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Probiotic bacteria, including the Enterococcus faecium strain, can improve intestinal mucosal health by several mechanisms, including modulation of the immune response, as well as by improving the protective function of the epithelial barrier. In this study, we tested the effect of Enterococcus faecium AL41 on the acute phase proteins response (blood), gene expression of selected molecules of mucosal immunity (immunoglobulin A, mucin-2, insulin-like growth factor 2) and mucus production (all parts of the small intestine) in broilers. Eighty broiler chicks were divided into two groups: a control and E. faecium AL41 (birds were inoculated with AL41 for 7 days) group. The whole experiment lasted 11 days. Our results revealed that the administration of E. faecium AL41 had no substantial effect on the concentrations of acute phase proteins, but we recorded a significant increase in β- and γ-globulin fractions at the end of the experiment, which may indicate an improvement in the immune status. A significant prolonged stimulatory effect of E. faecium AL41 on the relative expression of molecules (immunoglobulin A, mucin-2) as well as on the dynamic of mucus production in the chicken intestine was observed. In addition, AL41 significantly reduced the total number of enterococci in the cecum and faeces.

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.

To find natural feed additives with a beneficial effect on rabbit health, thymol alone and in combination with Enterocin M were administered in drinking water for 42 days (35 -77 days of age). A total of 48 rabbits based on their weight were randomly divided into four experimental groups: C – control (basal diet), T – thymol (250 mg/L), E – Enterocin M (Ent M) (50 μL/animal/day), T + E (thymol with Ent M). Ent M (p < .05) and thymol (p < .01) separately decreased malondialdehyde in the liver. Thymol separately and in combination significantly increased phagocytic activity in the blood (p = .0051) and lactic acid in the caecum (p = .0142) and decreased coagulase-positive staphylococci in the caecum (p = .0329). Ent M separately and in combination increased immunoglobulin A content in the jejunal wall (p = .002) and decreased coliform bacteria in faeces (p = .0002). Thymol and Ent M application separately or in combination improved the antioxidant and immune response of rabbits and demonstrated an antibacterial effect.

The use of antibiotics in farm animals is one of the main reasons for the development of resistant bacterial strains (e.g., zoonotic pathogens). Therefore, save alternatives are needed. Here, we examined how post-hatch application (day one to seven of life) of the probiotic Enterococcus faecium AL41 (EF) affects the development and tissue properties of the broiler pectoralis major muscle (PM). Expression of regulators, namely IGF-1, PAX7, and MYF5, was also investigated. At day 1 (n = 6), and days 5, 8, and 12 (n = 10), muscle samples were taken from control and EF supplemented chicks. From day 5 on, myonuclei number per fiber was elevated in EF chicks. Improved capillarization (from day 8), larger myofibers, increased body and PM weights (day 12) were found in the EF group. Part of our findings is explainable by higher intramuscular expression of IGF-1 and lower MYF5 expression in EF birds. In both groups IGF-1 expression decreases with age, thereby increasing the cellular myogenic potential. However, a strong increase in PAX7 expression and more PAX7-positive nuclei were found in EF chicks at day 12. We conclude that EF supplementation improves PM growth and health due to positive effects on bioavailability and fusion capacity of SATC progeny and better tissue perfusion.

Oral glucose supply is important for neonatal calves to stabilize postnatal plasma glucose concentration. The objective of this study was to investigate ontogenic development of small intestinal growth, lactase activity, and glucose transporter in calves (n = 7 per group) that were born either preterm (PT; delivered by section 9 d before term) or at term (T; spontaneous vaginal delivery) or spontaneously born and fed colostrum for 4 days (TC). Tissue samples from duodenum and proximal, mid, and distal jejunum were taken to measure villus size and crypt depth, protein concentration of mucosa and brush border membrane vesicles (BBMV), total DNA and RNA concentration of mucosa, mRNA expression and activity of lactase, and mRNA expression of sodium-dependent glucose co-transporter-1 (SGLT1) and facilitative glucose transporter 2 (GLUT2) in mucosal tissue. Additionally, protein expression of SGLT1 in BBMV and GLUT2 in crude mucosal membranes and immunochemical localization of GLUT2 in the enterocytes were determined. Villus height in distal jejunum was lower in TC than in T. Crypt depth in all segments was largest and the villus height/crypt depth ratio in jejunum was smallest in TC calves. Concentration of RNA was highest in duodenal mucosa of TC calves, but neither lactase mRNA and activity nor SGLT1 and GLUT2 mRNA and protein expression differed among groups. Localization of GLUT2 in the apical membrane was greater, whereas in the basolateral membrane was lower in TC than in T and PT calves. Our study indicates maturation processes after birth for mucosal growth and trafficking of GLUT2 from the basolateral to the apical membrane. Minor differences of mucosal growth, lactase activity, and intestinal glucose transporters were seen between PT and T calves, pointing at the importance of postnatal maturation and feeding for mucosal growth and GLUT2 trafficking.

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.

Nowadays, the use of probiotics, postbiotics, parabiotics and other beneficial natural substances dominate health status sustainability in animals. Mundticin-like substance EM 41/3 (MLS) is a thermo-stable substance of proteinaceous character—a bacteriocin (postbiotic) with a broad antimicrobial (inhibitory) spectrum produced by the horse-derived strain Enterococcus mundtii EM 41/3. The aim of this study was to find if this new one bacteriocin can have an effect and/or a similar effect in rabbits as was achieved previously by using enterocins detected and characterized in our laboratory. In addition, these tests have been conducted to secure the health of rabbits as food-derived animals. Moreover, the aim of this study was also to test the effect of MLS EM 41/3 in broiler rabbits as an animal model before its use in horses. The parameters tested were analyzed by validated methods. Administration of MLS EM 41/3 lead to a significant increase in the non-specific immunity parameter (phagocytic activity, p < 0.05), while microbiota in feces, caecum and appendix were not negatively influenced. However, the total enterococcal count was significantly increased in the EG compared to the CG (p < 0.001). Application of MLS EM 41/3 did not have a negative impact on blood biochemistry. No oxidative stress was noted. However, higher growth parameters were noted and also hydrolytic activity was increased after MLS EM 41/3 application. Finally, to check physico-chemical parameters, no negative influence on rabbit meat quality was noted. Postbiotic MLS EM 41/3 administration seems to be a contributing factor in the feeding strategy for animal health status sustainability.

IntroductionDue to the intensification of fish farming and the associated spread of antimicrobial resistance among animals and humans, it is necessary to discover new alternatives in the therapy and prophylaxis of diseases. Probiotics appear to be promising candidates because of their ability to stimulate immune responses and suppress the growth of pathogens.MethodsThe aim of this study was to prepare fish feed mixtures with various compositions and, based on their physical characteristics (sphericity, flow rate, density, hardness, friability, and loss on drying), choose the most suitable one for coating with the selected probiotic strain Lactobacillus plantarum R2 Biocenol™ CCM 8674 (new nom. Lactiplantibacillus plantarum). The probiotic strain was examined through sequence analysis for the presence of plantaricin- related genes. An invented coating technology based on a dry coating with colloidal silica followed by starch hydrogel containing L. plantarum was applied to pellets and tested for the viability of probiotics during an 11-month period at different temperatures (4°C and 22°C). The release kinetics of probiotics in artificial gastric juice and in water (pH = 2 and pH = 7) were also determined. Chemical and nutritional analyses were conducted for comparison of the quality of the control and coated pellets.Results and discussionThe results showed a gradual and sufficient release of probiotics for a 24-hour period, from 104 CFU at 10 mi up to 106 at the end of measurement in both environments. The number of living probiotic bacteria was stable during the whole storage period at 4°C (108), and no significant decrease in living probiotic bacteria was observed. Sanger sequencing revealed the presence of plantaricin A and plantaricin EF. Chemical analysis revealed an increase in multiple nutrients compared to the uncoated cores. These findings disclose that the invented coating method with a selected probiotic strain improved nutrient composition and did not worsen any of the physical characteristics of pellets. Applied probiotics are also gradually released into the environment and have a high survival rate when stored at 4°C for a long period of time. The outputs of this study confirm the potential of prepared and tested probiotic fish mixtures for future use in in vivo experiments and in fish farms for the prevention of infectious diseases.

Intensive fish farming is associated with a high level of stress, causing immunosuppression. Immunomodulators of natural origin, such as probiotics or phytoadditives, represent a promising alternative for increasing the immune function of fish. In this study, we tested the autochthonous trout probiotic strain L. plantarum R2 in a newly developed, low-cost application form ensuring the rapid revitalization of bacteria. We tested continuous and cyclic feeding regimes with regard to their effect on the intestinal immune response and microbiota of rainbow trout. We found that during the continuous application of probiotic feed, the immune system adapts to the immunomodulator and there is no substantial stimulation of the intestinal immune response. During the cyclic treatment, after a 3-week break in probiotic feeding and the reintroduction of probiotics, there was a significant stimulation of the gene expression of molecules associated with both cellular and humoral immunity (CD8, TGF-β, IL8, TLR9), without affecting the gene expression for IL1 and TNF-α. We can conclude that, in aquaculture, this probiotic feed can be used with a continuous application, which does not cause excessive immunostimulation, or with a cyclic application, which provides the opportunity to stimulate the immunity of trout, for example, in periods of stress.

Forty-eight rabbits aged five weeks (Hycole breed, both sexes) were divided into experimental (EG) and control (CG) groups, 24 animals in each, and fed a commercial diet with access to water ad libitum. Rabbits in EG had Enterococcus faecium EF9a probiotic strain added to their drinking water (1.0×109 colony forming units/mL 500 μL/d/animal) for 28 d (between 35 and 63 d). The experiment lasted for 42 d. The animals remained in good health condition throughout the experiment, and no morbidity and mortality was noted. There was a higher live weight at 63 d of age (+34 g; P

The effect of inorganic zinc and Ascaridia galli infection was studied on MUC1, MUC2 (mucin), sIgA (secretory immunoglobulin A), and metallothionein in the intestines of broilers. Thirty-five-day-old chickens (n = 24), COBB 500 breed, were included in a 14-day experiment. Chickens were divided into 4 groups of 6 chickens each: control ©, Ascaridia galli (AG), Zinc group (Zn), and combined group (AG + Zn). Samples from the intestine for determination of MUC1, MUC2, sIgA, and metallothionein were taken at 7 and 14 days during necropsy. Samples from the jejunum for determination of MUC1, MUC2, sIgA, and metallothionein were taken at 7 and 14 days during necropsy. The results demonstrated that 12 days’ administration of inorganic zinc increased production of MUC1 (p < 0.0001) and MUC2 (p < 0.001) in the Ascaridia galli-infected group (Ag + Zn) in comparison to control (C). The beneficial effect of zinc was also revealed in the production of sIgA (p < 0.0001) in the combined group (AG + Zn) at 7 days. The concentration of metallothionein increased mainly in the zinc group (p < 0.01) of first sampling and was upregulated in Zn and AG + Zn groups. The obtained data indicate the use of inorganic zinc as a suitable immunomodulator of intestinal immunity in Ascaridia galli-infected chickens.