Evaluation of the in vitro and in vivo efficiency of in-feed bacteriophage cocktail application to control Salmonella Typhimurium and Salmonella Enteritidis infection in broiler chicks.

RESEARCH HIGHLIGHTS: Bacteriophage (BP) cocktail was partially resistant to different temperatures and pH values.The BP cocktail showed lytic effects on different Salmonella isolates.The BP cocktail reduced Salmonella colonization in the internal organs of broilers.

Effects of in ovo injection of bacterial peptides and CpG-ODN on Salmonella enterica serovar Heidelberg infection in specific pathogen-free (SPF) chicks.

RESEARCH HIGHLIGHTS: Peptides + CpG-ODN reduced SH in caeca at the first week post-infection.Administered formulations did not reduce SH-faecal excretion.Levels of intestinal IgA were similar between all groups.CpG-ODN improved some parameters associated with chick intestinal health.

Oral vaccination of young broilers with a live Salmonella Typhimurium vaccine reduces caecal and internal organ colonization following a Salmonella Infantis challenge in a seeder-bird model.

Poultry products are an important source of foodborne Salmonella infections in humans. Amongst these, the prevalence of S. Infantis is rising. In this study, the protection efficacy of an authorized live-attenuated S. Typhimurium vaccine against S. Infantis, was examined using a seeder-bird model in broilers. Vaccinated birds displayed a significantly lower colonization of S. Infantis bacteria in the caeca compared to the non-vaccinated counterparts (P = 0.017), with no significant differences observed in the spleen among the groups, three days post-infection. Thirty-two days post-infection, the disparity in average S. Infantis concentration between all-vaccinated and non-vaccinated birds was significant in both caeca (P = 0.0003) and spleen (P = 0.0002). Interestingly, a third group, consisting of seeder birds that were not vaccinated but housed with vaccinated penmates, exhibited significantly lower S. Infantis levels in both caeca (P = 0.0014) and spleen (P < 0.0001) compared to the non-vaccinated group. These findings underscore the potential of a live-attenuated S. Typhimurium vaccine administered to 2-day-old chicks in conferring protection against S. Infantis in broilers up to slaughter age.

Social network analysis reveals the failure of between-farm movement restrictions to reduce Salmonella transmission.

An increasing number of countries are investigating options to stop the spread of the emerging zoonotic infection Salmonella Dublin (S. Dublin), which mainly spreads among bovines and with cattle manure. Detailed surveillance and cattle movement data from an 11-yr period in Denmark provided an opportunity to gain new knowledge for mitigation options through a combined social network and simulation modeling approach. The analysis revealed similar network trends for noninfected and infected cattle farms despite stringent cattle movement restrictions imposed on infected farms in the national control program. The strongest predictive factor for farms becoming infected was their cattle movement activities in the previous month, with twice the effect of local transmission. The simulation model indicated an endemic S. Dublin occurrence, with peaks in outbreak probabilities and sizes around observed cattle movement activities. Therefore, pre- and postmovement measures within a 1-mo time window may help reduce S. Dublin spread.

Combined Quantification and Deep Serotyping for Salmonella Risk Profiling in Broiler Flocks.

Despite a reduction of Salmonella contamination on final poultry products, the level of human salmonellosis cases attributed to poultry has remained unchanged over the last few years. There needs to be improved effort to target serovars which may survive antimicrobial interventions and cause illness, as well as to focus on lessening the amount of contamination entering the processing plant. Advances in molecular enumeration approaches allow for the rapid detection and quantification of Salmonella in pre- and postharvest samples, which can be combined with deep serotyping to properly assess the risk affiliated with a poultry flock. In this study, we collected a total of 160 boot sock samples from 20 broiler farms across four different integrators with different antibiotic management programs. Overall, Salmonella was found in 85% (68/80) of the houses, with each farm having at least one Salmonella-positive house. The average Salmonella quantity across all four complexes was 3.6 log10 CFU/sample. Eleven different serovars were identified through deep serotyping, including all three key performance indicators (KPIs; serovars Enteritidis, Infantis, and Typhimurium) defined by the U.S. Department of Agriculture-Food Safety and Inspection Service (USDA-FSIS). There were eight multidrug resistant isolates identified in this study, and seven which were serovar Infantis. We generated risk scores for each flock based on the presence or absence of KPIs, the relative abundance of each serovar as calculated with CRISPR-SeroSeq (serotyping by sequencing the clustered regularly interspaced palindromic repeats), and the quantity of Salmonella organisms detected. The work presented here provides a framework to develop directed processing approaches and highlights the limitations of conventional Salmonella sampling and culturing methods. IMPORTANCE Nearly one in five foodborne Salmonella illnesses are derived from chicken, making it the largest single food category to cause salmonellosis and indicating a need for effective pathogen mitigation. Although industry has successfully reduced Salmonella incidence in poultry products, there has not been a concurrent reduction in human salmonellosis linked to chicken consumption. New efforts are focused on improved control at preharvest, which requires improved Salmonella surveillance. Here, we present a high-resolution surveillance approach that combines quantity and identity of Salmonella in broiler flocks prior to processing which will further support improved Salmonella controls in poultry. We developed a framework for this approach, indicating that it is possible and important to harness deep serotyping and molecular enumeration to inform on-farm management practices and to minimize risk of cross-contamination between flocks at processing. Additionally, this framework could be adapted to Salmonella surveillance in other food animal production systems.

Assessing pig farm biosecurity measures for the control of Salmonella on European farms.

Salmonella spp. is a common zoonotic pathogen, causing gastrointestinal infections in people. Pigs and pig meat are a major source of infection. Although farm biosecurity is believed to be important for controlling Salmonella transmission, robust evidence is lacking on which measures are most effective. This study enrolled 250 pig farms across nine European countries. From each farm, 20 pooled faecal samples (or similar information) were collected and analysed for Salmonella presence. Based on the proportion of positive results, farms were categorised as at higher or lower Salmonella risk, and associations with variables from a comprehensive questionnaire investigated. Multivariable analysis indicated that farms were less likely to be in the higher-risk category if they had '<400 sows'; used rodent baits close to pig enclosures; isolated stay-behind (sick) pigs; did not answer that the hygiene lock/ anteroom was easy to clean; did not have a full perimeter fence; did apply downtime of at least 3 days between farrowing batches; and had fully slatted flooring in all fattener buildings. A principal components analysis assessed the sources of variation between farms, and correlation between variables. The study results suggest simple control measures that could be prioritised on European pig farms to control Salmonella.

Metabolomic and microbiome analysis of the protective effects of Puerarin against Salmonella Enteritidis Infection in chicks.

BACKGROUND: Salmonella Enteritidis is a zoonotic pathogen and poses a substantial risk to human health, as well as significant financial losses to the livestock and poultry industries. It is currently urgent to identify alternatives to antibiotic treatment. RESULTS: In this study, we explored the influence of Puerarin on the immunological response, intestinal flora, serum metabolome, and growth performance of chicks infected with Salmonella Enteritidis. Chicks were weighed at specific time points and the average daily gain (ADG) was calculated. Serum, intestinal, and cecal content samples were collected on days 10 and 17. The results showed that 100 mg/kg of Puerarin significantly suppressed inflammation and enhanced immune function. Metabolomic analysis showed significant differences in serum metabolites after Puerarin treatment and suggested that Puerarin may regulate abnormal amino acid and lipid metabolism after Salmonella Enteritidis infection through the autophagic and ABC transporter pathways. In addition, Puerarin suppressed Salmonella Enteritidis-induced intestinal flora dysbiosis through modulation of the microbial community structures (increased Lactobacillus, Faecalibacterium, and Subdoligranulum), as demonstrated by 16S rRNA analysis. CONCLUSIONS: In conclusion, Puerarin can improve growth performance in chicks, suppress the inflammatory response in vivo, enhance immunity, and regulate lipid and amino acid metabolism and the intestinal flora.

Nigella sativa-chitosan nanoparticles: Novel intestinal mucosal immunomodulator controls and protects against Salmonella enterica serovar Enteritidis infection in broilers.

BACKGROUND: Salmonella Enteritidis (SE) propagates in chickens' gastrointestinal surfaces and is transmitted to humans, causing food poisoning. Oral supplementation with natural nanoparticles can overcome the harsh gastrointestinal conditions facing oral vaccines and requires no antibiotic administration to protect against microbial infection. This study was designed to study Nigella sativa-chitosan nanoparticles (CNP-NS) prophylactic immunomodulatory efficacy against SE infection in broiler chicks. The CNP-NS was prepared and characterized, and its in vivo immunomodulatory activities against an avian virulent-MDR SE-induced challenge in chicks were investigated. RESULT: To verify the immune-protective activities of the CNP-NS, colony forming units (CFU) in the liver and fecal droppings; intestinal histopathological alterations and immune cell recruitment; MUC-2, TLR-4, cecal cytokines, and specific IgA gene expression levels were assessed. On the 7th and 12th days after the SE challenge, the CNP-NS supplemented chicks showed complete clearance of SE CFU in livers and fecal droppings, as well as an improvement in food conversion rate compared to non-supplemented CNP-NS that revealed the presence of the challenge SE CFU on the same days. A prominent influx of antigen presenting cells and lymphoid aggregates into the intestinal wall, spleen, and liver was detected with improvements in the intestinal villi morphometry of the CNP-NS-supplemented chicks. The changes of INF-γ, IL-1ß, and IL-4 cecal cytokines, as well as TLR-4, MUC-2, and IgA mRNA expression levels, confirm CNP-NS immunomodulatory activities and provide a mechanism(s) for its protective actions against the induced SE challenge of the tested chickens. CONCLUSION: These findings suggest promising useful insights into CNP-NS supplementation as a safe food additive for poultry meat consumers' and a protective immunomodulator of the chickens' mucosal immune systems. It could be recommended for epidemiological purposes to reduce the risk of SE food poisoning and transmission to humans.

The Importance of Hazard Analysis by Critical Control Point for Effective Pathogen Control in Animal Feed: Assessment of Salmonella Control in Feed Production in Sweden, 1982-2005.

This study is the first to show that Hazard Analysis by Critical Control Point (HACCP)-based monitoring can be an effective tool for ensuring Salmonella-safe feed, by virtually eliminating feedborne Salmonella infection even in broiler production. Data from the control of Salmonella in feed and food animal production during 1982-2005, showed that conventional endpoint testing in feed mills did not ensure a Salmonella-safe feed, and in one feed mill failed to detect Salmonella contamination, resulting in the feed infecting 80 out of 197 (40.6%) recipient broiler flocks. Following implementation in 1991 of a HACCP-based control in feed mills, the annual number of samples tested at specified critical control points during a 15-year period increased from ∼4400 to 10,000, while the proportion of Salmonella-contaminated samples decreased from 2.0% to 0.3%. Thus, introduction of HACCP was followed by a dramatic decrease, from 40 to <5, in the annual number of Salmonella-infected broiler flocks identified by preslaughter monitoring. Incidence has generally remained at that low level, despite production since 1980 increasing from 39 to 112 million chickens per year. Feed mills start using soymeal with an unsafe Salmonella status and possibly with a suboptimal HACCP control, increased their level of Salmonella-contaminated HACCP samples, and their feed subsequently infected 78 swine-producing herds. The results also show that the HACCP concept can be an effective tool to supply feed mills with Salmonella-safe feed ingredients as demonstrated for a soybean crushing plant, which produced Salmonella-safe soymeal over a 19-year period despite frequent (34%) and highly varied (92 different serovars) Salmonella contamination in samples from incoming soybean. Similar results are reported for a plant producing rapeseed meal. It is emphasized that the achievements described through use of the HACCP required interventions of relevant preventive biosecurity measures and corrective actions when the HACCP-based monitoring identified Salmonella contamination.

Polyimidazolium Protects against an Invasive Clinical Isolate of Salmonella Typhimurium.

Frequent outbreaks of Salmonella Typhimurium infection, in both animal and human populations and with the potential for zoonotic transmission, pose a significant threat to the public health sector. The rapid emergence and spread of more invasive multidrug-resistant clinical isolates of Salmonella further highlight the need for the development of new drugs with effective broad-spectrum bactericidal activities. The synthesis and evaluation of main-chain cationic polyimidazolium 1 (PIM1) against several Gram-positive and Gram-negative bacteria have previously demonstrated the efficacy profile of PIM1. The present study focuses on the antibacterial and anti-biofilm activities of PIM1 against Salmonella in both in vitro and in ovo settings. In vitro, PIM1 exhibited bactericidal activity against three strains of Salmonella at a low dosage of 8 µg/mL. The anti-biofilm activity of PIM1 was evident by its elimination of planktonic cells within preformed biofilms in a dose-dependent manner. During the host cell infection process, PIM1 reduces the extracellular bacterial load, which reduces adhesion and invasion to limit the establishment of infection. Once intracellular, Salmonella strains were tolerant and protected from PIM1 treatment. In a chicken egg infection model, PIM1 exhibited therapeutic activity for both Salmonella strains, using stationary-phase and exponential-phase inocula. Moreover, PIM1 showed a remarkable efficacy against the stationary-phase inocula of drug-resistant Salmonella by eliminating the bacterial burden in >50% of the infected chicken egg embryos. Collectively, our results highlight the potential for PIM1 as a replacement therapy for existing antibiotic applications on the poultry farm, given the efficiency and low toxicity profile demonstrated in our agriculturally relevant chicken embryo model.

Protective effects of E. coli Nissle 1917 on chickens infected with Salmonella pullorum.

The probiotic E. coli Nissle 1917 (EcN) plays an important role in regulating the microbial components of the gut and preventing inflammation of the gastrointestinal tract. Currently, the long-term use of antibiotics for the treatment of lethal white diarrhea in chicks caused by Salmonella has led to increased morbidity and mutation rates. Therefore, we want to use EcN as an antibiotic alternative as an alternative approach to prevent Salmonella-induced white diarrhea in chickens. To date, there are no reports of EcN being used for the prevention and control of Salmonella pullorum (S. pullorum) in chickens. In vitro, pretreatment with EcN significantly decreased the cellular invasion of S. pullorum CVCC533 in a chicken fibroblast (DF-1) cell model. Then, 0-day-old egg-laying chickens were orally inoculated with EcN at a dose of 109 CFU/100 µL at either Day 1 (EcN1) or both Day 1 and Day 4 (EcN2). Then, S. pullorum CVCC533 was used to challenge the cells at a dose of 1.0 × 107 CFU/100 µL on Day 8. Next, the body weights and survival rates were recorded for 14 consecutive days, and the colonization of S. pullorum in the spleen and liver at 7 days post-challenge (dpc) was determined. Chicken feces were also collected at 2, 4, 6 and 8 dpc to evaluate the excretion of pathogenic bacteria in feces. The liver, duodenum and rectum samples were collected and analyzed by pathological histology at 7 dpc to evaluate the protective effect of EcN on the mucosa, villi and crypts of the small intestine. The spleen and bursa were collected, and the immune organ index was calculated. In addition, the contents of the cecum of chicks were collected at 7 dpc for 16S rRNA sequencing to detect the distribution of microbial communities in the intestine. The results showed that EcN was able to protect against CVCC533 challenge, as shown by decreased body weight loss, mortality and shedding of pathogenic bacteria in fecal samples in the EcN1 plus Salmonella challenge group (EcN1S) but not the EcN2 plus Salmonella challenge group (EcN2S). The pathogenic changes in the liver, duodenum and rectum also demonstrated that one dose but not two doses of EcN effectively prolonged the length of the pilus with decreased crypt depth, indicating its protective effects against S. pullorum. In addition, the 16S rRNA sequencing results suggested that EcN could enlarge the diversity of intestinal flora, decrease the abundance of pathogenic bacteria and increase the abundance of beneficial bacteria, such as Lactobacillus. In conclusion, EcN has shown moderate protection against S. pullorum challenge in chickens.

Assessment of maternal immunity against Salmonella enterica serovar Heidelberg in progeny of broiler breeders vaccinated with different formulations of bacterins.

RESEARCH HIGHLIGHTSProgeny from vaccinated 30-week-old breeders presented less SH in caecal content compared to the control.High titres of maternal anti-Salmonella IgY could be associated with lower SH faecal shedding.

High specificity of the Salmonella Pullorum/Gallinarum rapid plate agglutination test despite vaccinations against Salmonella Enteritidis and Salmonella Typhimurium.

In Europe, monitoring of breeding stock for Salmonella Pullorum (SP) or Salmonella Gallinarum (SG) infections is compulsory at the point of lay. Vaccinations against Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST) are increasingly administered in Europe. These vaccines might induce cross-reactions in the rapid plate agglutination (RPA) SP/SG test due to shared O-antigens, possibly resulting in a lower test specificity. The extent to which the specificity of SP/SG serological tests is influenced by SE and/or ST vaccinations in the field has not been reported. In this paper, we report the diagnostic and flock specificity of the commercially available RPA SP/SG test using 1:2-1:16 serum dilutions on four panels of sera: SPF sera, field sera from flocks of varying age and SE/ST vaccination status, and reference sera from an international proficiency testing scheme. The results showed that the use of live SE/ST vaccines did not influence the specificity of the RPA SP/SG test. Inactivated vaccines showed a drop of the diagnostic specificity to 96.54% and a flock specificity of 34.1% when the 1:2 serum dilution was used. The 1:8 serum dilution showed a diagnostic specificity of 99.41% and a flock specificity of 86.4%. In conclusion, the use of SE/ST vaccines has either no effect or a modest effect on the specificity of the RPA SP/SG test used to monitor flocks. The main factors are the type of vaccine, and the serum dilution used for testing and a cut-off.

Immunological characterization of the chemically prepared ghosts of Salmonella Typhimurium as a vaccine candidate.

BACKGROUND: Bacterial ghosts are the evacuated bacterial cellular membranes from most of the genetic and protein contents which preserved their surface characters. Recently, bacterial ghosts exploited for different biomedical applications, for instance, vaccination. The purpose of this study is to measure the immunogenic protective response of bacterial ghosts of Salmonella Typhimurium in animals and to allow future testing this response in humans. The immunologic response was qualitatively, quantitatively, and functionally measured. We have measured the humoral and cellular immune responses, such as immunoglobulins elevation (IgG), increased granulocytes, serum antibacterial activity, clearance of virulence in feces and liver, and the survival rate. RESULTS: The bacterial ghosts' vaccine was able to protect 100% of subcutaneously vaccinated rats and 75% of adjuvant subcutaneously vaccinated rats. The lowest survival rate was in the orally vaccinated group (25%). The maximum level of serum IgG titers, as well as serum and feces bactericidal activity (100% eradication), was exhibited in the subcutaneously vaccinated group with adjuvant vaccines followed by the subcutaneously vaccinated one. Additionally, the highest granulocytes' number was observed in the adjuvant vaccine subcutaneously immunized group. The bacterial load in liver homogenate was eliminated in the subcutaneously vaccinated rats after the virulence challenge. CONCLUSIONS: The bacterial ghosts of Salmonella enterica serovar Typhimurium that prepared by Tween 80 Protocol showed an effective vaccine candidate that protected animals, eliminated the virulence in feces and liver. These findings show that chemically induced bacterial ghosts of Salmonella Typhimurium can be a promising vaccine.

Prebiotics can restrict Salmonella populations in poultry: a review.

Antibiotics were over the years, the common supplement used for poultry production. There is a global trend to lessen antibiotics' use due to the contamination of consumed meat with antibiotic residues. Also, there is a concern that human treatments might be jeopardized due to the emergence of antibiotic-resistant bacteria. Prebiotics are attractive supplements, particularly in poultry production, because of the diversity of their effects, including pH amendments, production of short-chain fatty acids (SCFA) and the inhibition of pathogens' growth. The commonly used prebiotics are carbohydrate sources that cannot be easily broken down by chickens. However, they can efficiently be utilized by the intestinal tract's microflora. Oligosaccharides, polysaccharides and lactose are non-digestible carbohydrate sources that are typically used in poultry diets as prebiotics. This review covers current applications and prospects for using prebiotics to improve poultry performance and reduce pathogens, particularly Salmonella, in gastrointestinal tract.

Salmonella Pullorum lacking srfA is attenuated, immunogenic and protective in chickens.

Sallmonella Pullorum is a host-restricted pathogen for poultry and causes severe economic importance in many developing countries. The development of novel vaccines for Salmonella Pullorum is necessary to eradicate the prevalence of the pathogen. In our study, a srfA deletion mutant (C79-13ΔsrfA) of Salmonella Pullorum was constructed, and then the biological characteristics and protective efficacy of the mutant were evaluated. The mutant C79-13ΔsrfA was much less virulent than its parental strain C79-13 in one-day-old HY-line white chickens, immunization with C79-13ΔsrfA (4 × 107 CFU) through oral pathway induced highly specific humoral and cellular immune responses, the growth performance of vaccinated chickens was consistent with that of unvaccinated chickens. The survival percentages of vaccinated chickens reached 90% and 80%, after challenge with Salmonella Pullorum strain C79-13 and Salmonella Gallinarum strain SG9 at 10 days post-immunization (dpi), respectively. Collectively, our results indicate that C79-13ΔsrfA is a live attenuated vaccine candidate.

Phloretin is protective in a murine salmonella enterica serovar typhimurium infection model.

Salmonella, an important zoonotic pathogen, causes significant morbidity and mortality in both humans and animals. Phloretin mainly isolated from strawberries and apples has the effects of treating inflammation and pathogenic bacteria, but its protective efficacy and mechanism of action against Salmonella spp. are less clear. In this study, we found that phloretin alleviated body weight loss, colon length shortening, and colonic pathological damage caused by S. Typhimurium. Phloretin also decreased S. Typhimurium translocation to the mesenteric lymph nodes (MLN) and spleen. Further mechanism studies showed that phloretin significantly inhibited inflammation and oxidative stress levels in the colonic tissue. Phloretin also prevented S. Typhimurium-mediated impairment in the colon epithelium barrier by the regulation ZO-1 and occludin levels. Interestingly, phloretin did not inhibit S. typhimurium growth in vitro, but reduced the internalization of S. Typhimurium into Caco-2 cells. Taken together, these findings indicated that phloretin may be a new dietary strategy to combat the disease.

Bacillus subtilis BSH has a protective effect on Salmonella infection by regulating the intestinal flora structure in chickens.

Salmonellosis is a worldwide zoonotic disease that poses a serious threat to the reproduction of livestock and poultry and the health of young animals. Probiotics including Bacillus species, have received increasing attention as a substitute for antibiotics. In this study, chicks infected with Salmonella were fed feed supplemented with the BSH to observe the pathological changes in the liver, detect the number of viable bacteria in the liver and spleen, and record the death of the chicks. The results showed that BSH could reduce the pathological changes in the liver and the invasion of Salmonella into the liver and spleen of chicks. In addition, the survival rate of chicks in the BSH experimental group was 60%, while that in the infected control group was 26%, indicating that BSH had a protective effect on chicks infected with Salmonella. Finally, the fecal microflora of 9-day-old chicks was analyzed by 16S rRNA high-throughput sequencing. The results showed that Salmonella infection could cause intestinal flora changes, while BSH could alleviate this change. In addition, BSH also promoted the proliferation of Lactobacillus salivarius in the cecum of chick. This study emphasized that BSH has anti- Salmonella infection effects in chickens and can be used as a candidate microecological preparation strain.

Antibacterial activity of ZnO quantum dots and its protective effects of chicks infected withSalmonella pullorum.

In light of emerging antibiotic resistance, synthesis of active, environmental friendly antimicrobial alternatives becomes increasingly necessary. In this study, ZnO quantum dots (ZnO QDs) were developed by the sol-gel method and characterized. The antibacterial activities of ZnO QDs againstEscherichia coli(E. coli),Staphylococcus aureus(S. aureus) andSalmonella Pullorum(S. Pullorum) were systematically investigated. Moreover, the protective effects of ZnO QDs on Salmonella-caused pullorosis in chicks were also explored. The results indicated that the size range of ZnO QDs was 3-6 nm. Antibacterial results showed that ZnO QDs treatment inhibited the growth ofE. coli,S. aureus, andS. Pullorumin the rate of 87.06 ± 0.98%, 94.75 ± 2.28%, and 85.55 ± 1.15%, respectively. Its excellent antibacterial property was manifested with the minimum inhibitory concentration of 0.7812, 0.0976, and 0.1953 mg ml-1, which may be attributed to the production of reactive oxygen species, the dissolution of Zn2+ions, and the loss of cell integrity. Furthermore, in thein vivotest, the ZnO QDs effectively reduced the mortality of chicks infected withS. Pullorumvia regulating the balance of the intestinal flora, protecting liver and intestine, and modulating the balance of antioxidation systems. This study reveals that ZnO QDs exerts remarkably antibacterial activityin vitroand anti-pullorosis effect in chicks.

Efficacy of chestnut and quebracho wood extracts to control Salmonella in poultry.

AIMS: The study was aimed to evaluate the antibacterial activity and efficacy of chestnut and quebracho wood extracts against Salmonella by in vitro assays and in vivo trials. METHODS AND RESULTS: The extracts showed inhibitory activity against Salmonella determined by the minimum inhibitory concentration method as well as on the adhesion and invasion of S. Gallinarum (SG) and S. Enteritidis (SE) in Caco-2 cells. Also, transmission electron microscopy revealed that extract-treated Salmonella showed disruption of cell walls and membranes, damage of the cytoplasm and tannin-protein aggregations. In addition, efficacy of the extracts to control SG and SE was evaluated in experimental infection trials in laying hens and broilers respectively. SE excretion was significantly reduced on days 5 (P < 0·01) and 12 (P < 0·025) only in the quebracho group. In the fowl typhoid infection model, hens that received the chestnut extract showed a significantly reduced mortality (P < 0·05). CONCLUSIONS: Our results evidence that these alternative natural products may be a useful tool to control Salmonella in poultry. SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella is a zoonotic pathogen usually associated with poultry production. This study provides information about the mechanism of antibacterial effects of chestnut and quebracho wood extracts to control Salmonella in poultry.