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A great interest is placed on the influence of probiotic, prebiotic and synbiotic preparations on animals in accordance with the principle of One health. The small intestine mucosa represents a complex ecosystem ensuring the homeostasis of the animal organism. The effect of Lacto-Immuno-Vital synbiotic preparation on the quantity of mucin produced in the broiler chicken small intestine was studied. The chickens (7 days old Hybrid ROSS 308) were divided into 3 equal size (n = 16) groups, housed in separate halls: control group (CG), and two experimental groups that received syn-biotic preparation Lacto-Immuno-Vital, – one with continuous synbiotic administration (EGC), and another with pulsed synbiotic administration (EGP). The preparation was administered to EGC group from the experimental day 1 to day 7 continuously every day (500 g per 1000 l of drinking water.day−1), and to EGP group from experimental day 8 to day 22 in a pulsed manner (every third day) at a dose of 300 g per 1000 l of drinking water. The experiment lasted 22 days. A significant effect on mucus production quantity was found in the duodenum (P < 0.001), in EG after both types of synbiotic supplementation compared to CG. The comparison of continuous and pulsed supplementation was as follows: a significant effect (P < 0.001) was observed after continuous supplementation of the synbiotic preparation, compared to pulsed supplementation in EG.

In this case report, we described a rare complication—lung torsion after esophageal atresia repair in a newborn. Torsion of the lung is a critical condition in which either the entire lung or a lung lobe twists, leading to occlusion of blood vessels and airways. The patient's clinical condition was poor after the primary operation. An emergency thoracotomy showed 180° torsion of the right upper lobe (RUL) and right middle lobe (RML). After detorsion, perioperatively, the lung was gradually reperfused and had a normal appearance. After surgery, the patient was unstable, which culminated in a fatal end (bradycardia, reperfusion injury). Immediate intervention can preserve the affected lung or lung lobe. However, pulmonary torsion typically has a poor prognosis due to misdiagnosis and delayed treatment. Additionally, diagnosis in the neonatal period is even more challenging because the clinical symptoms are nonspecific. In any case, the question is whether detorsion is the right solution or whether a segmentectomy is necessary. [ABSTRACT FROM AUTHOR]

In our previous studies, Lactobacillus reuteri B1/1, which was renamed Limosilactobacillus reuteri (L. reuteri), was able to modulate the production of pro-inflammatory cytokines and other components of the innate immune response in vitro and in vivo. In this study, we evaluated the effect of Lactobacillus reuteri B1/1 in two concentrations (1 × 107 and 1 × 109 CFU) on the metabolic activity, adherence ability and relative gene expression of pro-inflammatory interleukins (IL-1β, IL-6, IL-8, IL-18), lumican and olfactomedin 4 produced by non-carcinogenic porcine-derived enterocytes (CLAB). CLAB cells were cultured in a 12-well cell culture plate at a concentration of 4 × 105 cells/well in DMEM medium in a controlled humidified atmosphere for 48 h. A 1 mL volume of each probiotic bacterial suspension was added to the CLAB cells. Plates were incubated for 2 h and 4 h. Our results revealed that L. reuteri B1/1 was able to adhere to CLAB cells in sufficient numbers in both concentrations. In particular, the concentration of 109 L. reuteri B1/1 allowed to modulate the gene expression of pro-inflammatory cytokines, as well as to increase the metabolic activity of the cells. In addition, administration of L. reuteri B1/1 in both concentrations significantly stimulated gene expression for both proteins in the CLAB cell line after 4 h of incubation. [ABSTRACT FROM AUTHOR]

This study's objective was to determine the impact of dietary humic substances on immune response and blood profiles in piglets. A total of 24 crossbred piglets (Slovakian White × Landrace; 35 days old; average body weight of 11.67 kg) were allotted to two dietary groups with (experimental; 5 g·kg−1) or without (control; 0 g·kg−1) natural humic substances supplementation. In this study, we observed a significant increase of the proportion of CD4+CD8- lymphocytes (p < 0.001) in the experimental group. The results also showed a tendency for an increase of the phagocytic activity and the engulfing capacity of phagocytes and the numbers of the other monitored lymphocyte subpopulations (CD3+, CD21+, CD4-D8+, CD4+CD8+, CD4+CD25+) in piglets in the experimental group compared to the control group. Supplementation of humic substances increased serum alkaline phosphatase compared to the control group (p < 0.05). Other monitored blood parameters were not significantly affected by dietary treatment. It concluded that inclusion of humic substances in the diet of piglets could have a stimulating effect on cellular immunity, without a negative effect on haematological and biochemical parameters. [ABSTRACT FROM AUTHOR]

The anticancer potential of silymarin is well known, including its anti-inflammatory as well as antiproliferative effect mediated by influencing the cell cycle, suppression of apoptosis, and inhibition of cell-survival kinases. However, less is known about silybin, the main component of the silymarin complex, where studies indicate its dual effect on the proliferation and immune response of various cell types in a dose-dependent manner. Moreover, there is a lack of studies comparing the effect of silybin on the same type of healthy and tumor cells, especially intestinal ones. Therefore, our study aimed to investigate the concentration-dependent effect of silybin on the normal intestinal porcine epithelial cell line-1 (IPEC-1) and the human epithelial colorectal adenocarcinoma cell line (CaCo-2). The metabolic viability, cell cycle, mitochondrial membrane potential, apoptosis, and the relative gene expression for pro- and anti-inflammatory cytokines were monitored in cells treated with silybin. Silybin stimulates metabolic viability as well as proliferation in IPEC-1 cells, protects the mitochondrial membrane, and thus exerts a cytoprotective effect, and has only a minimal effect on the gene expression of pro-inflammatory cytokines but significantly increases the expression of anti-inflammatory TGF-β. In contrast, it inhibits metabolic viability in tumor intestinal CaCo-2 cells, has an antiproliferative effect accompanied by increased apoptosis, and significantly reduces the expression of genes for pro-inflammatory interleukins as well as TGF-β. The antiproliferative and anti-inflammatory effect of silybin on tumor intestinal cells without a negative effect on healthy cells is a prerequisite for its potential use in the adjuvant therapy of colon cancer; however, further studies are necessary. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Due to the interest in using probiotic bacteria in poultry production, this research was focused on evaluating the effects of Lactobacillus fermentum Biocenol CCM 7514 administration on body weight gain and cytokine gene expression in chickens challenged with Campylobacter jejuni. One-hundred and eight 1-day old COBB 500 broiler chickens were equally assigned to four experimental groups at random. In the control group (C) chicks were left untreated, whereas in groups LB and LBCj a suspension of L. fermentum was administered. A suspension of C. jejuni was subsequently applied to groups Cj and LBCj. Body weight was registered, and the individuals were later slaughtered; cecum samples were collected at 12, 36 and 48 h post-infection (hpi). The entire experiment lasted seven days. Reverse transcription quantitative PCR (RT-qPCR) was used to determine expression levels of IL-1β, IL-15, IL-17, and IL-18 at each time point. Pathogen-infected individuals were observed to weigh significantly less than those fed with the probiotic. Significant differences were also found in transcript abundance; expression of IL-15 was downregulated by the probiotic and upregulated by C. jejuni. The effects of bacterial treatments were time-dependent, as the expression profiles differed at later stages. The present outcomes demonstrate that L. fermentum both reduces the impact of C. jejuni infection on chicken body weight and regulates positively pro-inflammatory cytokine expression, which ultimately increase bird well-being and improves production. [ABSTRACT FROM AUTHOR]

The aim of the present study was to monitor selected parameters of mucosal immunity in jejunum and ileum (immunoglobulin A [IgA], mucin 2 [MUC-2], and pro-inflammatory cytokines) in commercial broiler farm chicken after treatment with flubendazole (Flimabend®) and natural extract from chestnut wood (Farmatan®). A total of 24 forty-day-old Kalimero-Super Master hybrid chickens were divided into 4 groups (n = 6): the Fli group received Flimabend ® per os, 100 mg/g suspension in 1.43 mg of active substance/kg body weight during 7 d of experiment; the Far group received Farmatan ® per os at 0.2% concentration for 6 h/d during 5 d (experimental d 3 to 7); the Far + Fli group received a combination of doses administered in the same way as for the first two groups; and the C group represented control with no active substance administration. The concentrations of secretory IgA (sIgA) and MUC-2 and relative expression of selected immune parameters were evaluated. Our results show strong suppressive effect of the Farmatan® and Flimabend® combination on relative expression of IL-1β and IL-18 in selected parts of the intestine. On the other hand, administration of natural extract from selected chestnut wood (Farmatan®) increased expression of total IgA as well as concentration of sIgA in the studied parts of the chicken intestine. Moreover, expression and concentration of MUC-2 was positively affected by addition of Farmatan®. In contrast, 7-d administration of Flimabend® resulted in upregulation of pro-inflammatory cytokines and decrease in IgA and MUC-2 gene expression. In conclusion, for maintenance of mucosal immunity via activation of IgA and mucin production, the long-term preventive use of Farmatan® is a suitable choice. [ABSTRACT FROM AUTHOR]

Campylobacteriosis is mainly caused by infection with Campylobacter jejuni following consumption or handling of Campylobacter-contaminated poultry meat. The aim of this study was to investigate the effect of probiotic Enterococcus faecium AL41 on TGF-β4 and IL-17 expression and on immunocompetent cell distribution after C. jejuni infection in broiler chicken, as a second part of the previous study of Karaffová et al. (2017). Accordingly, day-old chicks were randomly divided into four experimental groups of 10 chicks each (n = 10): control (C), E. faecium AL41 (EFAL41), C. jejuni CCM6191 (CJ), and combined E. faecium AL41 + C. jejuni CCM6191 (EFAL41 + CJ). Samples from the caecum were collected on days 4 and 7 post Campylobacter infection (dpi), for the isolation of mRNA of TGF-β4, IL-17 and for immunohistochemistry. The relative mRNA expression of TGF-β4 was upregulated in the combined (EFAL41 + CJ) group compared to other groups during both samplings, but the expression of IL-17 was downregulated. Similarly, the highest density of CD3+ was detected in the combined group at 7 dpi, but the number of IgA+ cells was increased in both groups with EFAL41. It was concluded that the EFAL41 probiotic E. faecium strain can modulate the expression of selected cytokines (upregulation of TGF-β4 but downregulation of IL-17 relative expression), and activate IgA-producing cells in the caeca of chicks infected with C. jejuni CCM6191. [ABSTRACT FROM AUTHOR]

The study compared the effect of dietary supplementation with an inorganic or organic source of zinc (Zn) on mucin 2 (MUC-2) and IgA gene expression, the cytokines IL-17 and TGF-β4 and the secretory IgA content (sIgA) in broiler jejunum. One-day-old chickens were fed an unsupplemented basal diet (BD) or the same BD supplemented with 30 or 70 mg/kg of added Zn from ZnSO4·H2O or Zn chelate of glycine hydrate for 40 days. The highly expressed MUC-2 and IgA genes were observed in both groups supplemented with the low-dose Zn sources (30 mg/kg). A higher sIgA concentration was observed in both the ZnSO4 groups and the glycine-zinc/30 mg group. Our data indicate that the organic Zn chelate has better availability than the inorganic Zn source, and the low-dose Zn diets proved to be more beneficial to the maintenance of intestinal immune homeostasis. [ABSTRACT FROM AUTHOR]

The aim of this study was to delineate the ability of selected probiotic strains to reduce invasion byCampylobacter jejuniin peripheral blood mononuclear cells (PMBCs). PMBCs were isolated from chicken blood and exposedin vitroto probiotic strainsEnterococcus faeciumEF55,E. faeciumAL41,E. faeciumH31,Lactobacillus fermentumAD1, and infected withC. jejuniCCM6191. The mRNA expression levels for selected cytokines were determined at 24 and 48 h postinfection (pi) using reverse transcriptase-polymerase chain reaction. The level of expression of IL-1β, LITAF, and MIP1-β was upregulated (P< 0.05;P< 0.01;P< 0.001) in LAD + CJ (L. fermentumAD1 +C. jejuni) group when compared to control (C), CJ (C. jejuni), and other combined groups (EF55 + CJ, AL 41 + CJ, H31 + CJ) 24 h pi. Overall,L. fermentumAD1 andE. faeciumAL41 demonstrated the highest immunomodulatory activity against campylobacter infectionin vitro. [ABSTRACT FROM AUTHOR]

Quantitative RT-PCR was used to determine the mRNA expression of pro-inflammatory cytokines IL-15, IL-17, IL18, LITAF, iNOS, and LyTact chemokine at the caecum and spleen of experimentally infected chicks with S. enterica serovar Enteritidis SE147 preventively treated with Enterococcus faecium EF55. One-day-old ROSS 308 female chicks (100) were randomly divided into 4 groups (n=25). The chicks of the probiotic (EF) and Salmonella+ probiotic (EFSE) groups received per os E. faecium EF55 (109 CFU/day) from 1 to 7 days of the experiment. The birds of the Salmonella (SE) and EFSE groups were perorally infected with Salmonella Enteritidis SE147 in a single dose (108 CFU) on Day 4 of the experiment. The preventive administration of E. faecium EF55 showed higher, however no significant, mRNA levels of studied proinflammatory cytokines and LyTact chemokine except of iNOS in caecum of EFSE experimental group of chicks mainly at Day 1 after S. Enteritidis SE147 infection compared to other groups. In contrast, the dynamics of cytokine/chemokine gene specific responses in caecum are not correlated with responses in the spleen. [ABSTRACT FROM AUTHOR]

Simple Summary: The cranial cruciate ligament rupture is characterized by chronic inflammation, osteoarthritis of the stifle joint, and extracellular matrix degeneration of the ligament itself in dogs. Early pre-clinical cranial cruciate ligament alteration cannot be detected by clinical examination or standard radiography. Therefore, we assessed the possible relationship of inflammatory markers in peripheral blood and synovial fluid of affected stifle joints in comparison to a control. We also evaluated components of the extracellular matrix of ruptured ligaments and finally compared the tibial plateau angle and the anatomical-mechanical angle between groups. Some of the assessed inflammatory markers were significantly increased in both the peripheral blood and synovial fluid compared with the control, as were collagens. The tibial plateau angle was not significantly different; however, the anatomical-mechanical angle significantly increased in the ruptured ligaments. Our results suggest a possible positive relationship between inflammatory markers of blood and synovial fluid in cranial cruciate ligament deficient stifles compared to the control. These findings may support both local and systemic inflammation process at the same time during osteoarthritis progression. Based on this, it would be interesting to investigate the predictive osteoarthritis pathway of inflammatory cytokines, matrix metalloproteinases, and their effect on the extracellular matrix components of the cranial cruciate ligament in future studies. The cranial cruciate ligament rupture (CrCLR) is characterized by chronic inflammation and osteoarthritis (OA) of the stifle joint and extracellular matrix (ECM) degeneration of the ligament itself in dogs. Generally, OA may arise from chronic low-grade systemic inflammation. We assessed the possible relationship of inflammatory markers in the peripheral blood (PB) and synovial fluid (SF) of affected stifle joints in comparison to a control. Moreover, no study has shown the possible association between PB and SF levels of inflammatory markers in CrCLR stifles of dogs in veterinary medicine yet. We also evaluated components of ECM of CrCLR and finally compared the tibial plateau angle (TPA) and the anatomical-mechanical angle (AMA) between groups. Samples from PB and SF were examined for mRNA expression of interleukins, TNF-α and INF-γ. ECM components—collagen 1A1 and 3A1 and elastin—were examined for mRNA expression from SF. The level of relative expression for IL-1β, IL-8 and IFN-γ was significantly increased in both PB and SF in CrCLR stifles as compared with the control. Collagens were also significantly increased in CrCLR stifles. TPA was not significantly different; however, the AMA angle significantly increased in the CrCLR group. Our results suggest a possible relationship between PB and SF levels of inflammatory markers in CrCLR stifles of dogs. [ABSTRACT FROM AUTHOR]

Immune response of day-old chicks infected with Salmonella Enteritidis PT4 and preventive administration of Enterococcus faecium AL41 were studied using hematology and flow cytometry of immunocompetent cells in blood, cecum, bursa and spleen for 11 days, and included 220 animals divided into four groups (n = 55). E. faecium AL41 was administered for 7 days to EF and EFSE groups and on day 4 SE and EFSE groups were infected with Salmonella Enteritidis. Values of monocytes at 4 dpi significantly increased in EFSE and lymphocytes at 7 dpi in EF groups. Blood CD3, CD4, CD8 and IgM lymphocytes improved in EF and EFSE groups and IgA in EF group at 4 dpi. Phagocytic activity of probiotic groups was improved in both samples. Cecal IEL and LPL lymphocytes showed at 7 dpi stimulation of CD3, CD4 and CD8 subpopulations in probiotic groups, especially in EFSE group, IgA IEL and IgA with IgM LPL in EF groups. Bursa Fabricii at 7 dpi presented overstimulation of IgG subpopulation in SE group, spleen CD3 and CD8 in EF and EFSE groups. E. faecium AL41 revealed the protective effect and positive influence on the local and systemic immune response in Salmonella Enteritidis PT4 infected chickens. [ABSTRACT FROM AUTHOR]

Despite the fact that humic substances (HS) have been frequently studied in relation to their effects on livestock health, studies on their influence on egg production and quality, immunity, and intestinal microbiota of laying hens are limited. In this study, the influence of 0.5% HS supplementation on the specific production parameters of eggshell mineral quality, immune parameters (relative expression of IgA, IGF-2, MUC-2 gene in cecum; activity of phagocytes, percentage of selected lymphocyte subpopulations in the peripheral blood), and number of lactic acid bacteria and enterobacteria in the intestinal contents in laying hens was tested. The addition of 0.5% HS to the laying hen feed had a positive effect on egg laying rate, daily egg mass, egg weight, feed conversion and eggshell quality and also had an immunostimulatory effect manifested by increased phagocyte activity and B cell response. Concurrently, an increase in the number of enterobacteria in the intestinal contents and a decrease in the proportion of T lymphocytes (p < 0.05) was observed, which can be considered as a negative effect of HS. The results confirmed that HS can be used for the improvement of egg production and targeted immunostimulation, but their effect on the intestinal microbiota and T lymphocytes should be studied in more detail. [ABSTRACT FROM AUTHOR]

Ascaridiosis in poultry results in a reduction in body weight gain, egg production, as well as microelement levels. Infected poultry have higher demands on feed with the addition of essential elements including zinc. The effects of the infection by Ascaridia galli and the supplementation of inorganic zinc on the immune status of broilers were monitored through evaluation of the relative expression of selected genes (interleukins, IFN-γ, and TNF-α) by real-time PCR, haematology parameters by microscopy, and quantitative changes of lamina propria lymphocytes by flow cytometry in day 7 and day 14 of the study. We observed that the enrichment of the diet with inorganic zinc has a positive effect on the relative percentage of CD4+ lamina propria lymphocytes in the jejunum and on heterophil counts in blood. In addition, it was concluded that inorganic zinc has an anti-inflammatory effect (downregulation of TNF-α and IL-17) and activates IgA-producing cells in the jejunum of chicks infected with A. galli. [ABSTRACT FROM AUTHOR]

The health benefits of kefir consumption have been well-known for hundreds of years. The objective of this study was to investigate the effect of kefir milk and the probiotic strain Lacticaseibacillus paracasei Ž2 isolated from kefir grains on the immune response and selected parameters of the lipid and liver enzymatic profiles of mice. Mice fed with kefir milk showed significantly increased phagocytic activity and percentages of B cells in the blood and increased gene expression for mucins and percentages of CD8+ lymphocytes in the gut. By applying kefir, we achieved a significant reduction in serum LDL cholesterol and an LDL/HDL ratio that favored an increase in HDL cholesterol. Regarding the hepatic enzymes, in particular a significant reduction in ALT activity was observed. L. paracasei Ž2 alone stimulated the immune response more markedly compared with kefir milk. Regarding the systemic level, we observed increases in the proportion of all T cells (CD3+), CD4+ lymphocytes and the ratio of CD4+:CD8+ cells, and regarding the local intestinal level we noted a significant increase in gene expression for mucins (MUC-1 and MUC-2) and IgA. Moreover, we confirmed the formation of a biofilm on the surface of the forestomach only after the application of L. paracasei Ž2 alone, but not after kefir administration. The results confirmed the hypothesis that the final effect of the probiotic does not correspond with the effect of the individual strain but is the result of mutual interactions of the microorganisms presented in a preparation, and therefore in the case of multi-strain probiotics, in vivo testing of the complex preparation is necessary. [ABSTRACT FROM AUTHOR]

Simple Summary: High consumption of chicken meat and derived products has been associated with Campylobacter jejuni infections in humans. Probiotics have been exploited successfully with the aim of preventing colonization by unwanted microorganisms in birds. In this research, we investigated the effects of Lactobacillus fermentum Biocenol CCM 7514 supplementation on body weight, morphometry of the intestine and the cecal cytokine response. Probiotic-treated chickens showed higher body weight values than those exposed to C. jejuni or reared under control conditions. These differences in body weight were correlated to the overall characteristics of the small intestine, with larger villi and deeper crypts, observed in chickens administered with L. fermentum; such conditions are known to favor nutrient absorption. Likewise, body weight proved to be correlated to transcript abundance of IL-1β and IL-13. In probiotic-treated birds, such factors were upregulated in comparison to what was detected in C. jejuni-infected chickens; these interleukins are considered crucial in the response to invading pathogens. Clearly, these results show that administration of this probiotic strain lessens the negative effects elicited by C. jejuni and ultimately improves chicken body weight. This research was conducted to investigate if the administration of the probiotic Lactobacillus fermentum could influence body weight, intestinal morphometry and the cecal cytokine response in Campylobacter jejuni-infected chickens. Seventy-two 1-day old COBB 500 male chicks were allocated randomly into four experimental groups. (I) Control group (C), in which chicks were left untreated. (II) LB group, treated with L. fermentum. (III) Cj group, infected with C. jejuni and (IV) coexposure group in which both bacteria were administered. Body weight was registered and then all birds were slaughtered; samples from the small intestine and caecum were collected at 4- and 7-days post infection. The experiment lasted eleven days. Villi height and crypt depth ratios of the duodenum, jejunum and ileum were evaluated using appropriate software, while reverse transcription quantitative PCR (RT-qPCR) was utilized for assessing transcript levels of key cecal inflammatory cytokines (IL-1β, IL-18, IL-17, IL-15, IL13 and IL-4). Campylobacter-infected birds showed lower body weight values than those supplemented with the probiotic; these birds, in turn, proved to be heavier than those reared under control conditions. L. fermentum administration improved morphometrical parameters of the duodenum, jejunum and ileum; in general, villi were larger and crypts deeper than those identified in control conditions. Moreover, the negative effects elicited by C. jejuni were not observed in chickens exposed to the probiotic. Significant differences were also determined with regards to transcript abundance of all evaluated cytokines in the caecum. C. jejuni induced a downregulation of the studied interleukins; however, such a response was heightened by administration of L. fermentum, with an increase rate of transcription that promoted a more effective response to a C. jejuni infection. The effects of experimental treatments proved to vary between sampling points. Conclusively, these results demonstrate that L. fermentum lessens the negative effects elicited by C. jejuni on body weight by alleviating the impact on intestinal morphometry and cecal cytokine response, which ultimately improve chicken growth performance. [ABSTRACT FROM AUTHOR]

Within the EU, campylobacteriosis is a frequently reported zoonotic disease in humans. Campylobacter species are especially associated with human gastrointestinal diseases. C. jejuni and c. coli occurring in chickens are responsible for them. The intestinal microbiota are maintained in a dynamic relationship with the host, involving a complex 'trialogue' between microorganism, epithelium and mucosal immune system. Noncommensal and probiotic bacteria are able to induce a gut mucosal immune response. The mechanisms by which probiotic bacteria affect the immune system are attributed to an increase in the innate or acquired immune response. We studied by flow cytometry in chickens the cecal intraepithelial and lamina propria CD3+, CD4+, CD8+ and IgA+ lymphocyte subpopulations in C. jejuni and C. coli infection after preventive administration of probiotic strains E. faecium AL41 and Lactobacillus fermentum CCM 7514. Two experiments with different probiotic strains and campylobacter bacteria were done in one-day-old broiler chickens of COOB 500 hybrid. Seven days preventive administration of Enterococcus faecium AL41 and C. jejuni (108 CFU/0.2 ml PBS) challenge on day 4 were used as first, Lactobacillus fermentum CCM 7514 and C. coli as second. Individual perorally administration of bacteria were used in campylobacter (CJ, Cc), probiotic (EF, LB) and combine groups (EFCJ, LBCc) compared to negative group without any bacteria in two samplings on days 4 and 7 post infection (dpi). Isolated cecal IEL and LPL, flow cytometry, direct immunofluorescence and double staining of lymphocytes by labelled mouse antichicken monoclonal antibodies were used. Enterococcus faecium AL41 showed in combined EFCJ group the increase of CD3+ IEL and LPL on 4 dpi. Cecal CD4+ LPL elevated on day 4 and IEL on day 7 pi. CD8+ showed only IEL improvement on 7 dpi. On the other hand, while IgA+ IEL not overnumbered CJ group, LPL raised on 4 and 7 dpi. Lactobacillus fermentum CCM 7514 presented in combined LBCc group later increase of IEL CD3+ and CD4+ (7 dpi), but earlier improvement of IEL and LPL CD8+ together with IgA+ (4 dpi). Both probiotic strains showed beneficial effect to campylobacters presented mainly by IgA+ increase, showing the contribution of humoral immune response. Acknowledgements: Thank you to the Grant Agency for Science of Slovak Republic VEGA 1/0562/16, 1/0112/18 and Slovak Research and Developmental Agency APVV-15-0165. [ABSTRACT FROM AUTHOR]

Campylobacter jejuni and C. coli are identified as major causes of food-borne disease in humans worldwide. Chicken TLRs are important in the recognition of PAMPs to induce the production of antimicrobial peptides and the expression of co-stimulatory molecules that may initiate adaptive immunity responses. Beta-defensins are antimicrobial peptides, which represent one family of host defence peptides that are modulated during campylobacter infection. On the other hand, it has been reported that campylobacter usually significantly downregulated their production. Promising therapy appears to be use of probiotic strains such as Lactobacillus spp. The aim of this study was to determine whether probiotic-feeding affected the expression of TLR4 and selected defensins (AvBD2, AvBD6) in response to C. coli challenge in the cecum of broilers. Forty one day-old broilers were randomly divided into 4 experimental groups (n= 10): control (C), L. fermentum CCM7514 (LB), C. coli (CJ), and combined L. fermentum CCM7514 + C. coli (LBCJ). L. fermentum was administered daily per os to LB and LBCJ groups from day 1 to day 7 of the experiment. C. coli was administered orally on day 4 of the experiment by a single dose to CJ and LBCJ groups. Samples from cecum were collected on day 4 and 7 post- Campylobacter infection (dpi). Samples were homogenized and total of RNA was isolated. Specific primers were used. Amplification and detection of specific products were performed using CFX 96 RT system with predefined program. Relative mRNA expression of defensins was upregulated in combined group compared to other groups (P<0.001) at 7 dpi. However, the increase relative expression of TLR4 was recorded in LB group compare to other groups (P < 0.05,P < 0.01,P<0.001) during both samplings. These results suggest that probioticsfeeding may stimulate the immunodefence system mediated by defensins and TLR4 against infection by C. coli in the cecum of broilers. [ABSTRACT FROM AUTHOR]

IgA gene expression and quantification of mucous IgA+, IgM+ and CD4+ lymphocytes in the cecum of chicks was studied by qRT-PCR, immunohistochemistry and flow cytometry. A total of 220 1-day-old Salmonella -free chicks of Cobb 500 were divided into four groups ( n = 55). Group 1 served as control (C), group 2 was pretreated with probiotic bacterial strain Enterococus faecium AL41 (EFAL41), group 3 was infected with Salmonella Enteritidis PT4 (SE), and group 4 was pretreated with E. faecium AL41 and subsequently challenged with Salmonella Enteritidis PT4 (EFAL41 + SE). The relative mRNA expression of IgA was upregulated in the EFAL41 group ( P < 0.05) when compared to control group at 4 dpi. In comparison to the control, EFAL41 and SE group, the relative mRNA expression of IgA was also upregulated in EFAL41 + SE group at 7 dpi ( P < 0.001). Immunohistochemistry revealed, that the density of IgA+ cells was higher in EFAL41 + SE group comparing to the controls and SE groups ( P < 0.001). Significantly more CD4+ cells were present in the SE group than in EFAL41 ( P < 0.05), and EFAL41 + SE groups ( P < 0.001) at 4 dpi. In contrast, higher density of CD4 + cells at 7 dpi was seen in EFAL41 + SE group as compared with controls ( P < 0.05). Flow cytometry determined that relative percentage of intraepithelial lymphocytes (IEL) IgA+ cells was higher in EFAL41 than in SE and EFAL41 + SE groups ( P < 0.05). Comparing to controls the number IgM+ cells increased in SE group ( P < 0.05) at 7 dpi. The results demonstrated beneficial effect of E. faecium AL41 on the mRNA expression of IgA and number of IgA+ cells. Lamina propria lymphocytes (IgA+, IgM+) were not affected by EFAL41 intake or salmonella infection. Probiotic bacterial strain EFAL41 positively influenced the number of IEL during the first days of infection. [ABSTRACT FROM AUTHOR]

The protective effect of polysaccharide glucan in chickens fed low doses of T-2 toxin was assessed. The binder effect of ß-D-glucan on jejunal mucosa in relation to the expression of Na+/K+-ATPase, proliferative activity of enterocytes and number of goblet cells was investigated. A total of 40 one-day-old chickens were allocated to four groups: control (C), ß-D-glucan (G), T-2 toxin (T) and combined ß-D-glucan+T-2 toxin (GT). The chickens were individually administrated per os 1.0 mg/bird/day of ß-D-glucan derived from Candida albicans on days 11, 12, and 21 of the experiment (totally 3 mg per bird). T-2 toxin at a concentration of 1.45 μg·kg-1 was added to the feed from day 14 to day 28 of the experiment. The a subunitspecific anti-Na+/K+-ATPase antibody was used to identify the protein by immunofluorescence in the cell membrane of jejunal enterocytes. Higher expression of Na+/K+-ATPase was found in the jejunal epithelial cells and lamina propria in the chickens fed T-2 toxin and administered glucan (P < 0.05) compared to control. The number of proliferated enterocytes was higher in group T compared to group G and control (P < 0.001), as well group GT (P < 0.01). Goblet cell density did not present significant differences between groups of chickens, but group G showed the highest values. These data suggest that administration of pure T-2 toxin at low doses affects primarily the protein synthesis of actively dividing cells. Higher distribution of Na+/K+-ATPase in enterocytes of chickens in GT group suggests positive influence of glucan and mycotoxin on the ion pump. A binding effect of this immunomodulator on the digestive tract mucosa in the applied setup was not observed. [ABSTRACT FROM AUTHOR]