In vitro study of biological activities of anthocyanin-rich berry extracts on porcine intestinal epithelial cells.

BACKGROUND: Anthocyanins, compounds that represent the major group of flavonoids in berries, are one of the most powerful natural antioxidants. The aim of this study was to evaluate biological activities and comparison of anthocyanin-rich extracts prepared from chokeberry (Aronia melanocarpa), elderberry (Sambucus nigra), bilberry (Vaccinium myrtillus) and blueberry (V. corymbosum) on the porcine intestinal epithelial IPEC-1 cell line. RESULTS: The IC50 values calculated in the antioxidant cell-based dichlorofluorescein assay (DCF assay) were 1.129 mg L(-1) for chokeberry, 1.081 mg L(-1) for elderberry, 2.561 mg L(-1) for bilberry and 2.965 mg L(-1) for blueberry, respectively. We found a significant negative correlation (P < 0.001) between cyanidin glycosides content and IC50 values. Moreover, extracts rich in cyanidin glycosides stimulated proliferation of IPEC-1 cells and did not have cytotoxic effect on cells at an equivalent in vivo concentration. CONCLUSIONS: We found that the chokeberry and elderberry extracts rich in cyanidin glycosides possess better antioxidant and anticytotoxic activities in comparison to blueberry or bilberry extracts with complex anthocyanin profiles.

Dynamics of antibody response and viraemia following natural infection of porcine circovirus 2 (PCV-2) in a conventional pig herd.

Porcine circovirus 2 (PCV-2) is a primary agent of post-weaning multi-systemic wasting syndrome (PMWS), ubiquitous in pig herds. The course of viraemia and seroconversion in naturally infected pigs were investigated in piglets from the 2nd week of their life. Piglets were divided into seropositive (Ab(+)) and seronegative (Ab(-)) groups. Subsequently, after vaccination against PCV-2 (Ingelvac(®) CIRCOFLEX™, Böehringer Ingelheim), they were further divided into non-vaccinated seronegative (NVAC/Ab(-)) and seropositive (NVAC/Ab(+)), and vaccinated seronegative (VAC/Ab(-)) and seropositive (VAC/Ab(+)). PCV-2 colostral antibodies failed to prevent development of natural PCV-2 infection in conventional piglets; however, this occurred at a higher age in comparison with seronegative pigs. Neither colostral nor post-infection antibodies prevented development of viraemia, which persisted up to the end of the study (the 19th week), but without clinical signs of PMWS. Vaccination failed to prevent development of natural PCV-2 infection, but viraemia was limited to between the 8th and 10th week. The presence of colostral anti-PCV-2 antibodies did not show any untoward effect to vaccination; on the contrary, VAC/Ab(+) animals showed the lowest titre of viraemia.

The effects of sage extract feed supplementation on biochemical parameters, weight of internal organs and Salmonella counts in chickens.

We investigated the effects of dietary addition of sage extract on the biochemical parameters, weight of some body organs and changes in the counts of Salmonella Enteritidis PT4 (SE) in experimentally infected chickens. The following diets were used: basal diet, basal diet with addition of an extract of Salvia officinalis L. (S), basal diet and SE, and basal diet and S and SE (SSE). Compared to the SE group, sage extract in the SSE group decreased activities of ALP and ALT and concentrations of glucose and bilirubin on the 4th day post inoculation (p.i.). However, on the 18th day p.i., lower levels of bilirubin and ALT activity only were detected. Addition of sage extract to the diets decreased the counts of Salmonella in the liver, spleen and caecum at both sampling times, along with lower production of mucus in the chickens' intestines. Our results suggest that the addition of sage extract to the diet could be effective in protecting SE-infected chickens.

Leukocytic responses and intestinal mucin dynamics of broilers protected with Enterococcus faecium EF55 and challenged with Salmonella Enteritidis.

The protective effect of Enterococcus faecium EF55 in chickens challenged with Salmonella enterica serovar Enteritidis phage type 4 (SE PT4) was assessed. The antibacterial effect on the bacterial microflora in the small intestine in relation to white blood cell count, phenotyping of peripheral blood and intestinal lymphocytes, functional activity of lymphocytes and phagocytes and mucin quantitation were investigated. Day-old chicks (85) were randomly divided into four groups. The probiotic group (EF) and Salmonella+probiotic group (EFSE) received E. faecium EF55 (10(9) CFU - 3 g/group/day) for 21 days. The Salmonella group (SE) and EFSE group were infected with Salmonella Enteritidis (10(8) CFU in 0.2 ml PBS) in a single dose per os on day four of the experiment. The control group chicks (C) were fed a commercial diet without added bacteria. Supplementation of EF55 in the diet of the chickens in the EFSE group, challenged with S. Enteritidis, caused the density of the intestinal mucin layer to increase significantly in non-specific regions (duodenum and jejunum), but decrease significantly in target regions (caeca) for S. Enteritidis. Probiotic treatment also appeared to result in a significantly higher number of lymphocytes in peripheral blood and a tendency to increase CD3, CD4, CD8, and IgM positive cells 3 days post-infection with S. Enteritidis. The results demonstrated an antibacterial effect and suggested that EF55 had a moderating effect on intestinal mucin production and leukocytic response in the early phase of S. Enteritidis infection.

The effect of supplementation of flax-seed oil on interaction of Lactobacillus plantarum--Biocenol™ LP96 and Escherichia coli O8:K88ab:H9 in the gut of germ-free piglets.

We investigated the influence of administration of flax-seed oil on interaction of Lactobacillus plantarum - Biocenol™ LP96 and Escherichia coli O8:K88ab:H9 in the gut of germ-free piglets. When compared to animals supplemented with L. plantarum, the counts of lactobacilli in the jejunal and ileal mucosa and in the intestinal content were significantly higher in LMK group (p<0.0001). Inter-groups comparison of the counts of E. coli K88 adhering to the jejunal and ileal mucosa revealed a significantly decrease in LMK animals (p<0.001; p<0.05).

The effect of the probiotic strain Enterococcus faecium DSM 7134 in combination with natural humic substances on performance and health of broiler chickens.

The present study investigated the effect of the probiotic strain Enterococcus faecium DSM 7134 (2 x 10(7) cfu per g of feed) (probiotic group), natural humic substances (7g x kg(-1) feed) (HS group), and the combination of both (probiotic + HS group) on the performance, serum biochemistry, and biological activity of leucocytes of broiler chickens. Average body weight of chickens in different phases as well as throughout the observation period under different treatments did not differ significantly. Final body weight and feed conversion ratio (FCR) reached in 41 days of trial were 2542.5; 2568.9; 2610.5, 2516.6 g and 1.93; 1.81; 1.78; 1.88 kg for HS, probiotic, probiotic + HS and control group respectively. The best performance variables including the European efficiency index (EEI) were observed in the probiotic + HS group, where the effect of the probiotic supplementation was improved by HS. There were no significant differences among groups in the serum levels of total protein, glucose, cholesterol, aminotransferase and phosphorus. A significant decrease of total lipids in the probiotic group (P < 0.01) and probiotic + HS group (P < 0.001) groups, triglycerides levels in the HS (P < 0.01), probiotic and probiotic + HS (P < 0.001) groups on day 35, calcium level of the probiotic (P < 0.05) and probiotic + HS (P < 0.01) groups on day 41 and alkaline phosphatase levels in the probiotic group (P < 0.01) on day 14 were observed in comparison to the control group. Significantly higher values of indices of metabolic activity of phagocytes (IMA) after 14, 35 and 41 days of the experiment were observed in the probiotic + HS group (P < 0.01, P < 0.05) when compared with the control group. No adverse health effects were observed in experimental groups The results of the present study indicate that HS and E. faecium DSM 7134 had no significant effect on performance. However an improvement in FCR and EEI was observed which is believed to be due to the significant increase of indices of metabolic activity of phagocytes, caused by HS when combined with. E. faecium DSM 7134.

The influence of omega-3 polyunsaturated fatty acids (omega-3 pufa) on lactobacilli adhesion to the intestinal mucosa and on immunity in gnotobiotic piglets.

The effect of application of omega-3 polyunsaturated fatty acids (omega-3 PUFA) on intestinal colonization by Lactobacillus paracasei and on cellular immunity has been investigated in gnotobiotic pigs. The administration of polyunsaturated fatty acids positively affected the adhesion of Lactobacillus paracasei to the jejunal mucosa of gnotobiotic piglets. When compared to the control group, the number of Lactobacillus paracasei adhering to the jejunal mucosa was by 12% higher in piglets of the experimental group (5.10 log 10/cm2 vs. 4.55 log 10/cm2). The respective counts of Lactobacillus paracasei adhering to the ileal and colonic mucosa of 28 day old gnotobiotic piglets reached 4.45 and 5.05 log 10/cm2 in group C and 4.44 and 4.95 log 10/cm2 in group E. Omega-3 PUFA supplementation increased the phagocytic activity of neutrophils by almost 100% on day 28 of life as well as the subpopulations of lymphocytes (CD8) in the peripheral blood of germ-free piglets on day 21 of life. Our results indicate that the action of probiotics in the gut may be modulated by dietary PUFA. The stimulatory effect of PUFA upon adhesion of lactobacilli could be used for enhancing the effectiveness of probiotics in inhibiting digestive tract pathogens.