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.     

In vivo Testing of Functional Properties of Three Selected Probiotic Strains

Summary Lactobacillus acidophilus M92, Lactobacillus plantarum L4 and Enterococcus faecium L3 were previously selected as probiotic strains on the base of in vitro selection criteria. To investigate functional properties of these three probiotic strains in vivo, Swiss albino mice were used as animal model. Survival, competition, adhesion and colonization were monitored in the gastrointestinal tract, as well as the immunomodulating capability of L. acidophilus M92, L. plantarum L4 and E. faecium L3. During the feeding of mice with probiotic strains with daily dose of 2 × 10¹⁰ rifampicin-resistant cells, the number of lactic acid bacteria in the faeces increased and reduction of enterobacteria and sulphite-reducing clostridia was observed. Rifampicin-resistant colonies of probiotic strains could be reisolated from the faeces of mice fed with the rifampicin-resistant cells. The similar results were obtained in homogenates of small and large intestine of mice on the first and fourteenth days after feeding with L. acidophilus M92, L. plantarum L4 and E. faecium L3. The adherence of the probiotic strains obtained in vitro correlated with their capability to adhere to mouse ileal epithelial cells in vivo. After oral immunization of mice with viable cells of L. acidophilus M92, L. plantarum L4 and E. faecium L3 with a daily dose of 2 × 10¹⁰ cells, the concentrations of serum IgA, IgG and IgM antibodies from all groups of mice were significantly higher in comparison to the control.     

Changes in the phase variant spectra in the populations of lactic acid bacteria under antibiotic treatment

Effect of the antibiotics kanamycin and ampicillin on the growth and phase variation of the populations of four strains of lactic acid bacteria (Lactobacillus sp. M76AT, L. casei MB, Enterococcus faecium M, and E. faecium M3185) was studied. The presence of antibiotics in the medium resulted in a dose-dependent decrease in viable cell numbers and in partial or complete substitution of the dominant S variant with the minor Sm and Sb variants. The variants differed in colony morphology, as well as in some physiological, biochemical, biotechnological, and probiotic characteristics. The Sm type variants of all strains exhibited the highest resistance to antibiotics. High production of exopolysaccharides was found in Sb variants of lactobacilli and in S variants of enterococci. The highest antibacterial activity was found in Sm variants of lactobacilli, especially in Lactobacillus sp. M76AT. The latter is biotechnologically the most promising strain, since all variants fermented milk yielding the products with uniformly pronounced functional and organoleptic properties. These patterns are of importance for the understanding of the mechanisms of antibiotic resistance and for selection of the variants with desired properties, as well as for quality control of the lactic acid bacteria starter cultures.     

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.     

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

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.     

Elucidation of the Taxonomic Status of Industrial Strains of Thermophilic Lactic Acid Bacteria by Sequencing of 16S rRNA Genes

Both phenotypic characteristics and results of PCR tests for the presence of species-specific genes indicate that a number of strains of thermophilic lactic acid bacteria previously considered as belonging to Streptococcus thermophilus are actually closely related to enterococci. In the present study, partial (over 500 nucleotides) sequencing of 16S rRNA genes from 12 strains of thermophilic lactic acid bacteria used as starters for manufacturing sour milk products on the territory of the Commonwealth of Independent States (CIS) has been performed. According to the results of the sequencing, seven of the strains have been classified with Enterococcus durans. The earlier classification (based on PCR tests) of two of the strains as S. thermophilus and three of the strains as E. faecium has been confirmed. The data obtained demonstrate that the enterococci E. durans and E. faecium are widely used as thermophilic starters for manufacturing sour milk products on the territory of the CIS.__________Translated from Mikrobiologiya, Vol. 74, No. 4, 2005, pp. 520–525.Original Russian Text Copyright © 2005 by Botina, Lysenko, Sukhodolets.     

The Lactic Acid Enterococci Enterococcus faecium and Enterococcus durans: Nucleotide Sequence Diversity in 16S rRNA Genes

Among the strains used as starters for making sour milk products on the territory of the CIS, the bacteria Enterococcus faecium and Enterococcus durans are frequently found. In this work, we studied a new collection of lactic acid enterococci and also obtained more complete data on the nucleotide sequences of 16S rRNA genes in some strains studied earlier and found that most strains had certain distinctions in their 16S rRNA genes as compared with the E. durans and E. faecium genes available in the NCBI database. Based on these data, it is suggested that the strains of lactic acid enterococci represent new, earlier unknown taxa of enterococci that use milk as an ecological niche.     

Bacteriocin-producing strain of Enterococcus faecium EK 13 with probiotic character and its application in the digestive tract of rabbits

Enterococcus faecium EK 13 is a bacteriocin-enterocin A producing strain with probiotic properties. In this study its colonization, stability and effect on microflora in rabbits was studied as well as its influence on zootechnical parameters. Fifty rabbits of both sexes (HYPLUS, 30-day old; after weaning) were divided into control (CG) and experimental (EG) groups. They were fed a standard diet. Moreover, 25 rabbits in EG were fed daily (for 4 weeks) 15 g (separate doses ∼1.6 g) of lyophilized EK13 strain (rifampicin resistant variant — rif^R; 10⁹ cfu/g) dissolved in drinking water. After cessation of EK13 (rif^R) strain application, the rabbits in both groups were fed a standard diet for the next 2 weeks. Sampling was performed in double on day 0 (at the beginning of experiment), weekly during EK13 (rif^R) strain application as well as on week 1 and 2 after cessation of EK13 (rif^R) strain application. The counts of EK13 (rif^R) strain reached 7.1 ± 2.6 log₁₀ cfu/g after 4 weeks and even on week 2 after its cessation the counts 5.6 ± 2.3 log₁₀ cfu/g were determined. The total counts of enterococci in the rabbits were already increased in EG comparing with CG (p < 0.05); even 2 weeks after EK13 (rif^R) strain cessation, their counts in EG were 7.2 ± 2.6 log₁₀ cfu/g (p < 0.001). Enterococci in CG reached at the same time the value 3.7 ± 2.6 log₁₀ cfu/g. The counts of E. coli were significantly reduced in EG during 4 weeks (p < 0.05, p < 0.001). Even 2 weeks after EK13 (rif^R) strain cessation significant difference in E. coli counts between CG and EG was detected (p < 0.001). Enterobacteria in EG were significantly reduced (p < 0.001). Average daily gain in EG was 41.0 ± 3.83 in comparison to CG (40.6 ± 3.72); it means almost the same; although rabbits in EG showed higher feed intake per kg of gain than rabbits in CG. Preliminary results demonstrated that EK13 is a perspective probiotic candidate for rabbits. Presented at the Second Probiotic Conference, Košice, 15–19 September 2004, Slovakia.     

Feeding the Probiotic Enterococcus faecium Strain NCIMB 10415 to Piglets Specifically Reduces the Number of Escherichia coli Pathotypes That Adhere to the Gut Mucosa

Feed supplementation with the probiotic Enterococcus faecium for piglets has been found to reduce pathogenic gut microorganisms. Since Escherichia coli is among the most important pathogens in pig production, we performed comprehensive analyses to gain further insight into the influence of E. faecium NCIMB 10415 on porcine intestinal E. coli. A total of 1,436 E. coli strains were isolated from three intestinal habitats (mucosa, digesta, and feces) of probiotic-supplemented and nonsupplemented (control) piglets. E. coli bacteria were characterized via pulsed-field gel electrophoresis (PFGE) for clonal analysis. The high diversity of E. coli was reflected by 168 clones. Multilocus sequence typing (MLST) was used to determine the phylogenetic backgrounds, revealing 79 sequence types (STs). Pathotypes of E. coli were further defined using multiplex PCR for virulence-associated genes. While these analyses discerned only a few significant differences in the E. coli population between the feeding groups, analyses distinguishing clones that were uniquely isolated in either the probiotic group only, the control group only, or both groups (shared group) revealed clear effects at the habitat level. Interestingly, extraintestinal pathogenic E. coli (ExPEC)-typical clones adhering to the mucosa were significantly reduced in the probiotic group. Our data show a minor influence of E. faecium on the overall population of E. coli in healthy piglets. In contrast, this probiotic has a profound effect on mucosa-adherent E. coli. This finding further substantiates a specific effect of E. faecium strain NCIMB 10415 in piglets against pathogenic E. coli in the intestine. In addition, these data question the relevance of data based on sampling fecal E. coli only.     

Sensitivity to Enterocins of Biogenic Amine-Producing Faecal Enterococci from Ostriches and Pheasants

Enterococci are widespread bacteria forming the third largest genus among lactic acid bacteria. Some possess probiotic properties or they can produce beneficial proteinaceous antimicrobial substances called enterocins. On the other hand, some enterococci produce biogenic amines (BAs), so this study is focused on the sensitivity to enterocins of biogenic amine-producing faecal enterococci from ostriches and pheasants. Altogether, 60 enterococci isolated from faeces of ostriches and pheasants were tested for production of BAs. This target of the identified enterococci involved 46 strains selected from 140 ostriches and 17 from 60 pheasants involving the species Enterococcus hirae, E. faecium, E. faecalis, and E. mundtii. Although BAs histamine, cadaverine, putrescine, and tryptamine were not detected in the enterococci tested, in general high BA production by the tested enterococci was noted. The species E. hirae formed the majority of the enterococcal strains from ostrichs faeces (34 strains). High production of tyramine (TYM) was measured with an average amount of 958.16 ± 28.18 mg/ml. Among the enterococci from pheasants, the highest was production of TYM compared to phenylethylamine, spermidine, and spermine. Enterococci featured high BA production; however, they were sensitive to seven enterocins with inhibition activity ranging from 100 up to 25,600 AU/ml.     

The inhibitory activity of Lactobacillus spp. isolated from breast milk on gastrointestinal pathogenic bacteria of nosocomial origin

Milk acts as a mean for transporting many essential substances from the mother to the child. In human beings, milk includes several predominant bacteria, such as staphylococci, streptococci, micrococci, lactobacilli, enterococci, lactococci and bifidobacteria. Besides, its intake favors the predominance of bifidobacteria and lactobacilli in the child’s intestinal microbiota. The present work explores the isolation and selection of lactobacilli strains with probiotic potential, focusing in their degree of hydrophobicity and antagonism against important gastrointestinal nosocomial pathogens. 98 lactobacilli were isolated from 48 breast milk samples, with most strains belonging to the obligately homofermentative group (36.7%). 63% of the isolated strains showed a high degree of hydrophobicity when tested on three solvents and were selected for detecting antimicrobial activity against gastrointestinal pathogens, including Escherichia coli, Shigella spp, Pseudomonas spp and Salmonella spp strains. When applying the agar diffusion test, many isolated strains presented inhibitory activity against pathogenic strains. We observed that: Salmonella enteriditis was the most inhibited pathogen, and the strains with the most inhibitory power were AR2 and O1 (both highly hydrophobic lactic acid bacteria), which showed an opposing effect against all nosocomial pathogens tested. Although more in vitro, in vivo or clinical data would be needed before any conclusion on the probiotic properties of the strains can be drawn, our results demonstrate that some of the tested strains may have good probiotic potential for their inclusion in products targeting infants.     

Probiotics L. plantarum and L. curvatus in Combination Alter Hepatic Lipid Metabolism and Suppress Diet‐Induced Obesity

Objective: To determine the effects of naturally derived probiotic strains individually or combination on a short‐term diet‐induced obesity model. Design and Methods: C57BL/6J mice (n = 50) were randomly divided into five groups, then fed a high‐fat high‐cholesterol diet (HFCD), HFCD and Lactobacillus plantarum KY1032 (PL, 10¹⁰cfu/day), HFCD and Lactobacillus curvatus HY7601 (CU, 10¹⁰cfu/day), HFCD and in combination with PL+CU (10¹⁰cfu/day), or a normal diet (ND) for 9 weeks. Results: PL and CU showed distinct and shared metabolic activity against a panel of 50 carbohydrates. Fat accumulation in adipose tissue and liver was significantly reduced by probiotic strains CU or PL+CU. Probiotic strains CU or PL+CU reduced cholesterol in plasma and liver, while PL+CL had a synergistic effect on hepatic triglycerides. Probiotic strains PL+CU combination was more effective for inhibiting gene expressions of various fatty acid synthesis enzymes in the liver, concomitant with decreases in fatty acid oxidation‐related enzyme activities and their gene expressions. Conclusions: Multi‐strain probiotics may prove more beneficial than single‐strain probiotics to combat fat accumulation and metabolic alterations in diet‐induced obesity.     

The effects of submerged aquatic vegetation on the persistence of environmental populations of Enterococcus spp.

Enterococcus spp. are utilized worldwide as faecal indicator bacteria, but certain strains exhibit extended survival in environmental habitats and the factors influencing their persistence are poorly understood. We used flowing freshwater mesocosms to explore the effect of submerged aquatic vegetation (SAV) on the persistence of natural enterococci populations from a subtropical lake. The highest mean densities of culturable enterococci over 2 weeks occurred in SAV [8.6 × 10² colony‐forming units (cfu) per 100 g wet weight], followed by sediments (1.3 × 10² cfu per 100 g) and water (18 cfu per 100 ml). However, due to relative differences in the total mass of each substrate in the entire system (water > sediments > SAV), SAV‐associated enterococci represented only a minor proportion of the total population. Vegetated mesocosms harboured significantly higher mean cfu per mesocosm and cfu densities in sediments compared with their unvegetated counterparts, suggesting that SAV indirectly facilitates persistence in aquatic habitats. Populations were dominated (> 96%) by a single Enterococcus casseliflavus strain according to BOX‐PCR genotyping, which did not change over the 10‐month study and strongly suggests bacterial replication in the lake. The presence of such strains in the environment may represent highly competitive, naturalized and reproducing indicator bacteria populations that are not directly related to pollution events.     

牦牛源产细菌素屎肠球菌的分离鉴定和益生特性

【背景】抗生素的滥用导致牦牛肠道常见病原菌耐药性增加,益生菌作为对抗耐药性细菌的新型武器,应用前景广阔。【目的】获取益生特性优良的牦牛源益生菌。【方法】将20份牦牛粪便样本在含0.5%CaCO3的MRS培养基上分离纯化,以大肠杆菌和金黄色葡萄球菌为指示菌,用牛津杯法筛选有抑菌活性的菌株;排除酸和过氧化氢后,经耐酸耐热试验和蛋白酶敏感试验筛选产细菌素菌株,用形态学和16S rRNA基因序列分析鉴定;通过对大肠杆菌、沙门氏菌等腹泻病原菌体外抑菌试验、耐模拟胃肠液、测定自聚集能力和疏水性及抗生素敏感试验分析益生特性。【结果】从20份牦牛粪便样本中共分离出11株产生溶钙圈的菌株,其中6株对大肠杆菌和金黄色葡萄球菌抑菌效果显著,经复筛得到2株产细菌素的乳酸菌SC6和SC9,经鉴定均为屎肠球菌(Enterococcus faecium)。其中SC9对腹泻病原菌抑菌效果明显,有良好的耐受性和肠道黏附能力,对5种常用抗生素均敏感。【结论】屎肠球菌SC9有一定的抗逆性和潜在的益生能力,具备作为益生菌的潜力。     

Encapsulation of Lactobacillus plantarum 423 and its Bacteriocin in Nanofibers

Plantaricin 423, produced by Lactobacillus plantarum 423, was encapsulated in nanofibers that were produced by the electrospinning of 18% (w/v) polyethylene oxide (200 000 Da). The average diameter of the nanofibers was 288 nm. Plantaricin 423 activity decreased from 51 200 AU/ml to 25 600 AU/ml and from 204 800 AU/ml to 51 200 AU/ml after electrospinning, as determined against Lactobacillus sakei DSM 20017 and Enterococcus faecium HKLHS, respectively. Cells of L. plantarum 423 encapsulated in nanofibers decreased from 2.3 × 10¹⁰ cfu/ml before electrospinning to 4.7 × 10⁸ cfu/ml thereafter. Cells entrapped in the nanofibers continued to produce plantaricin 423. This is the first report on the encapsulation of a bacteriocin and cells of L. plantarum in nanofibers. The method may be used to design a drug delivery system for bacteriocins and the encapsulation of probiotic lactic acid bacteria. The technology is currently being optimized.     

Impact of the probiotic bacteria Enterococcus faecium NCIMB 10415 (SF68) and Bacillus cereus var. toyoi NCIMB 40112 on the development of serum IgG and faecal IgA of sows and their piglets

Abstract

To examine the influence of two different probiotic bacteria on the humoral immune system of swine, two animal studies were carried out with sows and their litters. The sows' feed was supplemented with either Enterococcus faecium NCIMB 10415 (SF68) or Bacillus cereus var. toyoi NCIMB 40112 beginning early in pregnancy. The total IgA content in the faeces as well as the total IgG concentration in the blood of the sows was recorded before and after weaning. The same parameters were determined in the blood and faeces of the piglets. In sows, only feed supplementation with B. cereus led to a clear increase in faecal IgA. Serum IgG levels were not significantly affected by any probiotic feeding in sows. In piglets, the group that was fed B. cereus showed significantly higher faecal IgA levels shortly before weaning, whereas in the E. faecium group, a significant decrease in IgA levels was observed one week after weaning. In both probiotic fed groups the post-weaning IgG levels were significantly decreased compared to the respective control groups. We conclude that B. cereus var. toyoi feed supplementation led to an increased intestinal IgA secretion both in sows and piglets. This effect could be related to a more successful mucosal defence which in turn led to a lower level in systemic IgG production in piglets after weaning.     

Inhibitory Influence of Enterococcus faecium on the Propagation of Swine Influenza A Virus In Vitro

The control of infectious diseases such as swine influenza viruses (SwIV) plays an important role in food production both from the animal health and from the public health point of view. Probiotic microorganisms and other health improving food supplements have been given increasing attention in recent years, but, no information on the effects of probiotics on swine influenza virus is available. Here we address this question by assessing the inhibitory potential of the probiotic Enterococcus faecium NCIMB 10415 (E. faecium) on the replication of two porcine strains of influenza virus (H1N1 and H3N2 strain) in a continuous porcine macrophage cell line (3D4/21) and in MDBK cells. Cell cultures were treated with E. faecium at the non-toxic concentration of 1×10⁶ CFU/ml in growth medium for 60 to 90 min before, during and after SwIV infection. After further incubation of cultures in probiotic-free growth medium, cell viability and virus propagation were determined at 48 h or 96 h post infection. The results obtained reveal an almost complete recovery of viability of SwIV infected cells and an inhibition of virus multiplication by up to four log units in the E. faecium treated cells. In both 3D4/21- and MDBK-cells a 60 min treatment with E. faecium stimulated nitric oxide (NO) release which is in line with published evidence for an antiviral function of NO. Furthermore, E. faecium caused a modified cellular expression of selected mediators of defence in 3D4-cells: while the expression of TNF-α, TLR-3 and IL-6 were decreased in the SwIV-infected and probiotic treated cells, IL-10 was found to be increased. Since we obtained experimental evidence for the direct adsorptive trapping of SwIV through E. faecium, this probiotic microorganism inhibits influenza viruses by at least two mechanisms, direct physical interaction and strengthening of innate defence at the cellular level.     

Antioxidant properties of potentially probiotic bacteria: in vitro and in vivo activities

Thirty-four strains of lactic acid bacteria (seven Bifidobacterium, 11 Lactobacillus, six Lactococcus, and 10 Streptococcus thermophilus) were assayed in vitro for antioxidant activity against ascorbic and linolenic acid oxidation (TAA_AA and TAA_LA), trolox-equivalent antioxidant capacity (TEAC), intracellular glutathione (TGSH), and superoxide dismutase (SOD). Wide dispersion of each of TAA_AA, TAA_LA, TEAC, TGSH, and SOD occurred within bacterial groups, indicating that antioxidative properties are strain specific. The strains Bifidobacterium animalis subsp. lactis DSMZ 23032, Lactobacillus acidophilus DSMZ 23033, and Lactobacillus brevis DSMZ 23034 exhibited among the highest TAA_AA, TAA_LA, TEAC, and TGSH values within the lactobacilli and bifidobacteria. These strains were used to prepare a potentially antioxidative probiotic formulation, which was administered to rats at the dose of 10⁷, 10⁸, and 10⁹ cfu/day for 18 days. The probiotic strains colonized the colon of the rats during the trial and promoted intestinal saccharolytic metabolism. The analysis of plasma antioxidant activity, reactive oxygen molecules level, and glutathione concentration, revealed that, when administered at doses of at least 10⁸ cfu/day, the antioxidant mixture effectively reduced doxorubicin-induced oxidative stress. Probiotic strains which are capable to limit excessive amounts of reactive radicals in vivo may contribute to prevent and control several diseases associated with oxidative stress.     

Effects of Enterococcus faecium NCIMB 10415 as probiotic supplement on intestinal transport and barrier function of piglets

Abstract

Many studies report positive effects of probiotic supplementation on the performance and health of piglets. The intention of this study was to describe the effects of Enterococcus faecium NCIMB 10415 on the transport and barrier functions of pig small intestine to improve our understanding of the underlying mechanisms of this probiotic. Ussing chamber studies were conducted with isolated jejunal epithelia of piglets at the age of 14, 28, 35 and 56 days. Jejunal tissues of the control group were compared with epithelia of piglets that had received a diet supplemented with the probiotic Enterococcus faecium NCIMB 10415. Transport properties (absorption and secretion) of the epithelia were examined by mucosal addition of glucose or L-glutamine or by serosal addition of PGE₂. Electrophysiology of the epithelia was continuously recorded and the change in short circuit current (I_sc) was determined. Paracellular permeability was measured by measuring the flux rates of mannitol. The increase of I_sc caused by mucosal addition of glucose was, at all glucose concentrations, higher in the probiotic group compared with the control group. However, the difference (up to 100% of the control) was not significant. The increase of I_sc after the mucosal addition of L-glutamine (12 mmol/l) was higher in the tissues of the probiotic group but did not reach significance. Serosal PGE₂ induced a significantly higher increase of I_sc in tissues of the probiotic group at the age of 28 days. No consistent differences were observed in mannitol transport rates between the feeding groups.

Significant age-dependent alterations of absorptive and secretory properties of the jejunal epithelium were observed; these were independent of the treatment. A probiotic supplementation seems to influence transport properties of small intestine epithelium. The increased absorption of glucose could be interpreted as a positive effect for the animal.     

Growth and survival of lactic acid bacteria in lucerne silage

A rifampicin-resistant variant of two strains of Lactobacillus plantarum, one strain of Pediococcus acidilactici, and one strain of Enterococcus faecium were used for the experimental production of lucerne silage. Laboratory silage without inoculants served as a control. Counts of total anaerobes, total lactic acid bacteria (LAB), lactobacilli, pediococci, and enterococci were determined on days 14, 21, 30, 49, and 60 of lucerne fermentation. LAB dominated in silage microflora, reaching a percentage between 59 and 95 % of total anaerobes. Lactobacilli were found as a predominant group of LAB during the whole study. Lactobacilli reached numbers 8.74 log CFU/g in treated silage and 8.89 log CFU/g in the control at the first observation. Their counts decreased to 4.23 and 4.92 log CFU/g in treated silage and the control, respectively, on day 63 of fermentation. Similar decreases were observed in all bacterial groups. The treated silage samples possessed lower pH (4.2 vs. 4.5 in control samples) and contained more lactic acid compared to control silage. The identity of re-isolated rifampicin-resistant bacteria with those inoculated to the lucerne was evaluated by fingerprinting techniques. The fingerprint profiles of re-isolated bacteria corresponded to the profiles of strains used for the treatment. It could be concluded that supplemented LAB dominated in laboratory silage and overgrew naturally occurring LAB.