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Influence of a Probiotic Strain of Enterococcus faecium on Salmonella enterica Serovar Typhimurium DT104 Infection in a Porcine Animal Infection Model
Authors:István Szabó  Lothar H Wieler  Karsten Tedin  Lydia Scharek-Tedin  David Taras  Andreas Hensel  Bernd Appel  Karsten N?ckler
Abstract: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.The problem of increasing microbial resistance to antibiotics resulting from years of overuse and the resulting ban on the use of antibiotics in animal production have led to increased interest in alternatives to antibiotics in animal production. In recent years, probiotic bacteria have been considered as an alternative means of reducing pathogen loads in animal breeding and production units. However, while a number of studies have focused on the mode of action of probiotics, the mode of action these bacteria is not fully understood yet.A recent interdisciplinary research study of the modes of action of probiotics in swine showed that Enterococcus faecium NCIMB 10415 reduced the pathogenic bacterial load of healthy piglets (20, 26, 30, 36). In vitro studies further demonstrated that this E. faecium probiotic strain decreased the rate of invasion of a porcine intestinal epithelial cell line by Salmonella enterica serovar Typhimurium. To determine whether probiotics also provide a measure of protection during infections, experimental challenge studies with pathogenic bacteria at a defined infectious dose and under comparable conditions seem to be necessary. Field studies could be more representative of the real situation; however, the infection pressure is too low and difficult to define, and systematic sampling cannot be done.Studies of larger domestic and production animals are rare. Most such studies deal with the mode of action of probiotics in the healthy host, and only a few studies have investigated the mode of action in the context of infections with pathogenic bacteria, such as Salmonella.In a related study, weaned piglets were fed a mixture of five probiotic strains (one Pediococcus strain and four Lactobacillus strains) and challenged with Salmonella serovar Typhimurium (7). In that study, reduced incidence, severity, and duration of diarrhea and a reduced microbiological load of Salmonella were observed. Fedorka-Cray et al. (11) observed reduced numbers of Salmonella bacteria in cecal contents and at the ileocolic junction in S. enterica serovar Choleraesuis-challenged weaning piglets fed a competitive exclusion culture. In vitro investigations showed that Enterococcus strains have inhibitory effects on the growth of S. enterica serovar Enteritidis, and these effects were explained by both enterotoxin and nonenterotoxin factors (37). Other studies showed that E. faecium may be beneficial to the adhesion and colonization of Clostridium jejuni in the canine intestine (29) and reduced the rate of carryover infections with obligate intracellular pathogens from infected sows in piglets (26). E. faecium has also been shown to influence the composition of the bacterial community in the avian, swine, and canine gastrointestinal tracts (25, 29, 36).Infections with S. enterica are some of the most important sources of human gastroenteritis (39). In Germany, 52,563 human salmonellosis cases were reported in 2006 (http://www3.rki.de/SurvStat). The consumption of contaminated pork and pork products was found to be associated with 20% of human salmonellosis cases in Germany (33), indicating the importance of meat or meat products as a potential source of infection for consumers. Salmonella serovar Typhimurium, especially phage type DT104, is the Salmonella serotype most frequently isolated from pork (27), and it is of particular concern because of its acquisition of multiple antibiotic resistance (1, 38).In this study, we investigated the effect of E. faecium NCIMB 10415 on the infection dynamics of Salmonella serovar Typhimurium DT104, fecal shedding, and the patterns of Salmonella distribution in internal organs, as well as on the humoral immune response to Salmonella in weaning piglets. To the best of our knowledge, this is the first experimental study of the mode of action of a probiotic strain of E. faecium in which dissemination to different internal organs was investigated using weaned piglets experimentally infected with Salmonella.
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