry, and Salmonella exclusion, as members of a potential mixed probiotic supplement for poultry. The nucleotidic sequence from Lactobacillus crispatus P1, L. animalis L3, and Enterococcus faecium CRL 1385 (ex-J96) showed 100, 99.8, and 99.3% identity with L. crispatus DSM 20584 T, Ligilactobacillus salivarius ATCC 11741 T, and E. faecium ATCC 19434 T, respectively. These strains showed no resistance to relevant antibiotics usually administered to animals proposed by the European Food Safety Authority. They could endure the detrimental conditions of the gastrointestinal tract (pH 2.6 and oxgall 0.1 and 0.4% w/v). In an ex vivo assay, the LAB showed high adherence to the three sections of the GIT, reaching values higher than 70%. The adhesion to mucus was strain-dependent: L. crispatus CRL 1453 evidenced the highest adhesion (> 19%) while Lig. salivarius subsp. salivarius CRL 1417 and E. faecium CRL 1385 adhered to a lower extent (> 9 and 2%, respectively). Moreover, the LAB elicited remarkable anti-Salmonella activity, taking into account that they could inhibit elevated counts of different Salmonella serovars, especially the host-specific serovars S. Gallinarum and S. Pullorum (up to 8 log CFU/mL decrease in Salmonella counts).
