Cyperus esculentus L. Tubers (Tiger Nuts) Protect Epithelial Barrier Function in Caco-2 Cells Infected by Salmonella Enteritidis and Promote Lactobacillus plantarum Growth.

Cyperus esculentus L. tubers (tiger nuts) contain different compounds with several intestinal health-promoting properties. Here, we studied the capacity of tiger nuts from Valencia, Spain, to prevent epithelial barrier function disruption induced by Salmonella enteritidis in Caco-2 cell cultures. Paracellular permeability was assessed by transepithelial electrical resistance (TER) and tight junction protein immunolocalization. Moreover, the effect of tiger nuts on S. enteritidis agglutination, oxidative stress, and Lactobacillus plantarum growth was tested. Compared to controls, tiger nuts partially restored TER in S. enteritidis-infected cultures, an effect confirmed by immunolocalization of tight junction proteins ZO-1 and occludin. The results also revealed that this protective effect may be associated with the capacity to agglutinate the pathogen, restore TER in TNFα-stimulated cultures, and reduce reactive oxygen species in H2O2-stimulated cultures. Moreover, they favor L. plantarum growth. In conclusion, this study demonstrates that the tiger nut protects epithelial barrier function by reducing bacterial invasion, along with counteracting TNFα and H2O2 effects, thus giving an additional value to this tuber as a potential functional food.

The in vitro and in vivo anti-virulence activities of Cinnamomum bejolghota by inhibiting type three secretion system effector proteins of Salmonella.

The bark of Cinnamomum bejolghota (Buch.-Ham.) Sweet (C. bejolghota) is widely used as medicine to treat bacterial diarrhea in Myanmar. We previously reported that the bark extract of C. bejolghota significantly inhibited secretion effector proteins of the type three secretion system (T3SS) in Salmonella. This study is designed to investigate the anti-virulence potential of the C. bejolghota bark extract against Salmonella Typhimuriumin in in vivo and in vitro experiments. The results suggested that the polar fraction Fr.M1 inhibited the secretion of effector proteins SipA, SipB, SipC and SipD without affecting bacteria growth and the translocation of SipC into MDA-MB-231 cells. In addition, Fr.M1 alleviated inflammatory symptoms of mice in Salmonella-infected mouse model. Overall, the results provide evidence for medicinal usage of C. bejolghota bark to treat diarrhea in Myanmar.

Milk phospholipid supplementation mediates colonization resistance of mice against Salmonella infection in association with modification of gut microbiota.

Gut microbiota-mediated colonization resistance against enteropathogens is known to be greatly influenced by bioactive food compounds. This work aims to investigate the effects of milk phospholipid (MP) supplementation on the colonization resistance of mice to Salmonella enterica serovar Typhimurium (S. Typhimurium) infection, with the focus mainly on the change of gut microbiota. Comparative microbiota analysis based on 16S rRNA gene sequence data of mice under different MP supplementation situations allowed us to identify specific microbiota characteristics associated with the varying degree of susceptibility to S. Typhimurium infection. We found that a moderate dietary intake of MPs (0.05 wt%) significantly increased the relative abundance of Bacteroides spp. (p < 0.05) and the propionate level (p < 0.05) in the mouse colon and enhanced colonization resistance against S. Typhimurium infection, when compared with the un-supplemented S. Typhimurium-infected mice, whereas excessive MP supplementation (0.25 wt%) did not significantly change the level of Bacteroides spp. (p > 0.05) and propionate (p > 0.05) and even enhanced the susceptibility and severity of S. Typhimurium infection. Furthermore, the inhibitory effects of Bacteroides spp. and propionate on S. Typhimurium intestinal colonization were verified in an ex vivo S. Typhimurium-infected 3D colonoid culture system. Our results showed that the supplementation of nutraceuticals may not always be the more the better, particularly under specific pathological conditions, and identification of specific gut microbiota characteristics may have the potential to become an indicator of appropriate supplementation in specific cases.

Epithelium intrinsic vitamin A signaling co-ordinates pathogen clearance in the gut via IL-18.

Intestinal epithelial cells (IECs) are at the forefront of host-pathogen interactions, coordinating a cascade of immune responses to protect against pathogens. Here we show that IEC-intrinsic vitamin A signaling restricts pathogen invasion early in the infection and subsequently activates immune cells to promote pathogen clearance. Mice blocked for retinoic acid receptor (RAR) signaling selectively in IECs (stopΔIEC) showed higher Salmonella burden in colonic tissues early in the infection that associated with higher luminal and systemic loads of the pathogen at later stages. Higher pathogen burden in stopΔIEC mice correlated with attenuated mucosal interferon gamma (IFNγ) production by underlying immune cells. We found that, at homeostasis, the intestinal epithelium of stopΔIEC mice produced significantly lower amounts of interleukin 18 (IL-18), a potent inducer of IFNγ. Regulation of IL-18 by vitamin A was also observed in a dietary model of vitamin A supplementation. IL-18 reconstitution in stopΔIEC mice restored resistance to Salmonella by promoting epithelial cell shedding to eliminate infected cells and limit pathogen invasion early in infection. Further, IL-18 augmented IFNγ production by underlying immune cells to restrict pathogen burden and systemic spread. Our work uncovers a critical role for vitamin A in coordinating a biphasic immune response to Salmonella infection by regulating IL-18 production by IECs.

Beneficial effect of immunobiotic strains on attenuation of Salmonella induced inflammatory response in human intestinal epithelial cells.

Probiotic bacteria have the ability to modulate host immune responses and have potent therapeutic functional effects against several diseases, including inflammatory diseases. However, beneficial effects of probiotics are strain specific and their interactions with host immune cells to modulate inflammatory response are largely unknown. Intestinal epithelial cells (IECs), which are the first line of defense against invading pathogens, and connects between commensals/probiotics and immune system; therefore, in this study, we used human IECs to assess the probiotic effects of three selected Lactobacillus strains in vitro. An HT-29 colonic epithelial cell and HT-29/blood mononuclear cells co-culture system were stimulated with Lactobacillus followed by Salmonella for different hours, after which the mRNA level of cytokines, ß-defensin-2 and negative regulators for TLR signaling and protein levels of ZO-1 and IκB-α were analyzed by real-time polymerase chain reaction and western blot analysis. L. brevis decreased Salmonella induced IL-6, IL-8, MCP-1 and IL-1ß levels, whereas L. pentosus suppressed IL-6 and MCP-1 in HT-29 cells. Moreover, L. brevis was able to increase the mRNA levels of A20, Tollip, SIGIRR and IRAKM, while L. pentosus reduced the levels of A20, and IRAKM in response to Salmonella. In addition, decrease in protein level of TNF-α and increase in mRNA level of IL-10 was observed in L. brevis and L. pentosus treated HT-29 cells. Lactobacillus strains were differentially modulated ZO-1 and p-IκB-α in HT-29 cells treated with Salmonella. Overall, the results of this study indicate that Lactobacillus strains attenuate Salmonella induced inflammatory responses through beneficial modulation of TLR negative regulators and the NF-κB pathway.

Dietary Supplementation of n-3 LCPUFAs Prevents Salmonellosis in a Murine Model.

Salmonellosis is a world-wide epidemic, and n-3 long chain polyunsaturated fatty acids (LCPUFAs) possess various health benefits. This study is aimed to investigate the preventive effects of n-3 LCPUFAs against Salmonella infection. By pretreatment with n-3 LCPUFAs, but not n-6 LCPUFAs, the survival rate of the infected mice was increased. Further studies showed that n-3 LCPUFAs significantly increased the fecal contents of short-chain fatty acids (SCFAs). The cytokine expression in the liver and production in serum were both modulated by n-3 LCPUFAs into an anti-inflammatory profile against infection. Moreover, the changes in gut microbiota by n-3 LCPUFAs favored the host against pathogens, closely related to the modified SCFA production and immune responses. In conclusion, n-3 LCPUFAs prevented Salmonella infection through multiple mechanisms, especially by the interaction with gut microbiota and host immunology. Our results suggested great perspectives for n-3 LCPUFAs and their related products to control the prevalence of Salmonella, a most predominant food-borne pathogen.

Biofilm formation by Salmonella sp. in the poultry industry: Detection, control and eradication strategies.

Salmonella represents an important global public health problem and it is an emerging zoonotic bacterial threat in the poultry industry. Diverse registered human cases of salmonellosis shown poultry origins. Various control measures have been employed both at the farming and processing levels to address it. This review focuses on traditional and new detection techniques of biofilm formation by Salmonella spp. and different approaches that can be used to prevent and/or control biofilm formation by these bacteria. A number of methodologies based on different approximations have been recently employed to detect and evaluate bacteria attached to surfaces, including real-time polymerase chain reaction (PCR), confocal laser scanning microscopy and Optical Coherence Tomography. Due to persistence of Salmonella biofilm in food processing environments after cleaning and sanitation, control and eradication strategies in poultry industry should be constantly studied. In this sense, the use of several alternatives to control Salmonella biofilm formation, such as lactic acid bacteria, phagetherapy, extracts from aromatic plants, quorum sensing inhibitors, bacteriocins and nanomaterials, have been successfully tested and will be reviewed.

Efficacy of plant-derived antimicrobials for controlling Salmonella Schwarzengrund on dry pet food.

Salmonella enterica is a major human pathogen that is responsible for 23,000 hospitalizations annually in the United States. Contact with contaminated pet food and infected companion animals can transmit salmonellosis to humans. Recent multistate human outbreaks of salmonellosis linked to commercial contaminated dry dog foods underscore the need for controlling the pathogen in pet foods for protecting pet and public health. In this study, the efficacy of five Generally Recognized as Safe (GRAS) status, plant-derived antimicrobials (PDAs), namely trans-cinnamaldehyde (TC), carvacrol (CR), thymol (TY), eugenol (EG), and caprylic acid (CA) applied as a vegetable oil or chitosan based antimicrobial spray on dry pet food for reducing Salmonella Schwarzengrund was investigated. Three hundred gram portions of a commercial dry dog food were inoculated with a two-strain mixture of nalidixic acid (NA) resistant S. Schwarzengrund (~6 log CFU/g), followed by a spray treatment with 0%, 0.5%, 1% or 2% of TC, CR, TY, EG or CA in combination with 5% vegetable oil or 1% chitosan as a carrier. The control and treated dog food samples were stored at 25 °C for 28 days. On days 0, 1, 3, 5, 7, 14, 21, and 28, Salmonella on pet food was enumerated by serial dilution and plating on xylose lysine desoxycholate (XLD) agar. All PDAs at 1% and 2% applied in vegetable oil or chitosan reduced S. Schwarzengrund by at least ~2 log CFU/g on day 3 of storage when compared to control (P < 0.05). No significant reductions in Salmonella were observed on feed sprayed with only vegetable oil or chitosan (P > 0.05). Overall, 2% TC in vegetable oil or chitosan was the most effective treatment, where at least 3 to 3.5 log CFU/g reduction in bacterial populations was observed during storage (P < 0.05). Results suggest that the aforementioned PDAs could potentially be used as an antimicrobial spray to reduce S. Schwarzengrund on dry dog food. However, further studies on the acceptance of PDA-treated dry food by dogs are needed.

Laser micro-structured Si scaffold-implantable vaccines against Salmonella Typhimurium.

Salmonella Typhi is responsible for typhoid fever in humans. Despite the efforts, the development of long-lasting vaccines has failed and the available vaccines display only moderate activity, being considered as "international traveler's" vaccines. Taking advantage of the previously described implantable vaccine technology consisting on 3D laser-microstructured Si scaffolds loaded with antigen-seeded macrophages, the present study aimed to apply an antigenic stimulus of whole extracts of S. Typhimurium, which is the mouse analogue of the human Salmonella Typhi, and examine its ability to mount specific antibody response. After defining the experimental conditions for specific anti-S. Typhimurium IgG production in vitro, antigen-seeded macrophages loaded onto the 3D Si-scaffolds were implanted to mice, while parallel experiments used conventional Freund-complete-adjuvant vaccination protocols. The results showed that only the implantable vaccine protocol could mount a specific antibody response 14 days after implantation. The cytokine profile showed increase of IL-10 and IFN-γ in the case of implantable and conventional vaccination respectively, 7 days after implantation. Morphological studies on the excised scaffolds 14 days after implantation, showed the development of a well-structured adherent monolayer, establishing multiple contacts with lymphocytes in favor to immune response development. Based on the hypothesis that both stimulatory and suppressive components in the vaccination preparation, could affect the overall activity, peptidoglycan was applied as an antigen to the vaccination protocols. Surprisingly, peptidoglycan was shown to induce a mitogenic rather than specific immunogenic response. In this case, histological analysis of the excised scaffolds showed a restricted layer of adherent cells with cytoplasmic extensions, but hard to distinguish cell contacts with lymphocytes. Finally, the presented results showed a differential behavior of antigen presenting cells in accordance to the antigenic stimulus and consequently the activation state of the cells. Tailoring the micro/sub-micron 3D structures and chemistry of Si scaffolds, could control cell behavior according to the user's needs.

Salmonella infection - prevention and treatment by antibiotics and probiotic yeasts: a review.

Global Salmonella infection, especially in developing countries, is a health and economic burden. The use of antibiotic drugs in treating the infection is proving less effective due to the alarming rise of antibiotic-resistant strains of Salmonella, the effects of antibiotics on normal gut microflora and antibiotic-associated diarrhoea, all of which bring a growing need for alternative treatments, including the use of probiotic micro-organisms. However, there are issues with probiotics, including their potential to be opportunistic pathogens and antibiotic-resistant carriers, and their antibiotic susceptibility if used as complementary therapy. Clinical trials, animal trials and in vitro investigations into the prophylactic and therapeutic efficacies of probiotics have demonstrated antagonistic properties against Salmonella and other enteropathogenic bacteria. Nonetheless, there is a need for further studies into the potential mechanisms, efficacy and mode of delivery of yeast probiotics in Salmonella infections. This review discusses Salmonella infections and treatment using antibiotics and probiotics.

Strain-specific properties of Lactobacillus plantarum for prevention of Salmonella infection.

Salmonella is a common food-borne pathogen; since lactobacilli show great potential for protecting against Salmonella infections, they are used as dietary supplements in functional foods. The aim of this study is to investigate the strain-specific properties and the involved mechanisms of action of Lactobacillus plantarum towards prevention of Salmonella infection. Mice were pretreated with mixed strains or single strain of Lactobacillus plantarum for 10 d prior to infection with Salmonella typhimurium SL1344, and the survival rates showed that lactobacilli exhibited strain-specific properties for preventing Salmonella infection. Then, in vitro and in vivo studies were carried out to investigate the involved mechanism of the strain-specific properties. The results showed that different Lactobacillus plantarum strains had different effects on inhibiting Salmonella growth, thus preventing adhesion to and invasion of epithelial cells by pathogens and enhancing immune responses. The present study demonstrated strain-specific properties of probiotics to prevent Salmonella infection and elucidated their underlying mechanisms.

Combination of Antimicrobials and Essential Oils as an Alternative for the Control of Salmonella enterica Multiresistant Strains Related to Foodborne Disease.

Due to the increase in bacterial resistance to antimicrobials (AMBs) commonly used in veterinary and human medicine, the new strategies for controlling zoonoses focus on the study of natural products with demonstrated AMB activity, such as essential oils (EOs). The aim of this study was to evaluate the in vitro effect of the combination of enrofloxacin (ENR), ceftiofur (CEF), and trimethoprim-sulfamethoxazole (SXT) with cinnamon, clove, oregano, and red thyme EOs against multiple drug-resistant strains of Salmonella enterica. The minimum inhibitory concentration (MIC) of each product was determined by microdilution and "Checkerboard" methods and their combined effect was evaluated against 15 strains of S. enterica. The results were interpreted by the calculation of fractional inhibitory concentration (FIC) and their respective indexes (FICI). Significant susceptibility of all strains to the four EOs was observed. The results showed a synergistic effect between EOs and AMBs tested, highlighting the upper percentage of total synergies of the SXT with the four EOs (FICI ≤0.5 in 60% of assays), and the most effective combination being the one of ENR and cinnamon. The MIC of cinnamon was reduced from 1250 to 312.5 µg/mL and the MIC of ENR from 2 to 0.031 µg/mL. There was no antagonism in the tested combinations (AMBs-EOs). Our results support the combined use of EOs and AMBs for the control of multiresistant strains of S. enterica with a reduction of the minimum effective dose of AMBs and their adverse effects.

Changes in thermo-tolerance and survival under simulated gastrointestinal conditions of Salmonella Enteritidis PT4 and Salmonella Typhimurium PT4 in chicken breast meat after exposure to sequential stresses.

This study assessed changes in thermo-tolerance and capability to survive to simulated gastrointestinal conditions of Salmonella Enteritidis PT4 and Salmonella Typhimurium PT4 inoculated in chicken breast meat following exposure to stresses (cold, acid and osmotic) commonly imposed during food processing. The effects of the stress imposed by exposure to oregano (Origanum vulgare L.) essential oil (OVEO) on thermo-tolerance were also assessed. After exposure to cold stress (5°C for 5h) in chicken breast meat the test strains were sequentially exposed to the different stressing substances (lactic acid, NaCl or OVEO) at sub-lethal amounts, which were defined considering previously determined minimum inhibitory concentrations, and finally to thermal treatment (55°C for 30min). Resistant cells from distinct sequential treatments were exposed to simulated gastrointestinal conditions. The exposure to cold stress did not result in increased tolerance to acid stress (lactic acid: 5 and 2.5µL/g) for both strains. Cells of S. Typhimurium PT4 and S. Enteritidis PT4 previously exposed to acid stress showed higher (p<0.05) tolerance to osmotic stress (NaCl: 75 or 37.5mg/g) compared to non-acid-exposed cells. Exposure to osmotic stress without previous exposure to acid stress caused a salt-concentration dependent decrease in counts for both strains. Exposure to OVEO (1.25 and 0.62µL/g) decreased the acid and osmotic tolerance of both S. Enteritidis PT4 and S. Typhimurium PT4. Sequential exposure to acid and osmotic stress conditions after cold exposure increased (p<0.05) the thermo-tolerance in both strains. The cells that survived the sequential stress exposure (resistant) showed higher tolerance (p<0.05) to acidic conditions during continuous exposure (182min) to simulated gastrointestinal conditions. Resistant cells of S. Enteritidis PT4 and S. Typhimurium PT4 showed higher survival rates (p<0.05) than control cells at the end of the in vitro digestion. These results show that sequential exposure to multiple sub-lethal stresses may increase the thermo-tolerance and enhance the survival under gastrointestinal conditions of S. Enteritidis PT4 and S. Typhimurium PT4.

A New Way to Beat Intestinal Pathogens.

In the gastrointestinal tract, the tug of war for iron may provide a new way to vaccinate. Recent work shows that immunizing mice with siderophores (small molecules that microbes produce to capture iron) foils pathogen colonization and may instead allow a commensal to expand.

Animal Models for Salmonellosis: Applications in Vaccine Research.

Salmonellosis remains an important cause of human disease worldwide. While there are several licensed vaccines for Salmonella enterica serovar Typhi, these vaccines are generally ineffective against other Salmonella serovars. Vaccines that target paratyphoid and nontyphoidal Salmonella serovars are very much in need. Preclinical evaluation of candidate vaccines is highly dependent on the availability of appropriate scientific tools, particularly animal models. Many different animal models exist for various Salmonella serovars, from whole-animal models to smaller models, such as those recently established in insects. Here, we discuss various mouse, rat, rabbit, calf, primate, and insect models for Salmonella infection, all of which have their place in research. However, choosing the right model is imperative in selecting the best vaccine candidates for further clinical testing. In this minireview, we summarize the various animal models that are used to assess salmonellosis, highlight some of the advantages and disadvantages of each, and discuss their value in vaccine development.

A polysaccharide isolated from the liquid culture of Lentinus edodes (Shiitake) mushroom mycelia containing black rice bran protects mice against a Salmonella lipopolysaccharide-induced endotoxemia.

Endotoxemia (sepsis, septic shock) is an inflammatory, virulent disease that results mainly from bacterial infection. The present study investigates the inhibitory effect of a bioprocessed polysaccharide (BPP) isolated from the edible Lentinus edodes liquid mycelial mushroom culture supplemented with black rice bran against murine endotoxemia induced by the Salmonella lipopolysaccharide and d-galactosamine (LPS/GalN). BPP was obtained after dialysis against water using a cellulose tube with a molecular weight cutoff of 10000. BPP eluted as a single peak on an HPLC chromatogram. Acid hydrolysis of BPP showed the presence of the following sugars: fucose, galactose, galactosamine, glucose, glucosamine, mannose, rhamnose, and xylose. Treatment of BPP with ß-glucanase reduced its immunostimulating activity, suggesting that the polysaccharide has a ß-glucan structure. Pretreatment of mice with BPP via oral or intraperitoneal (ip) administration for 2 weeks resulted in the suppression of LPS/GalN-induced catalase, superoxide dismutase (SOD), and transaminase (GOT/GPT) liver enzymes, amelioration of necrotic liver lesions, and reduction of tumor necrosis factor α (TNF-α) and nitrite serum levels as well as myeloperoxidase (MPO) activity, an index of necrotic injury. Immunostimulating macrophage activity was up to 5.4-fold greater than that observed with the culture without the rice bran. BPP also extended the lifespan of the toxemic mice. These positive results with inflammation biomarkers and lifespan studies suggest that the BPP can protect mice against LPS/GalN-induced liver, lung, and kidney injuries and inflammation by blocking oxidative stress and TNF-α production, thus increasing the survival of the toxic shock-induced mice. The polysaccharide has the potential to serve as a new functional food.

A novel adjuvant, mixture of alum and the beta-adrenergic receptor antagonist propranolol, elicits both humoral and cellular immune responses for heat-killed Salmonella typhimurium vaccine.

OBJECTIVE: To determine the efficacy of the mixture of propranolol (PRP), a beta-adrenergic receptor antagonist, and alum, as a new adjuvant, in the induction of humoral and cellular immunity in response to heat-killed Salmonella typhimurium (S. typhimurium) (HKST) as a model vaccine. METHODS: BALB/c mice were divided into five groups. Mice in the experimental groups received either the HKST vaccine alone or in combination with the adjuvant alum, PRP or the alum-PRP mixture. Mice in the negative control group received phosphate-buffered saline. All mice were immunized two times on days 0 and 14. Two weeks after the last immunization, immune responses to S. typhimurium were assessed. RESULTS: Administration of the alum-PRP mixture as an adjuvant increased the ability of the HKST vaccine to enhance lymphocyte proliferation, shifted the immune response towards a T-helper (Th) 1 pattern and increased S. typhimurium specific IgG, IgG2a and IgG1. This resulted in improved protective immunity against S. typhimurium. CONCLUSION: Administration of the alum-PRP mixture as an adjuvant in combination with the HKST vaccine, can enhance both humoral and cellular immunity and shift the immune responses to a Th1 pattern.

Opportunities for mitigating pathogen contamination during on-farm food production.

Fruits, vegetables, and meat are susceptible to contamination by foodborne pathogens at many points from production through preparation in the home. This review will largely highlight approaches and progress made in the last five years to address strategies to reduce pathogen contamination in animal production but will also touch on the emerging field of preharvest produce food safety. Mitigation strategies can be divided into those that address pathogen reduction in the environment and those that target reduction/elimination of pathogen contamination in animals or plants. The former strategy has been encompassed in studies evaluating sanitation treatments of facilities as well as in numerous epidemiologic risk assessment studies (both on-farm assessments and computer simulation models) that identify management practices that impact pathogen prevalence in animals. Interventions to significantly reduce pathogen exposure via feed or water are dependent on their role as a significant contributor to pathogen contamination in the animal production system. In addition, inconsistent results obtained with interventions of dietary additives or formulation modifications (grain versus forage; inclusion of distiller's grains) on pathogen prevalence in animals have been attributed to a range of factors including target organism, grain type, level of inclusion, the animal's health or stress level, and ability to survive the gastric acidic conditions. Recent attempts to microencapsulate organic acids or bacteriophage within feed have met with only marginal improvements in reducing pathogen carriage in animals but this approach may have greater potential with other antimicrobial additives (i.e., essential oils). Bacteriophage therapy, in general, can significantly reduce pathogen carriage in animals but based on its transient nature and the potential for development of phage-resistant subpopulations, this approach should be administered to animals just prior to slaughter and preferably to animals that are suspected "super-shedders". Other promising on-farm intervention approaches have included breeding for pathogen resistance, vaccines, and dietary bacteriocins. To optimize interventions on a cost basis, studies have also determined that application of dietary interventions at specific time points in the animal's production cycle is a useful strategy to reduce pathogen carriage (e.g., probiotics to fertilized eggs and acidified feed to fattening swine). In conclusion, applicable management and intervention strategies may vary depending on the type of food under production; however, it is important to consider from a holistic view how any new intervention strategies will affect the overall production system in order to maintain a successful, efficient food production environment.

Ethnoveterinary application of Morinda citrifolia fruit puree on a commercial heifer rearing facility with endemic salmonellosis.

We have previously reported that Morinda citrifolia (noni) puree modulates neonatal calves developmental maturation of the innate and adaptive immune system. In this study, the effect of noni puree on respiratory and gastrointestinal (GI), health in preweaned dairy calves on a farm with endemic salmonellosis was examined. Two clinical trials were conducted whereby each trial evaluated one processing technique of noni puree. Trials 1 and 2 tested noni versions A and B, respectively. Puree analysis and trial methods were identical to each other, with the calf as the experimental unit. Calves were designated to 1 of 3 treatment groups in each trial and received either: 0, 15 or 30 mL every 12 hr of noni supplement for the first 3 weeks of life. Health scores, weaning age, weight gain from admission to weaning, and weaned by 6 weeks, were used as clinical endpoints for statistical analysis. In trial 1, calves supplemented with 15 mL noni puree of version A every 12 hr had a higher probability of being weaned by 6 weeks of age than control calves (P = 0.04). In trial 2, calves receiving 30 mL of version B every 12 hr had a 54.5% reduction in total medical treatments by 42 days of age when compared to controls (P = 0.02). There was a trend in reduced respiratory (61%), and GI (52%) medical treatments per calf when compared to controls (P = 0.06 and 0.08, respectively). There were no differences in weight gain or mortality for any treatment group in either trial.

Efficacy of various dietary calcium salts to improve intestinal resistance to Salmonella infection in rats.

Previous animal and human studies have shown protective effects of Ca on the resistance to enteropathogenic infections. Most interventions were performed with calcium phosphate and little is known about the protective effect of other dietary sources of Ca. Therefore, we investigated the efficacy of several Ca salts to enhance intestinal resistance to Salmonella enteritidis infection. Rats (n 7-8 per group) were fed a high-fat, Western human-style, purified diet with a low Ca content (20 mmol calcium phosphate/kg; negative control group) or the same diet supplemented with either (extra) calcium phosphate, milk Ca, calcium chloride or calcium carbonate (total of 100 mmol Ca supplement/kg). Diets contained Cr-EDTA for assessment of incremental changes in intestinal permeability. After an adaptation period of 2 weeks, animals were orally infected with S. enteritidis to mimic a human-relevant foodborne infection. Ca supplement-induced changes on faecal lactobacilli and enterobacteria were studied before infection. Changes in intestinal permeability were determined by measuring urinary Cr with time. Persistence of Salmonella was determined by studying faecal excretion of this pathogen in time. Overall, all Ca salts increased resistance towards Salmonella. After infection, body weight gain and food intake were higher in the calcium phosphate group. Calcium phosphate and milk Ca decreased faecal enterobacteria before infection. All Ca salts decreased infection-induced intestinal permeability and persistence of Salmonella. Calcium phosphate, milk Ca, calcium carbonate and calcium chloride are able to enhance the intestinal resistance to Salmonella in rats.