Effects of heat stress on the gut health of poultry.

Stress is a biological adaptive response to restore homeostasis, and occurs in every animal production system, due to the multitude of stressors present in every farm. Heat stress is one of the most common environmental challenges to poultry worldwide. It has been extensively demonstrated that heat stress negatively impacts the health, welfare, and productivity of broilers and laying hens. However, basic mechanisms associated with the reported effects of heat stress are still not fully understood. The adaptive response of poultry to a heat stress situation is complex and intricate in nature, and it includes effects on the intestinal tract. This review offers an objective overview of the scientific evidence available on the effects of the heat stress response on different facets of the intestinal tract of poultry, including its physiology, integrity, immunology, and microbiota. Although a lot of knowledge has been generated, many gaps persist. The development of standardized models is crucial to be able to better compare and extrapolate results. By better understanding how the intestinal tract is affected in birds subjected to heat stress conditions, more targeted interventions can be developed and applied.

Differential innate immune responses of bovine peripheral blood leukocytes to Salmonella enterica serovars Dublin, Typhimurium, and Enteritidis.

The majority of Salmonella serovars cause no clinical disease in cattle, while some are associated with severe disease. The objective of the current study was to determine the innate immune responses of bovine peripheral blood leukocytes exposed to Salmonella enterica serovar Dublin (bovine-specific), Salmonella typhimurium (murine adapted, but zoonotic), and Salmonella enteritidis (poultry host-adapted) in 3-week-old calves. All Salmonella exposures increased cell surface CD14 and CD18 regardless of serovar. The greatest CD14 marker mean fluorescence was in monocytes and the greatest mean fluorescent of the marker mean was in neutrophils. Phagocytosis increased with all serovars, but was not different among them. Neutrophils had the greatest marker mean fluorescence for phagocytosis, with all serovars being equal. Oxidative burst increased in all serovars compared to control cells, but were not different among the serovars. Neutrophils and monocytes were similar in the oxidative burst, with limited oxidative burst detected in the primarily lymphocyte population. mRNA expression of TNF-α, IL-8, and IL-12, increased above the control cells whereas none of these serovars affected mRNA expression of TLR4. TNF-α was greatest in S. enterica and S. typhimurium, compared to Salmonella dublin. In contrast, IL-8 was expressed more in S. dublin than S. typhiurium, with S. Enteriditus intermediary. These results show while cell surface markers, phagocytosis, and oxidative burst were largely unaffected by serovar, cytokine and chemokine expression differed among the Salmonella serovars. It appears that internal responses of the cells differ, rather than cell recognition, creating pathogenicity differences among of the serovars, even in the neonate with developing immunity.

Isolation, characterization, and application of bacteriophages for Salmonella spp. biocontrol in pigs.

Foodborne illness due to Salmonella-contaminated pork products is an important public health problem, causing significant economic losses worldwide. The use of bacteriophages is a potential intervention tool that has attracted interest for the control of foodborne pathogens. The objective of this study was to detect the presence of Salmonella in commercial pig farms and to isolate specific autochthonous bacteriophages against Salmonella Typhimurium, to characterize them and to evaluate their lytic capacity against Salmonella Typhimurium in vivo and in vitro. Salmonella was isolated on 50% (4/8) of the farms, with serotype Typhimurium being the most prevalent, detected in 48.2% of samples (13/27). The isolated Salmonella Typhimurium bacteriophages belong to the Podoviridae family, were active against serotypes Abony, Enteritidis, Typhi, and Typhimurium, but not against serotypes Arizonae, Cholerasuis, Gallinarum, and Pullorum. In in vitro tests, bacteriophage at 10(7) PFU/mL and 10(9) PFU/mL significantly reduced (p<0.05) Salmonella Typhimurium counts in 1.6 and 2.5 log10 colony-forming units (CFU)/mL, respectively, after 24 h. Before the in vivo treatment with bacteriophages, Salmonella was identified in 93.3% (28/30) of the fecal samples from the pigs inoculated with 10(6) CFU/mL, and only in 56.6% (17/30) after the treatment consisting of oral administration of the pool of the bacteriophages after the fasting period, simulating a common preslaughter practice. These results indicate that the pool of bacteriophages administered was capable of reducing the colonization of Salmonella in pigs.

Impact of Heat Stress on Poultry Production.

Understanding and controlling environmental conditions is crucial to successful poultry production and welfare. Heat stress is one of the most important environmental stressors challenging poultry production worldwide. The detrimental effects of heat stress on broilers and laying hens range from reduced growth and egg production to decreased poultry and egg quality and safety. Moreover, the negative impact of heat stress on poultry welfare has recently attracted increasing public awareness and concern. Much information has been published on the effects of heat stress on productivity and immune response in poultry. However, our knowledge of basic mechanisms associated to the reported effects, as well as related to poultry behavior and welfare under heat stress conditions is in fact scarce. Intervention strategies to deal with heat stress conditions have been the focus of many published studies. Nevertheless, effectiveness of most of the interventions has been variable or inconsistent. This review focuses on the scientific evidence available on the importance and impact of heat stress in poultry production, with emphasis on broilers and laying hens.

Direct feeding of microencapsulated bacteriophages to reduce Salmonella colonization in pigs.

Salmonella shedding often increases in pigs after transportation and/or lairage. We previously showed that administering anti-Salmonella bacteriophages to pigs by gavage significantly reduced Salmonella colonization when the pigs were exposed to a Salmonella-contaminated holding pen. Here we tested whether a microencapsulated phage cocktail would remain effective if the treatment was administered to pigs in the feed. Pigs (n=21) were randomly placed into three groups: feed, gavage, and control. The feed group was direct-fed a microencapsulated phage cocktail daily for 5 days. On the fifth day, the gavage group received the same phage cocktail by gavage, whereas control pigs received a mock treatment containing no phage. All pigs were then orally challenged with Salmonella enterica serovar Typhimurium. Fecal swab samples were collected every 2 h. At 6 h postchallenge, all pigs were euthanized, and ileal and cecal contents and mesenteric lymph nodes were collected and analyzed for the challenge organism. Pigs in the feed group were less likely to shed Salmonella Typhimurium at 2 h (38.1%) and 4 h (42.9%) postchallenge than pigs in both the gavage (2 h: 71.4%; 4 h: 81.1%) and control (2 h: 71.4%; 4 h: 85.7%) groups (p<0.05). Likewise, concentrations of Salmonella Typhimurium in ileal (2.0 log(10) colony forming units [CFU]/mL [contents]) and cecal (2.7 log(10) CFU/mL) contents from feed pigs were lower than ileal (3.0 log(10) CFU/mL) and cecal (3.7 log(10) CFU/mL) contents from control pigs. High concentrations of anti-Salmonella phages were detected in ileal and cecal contents from both feed and gavage pigs (feed ileal: 1.4×10(6); feed cecal 8.5×10(6); gavage ileal 2.0×10(4); gavage cecal: 2.2×10(3)). It is concluded that direct feeding of microencapsulated phages is a practical and effective means of reducing Salmonella colonization and shedding in pigs.

Immunological, physiological, and behavioral effects of Salmonella enterica carriage and shedding in experimentally infected finishing pigs.

Finishing pigs infected with Salmonella pose significant food safety risks by carrying the pathogen into abattoirs. This study was conducted to determine the dynamics of Salmonella infection in finishing pigs, and associated immunological, physiological, and behavioral alterations, by longitudinally comparing infected to noninfected pigs during 6 weeks postinfection (p.i.). Bacteriological data revealed that all inoculated pigs started shedding Salmonella within 2 h p.i., and persistently shed the bacteria up to the end of the study. Ileal and cecal contents, as well as mesenteric lymph node samples, were all positive throughout the study, containing 3-4 log(10) cfu/g of Salmonella at 24 h p.i., and 4-5 log(10) cfu/g of Salmonella up to 4 weeks p.i. Levels of Salmonella dropped markedly (p < 0.05) in all samples at 5 weeks p.i. There was no difference between groups for blood cell counts. Tumor necrosis factor-α was greater (p < 0.05) in infected pigs: (1) in the mesenteric lymph nodes by 48 h p.i.; (2) at 24 h and 3 weeks p.i. in the ileum; and (3) in the cecum and spleen at 3 weeks p.i. Interleukin-12, interleukin-1 and its antagonist, and a porcine-specific antimicrobial peptide RNA expression in tissues changed over time, but were not different between groups. Infected pigs spent more time in ventral recumbency, standing, and sitting than controls (p < 0.01). Infected pigs were also more active (p < 0.01), and approached a novel object more quickly than control pigs (p < 0.05). No treatment differences were detected for rectal temperature or plasma cortisol (p > 0.10). This study shows that finishing pigs can carry high levels of Salmonella for up to 4 weeks p.i. in the gastrointestinal contents and mesenteric lymph nodes, shedding high levels of the bacteria without developing clinical symptoms, but developing an immune response throughout the intestinal tract. Moreover, subtle behavioral changes measured as postures were detected, and therefore warrant additional investigation.

Phage therapy to reduce preprocessing Salmonella infections in market-weight swine.

Contamination of meat products with food-borne pathogens usually results from the carcass coming in contact with the feces of an infected animal during processing. In the case of Salmonella, pigs can become colonized with the organism during transport and lairage from contaminated trailers and holding pens, resulting in increased pathogen shedding just prior to processing. Increased shedding, in turn, amplifies the likelihood of carcass contamination by magnifying the amount of bacteria that enters the processing facility. We conducted a series of experiments to test whether phage therapy could limit Salmonella infections at this crucial period. In a preliminary experiment done with small pigs (3 to 4 weeks old; 30 to 40 lb), administration of an anti-Salmonella phage cocktail at the time of inoculation with Salmonella enterica serovar Typhimurium reduced Salmonella colonization by 99.0 to 99.9% (2- to 3-log reduction) in the tonsils, ileum, and cecum. To test the efficacy of phage therapy in a production-like setting, we inoculated four market-weight pigs (in three replicates) with Salmonella enterica serovar Typhimurium and allowed the challenged pigs to contaminate a holding pen for 48 h. Sixteen naïve pigs were randomly split into two groups which received either the anti-Salmonella phage cocktail or a mock treatment. Both groups of pigs were comingled with the challenged pigs in the contaminated pen. Treatment with the anti-Salmonella phage cocktail significantly reduced cecal Salmonella concentrations (95%; P<0.05) while also reducing (numerically) ileal Salmonella concentrations (90%; P=0.06). Additional in vitro studies showed that the phage cocktail was also lytic against several non-Typhimurium serovars.

Can stress in farm animals increase food safety risk?

All farm animals will experience some level of stress during their lives. Stress reduces the fitness of an animal, which can be expressed through failure to achieve production performance standards, or through disease and death. Stress in farm animals can also have detrimental effects on the quality of food products. However, although a common assumption of a potential effect of stress on food safety exists, little is actually known about how this interaction may occur. The aim of this review was to examine the current knowledge of the potential impact of stress in farm animals on food safety risk. Colonization of farm animals by enteric pathogens such as Escherichia coli O157:H7, Salmonella, and Campylobacter, and their subsequent dissemination into the human food chain are a major public health and economic concern for the food industries. This review shows that there is increasing evidence to demonstrate that stress can have a significant deleterious effect on food safety through a variety of potential mechanisms. However, as the impact of stress is difficult to precisely determine, it is imperative that the issue receives more research attention in the interests of optimizing animal welfare and minimizing losses in product yield and quality, as well as to food safety risks to consumers. While there is some evidence linking stress with pathogen carriage and shedding in farm animals, the mechanisms underlying this effect have not been fully elucidated. Understanding when pathogen loads on the farm are the highest or when animals are most susceptible to infection will help identifying times when intervention strategies for pathogen control may be most effective, and consequently, increase the safety of food of animal origin.

Split marketing as a risk factor for Salmonella enterica infection in swine.

On-farm reduction of Salmonella carriage prevalence in pigs requires the identification of risk factors to direct interventions development. This study was designed to determine if split marketing of finishing pigs constitutes a risk factor for Salmonella infections, by comparing Salmonella prevalence in the first group of pigs selected for harvest ("first pull") versus the prevalence in the last group of pigs selected for harvest ("close out") from multiple commercial finishing lots. Nine paired samplings were conducted consisting in matched groups of pigs from individual barns as the first pull and the close out with a 4-week interval between groups. From each group, fecal and meat samples were collected, on-farm and at harvest, respectively. Fecal samples were selectively enriched, and analyzed for the presence of Salmonella, whereas meat juice samples were analyzed for the presence of antibodies against Salmonella. In 7/9 (77.8%) of the studied barns, an increase in Salmonella prevalence was observed, based on both bacteriologic and serologic analysis. Overall, there was an increase of 9.2% (p < 0.05) in bacteriologic prevalence, and 31.3% (p < 0.05) in serologic prevalence from first pull to close out groups. This study demonstrates that a significant increase in Salmonella prevalence occurs between the first and the last group of pigs harvested from finishing lots, with close out groups of market pigs posing a higher risk for Salmonella contaminations.

Comparison of five culture methods for Salmonella isolation from swine fecal samples of known infection status.

The current study was conducted to evaluate 5 bacteriologic culture methods (methods 1, 2, 3, 4, and 5) for recovery of Salmonella enterica from swine feces, both for sensitivity of detection (ability to recover Salmonella from a positive sample) and for specificity (not to inadvertently identify an organism as a Salmonella species in a negative sample). Fifty-six negative samples and 46 positive samples were processed using each of the 5 methods, which differed primarily in the combinations of enrichment media used. All negative samples were negative for Salmonella when cultured by all 5 methods (100% specificity). Two of the methods (methods 1 and 4) resulted in the recovery of significantly less (P < 0.05) Salmonella when compared with the remaining 3 methods (methods 2, 3, and 5). No one method was successful in recovering Salmonella from all positive samples, although recovery with method 2 was statistically similar to the total number of positive samples analyzed (42 vs. 46 Salmonella-positive samples, P > 0.05). This study shows that culture methods differ significantly in their performance regarding the isolation of Salmonella from swine fecal samples.

Effect of preslaughter events on the prevalence of Salmonella in market-weight turkeys.

The goal of this study was to determine if preslaughter events, such as transport to and holding at the slaughterhouse, affect Salmonella prevalence in turkeys. Floors of transport crates were swabbed after loading and prior to transport at the farm (time 1, n = 100 swabs per trial) and after transport to and holding at the abattoir (time 2, n = 100 swabs per trial). In addition, environmental samples were taken at each of the six premises (n = 25 per premises) as well as in the holding shed at the abattoir (n = 25 samples per trial). At slaughter, the crops, ceca, and spleens were cultured (n = 50 each per flock). As shown from the culture of the crate floor swabs collected pre- and posttransport, when individual farms were analyzed, samples from only one premises exhibited a statistically significant change, as seen by the decline in Salmonella prevalence posttransport (P < 0.01). When the data from all farms were combined, Salmonella was recovered more frequently from swabs collected pretransport at loading on-farm (time 1, 47.6%) than from swabs collected after transport (time 2, 39.7%, P < 0.01). This suggests that transport to and holding at the abattoir do not increase the prevalence of Salmonella in turkeys. This observation contrasts with the increase in Salmonella prevalence reported for hogs and some broilers.

Culture methods differ on the isolation of Salmonella enterica serotypes from naturally contaminated swine fecal samples.

Four culture methods (A, B, C, and D) were comparatively evaluated for their ability to isolate Salmonella enterica from pooled swine fecal samples (n = 100). None of the methods was able to isolate Salmonella from all positive samples. The relative sensitivity of the culture methods evaluated was 82%, 94%, 95%, and 78% for methods A, B, C, and D, respectively. The comparison of sensitivities showed that methods B and C performed significantly better (P < 0.05) than methods A and D. Although relative sensitivities of methods B and C were equal, from the 89 positive samples concomitantly detected by both, 35 (39.3%) had different serotypes (no match) isolated by each method. On the basis of the results of this study, it was concluded that culture methods differ on the isolation of S. enterica serotypes from naturally contaminated swine fecal samples. Depending on the objective(s) of investigations on the ecology and epidemiology of S. enterica in swine populations, a method or a combination of methods should be considered for more reliable results.

Resting pigs on transport trailers as an intervention strategy to reduce Salmonella enterica prevalence at slaughter.

Recent research has shown that much preharvest Salmonella enterica infection in pigs occurs immediately before slaughter during this rest period in the contaminated abattoir holding pens. The objective of this study was to evaluate a potential intervention strategy to reduce the prevalence of S. enterica-positive pigs at slaughter, which consisted of resting pigs prior to slaughter on their transport vehicle, instead of in the abattoir holding pen. Additionally, the effect of transportation of pigs from farm to the abattoir on S. enterica prevalence was investigated. A total of 120 animals were included in the experiment, divided in four replicates (n = 30 pigs per replicate). Fecal samples were collected from each animal at the farm and at the abattoir, where 15 randomly chosen pigs were unloaded and moved to a holding pen, while the remaining 15 pigs stayed in the transport trailer. After approximately 1.5 h of resting, both groups were slaughtered. Samples collected included distal ileum portion, cecal contents, and ileocecal lymph node. The overall S. enterica prevalence (pigs positive in at least one of the samples collected at slaughter) was higher for pigs held in the abattoir pens (40.7% versus 13.3%, P < 0.05). There was no difference (P > 0.05) for the S. enterica prevalence before and after transportation from farm to abattoir (5.8% versus 0.8%, respectively). This study demonstrates that resting pigs on the transport vehicle has the potential to decrease S. enterica levels entering the abattoir.