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Abstract Background Less than half of U.S. hospitals meet guidelines for prompt treatment of ST-segment elevation myocardial infarction (STEMI). The Door-to-Balloon (D2B) Alliance is a collaborative effort of more than 900 hospitals committed to implementing a set of evidence-based strategies for reducing D2B time. This study presents data on (1) the prevalence of evidence-based strategies in U.S. hospitals that participated in the D2B Alliance and (2) identifies key hospital characteristics associated with their use. Methods We conducted a cross-sectional study of U.S. hospitals that joined the D2B Alliance through a Web-based survey about their current practices for patients with STEMI who received primary percutaneous coronary intervention (PCI). We used multivariate logistic regression to identify hospital characteristics associated with use of each strategy. Results Of the 915 U.S. hospitals enrolled in the D2B Alliance as of June 2007, 797 (87%) completed the survey. Only 30.4% of responding hospitals reported employing at least 4 of the 5 key strategies (emergency medicine activates catheterization laboratory, single-call activation, expectation that catheterization team is available in the laboratory within 20–30 minutes after page, prompt data feedback on D2B times, use of pre-hospital electrocardiograms to activate the laboratory while the patient is en route to the hospital); 9.3% employed none of the strategies. There was no clear pattern of correlation between hospital characteristics and reported strategies. Conclusion As of 2007, many hospitals had implemented few of the key strategies to reduce D2B time, suggesting substantial opportunity to improve care for patients with STEMI.

Maintenance of intestinal health is critical to successful poultry production and one of the goals of the poultry production industry. For decades the poultry industry has relied upon the inclusion of antibiotic growth promoters (AGP) to achieve this goal and improve growth performance. With the removal of AGPs, the emergence of chronic, low-level gut inflammation has come to the forefront of concern in the poultry industry with the diet being the primary source of inflammatory triggers. We have developed a dietary model of low-grade, chronic intestinal inflammation in broilers that employs feeding a high nonstarch polysaccharides (NSP) diet composed of 30% rice bran to study the effects of this inflammation on bird performance and physiology. For the present studies, we hypothesize that the low-grade chronic inflammation causes neurons in the intestinal enteric nervous system to secrete neurochemicals that activate immune cells that drive the inflammation and negatively affect bird performance. To test our hypothesis, 1-day-old broiler chickens were weighed and divided into 2 dietary regimes: a control corn-soybean diet and a group fed a high NSP diet (30% rice bran). At 7-, 14-, 21-, and 28-d posthatch (PH), birds were weighed, fecal material collected, and 5 birds were sacrificed and sections of duodenal and cecal tissues excised, and duodenal and cecal contents collected for ultra-high performance liquid chromatography analyses (UHPLC). UHPLC revealed 1000s-fold increase in the concentration of norepinephrine (NOR) in birds fed the high NSP diet compared to the control fed birds. Further, the fecal concentrations of NOR were also found to be significantly elevated in the birds on the NSP diet throughout all time points. There were no differences in weight gain nor feed conversion from 1 to 14 d PH, but birds fed the high NSP diet had significantly reduced weight gain and feed conversion from 14 to 28 d PH. The results revealed that a dietary-induced low-grade chronic inflammatory response increased NOR production in the gut which negatively affected bird performance. This study suggests that neuroimmune pathways may serve as a mechanistic target for the development of new interventions to decrease the incidence of chronic inflammation and thereby benefit performance., Competing Interests: DISCLOSURES The authors declare no conflict of interest with the data in this manuscript., (Published by Elsevier Inc.)

The avian immune system responds to Salmonella infection by expressing cytokines and chemokines. We hypothesized that the immune status of Salmonella Typhimurium (ST) challenged neonatal broilers would differ from the uninfected treatment. The objective of this experiment was to evaluate 12 cytokines. Day of hatch male chicks were randomly allocated into a control or ST challenged group. At day three of age, sterile diluent or 5.0 × 10 8  CFU of ST was given orally to each chick. Blood was obtained 24 h post challenge and serum separated for later analysis (n = 30 chicks/treatment). Significant ( p ≤ 0.05) increases in pro-inflammatory cytokines -interleukin-6 (IL-6), IL-16, and IL-21; anti-inflammatory cytokines - IL-10; chemokines -regulated on activation, normal T cell expressed and secreted (RANTES), macrophage inflammatory protein-1β (MIP-1β), and MIP-3α; colony stimulating factors -macrophage colony-stimulating factor (M-CSF); and growth factors -vascular endothelial growth factor (VEGF) were observed in the serum of the challenged chicks when compared to the control. No significant differences were observed in IL-2, interferon gamma (IFNγ), and IFNα. These data indicate the detection of mucosal immune responses in broiler chickens following ST infection. The heightened levels of pro-inflammatory cytokines, chemokines, and colony stimulating factors align with known inflammatory mechanisms, like the influx of immune cells. However, the elevation of IL-10 was unexpected, due to its immunoregulatory properties. Notably, the rise in VEGF levels is compelling, as it suggests the possibility of tissue repair and angiogenesis in ST infected birds., Competing Interests: Authors CL and MM were employed by the company Millipore Sigma. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Milby-Blackledge, Farnell, Zhao, Berghman, Laino, Muller, Byrd and Farnell.)

Human foodborne outbreaks with antibiotic-resistant Salmonella enterica associated with contaminated poultry products have recently involved serogroup C serovars Infantis and Hadar. The current study evaluated a commercially available Salmonella vaccine for cross-protection against Infantis and Hadar serovars in turkeys. The live, attenuated S. Typhimurium (serogroup B) vaccine significantly reduced colonization of intestinal tissues (cecum, cecal tonsils, and cloaca) by serovars Infantis (C1) and Hadar (C2) and significantly limited systemic dissemination to the spleen. S. Infantis, but not S. Hadar, disseminated to bone marrow in non-vaccinated turkeys, but vaccination prevented S. Infantis dissemination to the bone marrow. The S. Infantis challenge strain contained the pESI megaplasmid, and virulence mechanism(s) residing on this plasmid may support dissemination and/or colonization of systemic niches such as myeloid tissue. Collectively, the data indicate that vaccinating turkeys with the serogroup B S. Typhimurium vaccine limited intestinal colonization and systemic dissemination by serogroup C serovars Infantis and Hadar., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Shawn Bearson reports financial support was provided by USDA Food Safety and Inspection Service., (Copyright © 2023. Published by Elsevier Ltd.)

This study quantifies setup uncertainty in brain tumor patients who received image-guided proton therapy. Patients analyzed include 165 children, adolescents, and young adults (median age at radiotherapy: 9 years (range: 10 months to 24 years); 80 anesthetized and 85 awake) enrolled in a single-institution prospective study from 2020 to 2023. Cone-beam computed tomography (CBCT) was performed daily to calculate and correct manual setup errors, once per course after setup correction to measure residual errors, and weekly after treatments to assess intrafractional motion. Orthogonal radiographs were acquired consecutively with CBCT for paired comparisons of 40 patients. Translational and rotational errors were converted from 6 degrees of freedom to a scalar by a statistical approach that considers the distance from the target to the isocenter. The 95th percentile of setup uncertainty was reduced by daily CBCT from 10 mm (manual positioning) to 1-1.5 mm (after correction) and increased to 2 mm by the end of fractional treatment. A larger variation existed between the roll corrections reported by radiographs vs. CBCT than for pitch and yaw, while there was no statistically significant difference in translational variation. A quantile mixed regression model showed that the 95th percentile of intrafractional motion was 0.40 mm lower for anesthetized patients (p=0.0016). Considering additional uncertainty in radiation-imaging isocentricity, the commonly used total plan robustness of 3 mm against positional uncertainty would be appropriate for our study cohort.

Salmonella enterica is a group of facultative, gram-negative bacteria. Recently, new evidence indicated that Salmonella could reprogram the host metabolism to increase energy or metabolites available for intracellular replication. In this study, using a chicken-specific kinomic immunometabolism peptide array analysis, we found that infection by S. Enteritidis induced significant phosphorylation changes in many key proteins of the glycolytic pathway in chicken macrophage HD-11 cells, indicating a shift in glycolysis caused by Salmonella infection. Nitric oxide production and changes of glycolysis and mitochondrial oxidative phosphorylation (OXPHOS) represented by extracellular acidification rate (ECAR) and oxygen consumption rate (OCR), respectively, were measured in chicken macrophages infected with three Salmonella strains ( S . Enteritidis, S. Heidelberg, and S . Senftenberg). The infection reduced glycolysis and enhanced OXPHOS in chicken macrophages as indicated by changes of ECAR and OCR. Salmonella strains differentially affected macrophage polarization and glycolysis. Among three strains tested, S . Enteritidis was most effective in downregulating glycolysis and promoting M2 polarization as measured by ECAR, ORC, and NO production; while S . Senftenberg did not alter glycolysis and may promote M1 polarization. Our results suggested that downregulation of host cell glycolysis and increase of M2 polarization of macrophages may contribute to increased intracellular survival of S. Enteritidis.

Previously, the supplementation of a microencapsulated blend of organic acids and botanicals improved the health and performance of broiler breeders under non-challenged conditions. This study aimed to determine if the microencapsulated blend impacted dysbiosis and necrotic enteritis (NE) in broiler breeders. Day-of-hatch chicks were assigned to non-challenge and challenge groups, provided a basal diet supplemented with 0 or 500 g/MT of the blend, and subjected to a laboratory model for NE. On d 20-21, jejunum/ileum content were collected for microbiome sequencing (n = 10; V4 region of 16S rRNA gene). The experiment was repeated (n = 3), and data were analyzed in QIIME2 and R. Alpha and beta diversity, core microbiome, and compositional differences were determined (significance at p ≤ 0.05; Q ≤ 0.05). There was no difference between richness and evenness of those fed diets containing 0 and 500 g/MT microencapsulated blend, but differences were seen between the non-challenged and challenged groups. Beta diversity of the 0 and 500 g/MT non-challenged groups differed, but no differences existed between the NE-challenged groups. The core microbiome of those fed 500 g/MT similarly consisted of Lactobacillus and Clostridiaceae. Furthermore, challenged birds fed diets containing 500 g/MT had a higher abundance of significantly different phyla, namely, Actinobacteriota , Bacteroidota , and Verrucomicrobiota , than the 0 g/MT challenged group. Dietary supplementation of a microencapsulated blend shifted the microbiome by supporting beneficial and core taxa.

Addition of vitamins and antioxidants has been long associated with increased immunity and are commonly used in the poultry industry; however, less is known regarding their use in broiler breeder hens. The objective of this study was to determine if feeding a complex of protected biofactors and antioxidants composed of vitamins and fermentation extracts to broiler breeder hens conferred resistance against Salmonella enterica serovar Enteritidis (S. Enteritidis) in the progeny chicks. Three-day-old chicks from control- and supplement-fed hens were challenged with S. Enteritidis and necropsied 4- and 11-days postchallenge (dpc) to determine if there were differences in invasion and colonization. Serum and jejunum were evaluated for various cytokine and chemokine production. Fewer (P = 0.002) chicks from supplement-fed hens had detectable S. Enteritidis in the ceca (32.6%) compared to chicks from control-fed hens (64%). By 11 dpc, significantly (P < 0.001) fewer chicks from supplement-fed hens were positive for S. Enteritidis (liver [36%]; ceca [16%]) compared to chicks from the control hens (liver [76%]; ceca [76%]). The recoverable S. Enteritidis in the cecal content was also lower (P = 0.01) at 11 dpc. In additional to the differences in invasion and colonization, cytokine and chemokine production were distinct between the 2 groups of chicks. Chicks from supplement-fed hens had increased production of IL-16, IL-6, MIP-3α, and RANTES in the jejunum while IL-16 and MIP-1β were higher in the serum of chicks from the control-fed hens. By 11 dpc, production of IFN-γ was decreased in the jejunum of chicks from supplement-fed hens. Collectively, these data demonstrate adding a protected complex of biofactors and antioxidants to the diet of broiler breeder hens offers a measure of transgenerational protection to the progeny against S. Enteritidis infection and reduces colonization that is mediated, in part, by a robust and distinct cytokine and chemokine response locally at the intestine and systemically in the blood., (Published by Elsevier Inc.)

Poultry litter is a valuable crude protein feedstuff for ruminants, but it must be treated to kill pathogens before feeding. Composting effectively kills pathogens, but it risks losing ammonia to volatilization or leaching during degradation of uric acid and urea. Hops bitter acids also exert antimicrobial activity against certain pathogenic and nitrogen-degrading microbes. Consequently, the present studies were conducted to test if adding bitter acid-rich hop preparations to simulated poultry litter composts may improve nitrogen retention while simultaneously improving pathogen killing. Results from an initial study, testing doses of Chinook or Galena hops preparations designed to each deliver 79 ppm hops β-acid, revealed that, after nine days simulated composting of wood chip litter, ammonia concentrations were 14% lower ( p < 0.05) in Chinook-treated composts than untreated composts (13.4 ± 1.06 µmol/g). Conversely, urea concentrations were 55% lower ( p < 0.05) in Galena-treated than untreated composts (6.2 ± 1.72 µmol/g). Uric acid accumulations were unaffected by hops treatments in this study but were higher ( p < 0.05) after three days than after zero, six, or nine days of composting. In follow-up studies, Chinook or Galena hops treatments (delivering 2042 or 6126 ppm of β-acid, respectively) for simulated composts (14 days) of wood chip litter alone or mixed 3:1 with ground Bluestem hay ( Andropogon gerardii ) revealed that these higher dosages had little effect on ammonia, urea, or uric acid accumulations when compared to untreated composts. Volatile fatty acid accumulations measured in these later studies were affected by the hops treatments, with butyrate accumulations being lower after 14 days in hops-treated composts than in untreated compost. In all studies, beneficial effects of Galena or Chinook hops treatments were not observed on the antimicrobial activity of the simulated composts, with composting by itself decreasing ( p < 0.05) counts of select microbial populations by more than 2.5 log 10 colony forming units/g compost dry matter. Thus, while hops treatments had little effect on pathogen control or nitrogen retention within the composted litter, they did lessen accumulations of butyrate, which may prevent adverse effects of this fatty acid on palatability of litter fed to ruminants.

Using a previously characterized and described abdominal model to define the avian immune response to Salmonella intra-abdominal challenge in chickens, we have adapted this technique for the study of chickens' immune response to a Campylobacter intra-abdominal challenge. The intra-abdominal Campylobacter infection model facilitates the characterization of peripheral blood leukocyte dynamics and abdominal cell infiltrates. Day-of-hatch Leghorn chickens were injected intra-abdominally (IA) with Campylobacter jejuni [(CJ)1 × 10 8 colony-forming units (CFUs)]. Changes in peripheral blood leukocyte numbers and abdominal cell infiltrates were monitored at 0, 4, 8, and 24 h post-injection. Peripheral blood leukocyte numbers were also determined for 2 h post-injection. For mortality studies, birds were injected intra-abdominally with 1 × 10 8 CFUs CJ and mortalities were recorded for 72 h post-injection. In the peripheral blood of CJ-injected chicks, total white blood cell (WBC) numbers began increasing by 2 h post-injection, peaking at 4 h post-injection with the predominant cell type being polymorphonuclear leukocytes (heterophils). Total WBCs declined after 8 h and this decline continued at 24 h, with total WBC numbers approaching control values. The injection of CJ into the abdominal cavity caused a rapid rise in abdominal cell infiltrates with the predominant infiltrating leukocytes being heterophils. Peak abdominal heterophil infiltrates were observed at 8 h post-injection, declining only slightly by 24 h post-injection. Mortality in the CJ challenge groups reached 37%. Mortality in the Salmonella enteritidis positive control groups were greater than 50%. The data suggest that Campylobacter infection does stimulate the innate immune response in chickens when administered IA, however, the immune response and infection is not characterized with the high levels of mortality observed with a Salmonella infection. These data provide a basis for a more definitive characterization of chickens' immune response to Campylobacter and a model to evaluate intervention strategies to prevent the infection and colonization of poultry.

Poultry is a major source of human foodborne illness caused by broad host range Salmonella serovars (paratyphoid), and developing cost-effective, pre-harvest interventions to reduce these pathogens would be valuable to the industry and consumer. Host responses to infectious agents are often regulated through phosphorylation. However, proteomic mechanisms of Salmonella acute infection biology and host responses to the bacteria have been limited concentrating predominately on the genomic responses of the host to infection. Our recent development of chicken-specific peptide arrays for kinome analysis of host phosphorylation-based cellular signaling responses provided us with the opportunity to develop a more detailed understanding of the early (4-24 h post-infection) host-pathogen interactions during the initial colonization of the cecum by Salmonella. Using the chicken-specific kinomic immune peptide array, biological pathway analysis showed infection with S . Enteritidis increased signaling related to the innate immune response, relative to the non-infected control ceca. Notably, the acute innate immune signaling pathways were characterized by increased peptide phosphorylation (activation) of the Toll-like receptor and NOD-like receptor signaling pathways, the activation of the chemokine signaling pathway, and the activation of the apoptosis signaling pathways. In addition, Salmonella infection induced a dramatic alteration in the phosphorylation events of the JAK-STAT signaling pathway. Lastly, there is also significant activation of the T cell receptor signaling pathway demonstrating the initiation of the acquired immune response to Salmonella infection. Based on the individual phosphorylation events altered by the early Salmonella infection of the cecum, certain conclusions can be drawn: (1) Salmonella was recognized by both TLR and NOD receptors that initiated the innate immune response; (2) activation of the PPRs induced the production of chemokines CXCLi2 (IL-8) and cytokines IL-2, IL-6, IFN-α, and IFN-γ; (3) Salmonella infection targeted the JAK-STAT pathway as a means of evading the host response by targeting the dephosphorylation of JAK1 and TYK2 and STAT1,2,3,4, and 6; (4) apoptosis appears to be a host defense mechanism where the infection with Salmonella induced both the intrinsic and extrinsic apoptotic pathways; and (5) the T cell receptor signaling pathway activates the AP-1 and NF-κB transcription factor cascades, but not NFAT., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Kogut, Genovese, Byrd, Swaggerty, He, Farnell and Arsenault.)

Necrotic enteritis (NE) is a devastating disease that has seen a resurgence of cases following the removal of antibiotics from feed resulting in financial loss and significant animal health concerns across the poultry industry. The objective was to evaluate the efficacy of a microencapsulated blend of organic (25% citric and 16.7% sorbic) acids and botanicals (1.7% thymol and 1% vanillin [AviPlusP]) to reduce clinical NE and determine the signaling pathways associated with any changes. Day-of-hatch by-product broiler breeder chicks were randomly assigned to a control (0) or supplemented (500 g/MT) diet (n = 23-26) and evaluated in a NE challenge model (n = 3). Birds were administered 2X cocci vaccine on d 14 and challenged with a cocktail of Clostridium perfringens strains (10 7 ) on d 17 to 19. On d 20 to 21 birds were weighed, euthanized, and scored for NE lesions. Jejunal tissue was collected for kinome analysis using an immuno-metabolism peptide array (n = 5; 15/treatment) to compare tissue from supplement-fed birds to controls. Mortality and weight were analyzed using Student's t test and lesion scores analyzed using F-test two-sample for variances (P < 0.05). The kinome data was analyzed using PIIKA2 peptide array analysis software and fold-change between control and treated groups determined. Mortality in the supplemented group was 47.4% and 70.7% in controls (P = 0.004). Lesions scores were lower (P = 0.006) in supplemented birds (2.47) compared to controls (3.3). Supplement-fed birds tended (P = 0.19) to be heavier (848.6 g) than controls (796.2 g). Kinome analysis showed T cell receptor, TNF and NF-kB signaling pathways contributed to the improvements seen in the supplement-fed birds. The following peptides were significant (P < 0.05) in all 3 pathways: CHUK, MAP3K14, MAP3K7, and NFKB1 indicating their importance. Additionally, there were changes to IL6, IL10, and IFN- γ mRNA expression in tissue between control- and supplement-fed chickens. In conclusion, the addition of a microencapsulated blend of organic acids and botanicals to a broiler diet reduced the clinical signs of NE that was mediated by specific immune-related pathways., (Published by Elsevier Inc.)

Salmonella is a pathogen normally found in the gastrointestinal tract of poultry. The objective of this study was to determine changes in avian β-defensin (AvBD) and liver-enriched antimicrobial peptide 2 (LEAP2) mRNA following Salmonella challenge. Day of hatch chicks were challenged with 10 6 , 10 7 or 10 8 colony-forming units (cfu) of Salmonella typhimurium . There were dose-, tissue- and age-specific changes in AvBD and LEAP2 mRNA. At 1-day post-infection (dpi) there was a transient upregulation of AvBD1, 8, 10 and 12 mRNA in the 10 8 cfu group. At 5 dpi, all seven AvBD mRNA were downregulated in the ileum, while only AvBD1, 6, 10 and 11 mRNA were downregulated in the jejunum and AvBD6, 8, 10, 12 and 13 were downregulated in the cecum. At 7 dpi, there was downregulation of all seven AvBD mRNA in the duodenum and downregulation of selected AvBD in the jejunum, ileum and cecum. LEAP2 mRNA was downregulated at all doses of Salmonella in the cecum at 1 dpi and in the ileum at 5 dpi. In summary, Salmonella infection caused an initial upregulation followed by a downregulation of AvBD mRNA.

Salmonella Heidelberg (SH) on contaminated poultry causes economic and health risks to producers and consumers. We hypothesized that sodium bisulfate (SBS) would decrease SH biofilm on polyvinyl chloride (PVC) coupons and decrease the horizontal transfer of SH in broilers. Experiment 1: Salmonella Heidelberg biofilm was cultured with PVC coupons, which were treated with SBS at a pH of 3.5 for 10 min, 8 h, and 24 h. Experiment 2: Nine replicate pens per treatment were divided between two rooms. A seeder contact model was used to mimic a natural infection environment. Treatments consisted of tap water or sodium bisulfate in water at a pH of 3.5. Salmonella Heidelberg incidence and enumeration were measured in crops and ceca. Sodium bisulfate significantly reduced biofilm by 2.16 and 1.04 logs when treated for 8 and 24 h, respectively. Crop colonization was significantly decreased in trials 1 and 2 by 0.29 and 0.23 logs, respectively. Crop pH was significantly decreased in trial 2. Ceca colonization was significantly decreased in trial 1 by 0.39 logs. The results from the present study suggest that SBS may be administered to drinking water to decrease SH gut colonization and to reduce biofilm.

With the increasing demand for broiler meat, a thorough evaluation of the microbial community within the broiler houses and sites where litter is deposited is critical to animal and environmental wellbeing. However not much is known in this arena, so our work evaluates the litter bacterial microbial community within a house over a 2.5 year period through 11 flock rotations, a partial and a total cleanout, and the subsequent deposition of the litter as fertilizer on pastureland. The effects of both time and management practices correlated with alterations of the litter microbial community. The cleanout practices and introduction of new bedding had minimal influence on the house microbial community once it was established, which generally showed a consistent increase in the proportion of Actinobacteria and a decrease in Firmicutes over the 11 flock rotations. Analysis of the bacterial profile at the genus level gave increased resolution, revealing changes during the first and second flock rotation and after the total cleanout. The disturbance of the partial cleanout seemed to be buffered by the supporting conditions within the house while the total cleanout showed a small, but significant influence. The pastureland deposition of litter, however, was affected by time and abiotic factors that changed the litter microbial community structure weekly. The stockpiled litter had an increase in the phyla Actinobacteria and the class Bacilli that commonly have microbes utilizing nitrogen and decaying materials, in comparison to Native soil. Further, the soil beneath where the litter was stored for 20 weeks, lost diversity, indicating a possible effect of the litter stockpiling on environmental quality at that site. How management practices affect the composition of the microbial community within the litter of the broiler house is of interest in terms of bird health and environmentally for future utilization of spent litter., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Published by Elsevier B.V.)

Monitoring antimicrobial resistance of foodborne pathogens in poultry is critical for food safety. We aimed to compare antimicrobial resistance phenotypes in Salmonella isolated from poultry samples as influenced by isolation and antimicrobial susceptibility testing methods. Salmonella isolates were cultured from a convenience sample of commercial broiler ceca with and without selective broth enrichment, and resistance phenotypes were determined for 14 antimicrobials using the Sensititre ® platform and a qualitative broth breakpoint assay. The broth breakpoint method reported higher resistance to chloramphenicol, sulfisoxazole, and the combination of trimethoprim and sulfamethoxazole, and lower resistance to streptomycin as compared to the Sensititre ® assay in trial one. Selective enrichment of samples containing Salmonella in Rappaport-Vassiliadis broth reported lowered detectable resistance to amoxicillin/clavulanic acid, ampicillin, azithromycin, cefoxitin, ceftriaxone, nalidixic acid, and meropenem, and increased resistance to streptomycin and tetracycline than direct-plating samples in trial one. Using matched isolates in trial two, the Sensititre ® assay reported higher resistance to chloramphenicol and gentamicin, and lower resistance to nalidixic acid as compared to the broth breakpoint method. These results suggest methodology is a critical consideration in the detection and surveillance of antimicrobial resistance phenotypes in Salmonella isolates from poultry samples and could affect the accuracy of population or industry surveillance insights and intervention strategies.

Clostridium perfringens (Cp) is a Gram-positive anaerobe that is one of the causative agents of necrotic enteritis (NE) in chickens, which leads to high mortality. Owing to the ban of administering antibiotics in feed to chickens, there has been an increase in the number of NE outbreaks all over the world, and the estimated loss is approximately 6 billion U.S. dollars. The best alternative method to control NE without antibiotics could be vaccination. In this study, we exposed three different strains of Cp to electron beam (eBeam) irradiation to inactivate them and then used them as a killed vaccine to control the colonization of Cp in broiler chickens. The vaccine was delivered to 18-day old embryos in ovo and the chickens were challenged with the respective vaccine strain at two different time points (early and late) to test the protective efficacy of the vaccine. The results indicate that an effective eBeam dose of 10 kGy inactivated all three strains of Cp, did not affect the cell membrane or epitopes, induced significant levels of IgY in the vaccinated birds, and further reduced the colonization of Cp strains significantly ( p < 0.0001) in late challenge (JGS4064: 4 out of 10; JGS1473: 0 out of 10; JGS4104: 3 out of 10). Further studies are necessary to enhance the efficacy of the vaccine and to understand the mechanism of vaccine protection.

Foodborne illness is an ongoing problem worldwide and is caused by bacteria that invade the food chain from the farm, slaughter house, restaurant or grocery, or in the home and can be controlled by strategies using biocides (antiseptics and disinfectants). Susceptibility profiles were determined for 96 Campylobacter jejuni strains obtained in 2011-2012 from broiler chicken house environments to antimicrobials and disinfectants as per the methods of the Clinical and Laboratory Standards Institute and TREK Diagnostics using CAMPY AST Campylobacter plates. Low prevalence of antimicrobial resistance was observed in C. jejuni strains to tetracycline (TET; 21.9%), ciprofloxacin (CIP; 13.5%), and nalidixic acid (NAL; 12.5%). The resistance profiles had a maximum of 3 antimicrobials, CIP-NAL-TET, with TET being the main profile observed. No cross-resistance was observed between antimicrobials and disinfectants. The C. jejuni strains (99%) were resistant to triclosan, 32% were resistant to chlorhexidine, and they all were susceptible to benzalkonium chloride. The strains had low-level minimum inhibitory concentrations (MICs) to the disinfectants P-128, Food Service Sanitizer, F-25 Sanitizer, Final Step 512 Sanitizer, OdoBan, dioctyldimethylammmonium chloride, didecyldimethylammonium chloride (C10AC), benzyldimethyldodecylammonium chloride (C12BAC), and benzyldimethyltetradecylammonium chloride (C14BAC). Intermediate MICs against DC&R, cetylpyridinium bromide hydrate, hexadecylpyridinium chloride, ethylhexadecyldimethylammonium bromide, and hexadecyltrimethylammonium bromide with elevated intermediate MICs against Tek-Trol, benzyldimethylhexadecylammonium chloride, tris(hydroxylmethyl)nitromethane (THN), and formaldehyde. The highest MIC were obtained for povidone-iodine. The components THN and the benzylammonium chlorides C12BAC and C14BAC were responsible for the inhibition by DC&R. The components C10AC and C12BAC may act synergistically causing inhibition of C. jejuni by the disinfectant P-128. The formaldehyde component in DC&R was not effective against C. jejuni compared with the ammonium chloride components. Its use in disinfectants may result in additional unnecessary chemicals in the environment. Didecyldimethylammonium chloride is the most effective ammonium chloride component against C. jejuni., (Copyright © 2020. Published by Elsevier Inc.)

Poultry litter is a potentially valuable crude protein feedstuff for ruminants but must be treated to kill pathogens before being fed. Composting kills pathogens but risks losses of nitrogen due to volatilization or leaching as ammonia. Treatment of poultry litter with ethyl nitroacetate, 3-nitro-1-propionate, ethyl 2-nitropropionate (at 27 µmol/g), decreased numbers of experimentally-inoculated Salmonella Typhimurium (>1.0 log 10 compared to controls, 4.2 ± 0.2 log 10 CFU/g) but not endogenous Escherichia coli early during simulated composting. By day 9 of simulated composting, Salmonella and E. coli were decreased to non-detectable levels regardless of treatment. Some nitro-treatments preserved uric acid and prevented ammonia accumulation, with 18% more uric acid remaining and 17-24% less ammonia accumulating in some nitro-treated litter than in untreated litter (18.1 ± 3.8 µmol/g and 3.4 ± 1.4 µmol/g, respectively). Results indicate that nitro-treatment may help preserve uric acid in composted litter while aiding Salmonella control., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Necrotic enteritis (NE) is one of the most common and costly diseases in the modern broiler industry, having an estimated economic impact of $6 billion dollars annually. Increasing incidents of NE have resulted from restrictions on the use of antibiotic feed additives throughout the broiler industry. As such, finding effective antibiotic alternatives has become a priority. In this study, an experimental model of NE was used, comprising a commercial infectious bursal disease virus vaccine and Clostridium perfringens (C. perfringens) inoculation. Yeast cells wall (YCW) components, β-glucan (BG), and mannoproteins (MPTs) were evaluated for their effects on disease development. Chicken-specific immunometabolic kinome peptide arrays were used to measure differential phosphorylation between control (uninfected), challenged (infected), and challenged and treated birds in duodenal, jejunal, and ileal tissues. Treatment groups included crude YCW preparation, BG, MPT, or BG+MPT as feed additives. Data analysis revealed kinome profiles cluster predominantly by tissue, with duodenum showing the greatest relative signaling and jejunum showing the greatest response to treatment. BG, MPT, and BG+MPT cluster together, separate from controls and challenge birds in each tissue. Changes in signaling resulting from the treatments were observed in cell growth and survival responses as well as immune responses. None of the treatments of disease challenge returned the profiles to control-like. This is attributable to immune modulation and metabolic effects of the treatments generating distinct profiles from control. Importantly, all the treatments are distinct from the challenge group despite being challenged themselves. Only BG+MPT treatment had a significant effect on bird weight gain compared with the NE challenge group, and this treatment had the greatest impact on gut tissue signaling in all segments. The signaling changes elicited by BG+MPT during an NE challenge were increased cell growth and survival signaling, reducing cell death, apoptosis and innate inflammatory responses, and generating compensatory signaling to reduce disease severity., (Copyright © 2020. Published by Elsevier Inc.)

The microbiome within a poultry production house influences the attainment of physiologically strong birds and thus food safety and public health. Yet little is known about the microbial communities within the house and the effects on the soil microbes onto which the houses are placed; nor the effects of management practices on their equilibrium. This study looked at the soil bacterial microbiome before a broiler house was constructed, then through 11 flock rotations (2.5 years) that included a partial clean-out and a total clean-out within the management regimen. Major shifts were observed, occurring at the taxonomic class level, related to the introduction of bedding and birds on top of the soil. The partial clean-out of litter did not change the soil bacterial community in any substantial way, only prompting a temporary increase in some genera; however, the total litter clean-out caused a major increase in a cohort of Actinobacteria. The underlying soil contained bacteria beneficial for poultry metabolism, such as Lactobacillus , Faecalibacterium , Bacteriodes , and Ruminococcus . Additionally, management practices affected the class structure of the soil bacterial community beneath the poultry house. The scheduling of these practices should be leveraged to exploit maintenance of beneficial bacteria that maximize microbiome contributions to bird production processes, while minimizing possible antibiotic-resistant bacteria and environmental effects., (Copyright © 2019 Crippen, Sheffield, Singh, Byrd and Beier.)

Mechanically separated chicken (MSC) may be incorporated into other further processed foods, and has been identified as a transmission vehicle in human foodborne disease outbreaks involving the pathogen Salmonella enterica. Chickens possess pneumatic bones producing marrow, which may become infected with Salmonella and present a mechanism for Salmonella contamination of MSC. The primary objective of the current study was to ascertain whether chicken bone marrow was susceptible to Salmonella infection as a function of pathogen challenge method. Additionally, this study sought to determine the impact of Salmonella challenge site and inoculation on subsequent numbers of the microorganism in the cecum at 3, 6 and 9 d post-challenge. In this study, 30-day-old birds (Gallus domesticus) were separately challenged with S. enterica by one of the 6 experimental treatment methods: oral or endo-tracheal gavage, and transdermal scratch challenge on the breast or back muscles, with or without feathers (N = 150). Differing Salmonella recovery rates were detected in bone marrow from euthanized birds (P = 0.0015); oral and endo-tracheal gavage produced Salmonella recovery frequencies of 10% and 20%, respectively. Counts of Salmonella from chicken cecal samples statistically differed as a function of challenge method (P = 0.032); the highest numbers of Salmonella in cecal samples were obtained from orally gavaged birds (3.0 log10 cfu/g). Study results demonstrate the potential for chickens to contract short-term systemic infection following Salmonella challenge using methods of pathogen challenge simulating aerosol inhalation, ingestion, or wounding. Salmonella entry into chicken bone marrow may result in decreased likelihood of Salmonella performance standard compliance and increased pathogen transmission risk to consumers., (Published by Oxford University Press on behalf of Poultry Science Association 2019.)

Campylobacter jejuni is a bacterium that causes major diarrheal disease worldwide and is also one of the top five foodborne pathogens encountered in the United States. Poultry is a major source of C. jejuni , and a high-risk factor for contracting campylobacteriosis. Organic acids are used in the United States during food animal processing for removal of bacterial contamination from animal carcasses. Six organic acids were evaluated in inhibition studies of 96 C. jejuni strains obtained from shoe covers used in broiler chicken houses at different poultry farms in several states by determining the susceptibilities of the C. jejuni strains, along with the pH values at the molar minimum inhibitory concentrations (MIC M s). The undissociated and dissociated organic acid concentrations were calculated at the MIC M s with the Henderson-Hasselbalch equation. The results for the 96 C. jejuni strains were treated similarly for each different organic acid. Campylobacter jejuni inhibition did correlate with the dissociated organic acids, but did not correlate with pH or with the undissociated organic acids. When the concentrations of dissociated organic acids decreased, the C. jejuni strains were not disinfected. A carcass wash using organic acids should have the concentration of dissociated acid species carefully controlled. It is suggested to maintain a dissociated acid concentration for propionic, l-lactic, formic, citric, butyric, and acetic acids at 24, 40, 36, 21, 23, and 25 mM, respectively, and at these dissociated organic acid levels an acid wash would be expected to remove or inhibit 97% or more of the C. jejuni bacteria studied here. However, studies must be undertaken to confirm that the suggested concentrations of dissociated organic acids are adequate to remove C. jejuni bacteria in the field vs. the laboratory. Due to propionate, l-lactate, formate, butyrate, and acetate being utilized by C. jejuni , these organic acids may not be appropriate for use as a carcass wash to remove C. jejuni surface contamination. Of all tested organic acids, dissociated citric acid was the most efficient at inhibiting C. jejuni .

Cognitive remediation is aimed at reducing cognitive impairments in severe mental illnesses such as schizophrenia, but little is known about whether severity of cognitive impairment predicts benefit from this intervention. To address this question, this study aggregated data from five randomized controlled trials (N = 300) of a standardized comprehensive, multimodal outpatient cognitive remediation program, the Thinking Skills for Work program, and evaluated whether baseline level of cognitive impairment differentially predicted improvement in cognitive functioning following cognitive remediation vs. usual services. Using standardized scores of neuropsychological functioning to designate "low average," "moderate," and "severe" levels of cognitive impairment, participants with greater cognitive impairment were found to benefit differentially more from cognitive remediation than usual services compared to less cognitively impaired participants. The findings were unaffected by statistically controlling for participant demographic and clinical characteristics. The findings suggest that individuals with the greatest cognitive impairment, for whom cognitive remediation was developed, are also most likely to benefit from this intervention., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Campylobacter is a bacterium that colonizes the lower gastrointestinal tract of poultry and may influence the intestinal environment to promote its survival. The objective of this study was to characterize the effects of Campylobacter challenge on the mRNA abundance of nutrient transporters and host defense peptides (HDP), such as the avian β-defensins (AvBD) and liver expressed antimicrobial peptide 2 (LEAP2). On the day of hatch, broiler chicks were challenged with one of three (106, 107, 108 colony-forming units, cfu) levels of Campylobacter jejuni. Quantitative PCR analysis revealed that there were dose-, tissue-, and age-specific changes in gene expression for both nutrient transporters and HDP. Expression of zinc transporter 1 (ZnT1) mRNA increased on d 7 in the duodenum, ileum, and cecum of birds challenged with 106 cfu of C. jejuni. At d 14, there was upregulation of the amino acid transporter bo,+AT mRNA in the duodenum, jejunum, and ileum of birds challenged with 106 cfu of C. jejuni. Other transporters such as EAAT3, GLUT2, SGLT1, and ZnT1 showed upregulation of mRNA in the ileum of the 106 cfu challenged group. There was a delayed response of the HDP to the C. jejuni challenge, with only a few HDP changed at d 7 but all HDP changed at d 14. At d 7, there was upregulation of AvBD10 mRNA in the duodenum of the 106 cfu challenged group but downregulation of AvBD10 in the ileum and AvBD12 and LEAP2 in the cecum of the 108 cfu challenged group. At d 14, there was upregulation of AvBD1, AvBD6, AvBD8, AvBD10, AvBD11, AvBD12, and AvBD13 mRNA in the ileum and cecum of the 106 cfu challenged group but not the 107 and 108 cfu challenged groups compared to control. These results indicated that at a low dose (106 cfu) of C. jejuni, intestinal cells increased nutrient transporter and AvBD mRNA abundance to try to counter the infection, but that at higher doses the cellular response was suppressed.

Hatched male layer chicks are currently euthanized by maceration in the United States. Public concerns on the use of maceration have led to the search for alternative methods. We hypothesized that gas inhalation and low atmospheric pressure stunning (LAPS) are viable and humane alternatives to instantaneous mechanical destruction. The objective of this study was to evaluate the physiological and behavioral responses of recently hatched male layer chicks when subjected to carbon dioxide, nitrogen inhalation, or LAPS. The study consisted of seven treatments: breathing air (NEG), 25% carbon dioxide (CO₂), 50% CO₂, 75% CO₂, 90% CO₂, 100% nitrogen (N₂), or LAPS. Ten day-of-hatch, male layer chicks were randomly assigned to each treatment, and each treatment was replicated on ten different days. A custom-made vacuum system was used to reduce air pressure inside the chamber from 100.12 kPa to 15.3 kPa for the LAPS treatment. Serum corticosterone and serotonin levels were measured using commercially available competitive enzyme linked immunosorbent assay (ELISA). Latencies to loss of posture and motionlessness were determined from video recordings. The 25% and 50% CO₂ treatments were discontinued after the first replication, as the majority of the chicks recovered. The chicks in the negative (NEG) group had significantly higher levels of corticosterone than the other four euthanasia treatments. On the other hand, the serotonin levels of chicks in the NEG group was significantly lower when compared to the other four euthanasia treatments. The latencies to loss of posture and motionlessness of chicks exposed to 75% and 90% CO₂ were significantly shorter than those in the LAPS and N₂ inhalation treatments. These data suggest that the stress responses of chicks to the CO₂, N₂, and LAPS treatments do not differ among each other. However, the CO₂ inhalation method was faster in inducing loss of posture and motionlessness in chicks than the LAPS and N₂ inhalation treatments., Competing Interests: The authors declare no conflict of interest.

Reduction of stress is an important factor in improving poultry welfare, especially during periods of heat stress. A study was conducted to evaluate the effects of feeding the functional metabolites of Diamond V Original XPCTM to broilers reared under ambient or heat stress temperatures. Dietary treatments included: control feed (CON) and Original XPC fed continuously at 1.25 kg/MT (XPC). Half the birds in each dietary treatment were subjected to either no heat stress (24°C constant) or heat stress (35°C:24°C for 18:6 h daily) from 28 to 42 d. At the end of the heat stress period, blood was collected from 40 birds/treatment. Blood was analyzed for plasma corticosterone (CORT), plasma HSP70 (HSP70), and heterophil/lymphocyte ratios (H/L). At 42 d, bilateral metatarsal traits were also measured in 40 birds/treatment to assess physical asymmetry. Birds fed XPC had significantly lower CORT levels than CON (P < 0.001; 5,129 ± 617 vs. 8,433 ± 730, respectively). Physical asymmetry scores were also significantly higher in CON compared to XPC fed broilers (P < 0.001; 1.50 ± 0.13 vs. 0.54 ± 0.05, respectively). H/L ratios were significantly greater in CON than for XPC birds (P = 0.01; 0.81 ± 0.05 mm vs. 0.62 ± 0.05 mm, respectively). No differences were observed between CON and XPC fed broilers in HSP70. However, heat stress did increase (P < 0.0001) HSP70 compared to no heat stress birds (5.65 ± 0.12 vs. 4.78 ± 0.11 pg/mL, for heat stress and no heat stress, respectively). Feeding XPC to broiler chickens improved animal welfare via reduced stress indicators while under heat stress or no heat stress conditions. These results indicate that feeding XPC may improve poultry welfare by reducing heat stress susceptibility.

A study was conducted to evaluate the influence of the purification of yeast cell wall (YCW) preparations on broiler performance and immunogenic and metabolic pathways under microbial challenge. A total of 240 (day old) chicks were distributed among two battery brooder units (48 pens; 5 birds/pen; 8 replicates/treatment). A basal starter diet was divided into 5 batches to create 6 dietary treatments; non-challenge (NCh-C) and challenge (Ch-C) controls, semi-purified YCW containing cytosol contents (SPYCW; 250 mg/kg), purified YCW (PYCW; 250 mg/kg), 50% purified beta-glucan (BG; 130 mg/kg), and 99.9% purified mannan-oligosaccharide (MOS; 53 mg/kg). All birds were immunocompromised with infectious bursal disease vaccine (10× the recommended dose) on day 10 and then all birds except NCh-C birds were challenged with Clostridium perfringens (Cp) (107 cfu/mL) via oral gavage on days 16 and 17. On day 21, tissue samples were collected from the jejunum and duodenum for analysis with chicken-specific, peptide arrays to study the influence of YCW supplementation on immune and metabolic kinase pathways. On day 16, SPYCW had significantly lower body weight (BW) and weight gain (WG) than other treatments except BG (P < 0.05). The productivity index (PI) was lower in SPYCW and BG than in NCh-C, Ch-C, and PYCW. On day 21, after the Cp challenge, NCh-C was higher than Ch-C, SPYCW, and BG in BW, WG, and PI (P = 0.03). The PI of PYCW was similar to NCh-C. The addition of purified YCW to the starter broiler diets influenced the immune and metabolic pathways in the gut. A total of 459 and 367 peptides in the duodenum and jejunum, respectively, were changed due to the Cp challenge. The YCW treatments had different degrees of influence on these peptides for both the duodenum and jejunum. These results suggest that relative purification of YCW and specific fractions of the YCW can influence broiler performance differently during microbial challenges and can alleviate the impact of these stressors., (© 2017 Poultry Science Association Inc.)

The microbial quality of eggs entering the hatchery is an important critical control point for biosecurity, pathogen reduction, and food safety programs in poultry production. Developing interventions to reduce Salmonella contamination of eggs is important to improving the microbial food safety of poultry and poultry products. The hydrogen peroxide (H2O2) and ultraviolet light (UV) Advanced Oxidation Process (AOP) has been previously demonstrated to be effective in reducing Salmonella on the surface of experimentally contaminated eggs. The objective of this study was to evaluate the effect of treating eggs with an egg-sanitizing apparatus using the H2O2/UV AOP on Salmonella contamination during incubation, hatching, and in broiler chicks during grow-out. Experimentally contaminated eggs were treated using the automated H2O2/UV AOP egg sanitizer and incubated for 21 d. AOP sanitization reduced Salmonella up to 7 log10 cfu egg-1 (P < 0.05) from the surface of experimentally contaminated eggs and reduced the number of Salmonella positive eggs by up to 75% (P < 0.05) when treated 1 h post-inoculation. AOP treatment also reduced the number of Salmonella-positive eggs during incubation. Additionally, Salmonella was recovered from more chicks hatched from untreated eggs than from eggs treated using the H2O2/UV AOP egg sanitizer (P < 0.05) through 14 d posthatch. These data suggest reduction of Salmonella contamination on the surface of eggs using the H2O2/UV AOP egg sanitizer prior to incubation may reduce the gastrointestinal colonization of chicks by Salmonella., (© 2017 Poultry Science Association Inc.)

Under normal conditions, fungi are ignored unless a disease/syndrome clinical signs are reported. The scientific communities are largely unaware of the roles fungi play in normal production parameters. Numerous preharvest interventions have demonstrated that beneficial bacteria can play a role in improving productions parameters; however, most researchers have ignored the impact that fungi may have on production. The goal of the present study was to record fungi recovered from commercial broiler and layer houses during production. Over 3,000 cecal samples were isolated using conventional culture methodology and over 890 samples were further characterized using an automated repetitive sequence-based PCR (rep-PCR) methodology. Eighty-eight different fungal and yeast species were identified, including Aspergillus spp., Penicillium spp., and Sporidiobolus spp, and 18 unknown genera were separated using rep-PCR. The results from the present study will provide a normal fungi background genera under commercial conditions and will be a stepping stone for investigating the impact of fungi on the gastrointestinal tract and on the health of poultry., (Published by Oxford University Press on behalf of Poultry Science Association 2017.)

Non-typhoidal Salmonella enterica induces an early, short-lived pro-inflammatory response in chickens that is asymptomatic of clinical disease and results in a persistent colonization of the gastrointestinal (GI) tract that transmits infections to naïve hosts via fecal shedding of bacteria. The underlying mechanisms that control this persistent colonization of the ceca of chickens by Salmonella are only beginning to be elucidated. We hypothesize that alteration of host signaling pathways mediate the induction of a tolerance response. Using chicken-specific kinomic immune peptide arrays and quantitative RT-PCR of infected cecal tissue, we have previously evaluated the development of disease tolerance in chickens infected with Salmonella enterica serovar Enteritidis (S. Enteritidis) in a persistent infection model (4-14 days post infection). Here, we have further outlined the induction of an tolerance defense strategy in the cecum of chickens infected with S. Enteritidis beginning around four days post-primary infection. The response is characterized by alterations in the activation of T cell signaling mediated by the dephosphorylation of phospholipase c-γ1 (PLCG1) that inhibits NF-κB signaling and activates nuclear factor of activated T-cells (NFAT) signaling and blockage of interferon-γ (IFN-γ) production through the disruption of the JAK-STAT signaling pathway (dephosphorylation of JAK2, JAK3, and STAT4). Further, we measured a significant down-regulation reduction in IFN-γ mRNA expression. These studies, combined with our previous findings, describe global phenotypic changes in the avian cecum of Salmonella Enteritidis-infected chickens that decreases the host responsiveness resulting in the establishment of persistent colonization. The identified tissue protein kinases also represent potential targets for future antimicrobial compounds for decreasing Salmonella loads in the intestines of food animals before going to market.

The U.S. broiler meat market has grown over the past 16 years and destinations for U.S. broiler meat exports expanded to over 150 countries. This market opportunity has spurred a corresponding increase in industrialized poultry production, which due to the confined space in which high numbers of animals are housed, risks accumulating nutrients and pollutants. The purpose of this research was to determine the level of pollutants within poultry litter and the underlying soil within a production facility; and to explore the impact of spent litter deposition into the environment. The study follows a production facility for the first 2.5 years of production. It monitors the effects of successive flocks and management practices on 15 physiochemical parameters: Ca, Cu, electrical conductivity, Fe, K, Mg, Mn, moisture, Na, NO3(-)/N, organic matter, P, pH, S, and Zn. Litter samples were collected in-house, after clean-outs and during stockpiling. The soil before house placement, after the clean-outs and following litter stockpiling was monitored. Management practices markedly altered the physiochemical profiles of the litter in-house. A canonical discriminant analysis was used to describe the relationship between the parameters and sampling times. The litter profiles grouped into five clusters corresponding to time and management practices. The soil in-house exhibited mean increases in all physiochemical parameters (2-297 fold) except Fe, Mg, %M, and pH. The spent litter was followed after deposition onto a field for use as fertilizer. After 20 weeks, the soil beneath the litter exhibited increases in EC, Cu, K, Na, NO3(-)/N, %OM, P, S and Zn; while %M decreased. Understanding the impacts of industrialized poultry farms on the environment is vital as the cumulative ecological impact of this land usage could be substantial if not properly managed to reduce the risk of potential pollutant infiltration into the environment., (Published by Elsevier B.V.)

We developed a novel selection method based on an inherently high and low phenotype of pro-inflammatory mediators and produced "high" and "low" line chickens. We have shown high line birds are more resistant to Salmonella enterica serovar Enteritidis and Eimeria tenella compared to the low line. Clostridium perfringens is the fourth leading cause of bacterial-induced foodborne illness, and is also an economically important poultry pathogen and known etiologic agent of necrotic enteritis (NE). The objective of this study was to determine if high line birds were also more resistant to NE than low line birds using an established model. Birds were reared in floor pens and challenges were conducted twice (high line = 25/trial, 50 birds total; low line = 26/trial, 52 birds total). Day-old chicks were provided a 55% wheat-corn-based un-medicated starter diet. A bursal disease vaccine was administered at 10× the recommended dose via the ocular route at 14-d-of-age. Birds were challenged daily for 3 d beginning at 16-d-of-age by oral gavage (3 mL) with 10(7) colony forming units (cfu) of C. perfringens/mL then necropsied at 21-d-of-age. All birds had sections of the intestine examined and scored for lesions while the first 10 necropsied also had gut content collected for C. perfringens enumeration. Chickens from the high line were more resistant to C. perfringens-induced NE pathology compared to the low line, as indicated by reduced lesion scores. Ninety percent of the high line birds had lesions of zero or one compared to 67% of the low line birds. Wilcoxon rank sum test showed significantly higher lesion scores in the low line birds compared to the high line (P < 0.0001). There were no differences in the C. perfringens recovered (P = 0.83). These data provide additional validation and support selection based on elevated levels of pro-inflammatory mediators produces chickens with increased resistance against foodborne and poultry pathogens., (Published by Oxford University Press on behalf of Poultry Science Association 2015. This work is written by (a) US Government employee(s) and is in the public domain in the US.)

The gastrointestinal microbial community in broiler chickens consists of many different species of bacteria, and the overall microbiota can vary from bird to bird. To control pathogenic bacteria in broilers and improve gut health, numerous potential dietary amendments have been used. In this study, we used a pyrosequencing platform to evaluate the effect of sodium bisulfate on microbiota of the crop, cecum, and ileum of broiler chickens grown over several weeks. The diversity information in each digestive organ sample exhibited considerable variation and was clearly separable, suggesting distinct bacterial populations. Although no apparent microbial clustering occurred between the control and the dietary treatments, we did observe shifts in overall microbiota populations in the crop, ileum, and ceca as well as changes in specific microorganisms such as Bacteroides, Clostridium, and Lactobacillus species that were identified as birds became older., (© 2015 Poultry Science Association Inc.)

The effect dirt-floored broiler houses have on the underlying native soil, and the potential for contamination of the ground water by leaching under the foundation, is an understudied area. This study examines alterations in fifteen quantitative soil parameters (Ca, Cu, electrical conductivity, Fe, K, Mg, Mn, Na, NO3, organic matter, P, pH, S, soil moisture and Zn) in the underlayment of a newly constructed dirt-floored broiler house over the first two years of production (Native through Flock 11). The experiment was conducted near NW Robertson County, Texas, where the native soil is a fine, smectitic thermic Udertic Paleustalfs and the slopes range from zero to three percent. Multiple samples were collected from under each of three water and three feed lines the length of the house, in a longitudinal study during February 2008 through August 2010. To better define the relationship between the soil parameters and sampling times, a canonical discriminant analysis approach was used. The soil profiles assembled into five distinctive clusters corresponding to time and management practices. Results of this work revealed that the majority of parameters increased over time. The management practices of partial and total house clean-outs markedly altered soil profiles the house underlayment, thus reducing the risk of infiltration into the ground water near the farm. This is important as most broiler farms consist of several houses within a small area, so the cumulative ecological impact could be substantial if not properly managed.

Campylobacter jejuni is an important human food-borne pathogen that can contaminate meat and poultry during processing. Consequently, strategies are sought to reduce the carriage of C. jejuni in food animals before they arrive at the abattoir. Thymol is a natural product that reduces survivability of Campylobacter in vitro, but its rapid absorption from the proximal alimentary tract limits its bactericidal efficacy in vivo. Thymol-β-D-glucopyranoside is more resistant to absorption than free thymol, but its administration to chickens has not been reported. In the present studies, 1 mM thymol-β-D-glucopyranoside was shown to exhibit near equal anti-Campylobacter activity as 1 mM thymol when incubated anaerobically in avian crop or cecal contents in vitro, resulting in reductions of 1.10-2.32 log10 colony forming units mL(-1) in C. jejuni concentrations after 24 h incubation. In a follow-up live animal study, oral administration of thymol-β-D-glucopyranoside, but not free thymol, significantly lowered (>10-fold) recovery of Campylobacter from the crop of market-aged broilers when compared to placebo-treated controls (n = 6 broilers/treatment). Neither thymol-β-D-glucopyranoside nor thymol affected recovery of Campylobacter from cecal contents of the treated broilers. These results indicate that rapid absorption or passage of free thymol from the crop precluded its anti-Campylobacter activity at this site and throughout the entire gastrointestinal tract. Conversely, lower recovery of Campylobacter from the crop of birds treated with thymol-β-D-glucopyranoside indicates this conjugate was retained and able to be hydrolyzed to biologically active free thymol at this site as intended, yet was not sufficiently protected to allow passage of efficacious amounts of the intact glycoside to the lower gut. Nevertheless, these results warrant further research to see if higher doses or encapsulation of thymol-β-D-glucopyranoside or similar glycosides may yield an efficacious additive to reduce carriage of Campylobacter as well as other pathogens throughout the avian gut.

The gastrointestinal tract commensal microbiome is important for host nutrition, health and immunity. Little information is available regarding the role of these commensals at other mucosal surfaces in poultry. Tracheal mucosal surfaces offer sites for first-line health and immunity promotion in broilers, especially under stress-related conditions. The present study is aimed at elucidating the effects of feed supplementations with mannanoligosaccharides (MOS) prebiotic and a probiotic mixture (PM) on the caecal and tracheal microbiome of broilers kept under chronic heat stress (HS; 35 ± 2°C). Day-old chickens were randomly divided into five treatment groups: thermoneutral control (TN-CONT), HS-CONT, HS-MOS, HS-PM and HS synbiotic (fed MOS and PM). Caecal digesta and tracheal swabs were collected at day 42 and subjected to DNA extraction, followed by polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) and pyrosequencing. The PCR-DGGE dendrograms revealed significant (49.5% similarity coefficients) differences between caecal and tracheal microbiome. Tracheal microbiome pyrosequencing revealed 9 phyla, 17 classes, 34 orders, 68 families and 125 genera, while 11 phyla, 19 classes, 34 orders, 85 families and 165 genera were identified in caeca. An unweighted UniFrac distance metric revealed a distinct clustering pattern (analysis of similarities, P = 0.007) between caecal and tracheal microbiome. Lactobacillus was the most abundant genus in trachea and caeca and was more abundant in caeca and trachea of HS groups compared with the TN-CONT group. Distinct bacterial clades occupied the caecal and tracheal microbiomes, although some bacterial groups overlapped, demonstrating a core microbiome dominated by Lactobacillus. No positive effects of supplementations were observed on abundance of probiotic bacteria.

Two experiments were conducted to evaluate the effects of Arg, vitamin E (VE), and mannanoligosaccharide (MOS) on the immune response and clearance of Salmonella in broiler chickens. In each experiment, 1-d-old chicks (n = 160) were randomly distributed into 4 groups: antibiotic-free diet (negative control, CTL-), antibiotic-supplemented diet (positive control, CTL+), antibiotic free-diet plus Arg and VE (AVE), or antibiotic-free diet plus Arg, VE, and MOS (AVM). Birds were orally challenged with 10(6) cfu of a novobiocyn and nalidixic acid-resistant Salmonella enterica serovar Typhimurium strain at d 7 (experiment 1) or at d 3 (experiment 2). Heterophil- (HOB) and monocyte- (MOB) oxidative burst and lymphocyte proliferation (LPR), antibody titers, and Salmonella content in the ceca were measured at several intervals postinfection (PI). In experiment 1, both AVM and AVE decreased HOB compared with the controls 5 and 9 d PI, but increased LPR 9 d PI. In the same experiment, birds fed the AVE diet had higher MOB than birds fed CTL+ or the AVM diet at 7 d PI, whereas 9 d PI birds fed the AVM diet had the highest MOB. In experiment 2, birds fed the AVE diet had higher MOB, HOB, and LPR than birds in the other treatments 7 and 14 d PI, except at 7 d PI, when MOB was not different among treatments. Birds fed the AVM diet had the highest IgA antibody titer, and a higher IgM antibody titer than the CTL+ birds. In experiment 1, Salmonella Typhimurium content in the ceca was lower in birds fed the AVM diet compared with birds fed the CTL- diet 3 d PI, but later on (10 and 17 d PI), and in experiment 2 (7, 14, and 21 d PI), Salmonella Typhimurium concentrations were not different among treatments. Thus, Arg and VE improved immune response after a Salmonella Typhimurium challenge in young chicks, and although they did not reduce Salmonella Typhimurium concentrations in the ceca, they may improve bacterial resistance against other pathogens in commercial growing conditions.

Linalool is a natural plant-product used in perfumes, cosmetics, and flavoring agents. Linalool has proven antimicrobial and insect-repellent properties, which indicate it might be useful for control of enteropathogens or insect pests in poultry production. However, there are no published reports that linalool may be safely administered to or tolerated by chickens. Linalool was added to the diets of day-of-hatch chicks, and they were fed linalool-supplemented diets for 3 wk. We studied the effects of linalool on serum chemistry, gross pathology, feed conversion, and relative liver weights. Linalool had a dramatic negative dose-dependent effect on feed conversion at concentrations in the feed exceeding 2% linalool, but not on gross pathology. Liver weights were significantly increased in the 5% linalool-treated birds. There was a statistical effect on blood glucose, but this parameter remained below the cut-offs for elevated serum glucose, and the result is likely of no biological significance. Linalool caused serum aspartate aminotransferase (AST) levels to increase, but it did not increase serum gamma-glutamyl transferase levels. The linalool effect on AST was dose-dependent, but in linalool doses between 0.1 and 2% of the feed, AST was not elevated beyond normal parameters. Linalool at 2% or less may be safely added to chicken feed. We suggest future studies to evaluate the addition of linalool to the litter, where it may be used as an antimicrobial or an insect repellent or to produce a calming effect.

This pilot analysis was conducted with data from 52 conventional grow-out broiler flocks in a prospective field observational study in the southeastern United States during 2003-2006. Each flock was sampled for Salmonella 1 wk before the end of grow-out, upon arrival at the processing plant, and during processing (prior to and immediately after carcass chilling). The broiler litter was sampled on the day of bird harvest. The grow-out feeding programs, including the medications delivered in feed, were surveyed with questionnaires completed by the broiler managers and feedmill managers. Each detail of the feeding program was tested for statistical association with the frequency of Salmonella in the flock at each sampling point, after accounting for variation in Salmonella frequency between the farms, broiler complexes, and companies. Significant associations were found between Salmonella frequency in the broiler flock pre- and postharvest and the inclusion of feeds containing individual coccidiostats and other antimicrobial growth promoters, days on feed, and total consumption of feeds containing these products, as well as with practices such as a mash feed and a nonmedicated withdrawal feed. The analysis provided testable hypotheses for how broiler feed medications impact the frequency of Salmonella in the flocks.

Campylobacter is an important human pathogen, and consumption of undercooked poultry has been linked to significant human illnesses. To reduce human illness, intervention strategies targeting Campylobacter reduction in poultry are in development. For more than a decade, there has been an ongoing national and international controversy about whether Campylobacter can pass from one generation of poultry to the next via the fertile egg. We recognize that there are numerous sources of Campylobacter entry into flocks of commercial poultry (including egg transmission), yet the environment is often cited as the only source. There has been an abundance of published research globally that refutes this contention, and this article lists and discusses many of them, along with other studies that support environment as the sole or primary source. One must remember that egg passage can mean more than vertical, transovarian transmission. Fecal bacteria, including Campylobacter, can contaminate the shell, shell membranes, and albumen of freshly laid fertile eggs. This contamination is drawn through the shell by temperature differential, aided by the presence of moisture (the "sweating" of the egg); then, when the chick emerges from the egg, it can ingest bacteria such as Campylobacter, become colonized, and spread this contamination to flock mates in the grow house. Improvements in cultural laboratory methods continue to advance our knowledge of the ecology of Campylobacter, and in the not-so-distant future, egg passage will not be a subject continuously debated but will be embraced, thus allowing the development and implementation of more effective intervention strategies.

Reducing Salmonella in poultry has been a paramount goal of the poultry industry in order to improve food safety. Inclusion of high-fiber fermentable feedstuffs in chicken diets has been shown to reduce the incidence of Salmonella colonization in laying hens, but no work has been performed in growing birds. Therefore, the present study was designed to quantify differences in artificially inoculated cecal Salmonella Typhimurium populations in growing layer chicks (n = 60 in each of two replications) fed 0%, 25%, and 50% of their diet (w/w) replaced with alfalfa meal from day (d) 7 to d14 after hatch. Alfalfa supplementation reduced cecal populations of Salmonella by 0.95 and 1.25 log10 colony-forming unit per gram in the 25% and 50% alfalfa groups compared to controls. Alfalfa feeding reduced (p < 0.05) the number of cecal- and crop-positive birds compared to controls. Increasing levels of alfalfa increased (p < 0.05) total volatile fatty acids (VFA) and the proportion of acetate in the cecum. Surprisingly, alfalfa inclusion did not negatively impact average daily gain (ADG) in birds over the 7-d feeding period. Alfalfa inclusion at 50% of the diet increased (p < 0.05) the number of bacterial genera detected in the cecum compared to controls, and also altered proportions of the microbial population by reducing Ruminococcus and increasing Clostridia populations. Results support the idea that providing a fermentable substrate can increase gastrointestinal VFA production and bacterial diversity which in turn can reduce colonization by Salmonella via natural competitive barriers. However, further studies are obviously needed to more fully understand the impact of changes made in diet or management procedures on poultry production.