Purified galactooligosaccharide, derived from a mixture produced by the enzymic activity of Bifidobacterium bifidum, reduces Salmonellaenterica serovar Typhimurium adhesion and invasion in vitro and in vivo
The prebiotic Bimuno® is a mixture containing galactooligosaccharides (GOSs), produced by the galactosyltransferase activity of Bifidobacterium bifidum NCIMB 41171 using lactose as the substrate. Previous in vivo and in vitro studies demonstrating the efficacy of Bimuno® in reducing Salmonellaenterica serovar Typhimurium (S. Typhimurium) colonization did not ascertain whether or not the protective effects could be attributed to the prebiotic component GOS. Here we wished to test the hypothesis that GOS, derived from Bimuno®, may confer the direct anti-invasive and protective effects of Bimuno®. In this study the efficacy of Bimuno®, a basal solution of Bimuno® without GOS [which contained glucose, galactose, lactose, maltodextrin and gum arabic in the same relative proportions (w/w) as they are found in Bimuno®] and purified GOS to reduce S. Typhimurium adhesion and invasion was assessed using a series of in vitro and in vivo models. The novel use of three dimensionally cultured HT-29-16E cells to study prebiotics in vitro demonstrated that the presence of ∼5 mg Bimuno® ml−1 or ∼2.5 mg GOS ml−1 significantly reduced the invasion of S. Typhimurium (SL1344nalr) (P<0.0001). Furthermore, ∼2.5 mg GOS ml−1 significantly reduced the adherence of S. Typhimurium (SL1344nalr) (P<0.0001). It was demonstrated that cells produced using this system formed multi-layered aggregates of cells that displayed excellent formation of brush borders and tight junctions. In the murine ligated ileal gut loops, the presence of Bimuno® or GOS prevented the adherence or invasion of S. Typhimurium to enterocytes, and thus reduced its associated pathology. This protection appeared to correlate with significant reductions in the neutral and acidic mucins detected in goblet cells, possibly as a consequence of stimulating the cells to secrete the mucin into the lumen. In all assays, Bimuno® without GOS conferred no such protection, indicating that the basal solution confers no protective effects against S. Typhimurium. Collectively, the studies presented here clearly indicate that the protective effects conferred by Bimuno® can be attributed to GOS.
AgunosA.,
IbukiM.,
YokomizoF.,
MineY.2007; Effect of dietary β 1–4 mannobiose in the prevention of Salmonella enteritidis infection in broilers. Br Poult Sci 48:331–341[CrossRef]
AugeronC.,
LaboisseC. L.1984; Emergence of permanently differentiated cell clones in a human colonic cancer cell line in culture after treatment with sodium butyrate. Cancer Res 44:3961–3969
BaileyJ. S.,
BlankenshipL. C.,
CoxN. A.1991; Effect of fructooligosaccharide on Salmonella colonization of the chicken intestine. Poult Sci 70:2433–2438[CrossRef]
CarvalhoH. M.,
TeelL. D.,
GopingG.,
O'BrienA. D.2005; A three-dimensional tissue culture model for the study of attach and efface lesion formation by enteropathogenic and enterohaemorrhagic Escherichia coli
. Cell Microbiol 7:1771–1781[CrossRef]
DepeintF.,
TzortzisG.,
VulevicJ.,
l'AnsonK.,
GibsonG. R.2008; Prebiotic evaluation of a novel galactooligosaccharide mixture produced by the enzymatic activity of Bifidobacterium bifidum NCIMB 41171, in healthy humans: a randomized, double-blind, crossover, placebo-controlled intervention study. Am J Clin Nutr 87:785–791
FernandezF.,
HintonM.,
Van GilsB.2002; Dietary mannan-oligosaccharides and their effect on chicken caecal microflora in relation to Salmonella Enteritidis colonization. Avian Pathol 31:49–58[CrossRef]
GibsonG. R.,
ProbertH. M.,
Van LooJ.,
RastallR. A.,
RoberfroidM. B.2004; Dietary modulation of the human colonic microbiota: updating the concept of prebiotics. Nutr Res Rev 17:259–275[CrossRef]
GirardF.,
FrankelG.,
PhillipsA. D.,
CooleyW.,
WeyerU.,
DugdaleA. H. A.,
WoodwardM. J.,
La RagioneR. M.2008; Interaction of enterohemorrhagic Escherichia coli O157 : H7 with mouse intestinal mucosa. FEMS Microbiol Lett 283:196–202[CrossRef]
Höner zu BentrupK.,
RamamurthyR.,
OttC. M.,
EmamiK.,
Nelman-GonzalezM.,
WilsonJ. W.,
RichterE. G.,
GoodwinT. J.,
AlexanderJ. S.other authors2006; Three-dimensional organotypic models of human colonic epithelium to study the early stages of enteric salmonellosis. Microbes Infect 8:1813–1825[CrossRef]
La RagioneR. M.,
BestA.,
CliffordD.,
WeyerU.,
JohnsonL.,
MarshallR. N.,
MarshallJ.,
CooleyW. A.,
FarrellyS.other authors2006; Influence of colostrum deprivation and concurrent Cryptosporidium parvum infection on the colonization and persistence of Escherichia coli O157 : H7 in young lambs. J Med Microbiol 55:819–828[CrossRef]
LesuffleurT.,
RocheF.,
HillA. S.,
LacasaM.,
FoxM.,
SwallowD. M.,
ZweibaumA.,
RealF. X.1995; Characterization of a mucin cDNA clone isolated from HT-29 mucus-secreting cells. The 3′ end of MUC5AC?. J Biol Chem 270:13665–13673[CrossRef]
MeyerholzD. K.,
StabelT. J.,
AckermannM. R.,
CarlsonS. A.,
JonesB. D.,
PohlenzJ.2002; Early epithelial invasion by Salmonella enterica serovar Typhimurium DT104 in the swine ileum. Vet Pathol 39:712–720[CrossRef]
NaughtonP. J.,
MikkelsenL. L.,
JensenB. B.2001; Effects of nondigestible oligosaccharides on Salmonella enterica serovar Typhimurium and nonpathogenic Escherichia coli in the pig small intestine in vitro. Appl Environ Microbiol 67:3391–3395[CrossRef]
SatchithanandamS.,
Vargofcak-ApkerM.,
CalvertR. J.,
LeedsA. R.,
CassidyM. M.1990; Alteration of gastrointestinal mucin by fiber feeding in rats. J Nutr 120:1179–1184
SearleL. E. J.,
BestA.,
NunezA.,
SalgueroF. J.,
JohnsonL.,
WeyerU.,
DugdaleA. H.,
CooleyW. A.,
CarterB.other authors2009; A mixture containing galactooligosaccharide produced by the enzymic activity of Bifidobacterium bifidum , reduces Salmonella enterica serovar Typhimurium infection in mice. J Med Microbiol 58:37–48[CrossRef]
SmithH. W.,
HallS.1967; Observations by the ligated intestinal segment and oral inoculation methods on Escherichia coli infections in pigs, calves, lambs and rabbits. J Pathol Bacteriol 93:499–529[CrossRef]
SpringP.,
WenkC.,
DawsonK. A.,
NewmanK. E.2000; The effects of dietary mannanoligosaccharides on cecal parameters and the concentrations of enteric bacteria in the ceca of salmonella-challenged broiler chicks. Poult Sci 79:205–211[CrossRef]
TzortzisG.,
GoulasA. K.,
GeeJ. M.,
GibsonG. R.2005a; A novel galactooligosaccharide mixture increases the bifidobacterial population numbers in a continuous in vitro fermentation system and in the proximal colonic contents of pigs in vivo. J Nutr 135:1726–1731
TzortzisG.,
GoulasA. K.,
GibsonG. R.2005b; Synthesis of prebiotic galactooligosaccharides using whole cells of a novel strain, Bifidobacterium bifidum NCIMB 41171. Appl Microbiol Biotechnol 68:412–416[CrossRef]
WalesA. D.,
Clifton-HadleyF. A.,
CooksonA. L.,
Dibb-FullerM. P.,
La RagioneR. M.,
PearsonG. R.,
WoodwardM. J.2002; Production of attaching-effacing lesions in ligated large intestine loops of 6-month-old sheep by Escherichia coli O157 : H7. J Med Microbiol 51:755–763
Purified galactooligosaccharide, derived from a mixture produced by the enzymic activity of Bifidobacterium bifidum, reduces Salmonellaenterica serovar Typhimurium adhesion and invasion in vitro and in vivo