Sows and their piglets were fed a diet supplemented with or without the probiotic E. faecium NCIM... more 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 sam...
Backround: Vaginitis is among the most common conditions women are seeking medical care for. Alth... more Backround: Vaginitis is among the most common conditions women are seeking medical care for. Although these infections can easily be treated, the relapse rate is high. This may be due to inadequate use of the diagnostic potential.
Probiotic bacteria have been suggested to stimulate the host immune system. In this study we eval... more Probiotic bacteria have been suggested to stimulate the host immune system. In this study we evaluated the immunomodulatory effects of probiotic Bacillus cereus var. toyoi on the systemic immunity of piglets. A pool of 70 piglets was divided into a probiotic or control group. We determined the ratios of peripheral blood mononuclear cell (PBMC) subsets and measured proliferative responses and cytokine production of PBMCs and effects on vaccination responses. Blood samples of probiotic-treated piglets showed a significantly lower frequency of CD8 high /CD3+ T cells and CD8 low /CD3+ T cells and a significant higher CD4+/CD8+ ratio. IL-4 and IFN-g production of polyclonally stimulated PBMCs was on average higher in the probiotic group. Specific proliferative responses of PBMCs to Influenza vaccination antigens were significantly higher and antibody titers against H3N2 Influenza and Mycoplasma vaccination antigens were on average higher in the probiotic group. In conclusion, B. cereus var. toyoi therefore alters the immune status of piglets as indicated by changes in the ratios as well as functionalities of systemic immune cell populations. #
The influence of the probiotic bacterium Enterococcus faecium SF68 on the immune system and the i... more The influence of the probiotic bacterium Enterococcus faecium SF68 on the immune system and the intestinal colonization of pigs were determined in a feeding experiment with sows and piglets. Mucosal immunity of the developing piglets was monitored by isolation and detection of intestinal lymphocyte cell populations from the proximal jejunal epithelium and the continuous Peyers patches by the use of flow cytometry. The levels of intestinal IgA in both groups of piglets were compared, as well as total IgG in the serum of sows and piglets. Feces of the sows and intestinal contents of the piglets were taken for determination of total anaerobe and coliform bacterial counts in both probiotic and control groups. Villus length and depth of the crypts were measured in the jejunum of sacrificed piglets to monitor the development of the intestinal mucosal surface amplification.
Changing immune parameters during pregnancy have previously been reported in humans and cattle, a... more Changing immune parameters during pregnancy have previously been reported in humans and cattle, and have been suggested to contribute to increased susceptibility to infections. However, data regarding immune parameters during pregnancy in sows are rare. In this study, we investigated the peripartal immune status of sows using phenotypical (FACS analysis) as well as functional (proliferation assays, cytokine analysis) parameters of peripheral blood mononuclear cells (PBMCs) in pregnant sows. In previous studies, we reported a modulation of the immune system after feed supplementation of the probiotic Bacillus cereus var. toyoi in piglets [Schierack, P., Wieler, L.H., Taras, D., Herwig, V., Tachu, B., Hlinak, A., Schmidt, M.F., Scharek, L., 2007. Bacillus cereus var. toyoi enhanced systemic immune response in piglets. Vet. Immunol. Immunopathol. 118, 1-11]. Here, we extended these previous studies to include investigations of possible probiotic effects on the peripartal immune status of sows and their reproductivity. We show that immune parameters of sows change during pregnancy, the proliferative response of PBMCs to several bacterial antigens in control animals decreased from days 90 to 30 ante partum. Relative numbers (%) of CD3+CD8+, CD4+, cytotoxic T, CD14+ and CD21+ cells were reduced compared to non-pregnant sows. In contrast, the proliferative response of PBMCs of probiotic-treated sows increased during pregnancy. Bacterial antigens primarily stimulated the proliferation of naïve CD21+ cells and the relative CD21+ cell numbers were elevated in the probiotic group in the absence of effects on other immune cell populations. The clinical and microbial status of both control and probiotic sows was similar, excluding pre-existing health problems or infections as responsible for the immunological changes, and feed supplementation also had no significant effects on reproductivity. The results suggest that the probiotic B. cereus var. toyoi can alter the proliferative response of lymphocytes and affects the immune cell population ratios of pregnant sows. How and to what extent this may affect health and reproductivity should be the focus of further studies.
Due to its low digestibility in the small intestine, a major fraction of the polyol isomalt reach... more Due to its low digestibility in the small intestine, a major fraction of the polyol isomalt reaches the colon. However, little is known about effects on the intestinal microflora. During two 4-week periods in a double-blind, placebo-controlled, cross-over design, nineteen healthy volunteers consumed a controlled basal diet enriched with either 30 g isomalt or 30 g sucrose daily. Stools were collected at the end of each test phase and various microbiological and luminal markers were analysed. Fermentation characteristics of isomalt were also investigated in vitro. Microbiological analyses of faecal samples indicated a shift of the gut flora towards an increase of bifidobacteria following consumption of the isomalt diet compared with the sucrose diet (P, 0·05). During the isomalt phase, the activity of bacterial b-glucosidase decreased (P, 0·05) whereas b-glucuronidase, sulfatase, nitroreductase and urease remained unchanged. Faecal polyamines were not different between test periods with the exception of cadaverine, which showed a trend towards a lower concentration following isomalt (P¼0·055). Faecal SCFA, lactate, bile acids, neutral sterols, N, NH 3 , phenol and p-cresol were not affected by isomalt consumption. In vitro, isomalt was metabolized in several bifidobacteria strains and yielded high butyrate concentrations. Isomalt, which is used widely as a low-glycaemic and low-energy sweetener, has to be considered a prebiotic carbohydrate that might contribute to a healthy luminal environment of the colonic mucosa.
A strain of an unidentified strictly anoxic, Gram-postive, non-motile Ruminococcus-like bacterium... more A strain of an unidentified strictly anoxic, Gram-postive, non-motile Ruminococcus-like bacterium was isolated from a human faecal sample. The organism used carbohydrates as fermentable substrates, produced acetate, succinate, and hydrogen as the major products of glucose metabolism, and possessed a G + C content of 43.3 mol%. The morphological and biochemical characteristics of the organism were consistent with its assignment to the genus Ruminococcus but it did not correspond to any recognized species of this genus. Comparative 16S rRNA gene sequencing showed the unidentified bacterium represents a previously unrecognised sub-line within the Clostridium coccoides rRNA group of organisms. The nearest relative of the unknown bacterium corresponded to Ruminococcus obeum but a 16S rRNA sequence divergence value of > 3% demonstrated it represents a different species. Based on the presented findings a new species, Ruminococcus luti, is described. The type strain of Ruminococcus luti is BInIX T (DSM 14534 T , CCUG 45635 T ).
Inulin can stimulate the growth of the intestinal bacteria as well as alter the ratio among vario... more Inulin can stimulate the growth of the intestinal bacteria as well as alter the ratio among various short chain fatty acids (SCFA) produced. In the present study, we analyzed the effect of dietary inulin on the intestinal bacterial community as determined by denaturing gradient gel electrophoresis analysis of universal 16S rDNA after amplication with PCR and SCFA profile. Broilers were fed a diet primarily composed of corn-soybean meal or same diet with 1% inulin for 42 d. The relative weight of digesta-filled ceca of the inulin-fed group was higher (P<0.01) than in the control group. Amongst SCFA, only acetate could be detected in the jejunal digesta, which tended to be higher (P=0.09) in inulin-fed group compared with the control group. Inulin did not affect the total concentration of SCFA in the cecal digesta. The relative proportion of n-butyrate was elevated (P=0.05) with a concomitant decrease in the concentration of n-valerate (P<0.05) in the inulin-fed group compared with the control group. Dietary inulin did not affect the number of PCR-denaturing gradient gel electrophoresis bands nor their diversity in the jejunal and cecal digesta. Intragroup similarities were not different between the groups, nor were any differences between intra-and intergroup similarities in the jejunal and cecal samples. In conclusion, inulin altered the cecal microbial metabolic activity without any major impact on the composition of intestinal bacterial communities as measured by the present techniques.
ABSTRACT Two probiotics of different ecological origin, Enterococcus faecium NCIMB 10415 and Baci... more ABSTRACT Two probiotics of different ecological origin, Enterococcus faecium NCIMB 10415 and Bacillus cereus var. toyoi, were chosen as model organisms. Feed for sows during gestation/lactation and for piglets pre-/postweaning was supplemented with either of these probiotics. To evaluate the effect of different starting points of E. faecium NCIMB 10415 initiations the probiotic was administered to piglets of sows, which have not received the probiotic, from birth onwards or just postweaning. Here we report the impact of these variants on probiotic distribution in the gut, on the gut microbiota, on diarrhea and on performance. Both probiotic strains were detected immediately after the start of the supplementation in feces of sows and piglets. The vertical transfer of both probiotic strains with sow feces to piglets could be demonstrated already before suckling piglets had access to the supplemented diets. Both probiotics were recovered from all intestinal segments of piglets. The dominant autochthonous colonic microbiota of young piglets as revealed by denaturing gradient gel electrophoresis was more similar within than between treatment groups (control vs. probiotic). Both probiotics reduced the incidence of postweaning diarrhea (p
A new system, that allowed the monitoring of hydrogen (H2) excretion by gnotobiotic rats without ... more A new system, that allowed the monitoring of hydrogen (H2) excretion by gnotobiotic rats without affecting their defined microbial status, was developed. The system consists of an isolator containing a chamber for an experimental animal, and a life-support system (LSS). with a sampling port outside the isolator connected to it. H2 accumulation in the system was measured by analysing a defined volume of gas after removal. H2 concentrations were determined with an electrochemical cell or by gas chromatography. To validate this technique, H2 excretion by germ-free (GF) and mono-associated rats fed a chemically defined diet was measured after oral application of lactulose. Mono-associated rats had been obtained by colonizing GF rats with a Hrproducing Clostridium per!ringens type A strain isolated from human faeces of a healthy volunteer. Application of 50 mg lactulose to the monoassociated rats resulted in a significant increase in H2 excretion. The net H2 excretion was 7.82 ± 1.28 ml H2 in 12 h corresponding to a net maximal rate of 1.1± 0.3 ml H2/h. In contrast, in experiments with GF rats, less than 0.13 ml H2 were detectable within 12 h. The technique presented is a useful tool for studying bacterial H2 metabolism in vivo under gnotobiotic conditions.
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2005
Limit-dilution procedures were used to isolate seven, helically coiled bacterial strains from fae... more Limit-dilution procedures were used to isolate seven, helically coiled bacterial strains from faeces of swine that constituted two unidentified taxa. Comparative 16S rRNA gene sequence analysis showed highest similarity values with species of the genus Treponema indicating that the isolates are members of this genus. Strain 7CPL208 T , as well as five further isolates, and 14V28 T displayed the highest 16S rRNA gene sequence similarities with Treponema pectinovorum ATCC 33768 T (92?3 %) and Treponema parvum OMZ 833 T (89?9 %), respectively. Polar lipid profiles distinguished 7CPL208 T and 14V28 T from each other as well as from related species. Based on their phenotypic and genotypic distinctiveness, strains 7CPL208 T and 14V28 T are suggested to represent two novel species of the genus Treponema, for which the names Treponema berlinense sp. nov. and Treponema porcinum sp. nov. are proposed. The type strain for Treponema berlinense is 7CPL208 T (=ATCC BAA-909 T =CIP 108244 T =JCM 12341 T ) and for Treponema porcinum 14V28 T (=ATCC
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2008
A bacterium, designated strain lac18(T), was isolated in pure culture from the mucosal jejunum of... more A bacterium, designated strain lac18(T), was isolated in pure culture from the mucosal jejunum of a healthy pig, using a medium selective for anaerobic lactic acid bacteria and containing porcine gastric mucin as the main carbon and nitrogen source. Cells of this strain were coccus-shaped, arranged singly or in pairs and were Gram-stain-negative, oxidase-negative, non-spore-forming, anaerobic and microaerotolerant. An analysis based on 16S rRNA gene sequences indicated that strain lac18(T) should be assigned to the genus Veillonella, class Clostridia, phylum Firmicutes. 16S rRNA and dnaK gene sequence-based phylogenetic analyses both indicated that the most closely related species were Veillonella ratti ATCC 17746(T) (similarities of 96.6 and 84.5 %, respectively) and Veillonella criceti ATCC 17747(T) (similarities of 96.6 and 83.4 %, respectively). The results of DNA-DNA hybridizations between strain lac18(T) and these Veillonella species and the type species of the genus, Veillonella parvula DSM 2008(T), confirmed the genotypic distinctness of the novel isolate. Data from phenotypic studies also served to differentiate strain lac18(T) from related strains. Therefore strain lac18(T) represents a novel species of the genus Veillonella, for which the name Veillonella magna sp. nov. is proposed. The type strain is lac18(T) (=CCUG 55454(T)=CIP 109767(T)=DSM 19857(T)=JCM 15053(T)).
Trefoil factor family (TFF) peptides provide protective and reparative effects by enhancing epith... more Trefoil factor family (TFF) peptides provide protective and reparative effects by enhancing epithelial integrity and promoting mucosal restitution. TFF peptide expression is induced after mucosal damage. These processes are of central physiological relevance during the postnatal intestinal development and are strongly influenced during the weaning period. In piglets, weaning at early maturation stages frequently causes mucosal inflammation. The aim of this study was to evaluate postnatal intestinal TFF expression in a piglet probiotic trial. Low intestinal TFF2 expression was measured at early maturation stages. Weaning, however, was associated with a distinct response of increased TFF2 expression, indicating an important role in enhancing mucosal integrity. In the distal jejunum and ileum weaning could as well be associated with increased TFF3 mRNA lev-els. Differential TFF1 expression was not detected. Furthermore, TFF2 localization studies in different intestinal loci were performed by means of immunohistochemistry. Expression of selected genes (TGFA, EGFR, Cox-2) known to promote TFF signaling showed differential expression pattern as well, thereby providing further functional background. Furthermore, the expression patterns of EGFR observed in this study contribute to an advanced view of previous findings of EGFR regulation mainly obtained in rodents. An upregulated EGFR expression during early postnatal development suggests a local relevance to porcine intestinal maturation. However, a feed supplementation with the probiotic strain Enterococcus faecium did not influence TFF expression.
... DOI: 10.1080/17450390903052771 Lydia Scharek-Tedin a * , Matthias Filter b , David Taras c , ... more ... DOI: 10.1080/17450390903052771 Lydia Scharek-Tedin a * , Matthias Filter b , David Taras c , Paul Wrede b & Michael FG Schmidt a pages 343-355. ... Immune cells were isolated from the blood and the distal continuous Peyer's patch (PP) of the piglets. ...
As part of an interdisciplinary research project, we studied the performance response of sows and... more As part of an interdisciplinary research project, we studied the performance response of sows and their litters to the probiotic strain Bacillus cereus var. toyoi as well as feces consistency of piglets. Gestating sows (n ¼ 26) were randomly allotted into two groups. The probiotic B. cereus var. toyoi was administered by dietary supplementation to one group of sows and their respective litters (probiotic group) whereas the second group (control group) received no probiotic supplementation. The duration of the application was nearly 17 weeks for sows (day 90 ante partum until day 28 post partum) and six weeks for piglets (day 15 -56). Piglets were weaned after 28 days. Body weight and feed consumption were recorded weekly and fecal consistency of weaned piglets was studied daily. B. cereus var. toyoi was recovered from feces of sows and piglets as well as from digesta of piglets in the probiotic group, while being absent from all samples of control animals. In addition, the probiotic was detected in piglet feces and digesta before pre-starter feed was offered, indicating a second route of uptake besides diet. Sows of the probiotic group nursed numerically more piglets and supported a higher sum of total nursing days of all piglets within each litter than control sows ( p ¼ 0.04). In turn, body weight (BW) up to day 35 was greater for control piglets ( p 5 0.01), while average daily gain and gain to feed ratio (G:F) in weeks six and eight postweaning was higher in the probiotic group ( p 5 0.05). The overall G:F of the total postweaning period was 680 g/kg and 628 g/kg in the probiotic group and control group, respectively ( p ¼ 0.009). During the trial a high prevalence of liquid feces with its maximum in the second week after weaning was observed. Probiotic supplementation led to a reduction in the incidence of liquid feces and postweaning diarrhea by 38% and 59%, respectively ( p 5 0.001).
The intestinal bacterium Enterococcus faecium NCIMB 10415 (E. faecium SF68) has been used for mor... more 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 (1.8×10 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.
The beneficial effects of probiotic Enterococcus spp. in different hosts, such as mice and humans... more The beneficial effects of probiotic Enterococcus spp. in different hosts, such as mice and humans, have previously been reported in several studies. However, studies of large domestic animals, as well as challenge studies with pathogenic microorganisms, are very rare. Here, we investigated the influence of oral treatment of pigs with the probiotic bacterium Enterococcus faecium NCIMB 10415 on Salmonella enterica serovar Typhimurium DT104 infections in weaning piglets. Clinical symptoms, fecal excretion, the organ distribution of Salmonella, and the humoral immune response (immunoglobulin G [IgG], IgM, and IgA levels) in serum were examined. A pool of 89 piglets was randomly divided into probiotic and control groups. The probiotic group received a feed supplement containing E. faecium starting on day 14 postpartum prior to challenge with Salmonella serovar Typhimurium DT104 at 28 days postpartum. After challenge with Salmonella serovar Typhimurium DT104, piglets in both groups showed no severe clinical signs of salmonellosis. However, fecal excretion and colonization of Salmonella in organs were significantly greater in piglets fed E. faecium. Likewise, the humoral immune response against Salmonella (serum IgM and IgA levels) was significantly greater in the probiotic group animals than in control animals. The results of this study suggest that E. faecium NCIMB 10415 treatment enhanced the course of infection in weaning piglets challenged with Salmonella serovar Typhimurium DT104. However, the probiotic treatment also appeared to result in greater production of specific antibodies against Salmonella serovar Typhimurium DT104.
Page 1. mm mm L DiFFerence A Comparative View of the Role of the Internet in Election Politics eD... more Page 1. mm mm L DiFFerence A Comparative View of the Role of the Internet in Election Politics eDITeD BY STePHen warD, Diana owen, RiCHarD Davis, anD DaviD Taras Page 2. Page 3. Making a Difference Page 4. Lexington ...
Sows and their piglets were fed a diet supplemented with or without the probiotic E. faecium NCIM... more 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 sam...
Backround: Vaginitis is among the most common conditions women are seeking medical care for. Alth... more Backround: Vaginitis is among the most common conditions women are seeking medical care for. Although these infections can easily be treated, the relapse rate is high. This may be due to inadequate use of the diagnostic potential.
Probiotic bacteria have been suggested to stimulate the host immune system. In this study we eval... more Probiotic bacteria have been suggested to stimulate the host immune system. In this study we evaluated the immunomodulatory effects of probiotic Bacillus cereus var. toyoi on the systemic immunity of piglets. A pool of 70 piglets was divided into a probiotic or control group. We determined the ratios of peripheral blood mononuclear cell (PBMC) subsets and measured proliferative responses and cytokine production of PBMCs and effects on vaccination responses. Blood samples of probiotic-treated piglets showed a significantly lower frequency of CD8 high /CD3+ T cells and CD8 low /CD3+ T cells and a significant higher CD4+/CD8+ ratio. IL-4 and IFN-g production of polyclonally stimulated PBMCs was on average higher in the probiotic group. Specific proliferative responses of PBMCs to Influenza vaccination antigens were significantly higher and antibody titers against H3N2 Influenza and Mycoplasma vaccination antigens were on average higher in the probiotic group. In conclusion, B. cereus var. toyoi therefore alters the immune status of piglets as indicated by changes in the ratios as well as functionalities of systemic immune cell populations. #
The influence of the probiotic bacterium Enterococcus faecium SF68 on the immune system and the i... more The influence of the probiotic bacterium Enterococcus faecium SF68 on the immune system and the intestinal colonization of pigs were determined in a feeding experiment with sows and piglets. Mucosal immunity of the developing piglets was monitored by isolation and detection of intestinal lymphocyte cell populations from the proximal jejunal epithelium and the continuous Peyers patches by the use of flow cytometry. The levels of intestinal IgA in both groups of piglets were compared, as well as total IgG in the serum of sows and piglets. Feces of the sows and intestinal contents of the piglets were taken for determination of total anaerobe and coliform bacterial counts in both probiotic and control groups. Villus length and depth of the crypts were measured in the jejunum of sacrificed piglets to monitor the development of the intestinal mucosal surface amplification.
Changing immune parameters during pregnancy have previously been reported in humans and cattle, a... more Changing immune parameters during pregnancy have previously been reported in humans and cattle, and have been suggested to contribute to increased susceptibility to infections. However, data regarding immune parameters during pregnancy in sows are rare. In this study, we investigated the peripartal immune status of sows using phenotypical (FACS analysis) as well as functional (proliferation assays, cytokine analysis) parameters of peripheral blood mononuclear cells (PBMCs) in pregnant sows. In previous studies, we reported a modulation of the immune system after feed supplementation of the probiotic Bacillus cereus var. toyoi in piglets [Schierack, P., Wieler, L.H., Taras, D., Herwig, V., Tachu, B., Hlinak, A., Schmidt, M.F., Scharek, L., 2007. Bacillus cereus var. toyoi enhanced systemic immune response in piglets. Vet. Immunol. Immunopathol. 118, 1-11]. Here, we extended these previous studies to include investigations of possible probiotic effects on the peripartal immune status of sows and their reproductivity. We show that immune parameters of sows change during pregnancy, the proliferative response of PBMCs to several bacterial antigens in control animals decreased from days 90 to 30 ante partum. Relative numbers (%) of CD3+CD8+, CD4+, cytotoxic T, CD14+ and CD21+ cells were reduced compared to non-pregnant sows. In contrast, the proliferative response of PBMCs of probiotic-treated sows increased during pregnancy. Bacterial antigens primarily stimulated the proliferation of naïve CD21+ cells and the relative CD21+ cell numbers were elevated in the probiotic group in the absence of effects on other immune cell populations. The clinical and microbial status of both control and probiotic sows was similar, excluding pre-existing health problems or infections as responsible for the immunological changes, and feed supplementation also had no significant effects on reproductivity. The results suggest that the probiotic B. cereus var. toyoi can alter the proliferative response of lymphocytes and affects the immune cell population ratios of pregnant sows. How and to what extent this may affect health and reproductivity should be the focus of further studies.
Due to its low digestibility in the small intestine, a major fraction of the polyol isomalt reach... more Due to its low digestibility in the small intestine, a major fraction of the polyol isomalt reaches the colon. However, little is known about effects on the intestinal microflora. During two 4-week periods in a double-blind, placebo-controlled, cross-over design, nineteen healthy volunteers consumed a controlled basal diet enriched with either 30 g isomalt or 30 g sucrose daily. Stools were collected at the end of each test phase and various microbiological and luminal markers were analysed. Fermentation characteristics of isomalt were also investigated in vitro. Microbiological analyses of faecal samples indicated a shift of the gut flora towards an increase of bifidobacteria following consumption of the isomalt diet compared with the sucrose diet (P, 0·05). During the isomalt phase, the activity of bacterial b-glucosidase decreased (P, 0·05) whereas b-glucuronidase, sulfatase, nitroreductase and urease remained unchanged. Faecal polyamines were not different between test periods with the exception of cadaverine, which showed a trend towards a lower concentration following isomalt (P¼0·055). Faecal SCFA, lactate, bile acids, neutral sterols, N, NH 3 , phenol and p-cresol were not affected by isomalt consumption. In vitro, isomalt was metabolized in several bifidobacteria strains and yielded high butyrate concentrations. Isomalt, which is used widely as a low-glycaemic and low-energy sweetener, has to be considered a prebiotic carbohydrate that might contribute to a healthy luminal environment of the colonic mucosa.
A strain of an unidentified strictly anoxic, Gram-postive, non-motile Ruminococcus-like bacterium... more A strain of an unidentified strictly anoxic, Gram-postive, non-motile Ruminococcus-like bacterium was isolated from a human faecal sample. The organism used carbohydrates as fermentable substrates, produced acetate, succinate, and hydrogen as the major products of glucose metabolism, and possessed a G + C content of 43.3 mol%. The morphological and biochemical characteristics of the organism were consistent with its assignment to the genus Ruminococcus but it did not correspond to any recognized species of this genus. Comparative 16S rRNA gene sequencing showed the unidentified bacterium represents a previously unrecognised sub-line within the Clostridium coccoides rRNA group of organisms. The nearest relative of the unknown bacterium corresponded to Ruminococcus obeum but a 16S rRNA sequence divergence value of > 3% demonstrated it represents a different species. Based on the presented findings a new species, Ruminococcus luti, is described. The type strain of Ruminococcus luti is BInIX T (DSM 14534 T , CCUG 45635 T ).
Inulin can stimulate the growth of the intestinal bacteria as well as alter the ratio among vario... more Inulin can stimulate the growth of the intestinal bacteria as well as alter the ratio among various short chain fatty acids (SCFA) produced. In the present study, we analyzed the effect of dietary inulin on the intestinal bacterial community as determined by denaturing gradient gel electrophoresis analysis of universal 16S rDNA after amplication with PCR and SCFA profile. Broilers were fed a diet primarily composed of corn-soybean meal or same diet with 1% inulin for 42 d. The relative weight of digesta-filled ceca of the inulin-fed group was higher (P<0.01) than in the control group. Amongst SCFA, only acetate could be detected in the jejunal digesta, which tended to be higher (P=0.09) in inulin-fed group compared with the control group. Inulin did not affect the total concentration of SCFA in the cecal digesta. The relative proportion of n-butyrate was elevated (P=0.05) with a concomitant decrease in the concentration of n-valerate (P<0.05) in the inulin-fed group compared with the control group. Dietary inulin did not affect the number of PCR-denaturing gradient gel electrophoresis bands nor their diversity in the jejunal and cecal digesta. Intragroup similarities were not different between the groups, nor were any differences between intra-and intergroup similarities in the jejunal and cecal samples. In conclusion, inulin altered the cecal microbial metabolic activity without any major impact on the composition of intestinal bacterial communities as measured by the present techniques.
ABSTRACT Two probiotics of different ecological origin, Enterococcus faecium NCIMB 10415 and Baci... more ABSTRACT Two probiotics of different ecological origin, Enterococcus faecium NCIMB 10415 and Bacillus cereus var. toyoi, were chosen as model organisms. Feed for sows during gestation/lactation and for piglets pre-/postweaning was supplemented with either of these probiotics. To evaluate the effect of different starting points of E. faecium NCIMB 10415 initiations the probiotic was administered to piglets of sows, which have not received the probiotic, from birth onwards or just postweaning. Here we report the impact of these variants on probiotic distribution in the gut, on the gut microbiota, on diarrhea and on performance. Both probiotic strains were detected immediately after the start of the supplementation in feces of sows and piglets. The vertical transfer of both probiotic strains with sow feces to piglets could be demonstrated already before suckling piglets had access to the supplemented diets. Both probiotics were recovered from all intestinal segments of piglets. The dominant autochthonous colonic microbiota of young piglets as revealed by denaturing gradient gel electrophoresis was more similar within than between treatment groups (control vs. probiotic). Both probiotics reduced the incidence of postweaning diarrhea (p
A new system, that allowed the monitoring of hydrogen (H2) excretion by gnotobiotic rats without ... more A new system, that allowed the monitoring of hydrogen (H2) excretion by gnotobiotic rats without affecting their defined microbial status, was developed. The system consists of an isolator containing a chamber for an experimental animal, and a life-support system (LSS). with a sampling port outside the isolator connected to it. H2 accumulation in the system was measured by analysing a defined volume of gas after removal. H2 concentrations were determined with an electrochemical cell or by gas chromatography. To validate this technique, H2 excretion by germ-free (GF) and mono-associated rats fed a chemically defined diet was measured after oral application of lactulose. Mono-associated rats had been obtained by colonizing GF rats with a Hrproducing Clostridium per!ringens type A strain isolated from human faeces of a healthy volunteer. Application of 50 mg lactulose to the monoassociated rats resulted in a significant increase in H2 excretion. The net H2 excretion was 7.82 ± 1.28 ml H2 in 12 h corresponding to a net maximal rate of 1.1± 0.3 ml H2/h. In contrast, in experiments with GF rats, less than 0.13 ml H2 were detectable within 12 h. The technique presented is a useful tool for studying bacterial H2 metabolism in vivo under gnotobiotic conditions.
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2005
Limit-dilution procedures were used to isolate seven, helically coiled bacterial strains from fae... more Limit-dilution procedures were used to isolate seven, helically coiled bacterial strains from faeces of swine that constituted two unidentified taxa. Comparative 16S rRNA gene sequence analysis showed highest similarity values with species of the genus Treponema indicating that the isolates are members of this genus. Strain 7CPL208 T , as well as five further isolates, and 14V28 T displayed the highest 16S rRNA gene sequence similarities with Treponema pectinovorum ATCC 33768 T (92?3 %) and Treponema parvum OMZ 833 T (89?9 %), respectively. Polar lipid profiles distinguished 7CPL208 T and 14V28 T from each other as well as from related species. Based on their phenotypic and genotypic distinctiveness, strains 7CPL208 T and 14V28 T are suggested to represent two novel species of the genus Treponema, for which the names Treponema berlinense sp. nov. and Treponema porcinum sp. nov. are proposed. The type strain for Treponema berlinense is 7CPL208 T (=ATCC BAA-909 T =CIP 108244 T =JCM 12341 T ) and for Treponema porcinum 14V28 T (=ATCC
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2008
A bacterium, designated strain lac18(T), was isolated in pure culture from the mucosal jejunum of... more A bacterium, designated strain lac18(T), was isolated in pure culture from the mucosal jejunum of a healthy pig, using a medium selective for anaerobic lactic acid bacteria and containing porcine gastric mucin as the main carbon and nitrogen source. Cells of this strain were coccus-shaped, arranged singly or in pairs and were Gram-stain-negative, oxidase-negative, non-spore-forming, anaerobic and microaerotolerant. An analysis based on 16S rRNA gene sequences indicated that strain lac18(T) should be assigned to the genus Veillonella, class Clostridia, phylum Firmicutes. 16S rRNA and dnaK gene sequence-based phylogenetic analyses both indicated that the most closely related species were Veillonella ratti ATCC 17746(T) (similarities of 96.6 and 84.5 %, respectively) and Veillonella criceti ATCC 17747(T) (similarities of 96.6 and 83.4 %, respectively). The results of DNA-DNA hybridizations between strain lac18(T) and these Veillonella species and the type species of the genus, Veillonella parvula DSM 2008(T), confirmed the genotypic distinctness of the novel isolate. Data from phenotypic studies also served to differentiate strain lac18(T) from related strains. Therefore strain lac18(T) represents a novel species of the genus Veillonella, for which the name Veillonella magna sp. nov. is proposed. The type strain is lac18(T) (=CCUG 55454(T)=CIP 109767(T)=DSM 19857(T)=JCM 15053(T)).
Trefoil factor family (TFF) peptides provide protective and reparative effects by enhancing epith... more Trefoil factor family (TFF) peptides provide protective and reparative effects by enhancing epithelial integrity and promoting mucosal restitution. TFF peptide expression is induced after mucosal damage. These processes are of central physiological relevance during the postnatal intestinal development and are strongly influenced during the weaning period. In piglets, weaning at early maturation stages frequently causes mucosal inflammation. The aim of this study was to evaluate postnatal intestinal TFF expression in a piglet probiotic trial. Low intestinal TFF2 expression was measured at early maturation stages. Weaning, however, was associated with a distinct response of increased TFF2 expression, indicating an important role in enhancing mucosal integrity. In the distal jejunum and ileum weaning could as well be associated with increased TFF3 mRNA lev-els. Differential TFF1 expression was not detected. Furthermore, TFF2 localization studies in different intestinal loci were performed by means of immunohistochemistry. Expression of selected genes (TGFA, EGFR, Cox-2) known to promote TFF signaling showed differential expression pattern as well, thereby providing further functional background. Furthermore, the expression patterns of EGFR observed in this study contribute to an advanced view of previous findings of EGFR regulation mainly obtained in rodents. An upregulated EGFR expression during early postnatal development suggests a local relevance to porcine intestinal maturation. However, a feed supplementation with the probiotic strain Enterococcus faecium did not influence TFF expression.
... DOI: 10.1080/17450390903052771 Lydia Scharek-Tedin a * , Matthias Filter b , David Taras c , ... more ... DOI: 10.1080/17450390903052771 Lydia Scharek-Tedin a * , Matthias Filter b , David Taras c , Paul Wrede b & Michael FG Schmidt a pages 343-355. ... Immune cells were isolated from the blood and the distal continuous Peyer's patch (PP) of the piglets. ...
As part of an interdisciplinary research project, we studied the performance response of sows and... more As part of an interdisciplinary research project, we studied the performance response of sows and their litters to the probiotic strain Bacillus cereus var. toyoi as well as feces consistency of piglets. Gestating sows (n ¼ 26) were randomly allotted into two groups. The probiotic B. cereus var. toyoi was administered by dietary supplementation to one group of sows and their respective litters (probiotic group) whereas the second group (control group) received no probiotic supplementation. The duration of the application was nearly 17 weeks for sows (day 90 ante partum until day 28 post partum) and six weeks for piglets (day 15 -56). Piglets were weaned after 28 days. Body weight and feed consumption were recorded weekly and fecal consistency of weaned piglets was studied daily. B. cereus var. toyoi was recovered from feces of sows and piglets as well as from digesta of piglets in the probiotic group, while being absent from all samples of control animals. In addition, the probiotic was detected in piglet feces and digesta before pre-starter feed was offered, indicating a second route of uptake besides diet. Sows of the probiotic group nursed numerically more piglets and supported a higher sum of total nursing days of all piglets within each litter than control sows ( p ¼ 0.04). In turn, body weight (BW) up to day 35 was greater for control piglets ( p 5 0.01), while average daily gain and gain to feed ratio (G:F) in weeks six and eight postweaning was higher in the probiotic group ( p 5 0.05). The overall G:F of the total postweaning period was 680 g/kg and 628 g/kg in the probiotic group and control group, respectively ( p ¼ 0.009). During the trial a high prevalence of liquid feces with its maximum in the second week after weaning was observed. Probiotic supplementation led to a reduction in the incidence of liquid feces and postweaning diarrhea by 38% and 59%, respectively ( p 5 0.001).
The intestinal bacterium Enterococcus faecium NCIMB 10415 (E. faecium SF68) has been used for mor... more 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 (1.8×10 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.
The beneficial effects of probiotic Enterococcus spp. in different hosts, such as mice and humans... more The beneficial effects of probiotic Enterococcus spp. in different hosts, such as mice and humans, have previously been reported in several studies. However, studies of large domestic animals, as well as challenge studies with pathogenic microorganisms, are very rare. Here, we investigated the influence of oral treatment of pigs with the probiotic bacterium Enterococcus faecium NCIMB 10415 on Salmonella enterica serovar Typhimurium DT104 infections in weaning piglets. Clinical symptoms, fecal excretion, the organ distribution of Salmonella, and the humoral immune response (immunoglobulin G [IgG], IgM, and IgA levels) in serum were examined. A pool of 89 piglets was randomly divided into probiotic and control groups. The probiotic group received a feed supplement containing E. faecium starting on day 14 postpartum prior to challenge with Salmonella serovar Typhimurium DT104 at 28 days postpartum. After challenge with Salmonella serovar Typhimurium DT104, piglets in both groups showed no severe clinical signs of salmonellosis. However, fecal excretion and colonization of Salmonella in organs were significantly greater in piglets fed E. faecium. Likewise, the humoral immune response against Salmonella (serum IgM and IgA levels) was significantly greater in the probiotic group animals than in control animals. The results of this study suggest that E. faecium NCIMB 10415 treatment enhanced the course of infection in weaning piglets challenged with Salmonella serovar Typhimurium DT104. However, the probiotic treatment also appeared to result in greater production of specific antibodies against Salmonella serovar Typhimurium DT104.
Page 1. mm mm L DiFFerence A Comparative View of the Role of the Internet in Election Politics eD... more Page 1. mm mm L DiFFerence A Comparative View of the Role of the Internet in Election Politics eDITeD BY STePHen warD, Diana owen, RiCHarD Davis, anD DaviD Taras Page 2. Page 3. Making a Difference Page 4. Lexington ...
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