Modulation of lymphocyte subpopulations in the small intestine of mice treated with probiotic bacterial strains and infected with Trichinella spiralis.

AIMS: To study the local intestinal lymphocyte immunity in mice with trichinellosis affected by probiotic bacteria. METHODS AND RESULTS: Enterococcus faecium CCM8558, Enterococcus durans ED26E/7, Limosilactobacillus fermentum CCM7421 and Lactiplantibacillus plantarum 17 L/1 were administered daily (109  CFU ml-1 ) and mice were infected with Trichinella spiralis (400 larvae) on the 7th day of treatment. T. spiralis infection significantly inhibited lymphocyte subpopulations from 5 to 25 days postinfection (dpi). L. fermentum CCM7421 and L. plantarum 17 L/1 restored the CD4+ T cell numbers in the epithelium and lamina propria at the control level from 11 dpi. All strains stimulated the CD8+ T cells numbers in infected mice, which were restored in the lamina propria on 11 dpi and in the epithelium only on 32 dpi. B cells (CD19+ ) inhibition after T. spiralis infection was not affected by treatment till 25 dpi. CONCLUSIONS: The strain-specific immunomodulatory effect of tested bacteria was confirmed. L. fermentum CCM7421 and L. plantarum 17 L/1 showed the greatest immunomodulatory potential on CD4+ and CD8+ T lymphocytes in trichinellosis. E. faecium CCM8558 and E. durans ED26E/7 activated only CD8+ T cells in the lamina propria. SIGNIFICANCE AND IMPACT OF THE STUDY: Positive modulation of the gut lymphocyte immunity in T. spiralis infection with bacterial strains showed their beneficial effect with the host's antiparasitic defence.

Antimicrobial Susceptibility to Natural Substances of Campylobacter jejuni and Campylobacter coli Isolated from Italian Poultry.

Campylobacter spp. isolated from poultry are a major public health concern. Antimicrobial resistance is increasing in general, which is associated with limited treatment of diseases in animal husbandry and in human as well. The use of natural substances (enterocins and phytoadditives) seems to be a promising alternative, either in the prevention or treatment of the gastrointestinal tract infections. A total 28 broiler carcasses from Italian poultry were sampled and 39 Campylobacter strains were isolated. Twenty-four isolates were identified as Campylobacter jejuni and 15 strains were identified as Campylobacter coli by polymerase chain reaction. All Campylobacter strains were determined to be resistant to nalidixic acid, ciprofloxacin, tetracycline (E-test), cefotaxim, and ampicillin using the disk diffusion method. Interestingly, the study revealed multidrug resistance of the Campylobacter strains; however, the strains were susceptible to natural substances, such as enterocins, oregano, and sage. Eighty-five percent of the Campylobacter strains were susceptible to at least one enterocins, and 77% of the strains were susceptible to enterocin (Ent) 9296, 69% were susceptible to Ent EM41, and 69% were susceptible to Ent A(P). Sage inhibited the growth of 38% of C. jejuni strains and 53% C. coli strains, whereas oregano inhibited the growth of 50% of the C. jejuni strains and up to 67% C. coli strains. Owing to the One health concept, application of enterocins and herbal extracts is a more ecological approach for preventing bacterial contamination during production and processing of poultry, which also leads to the protection of human health.

Thymol in fattening rabbit diet, its bioavailability and effects on intestinal morphology, microbiota from caecal content and immunity.

To our knowledge, no study has been carried out to evaluate the effects of thymol sustained administration on gut health and immune response in terms of its bioavailability in the rabbit organism. A total of 48 rabbits were randomly divided at 35 days of age into two dietary treatment groups (C/control or T/thymol at 0.025% addition). Thymol was added for 21 days and then withdrawn for 7 days. Thymol content in faeces (p < 0.001) and caecal content (p < 0.05) was significantly higher than in plasma during the thymol addition and withdrawal (2442 ± 451.4, 881.9 ± 231.3 ng/g DM and 46.2 ± 28.4 ng/ml, respectively; 149.5 ± 40.54, 45.76 ± 12.44 ng/g DM and 2.73 ± 0.45 ng/ml, respectively). Increasing of villi height to crypt depth ratio in small intestinal wall (p < 0.01), phagocytic activity in blood (p < 0.0001) and lactic acid bacteria in caecal appendix (p < 0.01) and faeces (p < 0.05) was still presented after withdrawn of thymol. The thymol at this concentration demonstrated its biological properties and was able to positively affect gut health and immune response of rabbits.

Effect of enterocin M and durancin ED26E/7 supplementation on blood parameters, immune response and jejunal morphometry in rabbits.

Natural feed additives application in rabbit nutrition can help to control and prevent digestive disturbances and improve gut health and immunity around the critical weaning period. While probiotics are frequently used in rabbits, in vivo administration of bacteriocins is often limited. Therefore, the present study evaluates the effect of enterocin EntM, durancin EntED26E/7 and their combination on serum biochemistry, phagocytic activity and jejunal morphometry of rabbits. Eighty rabbits (aged 35 days, meat line M91, both sexes) were divided into experimental groups E (EntM; dose 50 µl/animal/day, activity 25,600 AU/ml), D (EntED26E/7; 50 µl/animal/day, 12,800 AU/ml), E + D (50 µl EntM + 50 µl EntED26E/7 /animal/day) and control group (C). Additives were administrated in drinking water for 21 days. Both enterocins positively influenced tested serum parameters, with emphasis on durancin EntED26E/7 administration, alone and/or in combination with EntM. Increased total proteins (E, D: p < 0.001), urea (D: p < 0.001), albumin (D: p < 0.05) and triglycerids (E, D, E+D: p < 0.001) were found. Hypocholesterolaemic effect of both additives was recorded (p < 0.001), with the lowest HDL concentration in E + D. The most of tested hepatic enzymes were positively influenced by enterocins combination (E + D; p < 0.001). The lowest AST was noted in group D (p < 0.001). Mineral profile was also improved (p < 0.001), with the highest values in D. Oxidative stress, was not evoked during enterocins application. Both additives showed a tendency to improve phagocytic activity (prolonged effect of EntED26D/7; D, E+D: p < 0.05) and jejunal morphometry parameters (increased villus cut surface; E, D, E+D; p < 0.001). Diet supplementation with EntM and mostly with EntED26E/7 can improve serum biochemistry, phagocytic activity and jejunal morphometry.

Oral administration of bacteriocin-producing and non-producing strains of Enterococcus faecium in dogs.

The current effort to incorporate microbial cultures in canine nutrition and thus intake of them on daily base increases our interest in careful and more complex study of their effects in dogs. Many of the commercially used strains have not been tested in dogs and are incorporated only on the base of beneficial effects observed in humans with specific disorders. Moreover, no information on the effects of bacteriocin-producing strains in dogs is available. Therefore, we decided to test and to compare overall effect of bacteriocin non-producing Enterococcus faecium DSM 32820 and enterocin B-producing E. faecium LMG 30881 strain (both of canine origin). Dogs were divided into three treatment groups of ten dogs each: control; DSM 32820 group; and LMG 30881 group, dosing 109 CFU/day/dog. The experiment lasted 35 days with a 14-day treatment period (sample collection at days 0, 7, 14, 35). Despite bacteriocin production is believed that may provide a competitive advantage over neighbouring sensitive strains within shared environment, results indicated somewhat better survival for the DSM 32820 compared to the LMG 30881 group. Furthermore, dogs of DSM 32820 group had optimal faecal consistency throughout the experiment, significantly stimulated phagocytic activity (days 7 and 14) and metabolic burst activity of leukocytes (days 14 and 35) and lower serum glucose concentration (day 35). In contrast, dogs of LMG 30881 group showed higher faecal count of Gram-negative bacteria (day 35), lower haemoglobin and glucose concentration (day 35), and higher metabolic burst activity (days 14 and 35). These results are further evidence of the existence of inter-strain differences in efficacy despite the same origin.

Health benefits observed after probiotic Lactobacillus fermentum CCM 7421 application in dogs.

The importance of the intestinal microbiota has attracted much interest in recent years particularly with respect to ways in which the microbiota can be manipulated in order to improve health. Improving gut health through the use of probiotic microorganisms has become an area of research activity in both human and animal nutrition. However, the amount of research using companion animals is insufficient. The present review evaluates and compares the effects achieved after application of canine-derived probiotic strain Lactobacillus fermentum CCM 7421 to healthy dogs as well as to dogs suffering from gastrointestinal disorders. The experiments involved varying duration of application (4 days-14 days), dosage (107-109 CFU), form of application (fresh culture or lyophilized from) or combination with natural substances. Results from nine independent studies show the ability of probiotic strains to establish themselves in the canine gastrointestinal tract, alter the composition of intestinal microbiota and metabolites (organic acids), and modulate the physiology (serum biochemical parameters) and immunity parameters in dogs. Almost all observed effects were irrespective of dose or duration of probiotic administration.

Sensitivity to Enterocins of Thermophilic Campylobacter spp. from Different Poultry Species.

Campylobacter spp. have been isolated from various animals, including poultry. They are rapidly transmitted throughout broiler sheds by the fecal-oral route. A promising strategy to reduce Campylobacter spp. in poultry may be done due to the beneficial properties of probiotic bacteria and their bacteriocins. In this study, inhibition spectrum/activity of different enterocins was evaluated against Campylobacter spp. (isolated from different poultry) to indicate further practical use of enterocins. Enterocins are antimicrobial proteinaceous substances produced mostly by enterococci. Feces from broiler chickens (10), laying hens (47), ostriches (140), and ducks (40) were screened. Altogether, 23 strains were allotted to the species Campylobacter jejuni and Campylobacter coli using MALDI TOF mass spectrometry and confirmed by genotyping (PCR method). In the feces of ostriches, Campylobacter spp. were not confirmed. Campylobacter spp. isolated from different poultry showed resistance to nalidixic acid, ciprofloxacin, and ampicillin. Interestingly, strains demonstrating antibiotic resistance revealed sensitivity to at least one of the nine enterocins used (except C. coli Kc1, SZ3, and C. jejuni 1/D). Almost 52% strains were inhibited by Ent A (P). Enterocins can therefore be used to prevent or reduce Campylobacter spp.; it is a basis for practical use.

Staphylococci Related to Farm Ostriches and Their Sensitivity to Enterocins.

In Slovakia, ostriches are reared mainly for their meat. There is still limited information related to microflora of ostriches, including staphylococci. Knowing the composition of microflora is very important for the recognition of potential pathogenic agents. Recently, a frequent problem in animals is the occurrence of bacteria resistant to antibiotics. The aim of this study was to detect staphylococcal species in feces of farm ostriches and to test their sensitivity to antibiotics and enterocins. Altogether 140 ostriches from three age groups were sampled (n = 18, faecal mixture samples from each group) on a farm in Slovakia or on Slovak farm. From 54 fecal samples, the staphylococcal count reached an average 4. 3 ± 0. 63 (log10) CFU/g. Twenty-four lactic acid producing strains were taxonomically classified to eight species of the genus Staphylococcus: Staphylococcus equorum, S. xylosus, S. epidermidis, S. haemolyticus, S. cohnii, S. succinus, S. warneri, and S. hominis. Strains were evaluated by secure probable species identification/probable species identification (score value up to 2.299) confirmed also by phenotypization. Most strains were sensitive to antibiotics. Four strains (S. haemolyticus SHae 111, S. haemolyticus SHAe 371, S. xylosus SX 2133, and S. warneri SW 292) were resistant to methicillin but sensitive to six or five of the seven enterocins tested (inhibitory activity 200-12,800 AU/mL). S. warneri SW 292 was sensitive to all enterocins (activity up to 12,800 AU/mL).

Experimental application of Lactobacillus fermentum CCM 7421 in combination with chlorophyllin in dogs.

Chlorophyll belongs in a larger class of phytochemical plant pigments currently receiving more attention as a physiologically active dietary component. Although most research has focused on its biological activities such as its antioxidant, antimutagenic, anti-inflammatory or apoptotic effects in humans or rodents, there is limited knowledge at this time about the combinative possibilities of chlorophyll with probiotic bacteria. Our aim was to test the growth characteristics of canine-derived probiotic strain Lactobacillus fermentum CCM 7421 in the presence of different concentrations of chlorophyllin in vitro. Antimicrobial activity of chlorophyllin against canine indicator bacteria was also detected. In the in vivo study, chlorophyllin, L. fermentum CCM 7421 and the combination of both additives on faecal microbiota, faecal organic acid concentrations, haematological and immunological parameters in dogs were tested. Forty dogs were divided into 4 treatment groups; control (C); receiving chlorophyllin (60 mg/day/dog, CH group); L. fermentum CCM 7421 (10(8) CFU/day/dog, LF group); and both additives (CH + LF group), 10 dogs in each group. The experiment lasted for 28 days with a 14-day treatment period (sample collection at days 0, 7, 14 and 28). Results showed no growth inhibition of strain CCM 7421 by 0.05-0.25 % of chlorophyllin in broth after 24 h. Reduced growth of staphylococci, Listeria monocytogenes and Citrobacter freundii was observed at 1 % chlorophyllin (P < 0.05). In dogs, lower coliform bacteria numbers and higher concentration of propionic acid in faeces of the CH group during the treatment compared to baseline were detected (P < 0.01). Phagocytic activity of leukocytes was stimulated in all three treated groups of dogs (P < 0.05).

Food-producing animals and their health in relation to human health.

The fields of immunology, microbiology, and nutrition converge in an astonishing way. Dietary ingredients have a profound effect on the composition of the gut microflora, which in turn regulates the physiology of metazoans. As such, nutritional components of the diet are of critical importance not only for meeting the nutrient requirements of the host, but also for the microbiome. During their coevolution, bacterial microbiota has established multiple mechanisms to influence the eukaryotic host, generally in a beneficial fashion. The microbiome encrypts a variety of metabolic functions that complements the physiology of their hosts. Over a century ago Eli Metchnikoff proposed the revolutionary idea to consume viable bacteria to promote health by modulating the intestinal microflora. The idea is more applicable now than ever, since bacterial antimicrobial resistance has become a serious worldwide problem both in medical and agricultural fields. The impending ban of antibiotics in animal feed due to the current concern over the spread of antibiotic resistance genes makes a compelling case for the development of alternative prophylactics. Nutritional approaches to counteract the debilitating effects of stress and infection may provide producers with useful alternatives to antibiotics. Improving the disease resistance of animals grown without antibiotics will benefit the animals' health, welfare, and production efficiency, and is also a key strategy in the effort to improve the microbiological safe status of animal-derived food products (e.g. by poultry, rabbits, ruminants, or pigs). This review presents some of the alternatives currently used in food-producing animals to influence their health in relation to human health.

Isolation and characterization of faecal bifidobacteria and lactobacilli isolated from dogs and primates.

Although bifidobacteria and lactobacilli have been suggested beneficial for the host and are components of many probiotics and competitive exclusion mixtures, the knowledge on abundance, metabolic and probiotic characteristics in isolates from dogs and monkeys is still limited. The present study was aimed to isolate Bifidobacterium and Lactobacillus strains (faeces of 22 dogs and of 5 primates: Cebus apella, Eulemur fulvus, Erythrocebus patas, Macaca fascicularis, Papio hamadryas) with the MALDI-TOF identification system Lactobacillus murinus, Bifidobacterium animalis and Pediococcus acidilactici were more frequently isolated species in dogs while Lactobacillus plantarum was isolated in several species of primates. Ten strains of 6 species were assayed for enzymatic activities (only Lactobacillus reuteri strains showed no undesirable enzymatic activity), antimicrobial susceptibility (detected higher minimum inhibitory concentration levels for tetracycline and gentamicin), and inhibitory activity against 15 indicator bacteria. All strains inhibited Gram-negative indicators while lactobacilli showed larger inhibition zones than bifidobacteria. L. reuteri II/3b/a (isolate from M. fascicularis) showed the best antimicrobial properties. Resistance to bile (0.3% w/v) was observed in all tested strains (no decrease of CFU/ml) whereas the decrease of 68.4-94.4% (after 90 min exposition) and 78.4-99.9% CFU/ml (after 180 min) depending on the strain was detected in the artificial gastric juice.

Effect of Bifidobacterium animalis B/12 administration in healthy dogs.

Bifidobacterium species constitute the most frequently used health-enhancing bacteria in functional foods or probiotic products, and most of their health benefits have been demonstrated in human or mice studies. However, knowledge of the effects of these bacteria in the canine organism is very limited. In this study, the canine-derived strain Bifidobacterium animalis B/12 (10(9) CFU) was tested for its effects on faecal microbiota, faecal characteristics, faecal organic acid concentrations, blood biochemistry, haematological and immunological parameters in healthy dogs (C-control, BA-B. animalis B/12 group, 10 dogs in each). The experiment lasted for 49 days with a 14-day treatment period (sample collection at days 0, 7, 14, 21, 28, and 49). A significantly higher population of lactic acid bacteria was detected (day 7) while the counts of coliform bacteria were lower in faeces of the BA group (days 14, 21, 28, 49) compared to control group C. Faecal concentrations of acetic (day 7, 21, 28, 49), acetoacetic (7-49) and valeric acid (14) were higher in contrast to formic acid (day 7-21), which was decreased after the treatment. In blood serum, significantly lower concentrations of triglyceride (day 14) and albumin (day 14, 28, 49) and significantly higher levels of alanine aminotransferase (day 14) and alkaline phosphatase (day 14, 28) were observed in the BA dogs. The phagocytic activity of leukocytes (especially of neutrophils) was higher in dogs after 14-day consumption of B/12 strain (day 14). The results show that many of these effects could also still be recorded several weeks after the treatment period.

Susceptibility to postbiotics - enterocins of methicillin-resistant Staphylococcus aureus strains isolated from rabbits.

There is a major problem with the rising occurrence of highly virulent and multiply-resistant strains, including methicillin-resistant Staphylococcus aureus (MRSA), because of their difficult treatment. This study aimed to evaluate the antibacterial and antibiofilm effect of new enterocins (Ent) against potential pathogenic MRSA strains isolated from rabbits. Staphylococci were identified with PCR and screened for methicillin/oxacillin/cefoxitin resistance (MR) using the disk diffusion method and the PBP2' Latex Agglutination Test Kit. Enzyme production, hemolysis, DNase activity, slime production, and biofilm formation were tested in MRSA strains. The susceptibility of MRSA to eight partially-purified enterocins (Ent) produced by E. faecium and E. durans strains was checked using agar spot tests. The antibiofilm activity of Ents was tested using a quantitative plate assay. Out of 14 MRSA, PBP testing confirmed MR in 8 strains. The majority of MRSA showed DNase activity and ß-hemolysis. Slime production and moderate biofilm formation were observed in all strains. MRSA were susceptible to tested Ents (100-12,800 AU/mL; except Ent4231). The antibiofilm effect of Ents (except Ent4231) was noted in the high range (64.9-97.0%). These results indicate that enterocins offer a promising option for the prevention and treatment of bacterial infections caused by biofilm-forming MRSA.

Antiparasitic Activity of Enterocin M and Durancin-like from Beneficial Enterococci in Mice Experimentally Infected with Trichinella spiralis.

Beneficial/probiotic strains protect the host from pathogens by competitive displacement and production of antibacterial substances, i.e., bacteriocins. The antiparasitic potential of bacteriocins/enterocins and their producing strains in experimental murine trichinellosis were tested as a new therapeutic strategy. Enterocin M and Durancin-like and their producers Enterococcus faecium CCM8558 and Enterococcus durans ED26E/7 were administered daily to mice that were challenged with Trichinella spiralis. Our study confirmed the antiparasitic effect of enterocins/enterococci, which reduced the number of adults in the intestine (Enterocin M-43.8%, E. faecium CCM8558-54.5%, Durancin-like-16.4%, E. durans ED26E/7-35.7%), suppressed the Trichinella reproductive capacity ex vivo (Enterocin M-61%, E. faecium CCM8558-74%, Durancin-like-38%, E. durans ED26E/7-66%), and reduced the number of muscle larvae (Enterocin M-39.6%, E. faecium CCM8558-55.7%, Durancin-like-15%, E. durans ED26E/7-36.3%). The direct effect of enterocins on Trichinella fecundity was documented by an in vitro test in which Durancin-like showed a comparable reducing effect to Enterocin M (40-60%) in contrast to the ex vivo test. The reducing activity of T.spiralis infection induced by Enterocin M was comparable to its strain E. faecium CCM8558; Durancin-like showed lower antiparasitic activity than its producer E. durans ED26E/7.

Fortification of Goat Milk Yogurts with Encapsulated Postbiotic Active Lactococci.

The species Lactococcus lactis is a bacterium extensively used in the dairy industry. This bacterium is Generally Recognized as Safe and was added to the European Food Safety Authority's Qualified Presumption of Safety list. The major functions of this species in dairy fermentation are the production of lactic acid from lactose, citric acid fermentation, and the hydrolysis of casein. But, the representatives of this species that produce bacteriocin substances can also exert an inhibitory effect against spoilage bacteria. The aims of this study were to test three lactococcal strains isolated from raw goat milk for their postbiotic activity and to test their stability in goat milk yogurts after their application in encapsulated form for their further application. To achieve these aims, validated methods were used. Three Lactococcus lactis strains (identified by Blastn 16S rRNA analysis) produced bacteriocin substances/postbiotics. These concentrated postbiotics inhibited the growth of enterococci and staphylococci (by up to 97.8%), reaching an inhibitory activity of up to 800 AU/mL. The encapsulated (freeze-dried) lactococci survived in the goat milk yogurts with sufficient stability. Strain MK2/8 fortified the yogurts in the highest amount (8.1 ± 0.0 cfu/g log 10). It did not influence the pH of the yogurt.

The effect of enterocin A/P dipeptide on growth performance, glutathione-peroxidase activity, IgA secretion and jejunal morphology in rabbits after experimental methicillin-resistant Staphylococcus epidermidis P3Tr2a Infection.

The increasing frequency of methicillin-resistant (MR) staphylococci in humans and animals need special attention for their difficult treatment and zoonotic character, therefore novel antimicrobial compounds on a natural base against antibiotic-resistant bacteria are requested. Currently, bacteriocins/enterocins present a new promising way to overcome this problem, both in prevention and treatment. Therefore, the preventive and medicinal effect of dipeptide enterocin EntA/P was evaluated against MR Staphylococcus epidermidis SEP3/Tr2a strain in a rabbit model, testing their influence on growth performance, glutathione-peroxidase (GPx) enzyme activity, phagocytic activity (PA), secretory (s)IgA, and jejunal morphometry (JM). Eighty-eight rabbits (aged 35 days, meat line M91, both sexes) were divided into experimental groups S (SEP3/Tr2a strain; 1.0 × 105 CFU/mL; dose 500µL/animal/day for 7 days, between days 14 and 21 to simulate the pathogen attack), E (EntA/P; 50 µL/animal/day, 25,600 AU/mL in two intervals, for preventive effect between days 0 and 14; for medicinal effect between days 28 and 42), E + S (EntA/P + SEP3/Tr2a; preventive effect; SEP3/Tr2a + EntA/P; medicinal effect) and control group (C; without additives). Higher body weight was recorded in all experimental groups (p < 0.001) compared to control data. The negative influence/attack of the SEP3Tra2 strain on the intestinal immunity and environment was reflected as decreased GPx activity, worse JM parameters and higher sIgA concentration in infected rabbits. These results suggest the promising preventive use of EntA/P to improve the immunity and growth of rabbits, as well as its therapeutic potential and protective role against staphylococcal infections in rabbit breeding.

Dairy-Derived and Bacteriocin-Producing Strain Lactiplantibacillus plantarum LP17L/1: An Assessment of Its Safety and Effect Using Broiler Rabbits.

BACKGROUND: Using bacterial (probiotic) strains can influence beneficial health statuses, e.g., through dairy products; however, they must be assessed as safe before consumption; broiler rabbits represent a suitable model for this purpose. This study evaluated the safety and effect (functionality) of the dairy-derived, plantaricin-producing strain Lactiplantibacillus plantarum LP17L/1 using broiler rabbits. The following parameters were assessed to evaluate safety and functionality: microbiota, phagocytic activity (non-specific immunity parameter), blood serum biochemistry, oxidative stress enzyme, growth performance, cecal hydrolytic activity, and jejunal morphometry. METHODS: Previously validated methods were used for the analyses. The appropriate administrations approved the experiments. RESULTS: Although only up to 1.0 colony forming unit (CFU)/g (log10) LP17L/1 reached the gastrointestinal tract of the rabbits, the total lactic acid bacteria and amylolytic streptococci were significantly increased (p < 0.001). The other microbiota were not influenced, meaning there was no negative influence on phagocytic activity, growth parameters, and biochemistry in the analyzed blood serum. Conversely, lower GPx values were measured in the experimental group than in the control group, meaning LP17L/1 did not induce oxidative stress. LP17L/1 caused an increase in villi length to crypt depth ratio, while hydrolytic activity was also significantly increased (p < 0.01, p < 0.001). CONCLUSIONS: L. plantarum LP17L/1 was confirmed as safe. Regarding the functionality, no negative influence on the microbiota was noted, and the tested parameters were not negatively influenced. Demonstrating that the LP17L/1 strain is safe provides further chances for its industrial application. Additional studies are being conducted.

Synbiotic administration of canine-derived strain Lactobacillus fermentum CCM 7421 and inulin to healthy dogs.

The canine-derived strain Lactobacillus fermentum CCM 7421 has been demonstrated to exert certain health benefits as a probiotic in dogs. Synbiotic combinations are widely used but are rarely studied in dogs. In this study the prebiotic inulin in combination with L. fermentum CCM 7421 was tested for its effects on faecal microbial populations, faecal characteristics, and blood biochemistry in canine experiments. Healthy adult dogs (n = 36) were randomly assigned to 1 of 3 treatment groups (12 dogs/treatment): (i) the control group (C), (ii) the probiotic group (LF group: L. fermentum CCM 7421, 10(8) CFU/mL of Ringer buffer, 0.1 mL/kg of body mass), and (iii) the synbiotic group (LF+I group: L. fermentum CCM 7421 + inulin (I; Raftifeed IPS) added as 1% of diet). The experiment lasted for 7 weeks with a 2-week treatment period. We detected a significant increase of lactic acid bacteria (LF versus C, day 7; LF versus C and LF versus LF+I, days 28 and 49), a decrease of clostridia (LF versus C, day 14), a lower pH value (LF versus LF+I, day 28), and a higher ammonia concentration (LF versus LF+I, days 14 and 49) in faecal samples. The synbiotic LF+I combination did not intensify the probiotic L. fermentum CCM 7421 efficacy, but its slight laxative effect can be useful to prevent constipation, e.g., in senior dogs.

Effect of thyme oil on small intestine integrity and antioxidant status, phagocytic activity and gastrointestinal microbiota in rabbits.

The effects of 0.5 g thyme oil per kg dry matter (DM) of diet on duodenal tissue integrity, antioxidant status, phagocytic activity and selected microbiota in the caecum and faeces of rabbits were studied. Twenty-four rabbits were divided into two groups and were fed a commercial granulated diet for growing rabbits (CD) with access to water ad libitum. The first group was fed the CD, while to the CD of the second group thyme oil was added. Intestinal integrity was tested by measuring the transepithelial electrical resistance (TEER). Thyme oil significantly increased the value of total antioxidant status (TAS) in the blood plasma and glutathione peroxidase (GPx) activity in the liver, and it decreased malondialdehyde (MDA) concentration in the duodenal tissue. Thyme oil resulted in strengthened intestinal integrity, as the essential oil supplementation significantly increased TEER values in the experiment. The faecal microbiota of rabbits was almost completely balanced in both groups, and only a slight decrease was found in the microbial population at day 42 of the trial. In both groups, the bacterial counts were generally lower in the caecum than in the faecal samples. In conclusion, dietary supplementation with 0.5 g/kg DM thyme oil may improve intestinal integrity, and it may have an antioxidant effect. A tendency was also found for thyme oil to stimulate the abundance of some microbes beneficial in the rabbit gut.

Enterococcus moraviensis EMo 1-1Nik of horse origin:characteristics and potential bacteriocin-producing strain.

Nowadays, developed more precisious identification techniques have allowed to validate newer enterococcal species. Among them, the species Enterococcus moraviensis was also validated, at first from surface waters. However, in this study, characteristics and potential to bacteriocin production by the strain E. moraviensis EMo 1-1Nik isolated from buccal mucosa of Slovak warm-blood horse breed has been studied. BLASTn analysis allotted this strain to the species E. moraviensis with percentage identity BLASTn 16S rRNA sequence in the strain up to 100% (99.93% similarity with E. moraviensis NR113937.1). The strain EMo 1-1Nik has been provided with GenBank accession number MW326085. It is hemolysis-negative (γ-hemolysis), deoxyribonuclease-negative and gelatinase-negative; absent of virulence factor genes, low-grade biofilm-positive (0.133 ± 0.36), mostly susceptible to tested antibiotics. Moreover, 60% of EMo1-1Nik colonies were found as bacteriocin-producing against the principal indicator Enterococcus avium EA5. The concentrated substance (CS, pH 4.5) of EMo1-1Nik showed the inhibitory activity against EA5 strain (800 AU/mL); CSs with pH 6.3 and 7.3 reached inhibitory activity 100 AU/mL against EA5 strain. CS was thermo-stable and it does not lost activity after enzymes treatment. Oppositelly, EMo 1-1Nik was susceptible to Mundticin EM 41/3 (800 AU/mL) produced by horse fecal strain E. mundtii EM 41/3 and enterocins (up to 51 200 AU/mL). In spite of the preliminary results, it has been shown a potential to produce bacteriocin substance of the safe strain E. moraviensis EMo1-1Nik. The additional studies are in processing.