Horses as a source of bioactive fecal strains Enterococcus mundtii.

Bacteriocin-producing bacteria with probiotic character are known as nutritional supplements mainly for livestock. Among those beneficial bacteria we also found enterococci. Because the species strains Enterococcus mundtii also can produce bacteriocins, this study was focused on fecal strains E. mundtii from horses and their bioactivity with a view to their possible future use in breeding. Rectal removal from 47 horses (40 mares and 7 stallions), the Norik breed from Murán were sampled in eastern Slovakia during November 2019 year. Horses age ranged from five months up to 23 years. Using MALDI-TOF mass spectrometry and 16S rRNA sequences analysis, 14 strains were allotted to the species E. mundtii. Bacteriocin substances produced by the strains EMKD 38/1, EMKD 40/2, EMKD 34/2 and EMKD 41/3 showed inhibitory activity against the most susceptible (principal) indicator strain Enterococcus avium EA5 and against listeriae as well (inhibitory activity from 100 up to 1 600 AU/mL). Only strain EMKD 41/3 possess Ent P and Mundticin KS genes and showed the broadest inhibitory activity. Ent B gene possessing strain EMKD 24/1 inhibited a growth of only indicator strain EA5. Identified E. mundtii tolerate low pH 3 and oxgall/bile. They were hemolysis, gelatinase and DNase negative and mostly susceptible to clinical antibiotics which are properties requested for application potential of strain. Substance from the strain with the broadest antimicrobial spectrum showed its practical/application potential, e.g. for optimizing the host microbiota which is important regarding the maintenance of animal`s health status.

Enterocin Ent7420 and sage application as feed additives for broiler rabbits to improve meat carcass parameters and amino acid profile.

The effect of enterocin (Ent)7420 and sage on rabbit meat carcass quality and amino acid (AA) conetnt was evaluated. Ninty-six Hyla male rabbits (35 days aged) were divided into experimental: E (Ent7420), S (sage), E + S (Ent7420 + sage) and control (C) groups. The additives were administrated in drinking water during 21 days. Time and time and treatment interaction effect were noted on carcass traits. The highest protein level was noted in rabbits LTL receiving Ent7420 in combination with sage. Essential (EAA) and non-essential amino acid (NEAA) levels increased in all experimental groups, with the highest EAAs in group E + S (E + S vs. E, S: P < 0.01; E + S, E, S vs. C: P < 0.001) and NEAAs in group E (E vs. S, E + S, C: P < 0.001). The dietary inclusion of Ent7420 alone and in combination with sage can improve the rabbit meat quality due to its higher protein, EAAs and NEAAs profile.

Enterocins as Novel Feed Additives in Rabbit Diet: Enterocin Ent M and Durancin Ent ED26E/7, Their Combination, and Effects on Microbiota, Caecal Fermentation, and Enzymatic Activity.

The present study investigates the effects of enterocin Ent M and durancin Ent ED26E/7 applied separately and in combination on the intestinal microbiota, caecal enzymatic activity, and fermentaion of rabbits. Eighty rabbits (M91 meatline, aged 5 weeks, both sexes) were divided into groups E (Ent M; 50 µL/animal/day), D (Ent ED26E/7; 50 µL/animal/day), E + D (Ent M + Ent ED26E/7), and control (C). The additives were administered in drinking water for 21 days. Antimicrobial activity of Ent M and Ent ED26E/7 on coliforms (E, E + D: P < 0.001) and pseudomonads (D: P < 0.05) in feces was noted, compared to C. Ent M and Ent ED26E/7 application stimulated caecal enzymatic activity in rabbits. Pectinolytic (E vs. D, E + D: P < 0.01), inulolytic (E vs. E + D: P < 0.01; E vs. C: P < 0.05), and amylolytic (E vs. D, E + D. P < 0.001; E vs. C: P < 0.01) activities were influenced by Ent M, while cellulolytic (D vs. E + D: P < 0.01) and inulolytic (D vs. E + D, C: P < 0.01) activities by Ent ED26E/7 treatment. The cellulolytic and pectinolytic acitivities changed with time. Treatment × time interaction was detected for cellulose and xylan degradation. During Ent M and Ent ED26E/7 treatment, increased ammonia, lactic, butyric and iso-valeric acid, and lower acetic, propionic, iso-butyric, valeric, and caproic acid concentrations were noted. It can be concluded that Ent M and Ent ED26E/7 application can improve rabbit health due to reduced spoilage microbiota and enhanced caecal enzymatic activity.

The effect of dietary supplementation of sage plant extract and Enterocin M on the mucus in the the small intestine and caecum in rabbits.

The aim of this study was to determine the beneficial effect of natural substances - enterocin M (Ent M; the proteinaceous substance produced by Enterococcus faecium CCM8558) and sage plant ( Salvia officinalis L.) extract on the production of mucus in the rabbits small intestine and caecum. Sixty four post-weaned rabbits (meat line M91) were divided into three experimental groups (EG - Ent M; SG - sage extract; ESG - combination Ent M with sage extract) and control group (CG). The experiment lasted for 35 days, the natural substances were administered during the first 21 days, Ent M in EG/ESG, sage extract in SG/ESG. The beneficial effect on mucus production quantity occured in the duodenum (p⟨0.001) and jejunum (p⟨0.01) in ESG compared to that found in CG on day 21, the prolonged effect in EG in the duodenum (p⟨0.001) compared to that observed in CG at the end of the experiment and to that in EG on day 21. The novelty of the study is in the application and monitoring the effect of non-rabbit-derived probiotic strain ( Enterococcus faecium CCM8558) bacteriocin - Enterocin M and sage plant extract on mucus quantity (expressed in gram) in different segments of the rabbit small intestine as well as the caecum. The results obtained indicate that supplementation of selected natural substances in the feed has the potent stimulatory effects on mucus production in the rabbit small intestine.

Probiotic Bacteria can Modulate Murine Macrophage's Superoxide Production in Trichinella Spiralis Infection.

The effect of probiotic strains (Enterococcus faecium EF55, E. faecium CCM7420, E. faecium CCM8558, E. durans ED26E/7, Lactobacillus fermentum CCM7421, L. plantarum 17L/1) on the production of superoxide anion (O2 -) in peritoneal macrophages of Trichinella spiralis infected mice was examined. E. faecium EF55 and E. faecium CCM8558 strains increased the O2 -production prior to parasitic infection,at the day7of application.A significant inhibition of the O2 - production caused by T. spiralis infection on day 5 post infection (p.i.) was prevented by all examined strains. Lactobacilli stimulated metabolic activity of macrophages during intestinal and early muscular phase (from day 5 to 25 p.i.) of trichinellosis. Enterococci increased the O2 - production in early intestinal phase (day 5 p.i.) and during the muscular phase of trichinellosis (days 25 and 32 p.i.). Respected increase in macrophage's metabolic activity induced by probiotic treatment in the intestinal phase of trichinellosis augmented the host antiparasite defence (damage and killing of newborn larvae with reactive oxygen species from macrophages).

Short communication: Antimicrobial potential of Lactobacillus plantarum strains isolated from Slovak raw sheep milk cheeses.

The objective of this study was to screen Lactobacillus plantarum strains isolated from the traditional Slovak raw sheep milk cheese for their inhibitory potential. Seventy-two strains were obtained from samples of raw sheep milk and raw sheep milk cheeses collected from April 2017 to September 2018, in 23 geographical areas of Eastern Slovakia, by inoculation of de Man, Rogosa, and Sharpe agar plates (Oxoid, Basingstoke, UK). Using both the genus- and species-specific PCR methods, 43 strains were identified as Lactobacillus spp., and 10 strains were confirmed as Lb. plantarum. First, the whole bacterial cultures of Lb. plantarum strains were tested by disc diffusion assay. All showed very good antibacterial activities against 6 selected foodborne pathogens, including Escherichia coli, Salmonella Enteritidis, Pseudomonas aeruginosa, Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus. Then, cell-free neutralized supernatants and partially purified bacteriocins were prepared from the 4 Lb. plantarum strains that exhibited the best antibacterial potential, and they were tested the same way as the whole bacterial cultures. Seven of the 10 Lb. plantarum strains harbored the plnEF gene, 3 strains harbored the plnD gene, and 2 strains possessed both the plnA and plnC genes that encode the production of the respective plantaricins. The presence of both plnR and plnL genes was only detected in a single Lb. plantarum isolate. Based on the results of this study, 4 strains of Lb. plantarum appeared to be suitable candidates for further testing in the dairy manufacturing sector, particularly in the production of raw sheep milk products.

Characterisation of Faecal Staphylococci from Roe Deer (Capreolus capreolus) and Red Deer (Cervus elaphus) and Their Susceptibility to Gallidermin.

Our current knowledge of microbiota in wild ruminants is limited. The goal of this study was to evaluate staphylococcal species in red and roe deer for various attributes (haemolysis, DNase, and urease activities; lactic acid and biofilm production; and antibiotic profile) and their susceptibility to gallidermin. Sixteen staphylococcal strains were identified from faeces of 21 free-living animals (9 adult female Cervus elaphus-red deer and 12 young female Capreolus capreolus-roe deer) sampled by the Polish colleagues in the Strzalowo Forest District, Piska Primaeval Forest. The variability in the species of staphylococci was determined. Seven species (Staphylococcus capitis, S. epidermidis, S. haemolyticus, S. hominis, S. pseudintermedius, S. vitulinus and S. warneri) and five clusters/groups of coagulase-negative staphylococci (CoNS) were identified. The strains were generally not haemolytic and Dnase negative; did not form biofilms or only produced low-grade biofilms; exhibited high levels of lactic acid; were urease positive; and were generally susceptible to antibiotics (only two strains were resistant to multiple antibiotics). However, all of the strains were susceptible to the lantibiotic bacteriocin gallidermin, with a minimal inhibitory concentration of 0.0156 µg (up to 6400 AU/ml in arbitrary units). This is the first study to perform a detailed study of the properties of CoNS from roe and red deer.

Enterocin M and Sage Supplementation in Post-weaning Rabbits: Effects on Growth Performance, Caecal Microbiota, Fermentation and Enzymatic Activity.

The effects of enterocin (Ent) M and sage extract applied separately and in combination were investigated. EntM (E 50 µL/animal/day in water) and sage extract (S 10 µL/animal/day in water) were applied individually and in combination (E+S) to rabbits during 21 days of treatment. The rabbits' growth was not significantly influenced by the additives. Lower feed conversion (FC) was noted in the experimental groups compared with controls, with the lowest data detected in E. The antimicrobial activity of EntM was noted (in E+S: lactic acid bacteria-P < 0.01; in E, E+S: enterococci, enterobacteria-P > 0.05; in E: clostridia-P > 0.05). The most significant changes in fermentation between weaned and older rabbits were noted in amylolytic activity at day 21 (E P < 0.05; E + S P < 0.05); prolonged reduction effect of sage extract on amylolytic activity was observed. The activity of cellulase, pectinase and xylanase was higher in older than in younger animals. Decrease in lactic acid and volatile fatty acids was noted during EntM administration, with significant effect on propionic acid concentration (E P < 0.05; E+S P < 0.001). The sage extract reduced propionic acid (S P < 0.001) and butyric acid levels (S P < 0.05) and increased the concentrations of butyric, iso-valeric, valeric, caproic acids and lactic acid (P < 0.001). It seems to be that EntM and sage supplementation may improve the economy of rabbit farms (increased FC) and the health status of rabbits (reduction of spoilage microbiota, enhanced enzymatic activities in caecum).

Domestic fowl of ducks, a source of faecal bioactive Enterococcus hirae strains.

Faecal Enterococcus hirae from domestic ducks were studied for their bioactivity to select bioactive strain for more detailed study with its probable use in poultry and also to bring novelty in basic research. After defecation, faeces (n=23, faecal mixture of 40 ducks) were sampled from domestic ducks in eastern Slovakia; birds were aged from eight to 14 weeks. E. hirae strains were identified using Matrix-assisted laser desorption/ionization time-of flight mass spectrometry with a highly probable species identification score (2.300-3.000) or a secure genus identification/ /probable species identification score (2.000-2.299), confirmed by polymerase chain reaction and phenotypization in accordance with the properties for the type strain E. hirae ATCC 9790. Strains were hemolysis negative (γ-hemolysis), and did not have active enzyme stimulating disorders. Enterocin genes were detected in three strains out of seven. Three out of four Enterocin genes were detected in Kc1/b (Ent A, P, L50A); the most frequently detected was the Ent P gene. The strains inhibited indicator strains E. faecalis, listeriae, but also Escherichia coli and Buttiauxiella strains. Lactic-acid producing E. hirae were mostly susceptible to antibiotics. Based on parameter evaluation, E. hirae Kc1/b, Kc6 can be additionally studied to select the type of bioactive substance.

Relation to enterocins and herbal extracts of fecal hemolytic Escherichia coli from domestic ducks detected with MALDI-TOF mass spectrometry.

Surveillance studies have generally reported an increase in Escherichia coli strains resistant to major classes of antibiotics used for animals' treatment. The aim of this study was to test the susceptibility of 25 strains (isolated from 30 domestic Mallard ducks-Anas platyrhynchos, both sex, aged 8 to 14 wk, taxonomically alloted to the species E. coli using MALDI TOF mass spectrometry system) to antimicrobials (antibiotics, enterocins, and herbal extracts). Testing was performed using the agar disc method and the agar diffusion method. A total of 19 E. coli strains were multiresistant to antibiotics; but 10 of those strains were susceptible to enterocins with an inhibition activity of 100 AU/mL. All strains were susceptible to herbal extracts. These results indicate further benefit application of enterocins and herbal extracts to prevent/reduce problems caused with E. coli. Moreover, additional studies are in process.

Sensitivity to antimicrobials of faecal Buttiauxella spp. from roe and red deer (Capreolus capreolus, Cervus elaphus)detected with MALDI-TOF mass spectrometry.

Wild ruminants are an interesting topic for research because only limited information exists regarding their microbiota. They could also be an environmental reservoir of undesirable bacteria for other animals or humans. In this study faeces of the 21 free-living animals was sampled (9 Cervus elaphus-red deer, adult females, 12 Capreolus capreolus-roe deer, young females). They were culled by selective-reductive shooting during the winter season of 2014/2015 in the Strzalowo Forest District-Piska Primeval Forest (53° 36 min 43.56 sec N, 21° 30 min 58.68 sec E) in Poland. Buttiauxella sp. is a psychrotolerant, facultatively anaerobic, Gram-negative rod anaerobic bacte- rial species belonging to the Phylum Proteobacteria, Class Gammaproteobacteria, Order Entero- bacteriales, Family Enterobacteriacae and to Genus Buttiauxella. Buttiauxella sp. has never previ- ously been reported in wild ruminants. In this study, identification, antimicrobial profile and sensitivity to enterocins of Buttiauxella strains were studied as a contribution to the microbiota of wild animals, but also to extend knowledge regarding the antimicrobial spectrum of enterocins. Five strains were identified using the MALDI-TOF identification system (evaluation score value was up to 2.224) and allotted to the genus Buttiauxella including the species Buttiauxella gaviniae, B. ferragutiae, B. agrestis. Strains were DNase negative, and they hydrolysed esculin; fermentation of L-arabinose, D-mannitol and D-mannose was positive. Dulcitol, inositol reaction, urea and indol were negative. Buttiauxella strains did not form biofilm. They were resistant to at least one of the 13 antibiotics tested. B. agrestis 2/109/1 was resistant to amdinocillin, clindamycin and pen- icillin. However, Buttiauxella strains were sensitive to the enterocins used (inhibition activity ranged from 100 to 25 600 AU/ml).

Assessment of biofilm formation by faecal strains of Enterococcus hirae from different species of animals.

Enterococcus hirae belongs in the Enterococcus faecium group within the genus Enterococcus. This species occurs naturally in the environment, commensally in the alimentary tracts of animals, and pathologically for example in humans with urinary infections. Some strains of E. hirae possess virulence factors, including biofilm formation. Biofilm growth protects bacteria against host de- fences; biofilm can be a source of persistent infection. Testing bacterial strains for their ability to form biofilm might therefore facilitate their treatment or prevention. This study focuses on bio- film formation by E. hirae strains derived from various animals. This kind of testing has never been done before. A total of 64 identified E. hirae from laying hens, ducks, pheasants, ostriches, rabbits, horses and a goat were tested by means of three methods; using Congo red agar, the tube method and microtiter plate agar. The majority of strains were found to form biofilm. 62.5% of strains were biofilm-forming, four categorized as highly positive (OD570 ≥1); most strains were low-grade biofilm positive (0.1 ≤ OD 570 ⟨ 1). Related to poultry, 55 E. hirae strains were tested and found to produce biofilm; 24 strains did not form biofilm, 31 strains were biofilm-forming; 27 strains showed low-grade biofilm formation, and four strains were highly biofilm-forming. Four strains from hens and ostriches reached the highest OD570 values, more than 0.500. Rabbit-de- rived E. hirae strains as well as strains isolated from horses and the goat were low-grade bio- film-forming. Microtiter plate assay proved to be the best tool for testing the in vitro biofilm for- mation capacity of E. hirae strains from different species of animals.

The Anti-parasitic Effect of Probiotic Bacteria via Limiting the Fecundity of Trichinella Spiralis Female Adults.

A potential protective effect of probiotic strains against zoonotic Trichinella spiralis infection was investigated in the framework of a new therapeutic strategy aimed at using probiotics to control parasitic zoonoses. The study was focused on the impact of six selected probiotic (bacteriocinogenic) strains on the intensity of T. spiralis infection and female fecundity ex vivo and in vitro. Bacterial strains of different origin (Enterococcus faecium EF55, Enterococcus faecium 2019 = CCM7420, Enterococcus faecium AL41 = CCM8558, Enterococcus durans ED26E/7, Lactobacillus fermentum AD1 = CCM7421, Lactobacillus plantarum 17L/1) were administered daily in a dose of 109 CFU/ml in 100 µl, and mice were infected with 400 T. spiralis larvae on day 7 of treatment. Female adults of T. spiralis were isolated on day 5 post infection (p.i.) and subsequently were used in fecundity test ex vivo. E. faecium CCM8558, E. faecium CCM7420 and E. durans ED26E/7 strains significantly reduced the number of adults in the intestine. The application of L. fermentum CCM7421, L. plantarum 17L/1, E. faecium CCM8558 and E. durans ED26E/7 caused a significant decrease in the number of muscle larvae. The treatment with E. faecium CCM8558 and E. durans ED26E/7 showed the highest inhibitory effect on female fecundity (94 %). The number of newborn larvae (NBL) was also significantly decreased after administration of L. fermentum CCM7421 and L. plantarum 17L/1 (80 %). A direct impact of probiotic strains on female reproductive capacity was examined in vitro in females isolated from untreated infected mice on day 5 p.i. A correlation was found between the inhibitory effect and the concentration of probiotic strains. The reduction effects of the strains manifested as follows: L. fermentum CCM7421 (93 %), E. faecium CCM8558, L. plantarum 17L/1, E. faecium EF55 (about 80 %), E. faecium CCM7420 and E. durans ED26E/7 (about 60 %).

Escherichia coli strains from ostriches and their sensitivity to antimicrobial substances.

Ostriches are bred especially for their high-quality meat. There is a lack of knowledge concerning the ostrich's microflora. Escherichia coli is a commensal microorganism of the poultry intestine, ostriches included. However, some strains may become pathogenic. This study was therefore undertaken to detect coliform bacteria in ostrich faeces and to test their antibiotic profile and sensitivity to enterocins. Faeces (n=54, 18 mixture samples from 3 different age groups of 140 ostriches) were sampled to isolate coliform bacteria. The counts of coliform bacteria varied from 5.69 ± 2.4 log10 CFU/g to 5.73 ± 2.4 CFU/g. Pure colonies were identified using MALDI-TOF MS mass spectrometry and confirmed by phenotypization. Seventy-one strains were allotted to the species E. coli. Sixty-four of those 71 strains caused hemolysis. They were mostly polyresistant to antibiotics. Thirty-two poly-resistant strains of E. coli were sensitive to enterocins. These strains were most sensitive to Ent 9296 (26 strains). Moreover, Ent EM41 produced by E. faecium EM41 (isolated from ostrich faeces) inhibited the growth of 20 strains, reaching activity of 100 AU/ml. Our results indicate the possibility of enterocins being used for prevention/reduction of coliforms. Of course, in vivo studies are also being processed.

Characteristic and susceptibility to enterocins of enterococci in pheasants possessing virulence factor genes.

With an increasing number of pheasants as gamebirds being reared each year, these species are becoming a more prominent part of the workload of many veterinary practices. Only limited information can be found concerning the microflora of common pheasants. A significant part of the obligate microflora consists of lactic acid bacteria, including enterococci. In this study, faeces were sampled from 60 pheasants aged 16-17 weeks. Enterococcal counts reached 5.48±1.9 (log10) CFU/g. Strains (17) were taxonomically classified to the genus Enterococcus using the Maldi-Tof identification system; they were allotted to the species E. hirae (58.8%), E. faecium (23.5%) and E. faecalis (17.7%) by highly probable species identification or by secure genus identification/probable species identication. Species allocation was also confirmed using conventional biochemical tests. Most strains formed ß-hemolysis. Gelatinase active phenotype was found in three E. faecalis strains. Enterococci were ß-glucuronidase negative, mostly trypsin negative with slight or moderate production of α-chymotrypsin. EH52b and EF42 strains possessed the highest potential for pathogenicity. Average value of lactic acid was 1.78±0.33 mmo/L. Most strains were tetracycline resistant (82.4%). Polyresistant E. faecalis strains with positive gelatinase phenotype and possessing virulence factor genes confirmed using PCR (gelE, efaAfs, ccf cob, cpd) were sensitive to enterocins (activity 1600-25,600 AU/mL).

Use of bacteriocin-producing, probiotic strain Enterococcus faecium AL41 to control intestinal microbiota in farm ostriches.

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.

Effect of thyme essential oil and selenium on intestine integrity and antioxidant status of broilers.

1. This study evaluated the duodenal wall integrity, antioxidant status as well as some immunological parameters of broiler chickens supplemented with 0.5 g Thymus vulgaris essential oil (EO)/kg diet and 0.4 mg Se/kg DM (dry matter) derived from sodium selenite. 2. A total of 192 one-d-old randomly divided chickens of both sexes (Ross 308 hybrid broilers) were divided into 4 treatment groups of 48 birds each. 3. The first group was fed on a nutritionally balanced basal diet (BD). The other three groups received BD supplemented with 0.5 g/kg thyme oil, or 0.4 mg Se/kg DM, or both feed additives together. 4. The results for the evaluated feed additives were (1) thyme oil - decreased malondialdehyde (MDA) concentration in duodenal mucosa and kidney, increased immunoglobulin A (IgA) concentration in duodenal mucosa, stimulated phagocytic activity in blood, improved intestinal barrier integrity (2) selenium - increased glutathione peroxidase (GPx) activity in blood and liver as well as thioredoxin reductase (TrxR) activity in duodenal mucosa, liver and in the kidney, (3) EO with selenium - increased thioredoxin reductase (TrxR) activity in duodenal mucosa. 5. These results demonstrated that thyme oil alone showed more effective potential to improve intestinal barrier integrity and antioxidant status as well as evoking an immune response in chickens, than if diets were supplemented with both thyme oil and selenium.

Effectivity of freeze-dried form of Lactobacillus fermentum AD1-CCM7421 in dogs.

The impact of probiotic supplementation of canine-derived strain Lactobacillus fermentum AD1-CCM7421 in freeze-dried form on quantitative composition of microbiota and short-chain fatty acid profile in feces of dogs was demonstrated by two independent studies (straightforward repeated-measures model; study I: a dose of 2 g per dog for 2 weeks, 10(8) CFU/g, n = 12; study II: 1 g per dog for 1 week, 10(7) CFU/g, n = 11. The results revealed a significant increase of lactic acid bacteria population persisting also after the cessation of probiotic application in both studies. A reduction of clostridia (study I, p (sum) < 0.01) and tested Gram-negative bacterial genera (coliforms, Aeromonas sp., Pseudomonas sp., study II, p < 0.05) was also detected. The strain AD1-CCM7421 colonized the canine digestive tract in sufficient numbers (10(5)-10(6) CFU/g) and it persisted in the majority of dogs after cessation of probiotic application. An increase of short-chain fatty acid concentrations (study I: butyric, succinic, valeric, formic acid) especially in the early post-treatment phase (p < 0.05) most likely led to a decrease of fecal pH value (p < 0.05) without negative influence on fecal consistency throughout the studies.

Effect of combined administration of enterocin 4231 and sage in rabbits.

Enterocin (Ent) 4231, produced by non-rabbit origin strain Enterococcus faecium CCM 4231 was used in combination with sage plant extract in rabbits with the aim to check their antimicrobial activity against microbiota, their effect on immunological, biochemical blood parameters, values of volatile fatty acids in caecum, Eimeria sp. oocysts occurrence and selected parameters of rabbits meat. The animals were divided into three experimental groups (EG1-Ent 4231; EG2- sage; EG3- Ent 4231 with sage) and control group (CG); 24 rabbits in each. Natural substances (NS) were administered for 21 days. The experiment lasted for 42 days. The reduction of microbiota in faeces was observed in EG3 at day 21 by a decrease in the numer of coagulase-positive staphylococci (P<0.01) in comparison with that determined in CG. The bacterial counts in the caecum were lower than those found in faeces. A decrease in the numer of Pseudomonas-like sp. in caeca of the experimental groups was observed at days 21 and 42 (difference in range 0.40-1.87 log cycles) comparing with that determined in CG. At day 21, a significant increase in phagocytic activity (PA, P<0.001) was found in blood of rabbits from EG2 comparing with that observed in CG. At day 42, a significant increase in PA (P<0.001) was determined in all experimental groups in comparison with CG. At day 21, in caecal content of EG3 significantly higher values of lactic acid were observed (P<0.05) in comparison with those found in CG. The reduction of Eimeria sp. oocysts was demonstrated after application of each of NS. Addition of NS did not influence biochemical parameters, meat quality of the animals and does not influence negatively the health status of rabbits.

Antibiotic resistance and virulence factors among Enterococci isolated from chouriço, a traditional Portuguese dry fermented sausage.

Enterococci are ubiquitous microorganisms, found as part of the normal intestinal microbiota of many animals. They can be present in food products, for example, the Portuguese dry fermented sausage chouriço. Twenty enterococci were isolated from chouriço in two processing units; after identification and typification by conventional-molecular methods, the isolates were screened for virulence factors and antibiotic resistance. Identification allocated all enterococci to the species Enterococcus faecalis, and PCR fingerprinting demonstrated that each isolate was specific to the processing unit and chouriço from which it was recovered. Regarding the screening for virulence factors, 1 strain produced cytolysin and 4 were gelatinase positive, but none produced lipase. The ace gene was detected in 1 enterococci, ebpABC and efaA(fs) in 16 isolates each, esp in 3, fsrB in 5, gelE in 7, and cylA in 1. A multiresistant phenotype was observed in 8 isolates, 6 belonging to factory A. The antibiotic resistance gene ere(B) was detected in 9 enterococci, whereas the genes tet(M), aac(6')-Ie-aph(2''), and vanA were detected in 8 isolates each. As some of the E. faecalis chouriço isolates present a multiresistant profile and harbor virulence and/or resistance genes, to assess further the safety of Portuguese dry sausages, a larger number of products and processing units must by analyzed.