Ameliorative effect of Bacillus subtilis, Saccharomyces boulardii, oregano, and calcium montmorillonite on growth, intestinal histology, and blood metabolites on Salmonella-infected broiler chicken.

This study evaluated the effects of Bacillus subtilis, Saccharomyces boulardii, oregano, and calcium montmorillonite on the physical growth, intestinal histomorphology, and blood metabolites in Salmonella-challenged birds during the finisher phase. In this study, a total of 600 chicks (Ross 308) were randomly distributed into the following dietary treatments: basal diet with no treatment; infected with Salmonella; T1, infected + avilamycin; T2, infected + Bacillus subtilis; T3, Saccharomyces boulardii; T4, infected + oregano; T5, infected + calcium montmorillonite. Our results indicated that feed consumption, body weight gain, total body weight, and feed conversion ratio increased significantly (P < 0.01) in T1 and T2. Villus width increased significantly (P < 0.01) in T1 while the total area was significantly (P < 0.01) higher in T1 and T2 among the treatment groups. Blood protein was significantly (P < 0.01) high in T3 and T4; however, the glucose concentration was significantly (P < 0.01) high in T2, T3, and T4. The treatments increased significantly (P< 0.01) in the treatment groups compared to the negative control. Aspirate aminotransferase (AST) was significantly (P < 0.05) low in T3 compared to the positive control. In conclusion, the results indicated that supplementation of Bacillus subtilis and calcium montmorillonite improved the production performance compared to other feed additives in broiler chicks infected with Salmonella during the finisher phase.

Effects of Pediococcus acidilactici, mannan-oligosaccharide, butyric acid and their combination on growth performance and intestinal health in young broiler chickens challenged with Salmonella Typhimurium.

This study compared the efficacy of Pediococcus acidilactici, mannan-oligosaccharide, butyric acid, and their combination on growth performance and intestinal health in broiler chickens challenged with S. Typhimurium. Ross 308 male broilers (n = 420) were randomly assigned to one of the 6 treatments, resulting in 5 replicate pens of 14 chicks per treatment. The treatments included a negative control [(NC), no additive, not challenged]; positive control [(PC), no additive, but challenged with S. Typhimurium at d 3 posthatch], and 4 groups whereby birds were challenged with S. Typhimurium at d 3 posthatch and fed diets supplemented with either probiotic [0.1 g/kg Pediococcus acidilactici (PA)], prebiotic [2 g/kg mannan-oligosaccharides (MOS)], organic acid [0.5 g/kg butyric acid (BA)], or a combination of the 3 additives (MA). The S. Typhimurium challenge decreased feed intake, body weight gain and increased feed conversion ratio and reduced jejunum villus height (VH) and VH to crypt depth (CD) ratio (P < 0.05). Birds on the MA treatment exhibited similar performance to birds on the NC treatment (P > 0.05) and had a lower population of Salmonella in the ceca compared with birds on the PC treatment, at d 14 and 21 post-challenge (P < 0.05). The lowest heterophil to lymphocyte ratio was observed in birds on the MA and NC treatments (P < 0.05). Birds fed diets supplemented with MA or PA had greater VH and VH: CD ratio than birds on the PC treatment at d 7, 14 and 21 d post-challenge (P < 0.05). Suppressed amylase and protease activity was observed as a result of the S. Typhimurium challenge; the enzyme levels were restored in birds fed the additive-supplemented diets, when compared to the birds on the PC treatment, particularly at d 21 post-challenge (P < 0.05). These results indicate that dietary supplementation with a combination of PA, BA, and MOS in broiler chickens could be used as an effective tool for controlling S. Typhimurium and promoting growth performance.

High Dietary Iron Differentially Influences the Iron Distribution in the Livers and the Spleens of Laying Hens After Salmonella Typhimurium Infection.

Salmonella and the host battle for iron (Fe), due to its importance for fundamental cellular processes. To investigate Fe redistribution of Salmonella-infected hens and the effects of high dietary Fe on it, Salmonella-free hens were randomly assigned to 1 of 4 treatments in 2 (two dietary Fe level) × 2 (Salmonella-inoculation or -noninoculation) factorial assignment. After feeding a basal diet supplemented with 60 (adequate, control) or 300 mg Fe/kg (high-Fe) for 4 weeks, 59-week-old Salmonella-free hens were orally inoculated with 5 × 107 colony-forming units of Salmonella Typhimurium (infection) or PBS (vehicle). Blood, spleen, and liver samples (n = 8) were collected at 14 days post-inoculation to determine Fe concentration and Fe transporters expression. Salmonella infection decreased (P < 0.05) hematocrit, serum Fe concentration, and splenic Fe concentration regardless of high-Fe or control hens, whereas increased (P < 0.05) Fe centration in the livers of high-Fe-treated hens. High dietary Fe increased hematocrit and serum Fe concentration, but did not affect (P = 0.11) splenic Fe concentration in Salmonella-infected hens. Salmonella infection did not influence (P = 0.31) liver Fe centration in control hens, but increased (P = 0.04) it in high-Fe-treated hens. High dietary Fe decreased (P < 0.01) the mRNA abundance of divalent metal transporter 1 and transferrin receptor, but increased (P < 0.02) ferroportin-1 (FPN1) mRNA and protein in the spleens and the livers regardless of Salmonella-infected or vehicle hens. Salmonella infection increased (P < 0.02) FPN1 mRNA and protein expression in the spleens, but did not influence its expression in the livers. These results suggested Salmonella infection and high dietary Fe differently influence the Fe distribution in the spleen and the liver of Salmonella-infected hens.

Effect of Organic Acids on Salmonella Shedding and Colonization in Pigs on a Farm with High Salmonella Prevalence.

This study builds on the results of a previous study in which six commercial feed products based on organic acids were evaluated with respect to Salmonella contamination of piglets in an artificially challenged seeder model. In the present study, the efficacy of three of these commercial products was assessed for Salmonella reduction in fattening pigs on one closed farm with a natural high Salmonella prevalence. In each of four fattening compartments, one of the following feed treatments was evaluated during two consecutive fattening rounds: (i) butyric acid (active ingredients at 1.3 kg/ton of feed; supplement A1), (ii) a combination of short-chain organic acids (mixture of free acids and salts) and natural extracts (2.92 kg/ton; supplement A4), (iii) a 1:1 blend of two commercial products consisting of medium-chain fatty acids, lactic acid, and oregano oil (3.71 kg/ton; supplement A5+A6), and (iv) a control feed. On the farm, the Salmonella status of the fattening pigs was evaluated by taking fecal samples twice during the fattening period. At the slaughterhouse, samples were collected from the cecal contents and the ileocecal lymph nodes. Salmonella isolates were serotyped and characterized by pulsed-field gel electrophoresis. This farm had a particularly high number of pigs shedding Salmonella with a wide variety of sero- and pulsotypes. Only the feed blend based on the medium-chain fatty acids was able to significantly reduce Salmonella prevalence both on the farm and at the slaughterhouse. With this combined supplement, the Salmonella reduction in the feces at slaughter age, in cecal contents at slaughter, and the lymph nodes was 50, 36, and 67%, respectively, compared with the control animals. This promising finding calls for further investigation including cost-efficiency of this combined feed product and its effect on the animals.

Low dietary iron intake restrains the intestinal inflammatory response and pathology of enteric infection by food-borne bacterial pathogens.

Orally administrated iron is suspected to increase susceptibility to enteric infections among children in infection endemic regions. Here we investigated the effect of dietary iron on the pathology and local immune responses in intestinal infection models. Mice were held on iron-deficient, normal iron, or high iron diets and after 2 weeks they were orally challenged with the pathogen Citrobacter rodentium. Microbiome analysis by pyrosequencing revealed profound iron- and infection-induced shifts in microbiota composition. Fecal levels of the innate defensive molecules and markers of inflammation lipocalin-2 and calprotectin were not influenced by dietary iron intervention alone, but were markedly lower in mice on the iron-deficient diet after infection. Next, mice on the iron-deficient diet tended to gain more weight and to have a lower grade of colon pathology. Furthermore, survival of the nematode Caenorhabditis elegans infected with Salmonella enterica serovar Typhimurium was prolonged after iron deprivation. Together, these data show that iron limitation restricts disease pathology upon bacterial infection. However, our data also showed decreased intestinal inflammatory responses of mice fed on high iron diets. Thus additionally, our study indicates that the effects of iron on processes at the intestinal host-pathogen interface may highly depend on host iron status, immune status, and gut microbiota composition.

Dietary calcium inhibits the intestinal colonization and translocation of Salmonella in rats.

BACKGROUND & AIMS: Dietary calcium decreases the cytotoxicity of intestinal contents and intestinal epitheliolysis by precipitating cytotoxic surfactants such as bile acids. A decreased luminal cytotoxicity might not only strengthen the barrier function of the gut mucosa but also reinforce the protective, endogenous microflora. We hypothesized, therefore, that dietary calcium increases the resistance to intestinal infections. METHODS: Rats on a low-, medium-, or high-calcium purified diet were orally infected with a single dose of Salmonella enteritidis. The kinetics of fecal Salmonella excretion was studied to determine the colonization resistance. Intestinal bacterial translocation was quantitated by measuring urinary oxidation products of nitric oxide (NOx) excretion and culturing bacteria from tissues. RESULTS: Compared with the low-calcium group, the medium- and high-calcium diet-fed rats had a substantially improved colonization resistance. Calcium supplementation also reduced translocation of Salmonella, considering the diminished urinary NOx excretion and viable Salmonella counts in the Ileal Peyer's patches and spleen. Dietary calcium decreased the bile acid concentration and cytotoxicity of fecal water. Several indicators of fecal bacterial mass were significantly increased by supplemental calcium. CONCLUSIONS: Dietary calcium improves the colonization resistance and reduces the severity of gut-derived systemic infections, which is probably attributable to its luminal cytoprotective effects.

Increasing the intestinal resistance of rats to the invasive pathogen Salmonella enteritidis: additive effects of dietary lactulose and calcium.

BACKGROUND AND AIMS: Lactulose fermentation by the intestinal microflora acidifies the gut contents, resulting in an increased resistance to colonisation by acid sensitive pathogens. The extent of fermentation should be controlled to prevent acid induced epithelial cell damage. Considering the buffering capacity of calcium phosphate and its intestinal cytoprotective effects, whether supplemental calcium phosphate adds to the increased resistance to intestinal infections by lactulose fermentations was studied. METHODS: In a strictly controlled experiment, rats were fed a purified low calcium control diet, a low calcium/lactulose diet, or a high calcium/lactulose diet, and subsequently infected orally with Salmonella enteritidis. RESULTS: Lactulose fermentation lowered the pH and increased the lactic acid concentration of the intestinal contents, which significantly reduced excretion of this pathogen in faeces; thus it improved the resistance to colonisation. This agreed with the high sensitivity of S enteritidis to lactic acid (main metabolite of lactulose fermentation) in vitro. Calcium phosphate decreased translocation of S enteritidis to the systemic circulation, an effect independent of lactulose. The unfavourable increased cytotoxicity of faecal water caused by lactulose fermentation was more than counteracted by supplemental calcium phosphate. Moreover, calcium phosphate stimulated lactulose fermentation, as judged by the reduced lactulose excretion in faeces and increased lactic acid, ammonia, and faecal nitrogen excretion. CONCLUSION: Extra calcium phosphate added to a lactulose diet improves the resistance to colonisation and translocation of S enteritidis. This is probably mediated by a calcium induced stimulation of lactulose fermentation by the intestinal microflora and reversion of the lactulose mediated increased luminal cytotoxicity, which reduces damage inflicted on the intestinal mucosa.

Effect of Salmonella gallinarum infection on zinc metabolism in chicks.

The effect of Salmonella gallinarum infection in chicks on serum, liver, and kidney zinc concentrations was studied. Within 48 h after intraperitoneal administration of the organism, serum zinc declined to approximately one-half the control value. In one experiment, the serum zinc concentration remained low for the 12 days of the experiment, whereas in a second experiment, the concentration gradually increased after 6 days postinoculation but never returned to the control value. Feeding as much as 500 ppm supplemental zinc did not prevent the infection-induced decline in the serum zinc concentration. The infection resulted in a sequestering of zinc in the liver; the kidney remained relatively unresponsive in this system. Fractionation of liver homogenates by gel filtration column chromatography revealed that the zinc in the livers of the infected animals eluted in a volume characteristic of metallothionein, whereas that of control animals was associated with high molecular weight proteins. Increasing the zinc content of the serum by repeated subcutaneous injections of zinc had no effect on mortality from this infection. Restricting feed consumption of uninfected chicks to that of infected animals did mimic the influence of infection of serum zinc and hepatic metallothionein concentrations.

In vivo inhibition of plasma protein leakage and Salmonella enteritidis-induced mortality in the rat by a specific paf-acether antagonist: BN 52021.

The effects of BN 52021, a new specific paf-acether receptor antagonist and the total Ginkgo Biloba extract (GBE 761) from which this product was isolated, were studied in the rat on paf-acether-induced permeability and cell number changes and on endotoxin-induced lethality. Their activities were compared to those of cyclooxygenase, 5-lipoxygenase and phospholipase A2 inhibitors. BN 52021 given s.c. or orally exerted a dose-related inhibition of paf-acether deleterious effects as well as of endotoxin lethality whereas the other drugs tested were poorly effective. These results strongly suggest paf-acether involvement in endotoxic and septic shock.

Effect of indomethacin on intestinal water transport in salmonella-infected rhesus monkeys.

Indomethacin, a nonsteroidal anti-inflammatory agent, will abolish salmonella-induced rabbit ileal secretion when given prior to the establishment of infection. In the present study, we assessed whether indomethacin can inhibit salmonella-induced intestinal secretion when administered after infection and net intestinal secretion are well established. A physiological model of salmonellosis, salmonella-infected rhesus monkeys, was used. This model also permitted an examination of the effects of indomethacin in both the small and large intestines. The effect of indomethacin in control monkeys was also studied. Indomethacin caused a striking enhancement of net intestinal water transport in the jejunum,, ileum, and colon of salmonella-infected monkeys. These effects occurred promptly and were of sufficient magnitude in the ileum and colon to cause a reversal in the direction of net transport from net secretion to net absorption. Indomethacin also enhanced net water transport in the jejunum ileum, and colon of normal animals. These data show that indomethacin markedly enhances net intestinal water transport in both the small and large intestines of salmonella-infected monkeys, even when administered after salmonella infection and intestinal secretion are well established. Similar enhancement also occurs in the normal intestine. The mechanism(s) by which indomethacin produces these effects is not known.

Salmonellosis pacifarin activity of enterobactin.

Salmonellosis pacifarin activity is detected by an increased survivorship of mice, doubly infected with avirulent and virulent Salmonella typhimurium, when heretofore unknown agents, found in certain natural foodstuffs and in the supernatants of certain bacterial cultures, are fed to the infected animals as dietary supplements. We now announce the identity of one of these agents: it is enterobactin, a cyclic trimer of 2,3-dihydroxy-N-benzoyl-L-serine. Basal diets enriched with as little as 2 mg of crystalline trimer per kg of diet show pacifarin activity to a statistically significant degree. Diets supplemented with as much as 100 mg of the monomer per kg of diet show no such activity.