Dietary Supplementation of Potential Probiotics Bacillus subtilis, Bacillus licheniformis, and Saccharomyces cerevisiae and Synbiotic Improves Growth Performance and Immune Responses by Modulation in Intestinal System in Broiler Chicks Challenged with Salmonella Typhimurium.

This study evaluates the effects of probiotics and synbiotics on the performance, immune responses, and intestinal morphology, and the expression of immunity-related genes of broiler chicks challenged with Salmonella typhimurium. Three hundred and sixty broiler chicks were divided into six groups, including broiler chicks challenged and non-challenged with S. typhimurium and fed with probiotic, synbiotic, and basal diet without additive. Growth performance (food intake, daily gain, feed conversion ratio, and mortality), immune responses (antibody titer against sheep red blood cells, immunoglobulins G and M), intestinal morphology, lactic acid bacteria population, and the expression of immunity-related genes (interferon-γ, interleukins 6 and 12, and tumor necrosis factor-α) were investigated. The administration of S. typhimurium decreased growth performance (P = 0.0001), immune responses (P = 0.0001), intestinal morphology (P = 0.0001), lactic acid bacteria population (P = 0.0001), and the expression of immunity-related genes (P = 0.0001) of broiler chickens. However, broiler chicks fed with probiotic (P = 0.001) and synbiotic (P = 0.0001) showed better growth performance, immune responses, intestinal morphology, lactic acid bacteria population, and the expression of immunity-related genes in comparison with infected broiler chicks fed with basal diet lack of probiotic and synbiotic. Feeding probiotics (P = 0.001) and synbiotics (P = 0.0001) showed positive effects for challenged and non-challenged broiler chicks. In sum, feeding synbiotic and probiotic alleviated the negative effects of S. typhimurium on growth and immunity of broiler chicks. It can be suggested to apply synbiotic and probiotics as benefit additive against infectious challenges, such as S. typhimurium.

Deciphering the interplay between the genotoxic and probiotic activities of Escherichia coli Nissle 1917.

Although Escherichia coli Nissle 1917 (EcN) has been used therapeutically for over a century, the determinants of its probiotic properties remain elusive. EcN produces two siderophore-microcins (Mcc) responsible for an antagonistic activity against other Enterobacteriaceae. EcN also synthesizes the genotoxin colibactin encoded by the pks island. Colibactin is a virulence factor and a putative pro-carcinogenic compound. Therefore, we aimed to decouple the antagonistic activity of EcN from its genotoxic activity. We demonstrated that the pks-encoded ClbP, the peptidase that activates colibactin, is required for the antagonistic activity of EcN. The analysis of a series of ClbP mutants revealed that this activity is linked to the transmembrane helices of ClbP and not the periplasmic peptidase domain, indicating the transmembrane domain is involved in some aspect of Mcc biosynthesis or secretion. A single amino acid substitution in ClbP inactivates the genotoxic activity but maintains the antagonistic activity. In an in vivo salmonellosis model, this point mutant reduced the clinical signs and the fecal shedding of Salmonella similarly to the wild type strain, whereas the clbP deletion mutant could neither protect nor outcompete the pathogen. The ClbP-dependent antibacterial effect was also observed in vitro with other E. coli strains that carry both a truncated form of the Mcc gene cluster and the pks island. In such strains, siderophore-Mcc synthesis also required the glucosyltransferase IroB involved in salmochelin production. This interplay between colibactin, salmochelin, and siderophore-Mcc biosynthetic pathways suggests that these genomic islands were co-selected and played a role in the evolution of E. coli from phylogroup B2. This co-evolution observed in EcN illustrates the fine margin between pathogenicity and probiotic activity, and the need to address both the effectiveness and safety of probiotics. Decoupling the antagonistic from the genotoxic activity by specifically inactivating ClbP peptidase domain opens the way to the safe use of EcN.

IL-22 Preserves Gut Epithelial Integrity and Promotes Disease Remission during Chronic Salmonella Infection.

The cytokine IL-22 is rapidly induced at barrier surfaces where it regulates host-protective antimicrobial immunity and tissue repair but can also enhance disease severity in some chronic inflammatory settings. Using the chronic Salmonella gastroenteritis model, Ab-mediated neutralization of IL-22 impaired intestinal epithelial barrier integrity and, consequently, exaggerated expression of proinflammatory cytokines. As disease normally resolved, neutralization of IL-22 caused luminal narrowing of the cecum-a feature reminiscent of fibrotic strictures seen in Crohn disease patients. Corresponding to the exaggerated immunopathology caused by IL-22 suppression, Salmonella burdens in the gut were reduced. This enhanced inflammation and pathogen clearance was associated with alterations in gut microbiome composition, including the overgrowth of Bacteroides acidifaciens Our findings thus indicate that IL-22 plays a protective role by limiting infection-induced gut immunopathology but can also lead to persistent pathogen colonization.

Immune-Stimulatory and Therapeutic Activity of Tinospora cordifolia: Double-Edged Sword against Salmonellosis.

The present study was aimed at determining the activity of aqueous and methanolic extracts of Tinospora cordifolia (AETC and METC) against Salmonella typhimurium. In vitro anti-Salmonella activity of T. cordifolia was determined through the broth dilution and agar well diffusion assays. The immune-stimulating potential of AETC or METC was determined by measuring the cytokine levels in the culture supernatants of treated murine J774 macrophages. Antibacterial activity of AETC or METC was determined by treating S. typhimurium-infected macrophages and BALB/C mice. The toxicity of AETC or METC was determined by measuring the levels of liver inflammation markers aspartate transaminase (AST) and alanine transaminase (ALT) and antioxidant enzymes. Macrophages treated with AETC or METC secreted greater levels of IFN-γ, TNF-α, and IL-1ß. METC showed greater activity against S. typhimurium infection in macrophages and mice as well. Treatment with METC resulted in increased survival and reduced bacterial load in S. typhimurium-infected mice. Moreover, METC or AETC treatment reduced the liver inflammation and rescued the levels of antioxidant enzymes in S. typhimurium-infected mice. The results of the present study suggest that the use of T. cordifolia may act as a double-edged sword in combating salmonellosis.

Administration of defined microbiota is protective in a murine Salmonella infection model.

Salmonella typhimurium is a major cause of diarrhea and causes significant morbidity and mortality worldwide, and perturbations of the gut microbiota are known to increase susceptibility to enteric infections. The purpose of this study was to investigate whether a Microbial Ecosystem Therapeutic (MET-1) consisting of 33 bacterial strains, isolated from human stool and previously used to cure patients with recurrent Clostridium difficile infection, could also protect against S. typhimurium disease. C57BL/6 mice were pretreated with streptomycin prior to receiving MET-1 or control, then gavaged with S. typhimurium. Weight loss, serum cytokine levels, and S. typhimurium splenic translocation were measured. NF-κB nuclear staining, neutrophil accumulation, and localization of tight junction proteins (claudin-1, ZO-1) were visualized by immunofluorescence. Infected mice receiving MET-1 lost less weight, had reduced serum cytokines, reduced NF-κB nuclear staining, and decreased neutrophil infiltration in the cecum. MET-1 also preserved cecum tight junction protein expression, and reduced S. typhimurium translocation to the spleen. Notably, MET-1 did not decrease CFUs of Salmonella in the intestine. MET-1 may attenuate systemic infection by preserving tight junctions, thereby inhibiting S. typhimurium from gaining access to the systemic circulation. We conclude that MET-1 may be protective against enteric infections besides C. difficile infection.

Influence of bacteriophage P22 on the inflammatory mediator gene expression in chicken macrophage HD11 cells infected with Salmonella Typhimurium.

This study was designed to evaluate the effects of bacteriophage on the intracellular survival and immune mediator gene expression in chicken macrophage-like HD11 cells. The invasive ability and intracellular survival of Salmonella Typhimurium (ST(P22-) ) and lysogenic S. Typhimurium (ST(P22+) ) in HD11 cells were evaluated at 37 °C for 24 h postinfection (hpi). The expression of inflammatory mediator genes was determined in ST(P22-) - and ST(P22+) -infected HD11 cells treated with and without bacteriophage P22 at 1 and 24 hpi using quantitative RT-PCR. The ability of ST(P22-) and ST(P22+) to invade HD11 cells was significantly decreased by bacteriophage P22 at 1 hpi. The numbers of intracellular ST(P22-) and ST(P22+) were significantly decreased from 2.39 to 1.62 CFU cm(-2) and from 3.40 to 1.72 CFU cm(-2) in HD11 cells treated with bacteriophage P22, respectively, at 24 hpi. The enhanced expression of inflammatory mediators was observed in ST(P22-) - and ST(P22+) -infected HD11 cells treated with and without bacteriophage P22. These results suggest that the application of bacteriophage could be an effective way to control the intracellular infection.

Interference of Bifidobacterium choerinum or Escherichia coli Nissle 1917 with Salmonella Typhimurium in gnotobiotic piglets correlates with cytokine patterns in blood and intestine.

The colonization, translocation and protective effect of two intestinal bacteria - PR4 (pig commensal strain of Bifidobacterium choerinum) or EcN (probiotic Escherichia coli strain Nissle 1917) - against subsequent infection with a virulent LT2 strain of Salmonella enterica serovar Typhimurium were studied in gnotobiotic pigs after oral association. The clinical state of experimental animals correlated with bacterial translocation and levels of inflammatory cytokines [a chemokine, interleukin (IL)-8, a proinflammatory cytokine, tumour necrosis factor (TNF)-α and an anti-inflammatory cytokine, IL-10] in plasma and intestinal lavages. Gnotobiotic pigs orally mono-associated with either PR4 or EcN thrived, and bacteria were not found in their blood. No significant inflammatory cytokine response was observed. Mono-association with Salmonella caused devastating septicaemia characterized by high levels of IL-10 and TNF-α in plasma and TNF-α in the intestine. Di-associated gnotobiotic pigs were given PR4 or EcN for 24 h. Subsequently, they were infected orally with Salmonella and euthanized 24 h later. Pigs associated with bifidobacteria before Salmonella infection suffered from severe systemic infection and mounted similar cytokine responses as pigs infected with Salmonella alone. In contrast, EcN interfered with translocation of Salmonella into mesenteric lymph nodes and systemic circulation. Pigs pre-associated with EcN thrived and their clinical condition correlated with the absence of IL-10 in their plasma and a decrease of TNF-α in plasma and ileum.

Mast cell-derived TNF can exacerbate mortality during severe bacterial infections in C57BL/6-KitW-sh/W-sh mice.

We used mast cell-engrafted genetically mast cell-deficient C57BL/6-Kit(W-sh/W-sh) mice to investigate the roles of mast cells and mast cell-derived tumor necrosis factor in two models of severe bacterial infection. In these mice, we confirmed findings derived from studies of mast cell-deficient WBB6F(1)-Kit(W/W-v) mice indicating that mast cells can promote survival in cecal ligation and puncture (CLP) of moderate severity. However, we found that the beneficial role of mast cells in this setting can occur independently of mast cell-derived tumor necrosis factor. By contrast, using mast cell-engrafted C57BL/6-Kit(W-sh/W-sh) mice, we found that mast cell-derived tumor necrosis factor can increase mortality during severe CLP and can also enhance bacterial growth and hasten death after intraperitoneal inoculation of Salmonella typhimurium. In WBB6F(1)-Kit(W-sh/W-sh) mice, mast cells enhanced survival during moderately severe CLP but did not significantly change the survival observed in severe CLP. Our findings in three types of genetically mast cell-deficient mice thus support the hypothesis that, depending on the circumstances (including mouse strain background, the nature of the mutation resulting in a mast cell deficiency, and type and severity of infection), mast cells can have either no detectable effect or opposite effects on survival during bacterial infections, eg, promoting survival during moderately severe CLP associated with low mortality but, in C57BL/6-Kit(W-sh/W-sh) mice, increasing mortality during severe CLP or infection with S. typhimurium.

Salmonella enterica subsp. arizonae bone and joints sepsis. A case report and literature review.

Osteoarticular infections caused by Salmonella enterica subsp. arizonae are rarely seen in humans but young children and immunocompromised adults are at particular risk of acquiring this bacteria. Reptiles and their by-products (e.g. meat preparations or medications) are particularly likely to harbor Salmonella. We report on a case of septic arthritis of the hip transmitted by a reptile in a 10-month-old child. We carry out a recall of the complex nomenclature of Salmonella, a review of the literature and provide information on the recommended precautions for reducing the risk of transmission of Salmonella from reptiles to humans.

Modulation of gastric hemorrhage and ulceration by oxidative stress and histamine release in Salmonella typhimurium-infected rats.

Infection with Salmonella typhimurium can produce multiple organ dysfunctions. However, document concerning with gastric hemorrhagic ulcers occur in this infectious disease is lacking. The aim was to study modulation of gastric hemorrhagic ulcer by oxidative stress and mast cell histamine in S. typhimurium-infected rats. Additionally, the protective effects of drugs, such as ofloxacin, lysozyme chloride, ketotifen, ranitidine, and several antioxidants, including exogenous glutathione (GSH), allopurinol and dimethylsulfoxide (DMSO) were evaluated. Male Wistar rats were injected intrajejunally with a live culture of S. typhimurium (1 x 10(10) colony-forming units/rat) and followed by deprivation of food for 36 h. Age-matched control rats received sterilized vehicle only. Rat stomachs were irrigated for 3 h with either normal saline or a simulated gastric juice containing 100 mM HCl, 17.4 mM pepsin and 54 mM NaCl. S. typhimurium caused aggravation of offensive factors, including enhancing gastric acid back-diffusion, mucosal lipid peroxide generation, histamine release, microvascular permeability and hemorrhagic ulcer, as well as an attenuation of defensive substances, such as mucosal GSH and mucus level. Intragastric irrigation of gastric juice caused further aggravation of these gastric biochemical parameters. This exacerbation of ulcerogenic factors was abolished by pretreatment of ofloxacin and lysozyme chloride. Antioxidants, such as reduced GSH, allopurinol and DMSO also produced significant (P < 0.05) amelioration of gastric damage in S. typhimurium infected rats. In conclusion, gastric oxidative stress and histamine play pivotal roles in the formation of hemorrhagic ulcers that were effectively ameliorated by ofloxacin, lysozyme chloride, ketotifen, ranitidine, diamine oxidase and various antioxidants in S. typhimurium-infected rats.

Concurrent Aelurostrongylus abstrusus infection and salmonellosis in a kitten.

A 14-week-old kitten had a history of vomiting, diarrhoea and pyrexia, all of which resolved without treatment. Three weeks later the kitten developed a violent non-productive dry cough. Thoracic radiographs revealed pneumothorax and nodular alveolar disease. Aelurostrongylus abstrusus larvae and intracellular Gram-negative bacilli were seen in bronchial wash fluid and pleural exudate, and Salmonella Typhimurium was cultured from both fluids but not from faeces. Therapy included unilateral closed-tube thoracostomy, enrofloxacin and fenbendazole. Historical signs were compatible with gastrointestinal salmonellosis and secondary broncho-pneumonia. Seeding of the lungs with salmonellae may have occurred as a result of migration of A abstrusus from a gastro-intestinal tract residually infected or colonised by S Typhimurium. Alternatively, the development of lungworm infection in the cat may have activated quiescent S Typhimurium pulmonary granulomata from bacteraemia secondary to gastro-intestinal salmonellosis. Two years after diagnosis the cat was reportedly in good health.

[The clinico-pathogenetic validation of intravascular laser therapy in the combined treatment of salmonellosis in young infants]. / Kliniko-patogeneticheskoe obosnovanie vnutrisosudistoi lazeroterapii v kompleksnom lechenii sal'monellezov u detei rannego vozrasta.

The clinico-laboratory study of the effectiveness of the intravascular ultraviolet laser therapy (IUVLT) in the complex treatment of 25 young children with prolonged severe salmonellosis was made. The study revealed that under the action of IUVLT the main symptoms of salmonellosis (intoxication and diarrhea) disappeared at a shorter period than after treatment by traditional methods. The pathogenetic basis of the effectiveness of IUVLT was the induction of the enzymes of the monooxygenase system of the liver and an increase in the activity of the enzymes of antiradical protection.

A clinical trial of probiotic administration for prevention of Salmonella shedding in the postoperative period in horses with colic.

The purpose of this study was to evaluate the effects of probiotic administration on the prevalence of fecal shedding of Salmonella, the prevalence of postoperative diarrhea, the length of antimicrobial therapy, and the length of the hospitalization stay during the postoperative period in horses with colic. Two commercially available probiotics for horses were used in a double-blind prospective study of 200 horses undergoing surgery for colic. Probiotic or placebo was administered PO once a day for 7 days postoperatively, and fecal cultures for Salmonella were obtained daily for 10 days. After selection of 186 patients completing the treatment protocol, the results indicated that the commercial probiotic formulations had no effect on Salmonella shedding, prevalence of diarrhea, length of antimicrobial therapy, or length of hospitalization (P > .05). Twenty percent of the horses yielded 1 or more positive fecal cultures for Salmonella; of these horses, 74% were classified as asymptomatic shedders. Twenty-six percent of all horses had fluid diarrhea postoperatively, with only 12% of these horses having positive fecal cultures for Salmonella. The most common isolate was Salmonella krefeld (24 of 39 isolates). Among the different gastrointestinal disorders, horses with feed and sand impactions appeared to be more prone to shed Salmonella.

Orally administered interferon-gamma but not tumor necrosis factor-alpha suppress infection with Salmonella typhimurium in a mouse model.

Intragastrically inoculated Salmonella typhimurium produces a systemic infection in mice with high mortality. We have examined the effect of interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha on the development of the disease. IFN-gamma reduced penetration of bacteria into the gut epithelial cells, reduced the development of bacteremia, reduced mortality and prolonged the length of survival of mice both after peroral and after intraperitoneal administration. On the other hand TNF-alpha had a similar effect only when given intraperitoneally but not by peroral route. These findings indicate that the mechanisms by which these two cytokines influence the development of S. typhimurium infection are different. This is the first observation that perorally administered cytokines may have local and systemic effects on bacterial infection.

[Gamma-interferon and the antibacterial activity of mouse macrophages in a Salmonella infection]. / Gamma-interferon i antibakterial'naia aktivnost' makrofagov myshei pri sal'monelleznoi infektsii.

The influence of natural and recombinant mouse gamma-interferon (gamma-IF) on Salmonella typhimurium was experimentally studied. The preparations of gamma-IF were introduced into animals in a dose of 250 units per mouse according to different schedules before and after Salmonella inoculation. In these experiments the capacity of the animals for survival, the elimination of the infective agent from the body and the functional activity of peritoneal exudate macrophages were studied. Natural and recombinant gamma-IF was found to stimulate the ingestion activity and oxidizing metabolism of macrophages of the infected animals. At the same time the preparations of gamma-IF were not capable of enhancing the intracellular killing of salmonellae and tumor cells. Recombinant gamma-IF did not prevent the lethal outcome of S. typhimurium infection in mice and did not increase the elimination of the infective agent from the body. Natural gamma-IF, when introduced into mice in a dose of 250 units per mouse on days 1, 3 and 6 after Salmonella inoculation, ensured the survival rate of more than 80% of the animals, but did not essentially influence the dynamics of the elimination of the infective agent from the host body.

Treatment of experimental gram-negative bacterial sepsis with murine monoclonal antibodies directed against lipopolysaccharide.

Monoclonal antibodies (MAbs) directed against gram-negative bacterial lipopolysaccharide (endotoxin, LPS) are currently being evaluated as an adjunctive form of therapy for lethal gram-negative bacterial sepsis and shock. The exact binding site within the LPS molecule against which antibody should be directed in order to maximize both cross-reactivity among bacterial strains and protective capacity has not been established. By developing a panel of MAbs that bound to various regions of the LPS molecule (O saccharide; outer, intermediate, and inner core; lipid A), we were able to determine that some epitopes in the inner core/lipid A region of LPS were broadly shared among different genera of gram-negative microorganisms, on the basis of immunoblot analysis of MAb binding to LPS. Pretreatment with lower doses of O saccharide-specific MAbs (2 micrograms per animal) provided protection against a lethal intraperitoneal challenge of viable Salmonella minnesota bacteria, compared with core LPS-specific MAbs, which required at least 1.0 mg of MAb per mouse to provide a similar degree of immunoprotection. Although inner core LPS-specific MAbs are less protective than O saccharide-specific MAbs, these MAbs will probably be more useful in the treatment of gram-negative sepsis because of their ability to bind to many types of LPS and enhance survival during infection, which is caused by a wide variety of gram-negative bacteria.

[Changes in the bactericidal activity of macrophages from mice with various degrees of resistance to Salmonella typhimurium infection as affected by interferon]. / Izmenenie bakteritsidnoi aktivnosti makrofagov myshei s razlichnoi ustoichivost'iu k infektsii Salmonella typhimurium pod deistviem interferona.

The bactericidal activity of mouse macrophages with different sensitivity to Salmonella infection has been studied. The sensitivity of BALB/c mice to S. typhimurium infection is associated with the low bactericidal activity of their macrophages. The introduction of interferon stimulates the bactericidal activity of macrophages sensitive to Salmonella infection of mice, which sharply enhances the resistance of the animals to this infection.