The present study investigated the effects of enterocin (Ent) M and sage extract alone and also in combination on the gut microflora, phagocytic activity, blood biochemistry, and morphometry of rabbits. Sixty-four rabbits (aged five weeks, M91 meatline, both sexes) were divided into three experimental groups: E (EntM; 50 µL/animal/day), S (sage; 10 µL/animal/day), and E + S (EntM + sage) groups and control group (C). The additives were administered in drinking water for a period of 21 days. Dietary supplementation of EntM and sage significantly reduced the coliforms (E: p < 0.001; S: p < 0.001; E + S: p < 0.001) in feces, while simultaneous addition of EntM and sage decreased enterococci (E + S: p < 0.0001), lactic acid bacteria (E + S: p < 0.01), and coagulase-positive staphylococci (E + S: p < 0.0001) in the appendix. Sage addition reduced HDL (S: p < 0.001) and LDL cholesterol (S: p < 0.001; E + S: p < 0.001), LDL/HDL ratio (S: p < 0.001; E + S: p < 0.01), and increased urea (S: p < 0.01; E + S: p < 0.001) and creatinine (S: p < 0.001; E + S: p < 0.001) in serum. EntM and sage application, alone or in combination, improve the jejunal morphometry (p < 0.0001) in rabbits.

Enterococci are widespread bacteria forming the third largest genus among lactic acid bacteria. Some possess probiotic properties or they can produce beneficial proteinaceous antimicrobial substances called enterocins. On the other hand, some enterococci produce biogenic amines (BAs), so this study is focused on the sensitivity to enterocins of biogenic amine-producing faecal enterococci from ostriches and pheasants. Altogether, 60 enterococci isolated from faeces of ostriches and pheasants were tested for production of BAs. This target of the identified enterococci involved 46 strains selected from 140 ostriches and 17 from 60 pheasants involving the species Enterococcus hirae, E. faecium, E. faecalis, and E. mundtii. Although BAs histamine, cadaverine, putrescine, and tryptamine were not detected in the enterococci tested, in general high BA production by the tested enterococci was noted. The species E. hirae formed the majority of the enterococcal strains from ostrichs faeces (34 strains). High production of tyramine (TYM) was measured with an average amount of 958.16 ± 28.18 mg/ml. Among the enterococci from pheasants, the highest was production of TYM compared to phenylethylamine, spermidine, and spermine. Enterococci featured high BA production; however, they were sensitive to seven enterocins with inhibition activity ranging from 100 up to 25,600 AU/ml.

Laukova A., Pogany Simonova M., Kubasova I., Gancarcikova S., Placha I., Scerbova J., Revajova V., Herich R., Levkut Sn. M., Strompfova V. (2017): Pilot experiment in chickens challenged with Campylobacter jejuni CCM6191 administered enterocin M-producing probiotic strain Enterococcus faecium CCM8558 to check its protective effect. Czech J. Anim. Sci., 62, 491-500.Campylobacter jejuni is one of the most common food-borne pathogens and chickens are the main source of these bacteria. Enterococcus faecium AL41, enterocin M-producing strain (deponed to the Czech Culture Collection of Microorganisms, Brno, Czech Republic -CCM8558) is our isolate previously applied e.g. in broiler rabbits with beneficial effect. In this study it was used in a 11-day experiment with chickens (1-day-old, breed Cobb 500, n = 40) challenged with C. jejuni. Birds had free access to feed and water; they were randomly divided into four groups per 10 chicks each: control (CG), E. faecium CCM8558 (EG1), CCM8558 + C. jejuni CCM6191 (EG2), CCM6191 (EG3). E. faecium CCM8558 (10 9 CFU/ml, 200 µl) in Ringer solution was administered daily per os to EG1 and EG2 for 7 days (from day 0-1). EG3 and EG2 were infected individually per os (day 4, CCM6191, 10 8 CFU/ml in Ringer solution, 200 µl). For microbiota evaluation, faecal mixtures (n = 5) were sampled on day 0-1 (1 st sampling), on day 7 (2 nd sampling), and on day 11 (3 rd sampling) (day 7 of CCM8558 application, day 4 post-infection; day 4 of CCM8558 cessation, day 7 post-infection). Five birds from each group were sacrificed. CCM8558 sufficiently colonized chickens. In faeces of EG2 a tendency to reduce Campylobacter spp. (day 7 of application, day 4 of infection) was noted compared to EG2 (day 11, day 4 of cessation, day 7 postinfection; difference 1.21 log cycles), while in EG3 CCM6191 strain was not reduced. Phagocytic activity (PA) values were significantly higher in infected groups compared to CG and EG1. A significant increase in PA was also noted in EG2 and EG3 at the end of experiment compared to CG or EG1. The strain additive did not evoke oxidative stress. Biochemical parameters were influenced to the reference levels.

Only limited information exists concerning the microbiota in beaver (Castor fiber). This study has been focused on the virulence factors genes detection in enterococci from beavers. In general, animals are not affected by enterococcal infections, but they can be a reservoir of, e.g. pathogenic strains. Moreover, detection of virulence factors genes in enterococci from beavers was never tested before. Free-living beavers (12), male and female (age 4-5 years) were caught in the north-east part of Poland. Sampling of lower gut and faeces was provided according to all ethical rules for animal handling. Samples were treated using a standard microbiological method. Pure bacterial colonies were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) identification system. Virulence factors genes-gelE (gelatinase), agg (aggregation), cylA (cytolysin A), efaAfs (adhesin Enterococcus faecalis), efaAfm (adhesin Enterococcus faecium) and esp (surface protein) were tested by PCR. Moreover, gelatinase and antibiotic phenotypes were tested. Species detected were Enterococcus thailandicus, E. faecium, E. faecalis and Enterococcus durans. In literature, enterococcal species distribution was never reported yet up to now. Strains were mostly sensitive to antibiotics. Vancomycin-resistant E. faecalis EE9Tr1 possess cylA, efaAfs, esp and gelE genes. Strains were aggregation substance genes absent. Adhesin E. faecium (efaAfm) gene was detected in two of three E. faecium strains, but it was present also in E. thailandicus. Esp gene was present in EE9Tr1 and E. durans EDTr92. The most detected were gelE, efaAfm genes; in EF 4Hc1 also gelatinase phenotype was found. Strains with virulence factors genes will be tested for their sensitivity to antimicrobial enterocins.

No abstract

Worldwide consumers increasingly demand traditional/local products, to which those made from ewe’s milk belong. In Slovakia, dairy products made from ewe’s milk have a long tradition. A total of seventeen farmhouse fresh ewe’s milk lump cheeses from various local farm producers in central Slovakia were sampled at farms and then analyzed. Based on the sequencing data analysis, the phylum Firmicutes dominated (60.92%) in ewe’s lump cheeses, followed with the phylum Proteobacteria (38.23%), Actinobacteria (0.38%) and Bacteroidetes (0.35%). The phylum Firmicutes was represented by six genera, among which the highest amount possessed the genus Streptococcus (41.13%) followed with the genus Lactococcus (8.54%), Fructobacillus (3.91%), Enterococcus (3.18%), Staphylococcus (1.80%) and the genus Brochotrix (0.08%). The phylum Proteobacteria in ewe’s lump cheeses involved eight Gram-negative genera: Pseudomonas, Acinetobacter, Enterobacter, Ewingella, Escherichia-Shigella, Pantoea and Moraxella. The phylum Bacteroidetes involved three genera: Bacteroides, Sphingobacterium and Chrysobacterium. Results presented are original; the microbiome of Slovak ewe’s milk lump cheese has been not analyzed at those taxonomic levels up to now.

Probiotic enterococci can produce bacteriocins. Enterococcus faecium AL41 is an Enterocin M-producing, probiotic strain which has previously shown beneficial effect in broiler chickens. In this study, it was used to control intestinal microbiota in farm ostriches in a 42-day experiment with an experimental group (EG, 40 ostriches) and a control group (CG, 46). In addition to feed mixture, the ostriches in EG received Ent. faecium AL41 (10(9) CFU ml(-1); by rifampicin-marked variant) 400 μl per animal per day in their drinking water for 21 days. Sampling was carried out at the start of the experiment (at day 0/1), at day 21 (after 21 days of AL41 application) and at day 42 (21 days after AL41 cessation). Faeces (mixture, n = 6) were treated using the standard microbiological dilution method and cultivated on selective media (ISO). The highest count of AL41 was found at day 42. Its identity was confirmed with PCR and Maldi-Tof. The ostriches were free of Salmonella and Campylobacter cells. At day 21, antimicrobial effect was demonstrated by significant reduction in coagulase-positive and negative staphylococci in EG compared to CG (P < 0·001) and coliforms, Enterobacteria and Pseudomonas-like bacteria (P < 0·001). We conclude that AL41 can be used to control intestinal microbiota in farm ostriches. Significance and impact of the study: Ostriches are excellent for high intensity farming in a wide range of climates, requiring only limited space and giving high yields per hectare. They are reared mainly for their meat. Although adult birds possess quite good immunity, young birds can be threatened by spoilage bacteria, especially when they are transferred from the nests to the farm area. Based on our previous results related to the beneficial effect of bacteriocin-producing, probiotic strain Enterococcus faecium AL41 in poultry or rabbits, we decided to test its ability to control intestinal microbiota in farming ostriches which has never been tested previously.

Lauková A., Pogány Simonová M., Chrastinová Ľ., Kandričáková A., Ščerbová J., Plachá I., Čobanová K., Formelová Z., Ondruška Ľ., Štrkolcová G., Strompfová V. (2017): Beneficial effect of bacteriocin-producing strain Enterococcus durans ED 26E/7 in model experiment using broiler rabbits. Czech J. Anim. Sci., 62, 168-177.From the aspect of probiotic properties and bacteriocins, Enterococcus faecium belongs to the most frequently studied species among enterococci. This study deals with testing the strain of the species Enterococcus durans ED 26E/7 in broiler rabbits. The strain ED 26E/7 isolated from ewes lump cheese produces an antimicrobial substance durancin. Forty-eight post-weaned rabbits (aged 5 weeks) of both sexes were divided into experimental group (EG) and control group (CG) per 24 animals each, and kept in standard cages, two animals per cage. EG group rabbits were additionally administered the ED 26E/7 strain (500 µl/animal/day) into water for 21 days. CG group rabbits were fed a commercial feed. The experiment lasted 42 days. Faeces and blood samples were taken on days 0-1 (experiment onset), 21 (after a 3-week application), and 42 (3 weeks after ED 26E/7 strain cessation). On days 21 and 42, rabbits were slaughtered and caeca and appendix were sampled. The rabbits' digestive tract was found to be sufficiently colonized by the strain ED 26E/7; the antimicrobial effect was demonstrated in caecum and appendix (e.g. decrease in coliforms). Reduction of Eimeria sp. oocysts in EG compared to CG rabbits was detected on day 21, when also a significant (P < 0.05) increase of phagocytic activity in EG was registered. Values of glutathioneperoxidase were lower in EG than in CG rabbits on day 21 implying that the ED 26E/7 application had not evoked oxidative stress. Biochemical blood parameters and quality of meat were not negatively influenced. First time tested in animals, E. durans ED 26E/7 seems to be a new candidate for use in rabbits husbandry.