Effects of Enterococcus faecium and dried whey on broiler performance,gut histomorphology and intestinal microbiota

Abstract

The experiment was conducted to study the effects of supplementing a broiler starter diet with the probiotic Enterococcus faecium NCIMB 10415 and dried whey (80% lactose) on chick performance, gut histomorphology and intestinal microbiota. One-day-old male Ross 308 strain broiler chickens were fed diets containing: (i) control feed, (ii) control + 3.5% dried whey, (iii) control + 0.2%E. faecium, and (iv) control + 3.5% dried whey + 0.2%E. faecium. Birds were maintained in battery brooders confined in an environmentally controlled experimental room. The experiment lasted for 21 days. Birds fed E. faecium or E. faecium + dried whey exhibited significantly improved weight gain and feed conversion rate (FCR). Weight gain and FCR of treatment groups 1 – 4 were 628.7, 657.8, 690.9, 689.3 and 1.218, 1.193, 1.107, 1.116, respectively. Lactic acid bacteria counts in both the ileal content and excreta were significantly affected by dietary treatment. Supplementation of the E. faecium and dried whey separately and in combination increased lactic acid bacteria colonization in the ileal content from 4.2 to 5.0, 7.8 and to 5.1 log cfu/g, respectively (treatments 1 – 4). Similarly, supplementation of dried whey and E. faecium separately and in combination increased lactic acid bacteria in the excreta from 5.3 to 5.5, 8.0 and to 7.2 log cfu/g, respectively. Addition of the probiotic E. faecium increased villus height in the ileum (p < 0.05). Thus, supplementation of E. faecium enhanced broiler chick performance with respect to weight gain and FCR. No additive effect of E. faecium and dried whey was detected. Further studies are needed to investigate the relationship between E. faecium and dried whey with respect to gut histomorphology.     

Eucalyptus leaves powder,antibiotic and probiotic addition to broiler diets: effect on growth performance,immune response,blood components and carcass traits

The study was conducted to investigate the effects of different levels of eucalyptus powder (EP), virginiamycin and probiotic on performance, immunity, blood components and carcass traits of broiler chickens. A total of 250, 1-day-old male broiler chickens (Ross 308) were randomly allocated to five treatments with five replicates and 10 chicks each, as a completely randomized design. The dietary treatments consisted of: basal diet (BD), BD+0.25% EP, BD+0.5% EP, BD+0.01% of diet probiotic (Protexin), BD+0.02% of diet antibiotic (virginiamycin). Dietary supplementation did not affect feed intake, BW gain (BWG) and feed conversion ratio (FCR) during starter and grower phases, but BWG and FCR were affected during the finisher and whole periods (P<0.05).The highest BWG and lowest FCR were obtained in birds fed with virginiamycin and 0.5% EP. Dietary supplementation significantly increased the relative weight of carcass and breast (P<0.05). Treatments had no effect on relative weights of internal organs and small intestine except for bursa that increased by treatments. Relative length of jejunum also increased by treatments (P<0.05). Antibody production against sheep red blood cells did not changed in primary titer (day 35), but it significantly increased in secondary titer (day 42) by 0.5% EP. White blood cell counts were increased and cholesterol decreased by dietary supplementation (P<0.05). In conclusion, the results of this study showed that 0.5% EP served as a useful replacement for antibiotic and would improve performance and immune response of broiler chickens.     

Influence of a Probiotic Enterococcus Faecium Strain on Selected Bacterial Groups in the Small Intestine of Growing Turkey Poults

A feeding trial was carried out with turkey poults, which were fed a diet containing 10¹⁰ viable probiotic E. faecium NCIB 10415 cells/kg feed. Samples of the intestinal tract were analyzed for lactate, colony forming units of total anaerobic bacteria, lactic acid bacteria, enterobacteria and enterococci. Furthermore, metabolic activity of total eubacterial, lactobacilli and enterococci was recorded in selected RNA-extracts with specific ribosomal RNA oligonucleotide probes. Animals fed the probiotic diet showed continously increasing lactate concentrations throughout the sampling period up to day 42 of life. No correlation was found for colony forming units (cfu) of lactic acid bacteria, but metabolic activity of lactobacilli showed very close relation to continously increasing lactate concentrations. Throughout the feeding trial, enterococci in the control group continously increased to a maximum of 10⁴ cfu/g wet weight, but 10-fold higher enterococci cfu were generally found in the treated group. However, rRNA content as measure for metabolic activity showed a drastic decline in both groups after high metabolic activities on day 7. This study shows that E. faecium NCIB 10415 (E. faecium SF68) stimulates other lactic acid bacteria in the small intestine, especially lactobacilli.     

Dietary essential oils improve feed efficiency and hepatic antioxidant content of broiler chickens

The aim of this study was to test the hypothesis of an improved growth, dietary nutrient availability and overall health of broiler chickens reared on recycled litter when fed a standardised combination of essential oils (EO; carvacrol, cinnamaldehyde and capsicum oleoresin). To assess the effect of dietary treatments, feed intake, weight gain, feed efficiency, availability of dietary nutrients and energy, villus morphometry, excreta sialic acid concentration, hepatic antioxidants and serum amyloid A (SAA) when fed to broiler chickens were evaluated. Counts of Eimeria spp. oocysts were also determined in excreta samples. Four experimental diets were offered, including two basal control diets based on either wheat or maize that contained 215 g CP/kg and 12.13 MJ/kg metabolisable energy and another two diets using the basal control diets supplemented with the EO combination at 100 mg/kg diet. Each diet was fed to eight floor pens, containing two birds each, following randomisation. Birds fed the EO-supplemented diets had an improved (P<0.05) feed conversion ratio (FCR). Birds fed maize-based diet had an improved daily weight gain and FCR (P<0.05) compared with wheat-fed birds. Wheat-based diet tended (P=0.056) to have greater N-corrected apparent metabolisable energy and had greater fat retention coefficient (P<0.05) compared with maize-based diets. No differences (P>0.05) were observed in villus morphometry, sialic acid secretion, number of oocysts and SAA. Feeding the EO improved (P<0.05) the retention of dietary Ca and Na. Compared with maize, feeding wheat-based diets improved the retention coefficients for Ca, P and Na (P<0.05). Feeding dietary EO improved (P<0.05) the concentrations of the hepatic antioxidants, including carotene, coenzyme Q₁₀ and total vitamin E. The hepatic concentration of carotene of the maize-fed birds was 55.6% greater (P<0.05) compared with the wheat-fed birds. These results demonstrated that the addition of a standardised combination of EO in wheat- and maize-based diets provided benefits in terms of feed efficiency, mineral retention and antioxidant status of the birds when reared on recycled litter.     

Effect of the probiotic Enterococcus faecium NCIMB10415 on cell numbers of total Enterococcus spp., E. faecium and E. faecalis in the intestine of piglets

Sows and their piglets were fed a diet supplemented with or without the probiotic E. faecium NCIMB10415 (also known as SF68). Piglets were sacrificed 14, 28, 35 and 56 days after birth and DNA from intestinal segments was extracted and purified. A real time PCR assay was used to distinguish Enterococcus spp. (16s rDNA based), E. faecium (Efaafm gene), E. faecalis (Efaafs gene) as well as the probiotic strain (unique plasmid sequence). Extracts of autoclaved sow feces inoculated with E. faecium and E. faecalis cultures were used to calibrate real time PCR results. The probiotic strain was detected in 14 day old suckling piglets before the piglets had access to the starter diet. In piglets of the probiotic group, probiotic E. faecium cell counts were always a significant proportion of total E. faecium cells in stomach digesta (4-20%), however only a small fraction of the total Enterococcus spp. cell number on day 14 and 28 in all intestinal segments (0.1-0.7%). Compared to control samples, the probiotic E. faecium strain significantly (p < or = 0.05) decreased the amount of total Enterococcus spp. and E. faecalis cells in the colon of 14 day old suckling piglets as well as in jejunum and colon samples one week after weaning. E. faecium cell counts were not modified on any sampling day or intestinal segment. This study showed that the presence of probiotic E. faecium NCIMB10415 coincided with reduced total E. faecalis, but not total E. faecium cell numbers in the intestine of piglets. In view of unchanged cell numbers and ratios in sow feces, modifications must have taken place within the intestine of suckling piglets.     

Investigation on the effect of different levels of dried sweet orange (Citrus sinensis) pulp on performance,carcass characteristics and physiological and biochemical parameters in broiler chicken

Utilization of agricultural by-products in animal nutrition is a matter of great concern. Dried sweet orange (Citrus sinensis) pulp (DCSP) is a potential source of valuable nutrients and natural antioxidants for poultry feed. In the experiment, a feeding trial was conducted in order to investigate the effect of different levels of dried orange residues in diet on broiler growth performance, carcass characteristics, blood metabolites, humoral immunity, and cecum microbial population. A total of 200 one day experimental broiler chicks were distributed into a completely randomized design (CRD) which included 5 dietary treatments with 4 replicates per each treatment and 10 birds fed in each replicate. The experimental treatments consist of a control group (without additive), 0.5%, 1.0%, 1.5%, and 2% of DCSP (residue) in diet. Weight gain, feed intake and feed conversion ratio (FCR) were measured. Blood parameters and carcass traits were measured in the postnatal 35th day. The highest level of dried orange residues in treatment 5 (T5) had significantly increased the feed intake and body weight of broilers in groups and overall during the rearing period (P > 0.05). Different levels of dried orange residues had no significant effect on chicken FCR. Using of dried orange residues significantly decreased the liver and abdominal fat of broilers (P < 0.05). T5 has also significantly lower level of triglyceride than the control (T1) and treatment 2 (T2) (P < 0.05). In conclusion, the use of dried orange residues improved some performance (e.g. feed intake and body weight gain), decreased liver and abdominal fat and also serum triglyceride level in broiler chicken.     

Specific enumeration of the probiotic strain Enterococcus faecium NCIMB 10415 in the intestinal tract and in faeces of piglets and sows

The intestinal bacterium Enterococcus faecium NCIMB 10415 (E. faecium SF68) has been used for more than a decade as a probiotic strain in animal nutrition as well as in the prevention and treatment of diarrhoea in humans. Beneficial effects have been shown in feeding and clinical trials. However, the strain has no selective growth markers and monitoring in the intestinal tract is impossible by cultivation. Using specific nucleotide sequences, in this study a probe for colony hybridization was constructed in order to quantify this probiotic strain in feed and intestinal and faecal samples from piglets and sows. The probiotic strain showed almost constant amounts in sow faeces (1.8 x 10(5) cfu/g wet weight), while contents in digesta and piglet faeces varied on a lower level depending on gut section and piglet age. The ratio of specific probiotic counts and total enterococci was much lower than in sow faeces however the strain could be detected reliably in faeces already on the 14th day of life. The application of the colony hybridization method enables for the first time the selective detection of the widely used probiotic E. faecium NCIMB 10415 strain among total Enterococcus spp. counts of digesta, faeces and feed. It is now possible to monitor the presence of the probiotic in the intestinal tract and faeces. Results of this study have implications for the proposed modes of action of probiotics in animal nutrition.     

Addition of fermented and unfermented grape skin in broilers’ diets: effect on digestion,growth performance,intestinal microbiota and oxidative stability of meat

Grape skin is a source of polyphenols with antioxidant and antimicrobial properties. Little information is available regarding its application in animal feeding. The present study investigated the effect of inclusion of fermented (FS) and unfermented (UFS) grape skin at two different doses (30 g/kg, FS30 and UFS30, and 60 g/kg, FS60 and UFS60) and 200 mg/kg vitamin E (α-tocopheryl acetate) in a corn–soybean diet on growth performance, ileal protein digestibility, ileal and excreta total extractable polyphenols content and digestibility, intestinal microbiota and thigh meat oxidation in broiler chickens. Growth performance was depressed in chickens fed UFS and FS diets. A reduction in ileal protein digestibility was also observed in birds fed UFS, being this effect more pronounced in those fed 60 g/kg. The dietary inclusion of grape skin increased both ileal and excreta polyphenols contents, being higher in birds fed UFS than in those fed FS. Excreta moisture content increased in birds fed UFS and FS diets. No effect of dietary inclusion of grape skin was observed on ileal counts of lactic-acid bacteria and Clostridium, but UFS inclusion in the diet reduced ileal count of Escherichia coli as compared with FS dietary inclusion. After 7 days of refrigerated storage, values of thiobarbituric acid reactive substances (TBARS) were lower in chicken meat when grape skin was added in the diet at 60 g/kg instead of 30 g/kg, and meat from birds fed 60 g/kg of grape skin reached TBARS values similar to those of birds supplemented with vitamin E. In conclusion, high doses of grape skin polyphenols depressed growth performance and protein digestibility, and increased excreta moisture content. Unfermented grape skin contained more polyphenols than FS, and its inclusion in the diet led to higher ileal and excreta polyphenols contents and to a lower ileal count of E. coli. Furthermore, the antioxidant potential of the polyphenols present in grape skin was observed after 7 days of meat storage, with the dose of 60 g/kg of grape skin being as effective as vitamin E supplementation in maintaining oxidative stability of meat.     

Antimicrobial susceptibilities of enterococci isolated from faeces of broiler and layer chickens

Enterococci were isolated from faecal droppings of chickens in broiler and layer farms and the susceptibilities to nine therapeutic antimicrobial agents and six growth-promoting antibiotics were determined by the agar dilution method. Resistance to therapeutic antimicrobial agents such as ampicillin, clindamycin, erythromycin, streptomycin, tetracycline or tylosin was more frequent in enterococcal isolates from broiler farms than in those from layer farms. Resistance to ofloxacin was rare, occurring in only one (0.7%) of the Enterococcus faecium isolates from broiler farms. Resistance to growth-promoting antibiotics such as avilamycin, salinomycin and virginiamycin was common among isolates from broiler farms. Of the E. faecium isolates from broiler farms, 12.4% were resistant to avilamycin and 27.4% were resistant to virginiamycin. Resistance to salinomycin was detected in all enterococcal species, ranging from 12.4% of E. faecium isolates to 50% of E. hirae isolates.     

Microbial status and rumen enzyme profile of crossbred calves fed on different microbial feed additives

AIMS: To test various microbial cultures as cattle feed additives. METHODS AND RESULTS: Four groups of newly born crossbred calves (average body weight 23.5 kg) were reared on green berseem and calf starter which was devoid of cereal grains. Milk was fed up to 8 weeks of age, starting with one tenths and gradually reducing to one twentieths of the body weight. One hundred millilitres of microbial feed additive or 100 g fermented feed was fed to the animals of group 2 (curd containing lactic acid bacteria at 10(8) cfu x ml(-1)), group 3 (Saccharomyces cerevisiae NCDC-49 at 10(6) cfu x ml(-1)) and group 4 (Lactobacillus acidophilus-15 at 10(8) cfu x ml(-1)). Group 1 served as control. The incidence and duration of diarrhoea was lower in the animals of probiotic fed groups as compared to control group. Out of three microbial feed additives, yeast feeding showed maximum suppression of diarrhoea followed by Lactobacillus and curd. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: There was no effect of probiotic feeding on the log number of cells of lactic acid bacteria, yeast and coliform bacteria in the faeces and rumen liquor at any age. The activities of carboxymethylcellulase, xylanase, beta-glucosidase, alpha-glucosidase, alpha-amylase, protease, urease and pH of the rumen liquor remained unaffected by probiotic feeding at all ages tested in this experiment.     

Effect of lactulose on growth performance and intestinal morphology of pre-ruminant calves using a milk replacer containing Enterococcus faecium

The synthetic disaccharide lactulose is known to improve the intestinal microflora by stimulating the growth of selected probiotic bacteria in the gut. In our experiment the effects of lactulose in combination with the probiotic bacteria Enterococcus faecium on growth performance and morphology of the bovine intestine were examined. Calves aged 39 ± 2 days were randomised to three feeding groups (no. = 14 each group): control (L0), fed milk replacer (MR) containing E. faecium; a lactulose group (L1) containing additional 1% lactulose and a second lactulose group (L3) containing 3% lactulose dry matter. The calves were weighed weekly. After 19 weeks the calves were slaughtered and tissues were collected for histological studies. The average daily live weight gain tended to be higher (P < 0.1) for L3 (1350 g/day) than L0 (1288 g/day). Compared with L0, a reduction (P < 0.001) of ileal villus height due to lactulose treatment of approximately 14% in group L1 and 20% in L3 was determined. A significant decrease in the depth of the crypts about 12% in L1 and 8% in L3 was detected in the caecum. The surface area of lymph follicles from Peyer's patches was decreased by lactulose treatment. Results show that lactulose has an effect on the morphology of intestine. A significant effect on growth performance can not be confirmed. However, results permit the conclusion that lactulose feeding has the tendency to increase growth performance.     

No beneficial effects evident for Enterococcus faecium NCIMB 10415 in weaned pigs infected with Salmonella enterica serovar Typhimurium DT104

Salmonella enterica serovar Typhimurium DT 104 is the major pathogen for salmonellosis outbreaks in Europe. We tested if the probiotic bacterium Enterococcus faecium NCIMB 10415 can prevent or alleviate salmonellosis. Therefore, piglets of the German Landrace breed that were treated with E. faecium (n = 16) as a feed additive and untreated controls (n = 16) were challenged with S. Typhimurium 10 days after weaning. The presence of salmonellae in feces and selected organs, as well as the immune response, were investigated. Piglets treated with E. faecium gained less weight than control piglets (P = 0.05). The feeding of E. faecium had no effect on the fecal shedding of salmonellae and resulted in a higher abundance of the pathogen in tonsils of all challenged animals. The specific (anti-Salmonella IgG) and nonspecific (haptoglobin) humoral immune responses as well as the cellular immune response (T helper cells, cytotoxic T cells, regulatory T cells, γδ T cells, and B cells) in the lymph nodes, Peyer's patches of different segments of the intestine (jejunal and ileocecal), the ileal papilla, and in the blood were affected in the course of time after infection (P < 0.05) but not by the E. faecium treatment. These results led to the conclusion that E. faecium may not have beneficial effects on the performance of weaned piglets in the case of S. Typhimurium infection. Therefore, we suggest a critical discussion and reconsideration of E. faecium NCIMB 10415 administration as a probiotic for pigs.     

Taxonomic and gene-centric metagenomics of the fecal microbiome of low and high feed conversion ratio (FCR) broilers

Individual weight gain in broiler growers appears to vary, which may in part be due to variation in their gut microbiota. In this paper we analyse the fecal microbiota of low and high feed conversion ratio (FCR) broilers. After shotgun sequencing of the fecal microbiome, we used the SEED database to identify the microbial diversity and metabolic potential in low and high FCR birds. The domain-level breakdown of our samples was bacteria (>95 %), eukaryotes (>2 %), archaea (>0.2 %), and viruses (>0.2 %). At the phylum level, Proteobacteria (78.83 % in low and 52.04 % in high FCR), Firmicutes (11.97 % in low and 27.53 % in high FCR) and Bacteroidetes (7.10 % in low FCR and 17.53 % in high FCR) predominated in the fecal microbial community. Poultry fecal metagenomes revealed the sequences related to 33 genera in both low and high FCR with significantly different proportion. Functional analysis revealed that genes for the metabolism of carbohydrates, amino acids and derivatives and protein metabolism were most abundant in SEED subsystem in both samples. Genes associated with stress, virulence, cell wall and cell capsule were also abundant. Indeed, genes associated with sulphur assimilation, flagellum and flagellar motility were over represented in low FCR birds. This information could help in developing strategies to improve feed efficiency and feed formulation for broiler chickens.     

In ovo peptide YY and epidermal growth factor administration and their effects on growth and yolk utilization in neonatal meat-type chickens (Gallus domesticus)

The effects of in ovo peptide YY (PYY) or epidermal growth factor (EGF) administration on chick growth, yolk absorption and yolk stalk function in posthatch (0–5 days) meat-type or broiler chicks were determined. At Day 18 of incubation, treated eggs were injected into the air cell with 100 μl of either PYY (Trial 1) or EGF (Trial 2) at a dosage of 600 μg/kg egg weight. Saline-treated control eggs were injected similarly with 0.9% saline. At hatch, 200 μl of ⁵¹Cr-labeled microspheres were injected into chick yolk sacs. Epidermal growth factor increased ileal wet weight adjusted for body weight as well as ileal serosal dry matter. Body weight, feed consumption and excreta weight per bird, and relative weights of the yolk sac, intestine and liver were significantly affected by age of the chick in both trials. Relative radioactivity of the yolk sac, yolk stalk, blood, liver, and kidneys were affected by bird age in Trial 2; however, there were no significant effects due to PYY or EGF treatments on relative radioactivity of the tissues and organs examined. These data suggest that PYY and EGF had no effect on yolk absorption or yolk stalk function through 5 days in the posthatch chick.     

Intramucosal bacteria in colon cancer and their elimination by probiotic strainEnterococcus faecium M-74 with organic selenium

Intraepithelial bacteria were isolated by the gentamicin protection assay (GPA) from biopsy samples obtained at colonoscopy (colon cancer, n = 10 patients; colonic adenoma, n = 20; control group, n = 20; cancer patients without gastrointestinal tract GIT malignancy, n = 10). After a three-month administration of E. faecium M-74 to patients with positive GPA biopsies, 172 biopsy specimens from 60 patients were examined with the GPA. The number of biopsies with intracellular bacteria was significantly higher in adenoma and carcinoma group than in control group (26 vs. 10%; p = 0.004); in cancer patients without GIT malignancy the difference was nonsignificant. E. faecium M-74 was also administered to 5 patients with colonic adenoma; according to a control colonoscopy the number of biopsies with intracellular bacteria was significantly lower after probiotic administration (48 vs. 16%; p = 0.03). A striking prevalence of intraepithelial bacteria was also showed in patients with large bowel adenoma and carcinoma. The administration of probiotic strain M-74 can thus be considered to be an effective and promising method for elimination of pathogenic bacteria in the case of inflammatory bowel disease and colon cancer.     

Enzyme supplementation of wheat-based diets for broilers: 2. Effect on apparent metabolisable energy content and nutrient digestibility

Two experiments were conducted to determine the effect of enzyme supplementation on the nitrogen-corrected apparent metabolisable energy (AME_n) and apparent nutrient digestibilities in ileum and excreta from male broiler chickens fed diets containing high amounts of wheat (>80%). Four different enzyme preparations were added to the wheat-based diets in varying levels and combinations. The difference between Experiments 1 and 2 consisted in the addition of different enzymes and enzyme levels. Excreta and ileal content were collected from broiler chickens at 3 and 6 weeks of age. At 3 weeks of age enzyme supplementation increased (P<0.0001) the AME_n from an average value of 13.86 MJ kg⁻¹ dry matter (DM) to an average of 14.60 MJ kg⁻¹ DM in the two experiments. The apparent digestibility of protein (APD) and fat (AFD) were improved significantly as a result of enzyme addition in both experiments. At 3 weeks of age, improvements (P<0.05) in the ileal AFD and APD were on average 13% and 6%, respectively. The effect of enzyme supplementation on AFD measured in excreta from broiler chickens showed the same pattern. The positive effect of enzyme addition on the overall nutrient digestibility and AME_n was reflected in weight gain and feed conversion efficiency (FCE). Significant improvements in AME_n, APD, and AFD were still present in most of the groups fed with enzyme-supplemented diets at 6 weeks of age. The effect of enzyme addition, however, was less pronounced, especially in Experiment 1 where the ileal APD of broiler chickens did not differ significantly from the control group. Apparent starch digestibility (ASD), measured in Experiment 2, was very high in all groups, including the control; however, enzyme supplementation increased ASD (P<0.001) in ileum and excreta at both 3 and 6 weeks of age. The digestibility of total non-starch polysaccharides (NSP) in excreta was improved significantly (P<0.01) as a result of enzyme supplementation (Experiment 2). In addition, pH of caeca content decreased (P<0.02) in broiler chickens fed with enzyme-supplemented diets when compared with the control group. Decreased pH could indicate microbial fermentation of unabsorbed NSP residues and nitrogenous compounds. Overall, the results demonstrated that the nutritive value of wheat-based diets to broiler chickens improve enzyme supplementation. The apparent digestibility of total NSP and of arabinose and xylose residues improved (P<0.02) in the enzyme-supplemented diets indicating that the enzymes were able to break down the cell wall NSP to a certain extent.     

Antigenotoxicity of probiotics and prebiotics on faecal water-induced DNA damage in human colon adenocarcinoma cells

Six strains of lactic acid producing bacteria (LAB) were incubated (1 x 10(8)cfu/ml) with genotoxic faecal water from a human subject. HT29 human adenocarcinoma cells were then challenged with the resultant samples and DNA damage measured using the single cell gel electrophoresis (comet) assay. The LAB strains investigated were Bifidobacterium sp. 420, Bifidobacterium Bb12, Lactobacillus plantarum, Streptococcus thermophilus, Lactobacillus bulgaricus and Enterococcus faecium. DNA damage was significantly decreased by all bacteria used with the exception of Strep. thermophilus. Bif. Bb12 and Lact. plantarum showed the greatest protective effect against DNA damage. Incubation of faecal water with different concentrations of Bif. Bb12 and Lact. plantarum revealed that the decrease in genotoxicity was related to cell density. Non-viable (heat treated) probiotic cells had no effect on faecal water genotoxicity. In a second study, HT29 cells were cultured in the presence of supernatants of incubations of probiotics with various carbohydrates including known prebiotics; the HT29 cells were then exposed to faecal water. Overall, incubations involving Lact. plantarum with the fructooligosaccharide (FOS)-based prebiotics Inulin, Raftiline, Raftilose and Actilight were the most effective in increasing the cellular resistance to faecal water genotoxicity, whereas fermentations with Elixor (a galactooligosaccharide) and Fibersol (a maltodextrin) were less effective. Substantial reductions in faecal water-induced DNA damage were also seen with supernatants from incubation of prebiotics with Bif. Bb12. The supernatant of fermentations involving Ent. faecium and Bif. sp. 420 generally had less potent effects on genotoxicity although some reductions with Raftiline and Elixor fermentations were apparent.     

Enterococcus faecium EK13--an enterocin a-producing strain with probiotic character and its effect in piglets

The experiment was conducted to determine the effects of the inoculation of the probiotic and enterocin A-producing strain Enterococcus faecium EK13 on selected parameters of metabolic profile, gut microflora, growth, and health in newborn piglets of Slovak White Improved. Piglets for study were divided into two groups: one group (EK13 group, n=8) received strain EK13 per os once daily for 7 days (2ml per piglet, 10(9)CFU/mL of saline buffer). The control group of piglets (n=7) was given placebo-saline buffer. The experiment lasted 14 days. After 7 days, strain EK13 reached 9.8 log(10) CFU/g in faeces of E. faecium EK13 treated piglets while counts of Escherichia coli were significantly lower (P<0.01) than in piglets of the control group. The concentrations of total serum protein, calcium, haemoglobin, haematocrit, red blood cell count and index of phagocytic activity of leukocytes were significantly higher after application of strain EK13. On the other hand, cholesterol was significantly lower in the EK13 group of animals. On day 14, piglets were killed and samples of intestinal contents were taken. Total counts of bacteria in the intestinal contents (jejunum, ileum, caecum, colon) were not significantly influenced. The pH value was significantly lower (P<0.05) only in duodenum of piglets receiving E. faecium EK13. There was a significant higher concentration of lactic acid (P<0.01) and propionic acid in the colon (P<0.001) of the EK13 group. Application of E. faecium EK13 did not influence the daily body weight gain significantly.     

Probiotic potential of enterococci isolated from canine feed

Enterococci isolated from 28 different commercially available feeds (10-1000 CFU/mL) were identified and their probiotic potential was determined. Species identification of 22 selected strains was performed by intergenic length-polymorphism analysis (tRNA-PCR); PCR products were analyzed using capillary electrophoresis. Six strains were allotted to the species Enterococcus faecium, four to E. faecalis, one to E. hirae; the remaining strains were not classed. The strains were sensitive to vancomycin, ampicillin, tetracycline and rifampicin. They were able to adhere to human as well as canine intestinal mucus. They produced lactic acid (0.99-1.04 mmol/L) and most of them were urease-positive with sufficient survival in 5 % Oxgall-bile. They did not show any inhibitory activity due to antimicrobial substances. Plasmid DNA was detected in 8 strains, the bands responding to small molecular size (10 kbp). Considering all probiotically important properties, E. faecium strain EE3 was suggested as potential feed additive.     

Neonatal small bowel epithelia: enhancing anti-bacterial defense with lactoferrin and Lactobacillus GG

BACKGROUND AND AIMS: Extremely preterm human infants have increased susceptibility to small bowel infection. We hypothesized that early colonization of the immature small intestine with Lactobacillus GG (LGG), and use of a recombinant lactoferrin (rhLF) to promote growth of LGG, would enhance gut defenses against enteroinvasive Escherichia coli. METHODS: Newborn rat pups were treated with nothing, intra-gastric LGG, or rhLF + LGG on days 3 and 4 of life. Gut colonization by LGG was quantified in lavaged jejunal and ileal fluid and gut wall homogenates on day 5 of life. Separate studies used similarly treated litters of newborn rats that were infected late on day 4 of life with E. coli [10(12) CFU/kg]. Sixteen hours later, the numbers of E. coli were measured in small bowel fluid and gut wall homogenates. RESULTS: Control pups initially had lactic acid bacteria colonize the bowel, but these bacteria were not LGG. Pups treated with LGG or rhLF + LGG had significantly higher numbers of LGG in the ileum versus jejunum. Contrary to our hypothesis, rhLF did not augment LGG colonization. After E. coli-related gut infection, planktonic [lavage fluid] and epithelia-adherent growth [gut wall homogenates] of E. coli in the small bowel were most effectively reduced by pre-treatment with rhLF and LGG (P < .05). CONCLUSION: Prophylactic therapy with recombinant human lactoferrin and the probiotic, Lactobacillus GG, act to enhance defenses against invasive E. coli in the nascent small intestine. We suggest that rhLF and LGG are therapeutic agents that may reduce necrotizing enterocolitis and gut-related sepsis in preterm human infants.