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PURPOSE/OBJECTIVES: The purpose of this article was to provide the clinical nurse specialist (CNS), practicing in settings across the healthcare delivery continuum, with information about physical activity, nutrition, and social support that is essential for optimal aging. BACKGROUND/RATIONALE: Lifestyle choices older adults make concerning physical activity, diet, and social support greatly impact how well they age, their quality of life, and their well-being. DESCRIPTION OF THE PROJECT: This article focuses on 3 key factors essential to successful aging: physical activity, nutrition, and social support. It provides information and assessment and intervention strategy tools that can be used by CNSs to promote optimal aging for older clients. IMPLICATIONS FOR NURSING PRACTICE: This article provides the CNS with resources (literature, assessment tools, and tools for intervention strategies) to promote optimal aging and wellness. CNSs can assist their clients in various settings to age more slowly, delay and prevent certain age-related diseases, and promote more healthful, productive longevity by guiding health-promotion and wellness activities for their older clients. [ABSTRACT FROM AUTHOR]

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.

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.)

Electron-beam (eBeam) irradiation technology has a variety of applications in modern society. The underlying hypothesis was that eBeam-inactivated Salmonella enterica serovar Enteritidis (SE) cells can serve as a vaccine to control SE colonization and shedding in poultry birds. An eBeam dose of 2.5 kGy (kilograys) was used to inactivate a high-titer (10(8) colony-forming units [CFU]) preparation of SE cells. Microscopic studies revealed that the irradiation did not damage the bacterial cell membranes. The vaccine efficacy was evaluated by administering the eBeam-killed SE cells intramuscularly (1 x 10(6) CFU/bird) into 50-wk-old single comb white leghorn hens. On day 14 postvaccination, the hens were challenged orally with live SE cells (1 x 10(9) CFU) and SE colonization of liver, spleen, ceca, and ovaries determined on day 23. Blood samples were collected on days 0, 14, and 23 postvaccination and the sera were analyzed to quantify SE-specific IgG titers. The vaccinated chickens exhibited significantly (P < 0.0001) higher SE-specific IgG antibody responses and reduced SE ceca colonization (1.46 ± 0.39 logi10 CFU/g) compared to nonvaccinated birds (5.32 ± 0.32 log10 CFU/g). They also exhibited significantly lower SE colonization of the ovaries (1/30), spleen (3/30), liver (4/30), and ceca (7/30) compared to nonvaccinated birds. These results provide empirical evidence that eBeam-based SE vaccines are immunogenic and are capable of protecting chickens against SE colonization. The advantages of eBeam-based vaccine technology are that it is nonthermal, avoids the use of formalin, and can be used to generate inactivated vaccines rapidly to address strain-specific infections in farms or flocks.

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.

The etiological agent of necrotic enteritis is Clostridium perfringens. Traditionally, necrotic enteritis is controlled with in-feed antibiotics. However, increasing consumer demand for drug-free poultry has fostered the search for nonantibiotic alternatives. Yeast extract contain nucleotides that are immunomodulatory and also essential for cellular functions. An experiment was conducted to evaluate the efficacy of NuPro yeast extract (Alltech Inc., Nicholasville, KY) in reducing intestinal C. perfringens levels in broiler chickens. One hundred ninety-two 1-d-old male broiler chicks were obtained and randomly assigned to 6 treatments in a battery cage trial. Treatment 1 consisted of chicks fed a corn-soybean meal basal diet (BD) without added bacitracin methylene disalicylate or NuPro. Treatment 2 consisted of chicks fed BD into which bacitracin methylene disalicylate was added at 0.055 g/kg. Treatment 3 consisted of chicks fed BD supplemented with NuPro at a 2% level for the first 10 d of the experiment. Treatments 4 (PX), 5, and 6 (PN) consisted of chicks that were challenged with 3 mL of the C. perfringens inoculum (~10(7) cfu/mL) on d 14, 15, and 16 of the experiment and fed diets similar to treatments 1, 2, and 3, respectively. On d 1 and 7 postchallenge, intestinal C. perfringens levels, lesion scores, and alkaline phosphatase activity were assessed. On d 1 postchallenge, C. perfringens level in treatment 5 (2.09 log(10) cfu/g) was lower (P < 0.05) compared with the PX treatment (4.71 log(10) cfu/g) but similar to the PN treatment (2.98 log(10) cfu/g). A similar trend was observed on d 7 postchallenge. NuPro supplementation enhanced alkaline phosphatase activity (P < 0.05) in C. perfringens-challenged chicks and appeared to reduce intestinal lesion scores. Although dietary supplementation of NuPro in the PN treatment reduced C. perfringens levels by 1.73 and 0.68 log(10) cfu/g compared with the PX treatment on d 1 and 7 postchallenge, respectively, these reductions were not significant. Extending the period of NuPro supplementation beyond the first 10 d of life should be considered for achieving significant reduction in intestinal C. perfringensg levels.

CD40 is mainly expressed by professional antigen-presenting cells (APCs). Its ligand, CD40L, is transiently expressed on activated CD4(+) T-cells. CD40-CD40L interactions mediate T-cell help to APCs and provide crucial signals for affinity maturation and B-cell class switching. In mammals, agonistic monoclonal anti-CD40 antibodies (mAbs) mimic the effects of CD40L on APCs, leading to enhanced T-cell priming and expansion, increased antibody production and class switching. In this study, we describe agonistic anti-chicken CD40 mAb 2C5. This mAb detected CD40 on primary chicken B-cells and macrophages, DT40 B-cells, and HD11 macrophages, induced NO synthesis in HD11 macrophages, and stimulated DT40 B-cell proliferation. These observations demonstrated at least partial functional equivalence of 2C5 to chicken CD154. This mAb may therefore constitute a new tool to study the role of CD40 in the chicken immune system, and its agonistic effects suggest that it could also be used as an adjuvant., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Induction of the innate immune response in newly hatched chickens is important for limiting infections with bacteria, such as Salmonella enterica serovar Enteriditis (SE). CpG oligodeoxynucleotides (CpG-ODN) can stimulate the innate immune response of young chickens. Therefore, we examined the effectiveness of CpG-ODN administered in ovo on intestinal colonization by SE and the ability to modulate the function of heterophils in young chickens. Heterophils were isolated from 2-day-old chickens and were stimulated with heat-killed SE (HK-SE) or PMA for oxidative burst and HK-SE or live SE for degranulation assays. CpG-ODN treatment had no effect on heterophil oxidative burst when stimulated with HK-SE or PMA. However, HK-SE and live SE increased degranulation (P<0.01) in heterophils from CpG-ODN-treated birds compared to PBS-treated controls. In a second experiment, chickens were orally infected with SE on day 10 post-hatch and cecal contents were collected 6 days later for assessment of SE intestinal colonization. CpG-ODN treatment reduced SE colonization by greater than 10-fold (P<0.001) compared to PBS-injected control birds. Overall, we show for the first time that CpG-ODN given in ovo stimulates innate immune responsiveness of chicken heterophils and increases resistance of young chickens to SE colonization.

Foodborne Salmonella continues to be a major cause of salmonellosis with Salmonella Enteritidis and S. Typhimurium considered to be responsible for most of the infections. Investigation of outbreaks and sporadic cases has indicated that food vehicles such as poultry and poultry by-products including raw and uncooked eggs are among the most common sources of Salmonella infections. The dissemination and infection of the avian intestinal tract remain somewhat unclear. In vitro incubation of Salmonella with mammalian tissue culture cells has shown that invasion into epithelial cells is complex and involves several genetic loci and host factors. Several genes are required for the intestinal phase of Salmonella invasion and are located on Salmonella pathogenicity island 1 (SPI 1). Salmonella pathogenesis in the gastrointestinal (GI) tract and the effects of environmental stimuli on gene expression influence bacterial colonization and invasion. Furthermore, significant parameters of Salmonella including growth physiology, nutrient availability, pH, and energy status are considered contributing factors in the GI tract ecology. Approaches for limiting Salmonella colonization have been primarily based on the microbial ecology of the intestinal tract. In vitro studies have shown that the toxic effects of short chain fatty acids (SCFA) to some Enterobacteriaceae, including Salmonella, have resulted in a reduction in population. In addition, it has been established that native intestinal microorganisms such as Lactobacilli provide protective mechanisms against Salmonella in the ceca. A clear understanding of the key factors involved in Salmonella colonization in the avian GI tract has the potential to lead to better approach for more effective control of this foodborne pathogen.

Laying hens are typically induced to molt to begin a new egg-laying cycle by withdrawing feed for up to 12 to 14 d. Fasted hens are more susceptible to colonization and tissue invasion by Salmonella enterica serovar Enteritidis. Much of this increased incidence in fasted hens is thought to be due to changes in the native intestinal microflora. An alternative to feed withdrawal involves feeding alfalfa meal crumble to hens, which is indigestible by poultry but provides fermentable substrate to the intestinal microbial population and reduces Salmonella colonization of hens compared with feed withdrawal. The present study was designed to quantify differences in the cecal microbial population of hens (n=12) fed a typical layer ration, undergoing feed withdrawal, or being fed alfalfa crumble by using a novel tag bacterial diversity amplification method. Bacteroides, Prevotella, and Clostridium were the most common genera isolated from all treatment groups. Only the ceca of hens undergoing feed withdrawal (n=4) contained Salmonella. The number of genera present was greatest in the alfalfa crumble-fed group and least in the feed withdrawal group (78 vs. 54 genera, respectively). Overall, the microbial diversity was least and Lactobacillius populations were not found in the hens undergoing feed withdrawal, which could explain much of these hens' sensitivity to colonization by Salmonella.

A multi-state outbreak investigation of Salmonella Typhimurim cases associated with pet snakes and the frozen vacuum-packed rodents used to feed them identified a Texas frozen feeder rodent facility (Supplier A) as the source of the Salmonella-infected frozen rodents. Texas authorities collected samples directly from Supplier A. Seven Salmonella-positive samples out of 49 environmental swabs were found and one adult mouse out of 88 frozen feeder rodents was Salmonella-positive by culture. No Salmonella strains were isolated from rodent feeds. The pulsed-field gel electrophoresis (PFGE) subtype patterns of S. Typhimurium isolates from feeder rodent and environment samples were indistinguishable from the outbreak strain isolated from humans. A follow-up investigation was performed on all additional feeder rodent facilities identified in Texas. Salmonella was isolated at one of four facilities; seven of 100 rodent samples were positive for Salmonella at this facility. The serotype S. I 4,[5],12:i:- was isolated from seven feeder rodent samples, and PFGE patterns of the seven isolates were indistinguishable. As observed in the initial outbreak investigation, no Salmonella were cultured from rodent feeds at any of the facilities. The feeder rodent industry is an insufficiently recognized industry in the United States. Outbreak investigation and testing of additional feeder rodent facilities in Texas indicate that further evaluation of feeder rodent facilities as a source of Salmonella for pet snakes and humans is warranted.

Since one of the costs in the commercial egg industry is that of replacement pullets, commercial egg layer managers have opted to induce molt older hens in order to extend their productive life for additional egg laying cycles. Conventional molt induction involves the complete removal of feed for several days. However, this management practice can lead to deleterious physiological responses by the hen and subsequent susceptibility to infection by pathogens. Consequently less stressful molting regimens involving the feeding of low energy diets such as alfalfa have been developed. In this study, 80 week old laying hens that were deprived of feed or fed alfalfa meal during a nine day induced molt. Full fed hens were used as the control. On day 8 serum triglycerides were quantified and on day 9 hens were euthanized and the liver, spleen, heart, intestine, pancreas, ovary, and kidney were collected and weighed. Intestinal weight were highest in the non-molted hens, lower in the hens fed alfalfa, and lower still in the hens deprived of feed. Molted hens exhibited reduced weights of liver, heart, ovary, and pancreas compared to the non-molted hens. Serum triglycerides were highest in the non-molted hens, less in feed deprived hens, and the lowest in alfalfa fed hens. These results suggest that a comparable molt could be achieved with feeding alfalfa meal to 80 week hens compared to feed deprivation.

A study was conducted to evaluate the effect of experimental chlorate product (ECP) feed supplementation on Salmonella Typhimurium (ST) in the crop and ceca of market-age broilers. In trial 1, 160 market-age broilers were randomly assigned to 8 treatment groups and replicated twice, with 20 broilers per pen for 1 wk. Trial 2 used the same design, but used 80 market-age broilers with 10 broilers per pen. Treatments were as follows: 1) control feed + double-distilled drinking water (dd H(2)O); 2) control + 18.5% experimental zeolite carrier with dd H(2)O; 3 to 7) control feed supplemented with 0.5, 1.0, 5.0, 10.0, or 18.5% of a feed grade ECP + dd H(2)O; 8) control feed + 1x ECP (0.16% w/v; containing 15 mM chlorate ion equivalent) added to dd H(2)O. Seven-week-old broilers were provided experimental treatments for 7 d, killed, and then ceca and crops were removed and evaluated for ST. Broilers fed 5 to 18.5% ECP or water ECP had a significantly lower (P < 0.05) incidence of ST in the crop (36 to 38% and 14%, respectively) when compared with the control (60%). Broilers fed 10% ECP or water ECP had significantly lower ST crop concentrations (1.03 log(10) and 0.38 log(10) ST/g, respectively) when compared with broilers fed a control diet (1.54 log(10) ST/g). Crop and ceca ST incidence (32 to 48%) and concentration (1.00 to 1.82 log(10) ST/g) were significantly lower in broilers fed 5 to 18.5% ECP as compared with the control (78%; 2.84 log(10) ST/g). Broilers fed 5% or greater ECP had significantly higher water consumption (380 to 580 mL water/d) and litter moisture (31 to 56%) when compared with the control (370 mL water/d; 23% moisture). Only broilers fed 18.5% ECP had significantly lower 7-wk BW (2.77 kg of BW) when compared with the controls (3.09 kg of BW). Average daily gains were significantly depressed in broilers fed 10 or 18.5% ECP compared with the controls. These results indicate broilers supplemented with feed

Molting is a natural process, which birds undergo to rejuvenate their reproductive organs. The US poultry egg production industry has used feed withdrawal to effectively induce molt; however, susceptibility of Salmonella Enteritidis has encouraged the development of alternative methods. Previous research conducted in our laboratory showed that alfalfa is effective at molt induction and provides equivalent postmolt production numbers and quality when compared with feed withdrawal. In the attempt to further increase the efficacy of alfalfa molt diet and decrease the chicken susceptibility to Salmonella Enteritidis during molt, fructooligosaccharide (FOS) was added to a combination of 90% alfalfa and 10% layer ration in 2 levels (0.750 and 0.375%). Ovary and liver colonization by Salmonella Enteritidis in 3 and 2 of the 4 trials, respectively, were reduced (P 0.05) the production of cecal total volatile fatty acids when compared with hens undergoing feed withdrawal. However, in all 3 alfalfa molt diets, the concentrations of lactic acid were greater (P 0.05) were observed among hens fed alfalfa combined with FOS and hens fed alfalfa/layer ration without FOS. Overall, given the similarities between hens fed 0.750% FOS (H) and 0.375% FOS (L), molt diets combined with the lower level of FOS should be sufficient.

The objective of this in vitro study was to evaluate the effects of combining a prebiotic with alfalfa on fermentation by laying hen cecal bacteria. Cecal contents from laying hens were diluted to a 1:3,000 concentration with an anaerobic dilution solution and added to serum tubes filled with ground alfalfa or a layer ration with or without fructooligosaccharide (FOS) prebiotic. Samples were processed in an anaerobic hood, pressurized by using a pressure manifold, and incubated at 37 degrees C. Volatile fatty acid (VFA) and lactic acid concentrations were quantified at 6 and 24 h of substrate fermentation. In this study, fermentation of alfalfa resulted in greater production of acetate, VFA, and lactic acid compared with the layer ration. Although with a relative inconsistency in data between trials, the amendment of FOS to both alfalfa and the layer ration appeared to further increase fermentation as demonstrated by overall higher propionate, butyrate, VFA, and lactic acid concentrations. The effect was more pronounced after 24 h of fermentation, implying time constraints for the optimal production of fermentation products in the chicken gastrointestinal tract. These data indicate that in vitro cecal fermentation can be enhanced by the addition of FOS.

Two studies were conducted to evaluate the effects of melatonin on Salmonella Enteritidis infection in experimentally challenged laying hens subjected to a forced molt. Leghorn hens (>50 wk of age) were randomly assigned to rooms, acclimated to a 16L:8D regimen, and provided ad libitum access to a nonmedicated mash layer diet and water. Birds in one room were molted (8L:16D; complete feed withdrawal), whereas birds in the second room served as nonmolted controls (CONT). Within each room, birds were randomly assigned to melatonin treatment (MEL; 12 birds/treatment), dosed orally commencing the same day as feed withdrawal for 10 d: (experiment I: 0 or 5 mg of melatonin; experiment II: 0, 10, or 20 mg of melatonin). Three days following feed withdrawal, all birds were experimentally infected with Salmonella Enteritidis, and after 10 d of feed withdrawal, all birds were killed and necropsied. In experiment I, concentrations of Salmonella Enteritidis in the cecal contents and the number of Salmonella Enteritidis-positive tissues from the crop, ceca, liver, spleen, and ovary were higher (P < 0.0001) in the MOLT compared with the CONT treatments. No differences (P > 0.10) were observed in any of the parameters examined due to MEL treatment. For experiment II, cecal concentrations of Salmonella Enteritidis were generally higher in the MOLT compared with the CONT treatment and within molted birds, cecal concentrations were higher in the MEL treatment (P < 0.05). Melatonin treatment in molted birds increased (P < 0.05) the percentage of positive crops in the MOLT+20 MEL treatment (P < 0.05). Salmonella-positive cecal tissue was increased (P < 0.001) in MOLT compared with CONT birds and was also higher in MOLT+10 MEL and MOLT+20 MEL birds compared with the MOLT-only treatment. Results from the current research suggest that dosage with high levels of melatonin may exacerbate Salmonella Enteritidis infection in layers subjected to forced molt.

Induced molting by feed withdrawal has been a common practice in the commercial layer industry and usually involves the removal of feed for a period of up to 14 d. However, this is a practice that is believed to adversely influence the welfare of the hens and there is a need to examine behavioral responses to alternative molt regimens. The behavioral patterns of hens on 90% alfalfa:10% layer ration, 80% alfalfa:20% layer ration, and 70% alfalfa:30% layer ration molt diets were compared with feed withdrawal (FW) hens, and fully fed (FF) hens. The White Leghorn laying hens were approximately 54 wk old and were placed in 3 identical climate-controlled rooms. The hens were individually housed in 2-tier wire battery cages and provided treatment rations and water ad libitum. Nonnutritive pecking, walking, drinking, feeder activity, preening, aggression, and head movement were quantified during two 10-min periods each day for 6 hens from each treatment. Over the 9-d treatment period, hens in the FW, 70% alfalfa:30% layer ration, and 80% alfalfa:20% layer ration groups spent significantly more time walking than hens in the 90% alfalfa:10% layer ration group. The FF and 70% alfalfa:30% layer ration hens spent half as much time preening, whereas the FW hens displayed nearly twice as much nonnutritive pecking when compared with other treatments. Most differences in head movements occurred at the beginning of the molt period, whereas during the last half of molt, alfalfa-fed hens exhibited feeder activity similar to FF hens, and all were significantly higher than that of FW hens. After some initial adjustment by the hens, consumption of alfalfa molt diets appeared to reduce nonnutritive pecking behavior, which is characteristically associated with FW hens.

Several dietary alternatives to feed withdrawal have been proposed to induce a molt in laying hens. This study compared the behavior of laying hens on an alfalfa crumble diet (ALC) to hens that were either on a conventional layer diet (FF) or hens that had feed withdrawn (FW) during a 9-d trial. Each treatment consisted of 24 hens (3 hens per battery cage), and treatment began after a 2-week acclimation period. Video cameras connected to a digital multiplexer recorded the behavior of the hens. The percentages of observations performing nonnutritive pecking, feeder activity, drinking, walking, preening, head movement, and aggression were quantified for two 10-min periods at daily intervals. The FF hens spent significantly more (P < or = 0.05) time drinking than the other treatments, whereas FW hens displayed the most head movements. From d 1 through 7, FW hens walked less than ALC hens except on d 8 when FW hens walked more than ALC and FF hens. On d 4 and 6, the FW hens spent an increased amount of time preening compared with FF hens until the last few days of the molt period. For the most part, FW hens generally displayed more nonnutritive pecking than ALC and FF hens throughout the molt period. However, FW hen visits to the feeders declined as the trial proceeded, whereas ALC and FF hens generally spent more time at the feeder. In summary, the ALC diet showed potential as an alternative to FW for inducing a molt in laying hens based on reduced nonnutritive pecking behavior, head movements, and greater feeding activity.

The practice of induced molting involves the restriction of light, feed removal and optionally water for 5-14 days. However, there is growing concern regarding feed removal and animal welfare issues. With this in mind, alternative diets have been developed to produce similar molting effects as that of feed deprivation. Alfalfa, which largely consists of insoluble fiber, can be used as a molting diet. In this study, heterophil and lymphocyte counts, serum chemistry, and organ weight parameters were evaluated in hens that were deprived of feed or fed alfalfa during a nine day induced molt. Full-fed hens were used as the control. Blood serum parameters assessed included calcium, magnesium, glucose, total protein, ketone bodies, uric acid, and cholesterol. White blood cells were counted and categorized by cell type. On the ninth day of the trial, the hens were euthanized and the liver, spleen, heart, intestine, pancreas, ovary, oviduct, and kidney were collected and weighed. On day 8 birds molted with alfalfa or by feed deprivation had significantly higher (P<0.05) levels of ketone bodies and cholesterol and lower levels of calcium, and magnesium compared to the full-fed hens while birds molted by feed deprivation exhibited significantly lower levels of uric acid. Birds molted by both methods exhibited significant reductions in ovary, oviduct, liver and pancreas weights and increased spleen weights when compared to the non-molted hens. On days 0, 2, and 6 there were no significant differences (P>0.05) in either heterophil or lymphocyte percentages. However, heterophil percentages were higher in feed withdrawal birds than full-fed birds on day 4 but lymphocyte percentages were higher in full-fed birds compared to feed withdrawal birds. On day 8 of the induced molt lymphocyte percentages were higher from full-fed birds when compared to feed withdrawal birds but no significant differences were detectable for heterophil percentages. Based on reproductive organ weight loss and changes in serum and immunological responses of birds during molt, it appears that alfalfa meal can be an effective molt induction alternative.

Immunological responses of molting hens either infected or not infected with Salmonella enterica serovar Enteritidis were compared in 2 trials with Single Comb White Leghorn hens >50 wk old. The hens were placed into 6 treatment groups with 12 hens per group: nonmolted Salmonella Enteritidis positive (FF+), non-molted Salmonella Enteritidis negative (FF-), feed withdrawal Salmonella Enteritidis positive (FW+), FW Salmonella Enteritidis negative (FW-), alfalfa Salmonella Enteritidis positive (ALC+), and ALC Salmonella Enteritidis negative (ALC-). Each hen in the Salmonella Enteritidis-positive groups was challenged on d 4 of the study with 1 mL of 10(6)-cfu Salmonella Enteritidis, and diets were administered for 12 d. Blood samples were collected on d 2, 5, 9, and 12, and blood smears were enumerated for heterophil to lymphocyte (H:L) ratios. Serum samples were also analyzed for alpha(1)-acid glycoprotein (AGP) levels and antibody level. On d 12, hens were euthanized and bile samples from the gall bladder and sections of the ileum and the ceca were collected, and an ELISA was used to determine the intestinal, serum, and bile antibody responses. The FW+ hens produced more (P

The objective of this study was to examine an alfalfa crumble diet as an alternative molt diet and compare the physiological response of hens to the responses of feed-deprived molted hens. Hens >50 wk old were placed into 6 treatment groups (12 hens per group in trial 1 and 10 hens per group in trial 2): nonmolted Salmonella enterica serovar Enteritidis positive (FF+), nonmolted Salmonella Enteritidis negative (FF-), feed withdrawal Salmonella Enteritidis positive (FW+), FW Salmonella Enteritidis negative (FW), alfalfa Salmonella Enteritidis positive (ALC+), and ALC Salmonella Enteritidis negative (ALC-). Each hen in the Salmonella Enteritidis-positive groups was challenged on the fourth day of the study with 1 mL of 10(6)-cfu Salmonella Enteritidis. Blood was collected on d 2, 5, 9, and 12 of the trial. Blood plasma was collected and metabolite concentrations were analyzed for glucose, calcium, cholesterol, uric acid, total protein, and triglycerides. The feed intakes of the FF hens were 4- to 6-fold greater (P

The objective of this study was to investigate the effect of combining a prebiotic with poultry feeds on the growth of Salmonella enterica serotype Typhimurium (ST) in an in vitro cecal fermentation system. Cecal contents from three laying hens were pooled and diluted to a 1:3000 concentration in an anaerobic dilution solution. The cecal dilution was added to sterile test tubes filled with alfalfa and layer ration with and without fructooligosaccharide (FOS). Two controls containing cecal dilutions and anaerobic dilution solution were used. The samples were processed in the anaerobic hood and incubated at 37 degrees C. Samples were inoculated with Salmonella at 0 and 24h after in vitro cecal fermentation and plated at 0 and 24h after inoculation with ST. Plates were incubated for 24h and colony forming units (CFU) enumerated. The samples immediately inoculated with ST without prior cecal fermentation did not significantly lower ST counts 24h later. However, samples pre-incubated for 24h with cecal microflora prior to ST inoculation exhibited reduced ST CFU by approximately 2 logarithms, with the most dramatic decreases seen in alfalfa and layer ration combined with FOS. The addition of FOS to feed substrate diets in combination with cecal contents acted in a synergistic manner to decrease ST growth only after ST was introduced to 24h cecal incubations.

The objective of this study was to examine microbial population shifts and short-chain fatty acid (SCFA) responses in the gastrointestinal tract of Salmonella Enteritidis-challenged molted and nonmolted hens fed different dietary regimens. Fifteen Salmonella-free Single Comb Leghorn hens (>50 wk old) were assigned to 3 treatment groups of 5 birds each based on diet in 2 trials: 100% alfalfa crumbles (ALC), full-fed (FF, nonmolted) 100% commercial layer ration, and feed withdrawal (FW). A forced molt was induced by either a 12-d alfalfa diet or FW. In all treatment groups, each hen was challenged by crop gavage orally 4 d after molt induction with a 1-mL inoculum containing 10(6) cfu of Salmonella Enteritidis. Fecal and cecal samples (d 4, 6, 8, 11, and necropsy on d 12) were collected postchallenge. Microbial population shifts were evaluated by PCR-based 16S ribosomal RNA gene amplification and denaturing gradient gel electrophoresis, and SCFA concentrations were measured. Total SCFA in fecal and cecal contents for FW molted hens were generally lower (P < or = 0.05) in the later stages of the molt period when compared to ALC and FF treatment groups. The overall trend of SCFA in cecal and fecal samples exhibited similar patterns. In trials 1 and 2, hens molted with ALC diet generally yielded more similar amplicon band patterns with the FF hens in both fecal and cecal samples by the end of the molting period than with FW hens. The results of these studies suggest that ALC molted hens supported microflora and fermentation activities, which were more comparable to FF hens than FW hens by the end of the molting period.

The objectives of this study were to enumerate Salmonella enterica serovar Enteritidis colonization in fecal, cecal, and internal organs, and to compare the level of virulence gene expression (hilA) of experimentally challenged laying hens fed different dietary molt-induction regimens. Twelve Salmonella-free Single Comb Leghorn hens (>50 wk old) hens were randomly assigned to each of 6 treatment groups designated based on diet in 2 trials: 1) feed withdrawal Salmonella Enteritidis-positive (FW+), 2) fully fed Salmonella Enteritidis-positive (FF+), 3) 100% alfalfa crumble Salmonella Enteritidis-positive (ALC+), 4) feed withdrawal Salmonella Enteritidis-negative, 5) fully fed Salmonella Enteritidis-negative, and 6) 100% alfalfa crumble Salmonella Enteritidis-negative. A forced molt was induced by a 12-d alfalfa diet and a feed-withdrawal regimen. On d 4 of the molt, all hens in groups 1, 2, and 3 were challenged by crop gavage with 1 mL of inocula containing approximately 10(6) cfu of nalidixic acid- and novobiocin-resistant Salmonella Enteritidis (phage type 13A). At the conclusion of both trials, all hens were euthanized and Salmonella Enteritidis colonization was enumerated in the cecal contents, liver, spleen, and ovaries. In addition, fecal (d 4 and 8) and cecal samples (necropsy at d 12) were collected postchallenge from treatment groups 1, 2, and 3 (Salmonella Enteritidis-positive) to quantify hilA expression by PCR. In both trials, all nonchallenged birds were Salmonella Enteritidis-negative; therefore, no further analysis was done. In trial 1, a 2-fold reduction in Salmonella Enteritidis colonization was observed in the ALC+ hens (log10 Salmonella Enteritidis of 1.99) compared with the FW+ hens (log(10) Salmonella Enteritidis of 3.89). In trial 2, a 4-fold reduction in Salmonella Enteritidis colonization was observed in the ALC+ hens (log(10) Salmonella Enteritidis of 1.27) compared with the FW+ hens (log(10) Salmonella Enteritidis of 5.12). In trial 2, Salmonella Enteritidis colonization in spleens was higher (P 0.05). In trial 2, hilA expression in FW+ hens was higher (P

Clostridium perfringens is the etiologic agent of necrotic enteritis (NE) and is ubiquitous in nature. The incidence of NE has increased in countries and commercial companies that have stopped using antibiotic growth promoters. The mechanisms of colonization of C. perfringens and the factors involved in onset of NE are not fully understood. Previously, our laboratory has demonstrated that lactose could potentially reduce Salmonella and C. perfringens in ceca of poultry. In the present investigation, we hypothesized that dietary lactose would reduce the clinical signs of NE and could be used as an alternative to antibiotics. In experiment 1, day-of-hatch broilers were fed either a nonlactose control diet, a diet with 2.5% lactose, or a diet with 4.5% lactose throughout the experiment. Birds were administered C. perfringens (10(7) cfu/mL) daily via oral gavage for 3 consecutive days starting on d 17. When evaluating the intestinal lesions associated with NE, birds fed 2.5% lactose had significantly lower (P < 0.05) lesion scores (0.70 +/- 0.52) compared with the control (1.55 +/- 0.52) or the 4.5% lactose (1.60 +/- 0.52). The data from the microbial analysis showed that the addition of lactose did not affect any bacterial populations when compared with the control birds that did not receive dietary lactose over the 21-d evaluation. The overall lesion scores in experiment 2 were significantly (P < 0.05) reduced in birds fed 2.5% lactose compared with the birds fed the control diet with mean lesion scores of 1.10 +/- 0.73 and 1.80 +/- 0.73, respectively. These experiments suggest that lactose could be used as a potential alternative to growth-promoting antibiotics to help control this costly disease.

An experimental chlorate product that targets the respiratory nitrate reductase enzyme of bacteria such as Salmonella and Escherichia coli has shown promising results in reducing concentrations of these bacteria in the gut of food animals. Because expression of the target enzyme is induced by nitrate, we administered short-duration, low level nitrate or nitroethane preconditioning treatments to finishing swine to see if these would enhance the ability of an experimental chlorate product to kill these bacteria. Results from these studies showed that preconditioning the gut microflora of swine with low levels of nitrate or nitrocompounds enhanced (more than tenfold) the ability of the chlorate product to kill Salmonella and E. coli, but not Campylobacter. Further studies are needed before these compounds can be fed as feed additives to animals, although it is likely that nitrate preconditioning may be more near to market than the nitrocompounds, which may require more comprehensive review by regulatory authorities.

Objectives: To examine the ability of flavophospholipol to inhibit bacterial conjugation between Escherichia coli donor and recipient pairs in vitro and in day-of-hatch chickens., Methods: In vitro donor cultures were incubated in the presence of 0, 2, 4, 8, 16, 32 and 64 mg/L flavophospholipol during primary overnight mono-cultures only, secondary conjugation cultures only, or throughout primary and secondary cultures. Transconjugants were selected using oxytetracycline and nalidixic acid. Treatment groups A-G (n=20) of day-of-hatch broiler chickens received 0, 2, 4, 8, 16, 32 and 64 g/ton flavophospholipol, respectively, in their feed throughout the experiment. On day 4, all treatment groups were given 0.25 mL of donor and recipient E. coli at 7.0 and 9.0 log10 cfu/mL, respectively. On day 10, the birds were euthanized and the caecal contents were cultured on selective medium (oxytetracycline and nalidixic acid)., Results: A dose-dependent reduction in transconjugant populations was observed in vitro when flavophospholipol was present in the secondary conjugation culture. The susceptibility profiles of transconjugants obtained from in vitro studies were identical to the predicted profile of the donor and recipient combination. There was no significant difference (P>or=0.05) in the number of transconjugants isolated from chickens among any of the flavophospholipol treatment groups when compared with the controls. The susceptibility profiles of chicken transconjugants suggested acquisition of naturally occurring plasmids., Conclusions: Flavophospholipol strongly inhibited conjugation in vitro, but did not prevent recipient E. coli from acquiring resistance determinants in vivo.

Feed deprivation is commonly used by the poultry industry to induce molting and stimulate multiple egg-laying cycles. However, feed deprivation has been observed experimentally to increase susceptibility of poultry to Salmonella infections. Previous studies indicated that alfalfa was efficacious in reducing Salmonella; the present investigation was designed to evaluate the efficacy of combined alfalfa and layer diets on Salmonella colonization. Leghorn hens over 50 wk of age were divided into 12 groups of hens and placed in individual laying cages. One week prior to dietary changes, hens were put on an 8L:16D photoperiod that continued for the 9-d experiment. Hens were challenged orally with 104 cfu of Salmonella Enteritidis (SE) on d 4 of treatment and cultured for SE at the termination of the 9-d study. Two independent experiments were conducted consisting of the following treatment groups: nonfed hens, full-fed standard commercial layer diet, 100% alfalfa meal diet, a 90% alfalfa meal/10% standard commercial layer diet, and a 70% alfalfa meal/30% standard commercial layer diet. When evaluating SE colonization in the ceca (Exp. 1), a reduction (P < 0.05) was seen in the 100% alfalfa meal diet and the 70% alfalfa meal/30% standard commercial layer diet treatment groups when compared with the controls with Log10 values of 0.54, 0.44, and 2.82, respectively. Evaluation of physiological parameters showed the alfalfa treatment groups had reductions (P < 0.05) in weight loss, ovary weight, and feed consumption when compared with the full-fed standard commercial layer diet hens, and these results were comparable with the nonfed hens. In Exp. 2, all of the treatment groups had a reduction (P < 0.05) in SE colonization of the ceca when compared with the controls. There were also similar physiological reductions in weight loss, ovary weight, and feed consumption when birds were fed the alfalfa diets in Exp. 2. These data suggest that alfalfa can potentially be combined with layer ration to limit SE infection and still induce a molt comparable with feed withdrawal.

Foodborne diseases caused by enterohemorrhagic Escherichia coli, Salmonella, and Campylobacter species are of public health and economic significance. Shedding of these pathogens during production and slaughter are risks for contamination of products for human consumption. Consequently, strategies are sought to prevent or reduce the carriage of these pathogens in food animals before slaughter. Experimental products containing chlorate salts have been proven efficacious in reducing concentrations of E. coli and Salmonella Typhimurium in the gut of cattle, sheep, swine, and poultry when administered as feed or water additives. Mechanistically, chlorate selectively targets bacteria expressing respiratory nitrate reductase activity, such as most members of the family Enterobacteriaceae, as this enzyme catalyzes the reduction of chlorate to lethal chlorite. Most beneficial gut bacteria lack respiratory nitrate reductase activity, and thus the technology appears compatible with many bacteria exhibiting competitive exclusion capabilities. More recently, select nitrocompounds have been investigated as potential feed additives, and although these nitrocompounds significantly reduce pathogens on their own, evidence indicates that they may most effectively be used to complement the bactericidal activity of chlorate. A particularly attractive aspect of the nitrocompound technology is that, as potent inhibitors of ruminal methanogenesis, they may allow producers the opportunity to recoup costs associated with their use. At present, neither chlorate nor the nitrocompounds have been approved as feed additives by the US Food and Drug Administration, and consequently they are not yet available for commercial use.

Clostridium perfringens (CP) is the etiologic agent of necrotic enteritis (NE). Clinical signs of this disease include depression, decreased appetite, diarrhea, and severe necrosis of the intestinal tract. Understanding the disease progression of NE has been difficult due to its complexity and the involvement of multiple factors (dietary components, immunosuppression, and mechanical irritation of the gut) that appear to contribute to this syndrome. In the present investigation, day-of-hatch broilers were fed a 55% wheat diet and randomly assigned to 1 of 8 groups. Treatments included positive control (CP challenge only), commercial coccidia vaccine (CCV), commercial bursal disease vaccine (CBDV), or the combination of CCV and CBDV, and an appropriate negative control for each (vaccinated and not challenged). Challenged treatment groups received 10(7) cfu of CP twice daily. When compared with controls, broilers in each treatment group had increased (P < or = 0.05) lesion scores, with mean scores of 1.05 and 2.05 in the CP and CBDV + CP treatments, respectively. When compared with controls, the incidence of CP increased (P < or = 0.05) in all treatment groups (73 and 100% in the CCV + CP and CBDV + CP treatment groups, respectively). Compared with controls, percentage mortality increased (P < or = 0.05) from 2% to 26 and 34% in the CP and CBDV + CP treatment groups, respectively. Results of this study indicate that the methodology used provides a good model for studying NE.

Previous reports have shown that some bacteria, including Salmonella, use a dissimilatory nitrate reductase enzyme pathway (NREP) in anaerobic environments. This enzyme reduces nitrate to nitrite and has been shown to cometabolize chlorate to cytotoxic chlorite. The present investigations were performed to evaluate the susceptibility of a competitive exclusion culture (CE) to the experimental chlorate product (ECP). A commercially available CE product was evaluated for its nitrate reductase activity and therefore its chlorate sensitivity. Individual isolates (in triplicate) were cultured in 10 mL of Viande Levure broth containing 5 mM sodium nitrate or 10 mM sodium chlorate. Bacterial growth (optical density at 625 nm) was measured and 1-mL aliquots were removed concurrently for colorimetric determination of nitrate content at 0, 3, 6, and 24 h. Of the 15 different facultative strains, 11 had slight NREP utilization, 3 had moderate NREP utilization, and the remainder were NREP negative (with slight and moderate NREP utilization: >0.1 to <1.0 mM and >1.0 mM nitrate used within 6 h, respectively). Of the obligate anaerobes evaluated, 3 had slight NREP utilization and the remainder were NREP negative. In vivo studies utilizing both products (CE and ECP) in a horizontal transmission challenge model (seeders + contacts) showed significant reductions in Salmonella from 5.37 to 1.76 log10 cfu/g and 3.94 to 0.07 log10 cfu/g, respectively. The combined effect of the CE culture and an ECP are effective in killing these food-borne pathogens.

Feed deprivation is the most common method used to induce molting and stimulate multiple egg-laying cycles in laying hens for commercial egg production. Unfortunately, an increased risk of Salmonella enteritidis (SE) colonization may result from the use of this method. Methods to stimulate multiple egg-laying cycles without increasing the risk of SE are needed. In each of 3 experiments, hens over 50 wk of age were divided into groups of 12 and placed in individual laying cages. One week before dietary changes, hens were put on an 8L:16D photoperiod that continued for the 9-d experimental period. Hens in all treatments were challenged orally with 10(4) cfu of SE on the fourth day. Treatments were full fed hens (nonmolted, NM), nonfed hens (molted, M), a zinc acetate diet (ZAC), and a zinc propionate diet (ZPR). The zinc diets contained 10,000 mg of zinc per kilogram of diet. Body weight losses were significantly higher in the M, ZPR, and ZAC treatments than in the NM treatment. Crop lactic acid decreased more in M, ZPR, and ZAC treatments than in NM hens in trial 2. Crop pH was significantly (P < 0.05) lower in NM hens than in M, ZAC, and ZPR hens in trial 2. Although cecal individual or total volatile fatty acids (VFA), and lactic acid were not significantly (P > 0.05) different between NM hens and M, ZAC and ZPR hens in trial 1, lactic acid was significantly (P < 0.05) higher in NM hens than in M, ZAC and ZPR hens (trial 2), and cecal total VFA were lower in M hens than in NM, ZAC and ZPR hens (trial 3). Colonization of SE in the crop and ceca was higher in the M and ZPR hens (trials 1 and 2). Liver, spleen, or ovary invasion by SE was higher in the M and ZPR hens (trials 1 and 2) than in NM hens. At the zinc concentration used in these studies, the zinc dietary regimens may be effective for reducing the risk of SE during induced molt.

Mature, healthy lactating dairy cattle were sampled on two farms in the southwestern United States to examine the effects of heat stress (Experiment I) and stage of lactation (Experiment II) on the fecal shedding of E. coli O157:H7 and Salmonella. To examine the effects of heat stress, fecal samples were collected from 45 cows at 7:00 AM (coolest part of the day) and 5:00 PM (hottest part of the day) in August 2002 on a 250 cow dairy. The study was replicated one month later (n = 170 total samples). A temperature-heat index (THI) was calculated for each sampling time. In Experiment II, stage of lactation was examined by sampling lactating dairy cattle early [< 60 days in milk (DIM)] and late (> 150 DIM) in the lactation cycle in the summer of 2001. The study was replicated the following summer (60 cows/group/replicate; n = 240 total samples). For Experiment I, THI averaged 75 and 82 for the AM and PM samplings, respectively, indicating the cows were beginning to experience heat stress in the morning and by afternoon were in severe heat stress. The shedding of E. coli O157:H7 tended to be higher (p = 0.09) in the afternoon sampling of the first replicate, however was not different in the second replicate or when both replicates were pooled (p > 0.10). Salmonella shedding was not different (p > 0.10) at any sampling time with nearly 100% of the cows positive. Stage of lactation had no effect on the number of cows shedding E. coli O157:H7 (p > 0.10). Salmonella shedding tended to be higher (p = 0.09) in early lactation cows in the first replicate, while in the second replicate more late lactation cows were shedding Salmonella (p < 0.05); however, there were no differences due to stage of lactation when replicates were pooled (p > 0.10). While further research is needed, results of this research highlight the variability in pathogen shedding in healthy dairy cattle and indicate that environmental factors and/or production demands may influence shedding patterns of E. coli O157:H7 and Salmonella.

Mature dairy cattle were sampled over a 2-year period (2001-2002) on six farms in New Mexico and Texas. Fecal samples (n = 1560) were collected via rectal palpation and cultured for Salmonella, and one isolate from each positive sample was serotyped. Three isolates of each serotype, with the exception of Salmonella Newport (n = 12), were examined for susceptibility to 17 antimicrobial agents. Twenty-two different serotypes were identified from a total of 393 Salmonella isolates. Montevideo was the predominant serotype (27%) followed by Mbandaka (15%), Senftenberg (11.4%), Newport (6.4%), Anatum (4.8%), and Give (4.8%). Salmonella Typhimurium and Dublin, two frequently reported serotypes, accounted for only 1% of the observed serotypes in this study. Sixty-four percent of the serotypes were susceptible to all 17 antimicrobials, 14% were resistant to a single agent, and 22% were multiresistant (2-11 types of resistance). All isolates tested were susceptible to amikacin, apramycin, imipenem, ceftriaxone, nalidixic acid, and ciprofloxacin. The most frequent types of resistance were to sulfamethoxazole, tetracycline, streptomycin, kanamycin, chloramphenicol, and ampicillin (ranging from 8.9 to 22.4%). Serotypes demonstrating multiple resistance included Dublin and Give (resistant to three or more antibiotics), Typhimurium (resistant to five antibiotics), and Newport (four and two isolates resistant to six and nine antibiotics, respectively). Class 1 integrons were present in only two Salmonella Dublin isolates and one Salmonella Newport isolate. The most prevalent resistance patterns observed in this study were toward antimicrobial agents commonly used in cattle, while all Salmonella isolates were susceptible to ceftriaxone and ciprofloxacin, antibiotics used in human medicine.

Aims: To examine the variability in faecal shedding of Salmonella and Escherichia coli O157:H7 in healthy lactating dairy cattle and to evaluate the genetic relatedness of Salmonella isolates., Methods: Faecal samples were obtained from lactating Holstein dairy cattle on four commercial farms in the southwestern US. All farms were within an 8-km radius and were sampled in August 2001, January 2002 and August 2002 (60 cows per farm per sampling; n = 720 total samples). Samples were cultured for E. coli O157:H7 and Salmonella and a portion of the recovered Salmonella isolates were examined for genetic relatedness using pulsed-field gel electrophoresis (PFGE)., Results: Faecal shedding of E. coli O157:H7 and Salmonella varied considerably between farms and at the different sampling times. Large fluctuations in the percentage of positive animals were observed from summer to summer for both of these pathogens. Similarly, Salmonella serotype and serotype prevalence varied from farm to farm and within farm from one sampling time to another. Multiple Salmonella genotypes were detected for a number of serotypes and identical genotypes were found on different farms with one genotype of Salmonella Senftenberg identified on three of the four farms., Significance and Impact of the Study: This study demonstrated the wide variability in pathogen shedding within and among dairy farms all located in a small geographical region and highlights the complexity of pathogen control at the farm level.

Aims: To examine factors affecting faecal shedding of the foodborne pathogens Escherichia coli O157:H7 and Salmonella in dairy cattle and evaluate antimicrobial susceptibility of these isolates., Methods: Faecal samples were obtained in replicate from lactating (LAC; n = 60) and non-lactating (NLAC; n = 60) Holstein cattle to determine influence of heat stress, parity, lactation status (LAC vs NLAC) and stage of lactation [60 days in milk (DIM)] and cultured for E. coli O157:H7 and Salmonella. A portion of the recovered isolates were examined for antimicrobial susceptibility using the broth microdilution technique., Results: No effects of heat stress were observed. Lactating cows shed more (P < 0.01) E. coli O157:H7 than NLAC cows (43% vs 32%, respectively). Multiparous LAC cows tended to shed more (P = 0.06) Salmonella than primiparous LAC cows (39% vs 27%, respectively). Parity did not influence (P > 0.10) bacterial shedding in NLAC cows. Cows 60 DIM. Seventeen Salmonella serotypes were identified with the most prevalent being Senftenberg (18%), Newport (17%) and Anatum (15%). Seventy-nine of the Salmonella isolates were resistant to at least one of the seven antibiotics. Escherichia coli O157:H7 isolates were resistant to 11 different antibiotics with multiple resistance to nine or more antibiotics observed in five isolates., Significance and Impact of the Study: This study demonstrated differences in the shedding patterns of foodborne pathogens due to the stage of the milk production cycle and may help identify times when on-farm pathogen control would be the most effective.

Previous published research has identified the crop as a source of Salmonella and Campylobacter contamination for broiler carcasses and reported that broiler crops are 86 times more likely to rupture than ceca during commercial processing. Presently, we evaluated leakage of crop and upper gastrointestinal contents from broilers using a fluorescent marker at commercial processing plants. Broilers were orally gavaged with a fluorescent marker paste (corn meal-fluorescein dye-agar) within 30 min of live hang. Carcasses were collected at several points during processing and were examined for upper gastrointestinal leakage using long-wavelength black light. This survey indicated that 67% of the total broiler carcasses were positive for the marker at the rehang station following head and shank removal. Crops were mechanically removed from 61% of the carcasses prior to the cropper, and visual online examination indicated leakage of crop contents following crop removal by the pack puller. Examination of the carcasses prior to the cropper detected the marker in the following regions: neck (50.5% positive), thoracic inlet (69.7% positive), thoracic cavity (35.4% positive), and abdominal cavity (34.3% positive). Immediately prior to chill immersion, 53.2% of the carcasses contained some degree of visually identifiable marker contamination, as follows: neck (41.5% positive), thoracic inlet (45.2% positive), thoracic cavity (26.2% positive), and abdominal cavity (30.2% positive). These results suggest that this fluorescent marker technique may serve as a useful tool for rapid identification of potential changes, which could reduce the incidence of crop rupture and contamination of carcasses at processing.

The crop is a known source of Salmonella and Campylobacter contamination. We evaluated the use of selected organic acids (0.5% acetic, lactic, or formic) in drinking water during a simulated 8-h pretransport feed withdrawal (FW). Salmonella typhimurium was recovered from 53/100 control crops and from 45/100 of crops from acetic acid-treated broilers. However, treatment with lactic acid (31/100) or formic acid (28/76) caused significant (P < 0.05) reduction in incidence. Reductions of recovered incidence were also associated with reduced numbers of S. typhimurium recovered (e.g., control, log 1.45 cfu/crop; lactic acid, 0.79 cfu/crop). In an additional commercial farm study, broilers were provided 0.44% lactic acid during a 10-h FW (4 h on the farm and 6 h transport) and pre-FW crop, post-FW crop, and pre-chill carcass wash samples were collected for Campylobacter and Salmonella detection. Crop contamination with Salmonella was significantly reduced by lactic acid treatment (6/175) as compared with controls (29/175). Importantly, Salmonella isolation incidence in prechill carcass rinses was significantly reduced by 52.4% with the use of lactic acid (26/175 vs. 55/176). Crop contamination with Campylobacter was significantly reduced by lactic acid treatment (62.3%) as compared with the controls (85.1%). Lactic acid also reduced the incidence of Campylobacter found on pre-chill carcass rinses by 14.7% compared with the controls. These studies suggest that incorporation of lactic acid in the drinking water during pretransport FW may reduce Salmonella and Campylobacter contamination of crops and broiler carcasses at processing.

Competitive exclusion (CE) cultures may offer alternatives to antimicrobial agents for disease prophylaxis in poultry. To avoid potential transfer of antibiotic resistance, safe and effective CE cultures must, by necessity, be highly sensitive to antimicrobial residues. The following studies evaluated the effect of maternal administration of selected antibiotics on the establishment of a licensed CE culture, PREEMPT. Selected antibiotics were administered to actively laying hens for a period of 7 days (experiment 1) or 9 days (experiment 2) in drinking water [sulfadimethoxine (0.05%), enrofloxacin (0.005%), and tylosin tartrate (0.05%)] or feed (sulfadimethoxine with ormetoprim, 250 ppm). In experiment 1, fertile eggs were collected daily and subjected to bioassay for detectable antimicrobial residues in yolk. Antimicrobial residues were not detected during the 7 days of treatment or the subsequent 3 days following cessation of treatment in the control, sulfadimethoxine, sulfadimethoxine with ormetoprim, or tylosin treatment groups. However, detectable residues were observed in eggs derived from enrofloxacin-treated hens on days 6 and 7 during antibiotic administration and also on days 2 and 3 post-antibiotic administration. In experiment 2, antimicrobial residues were also only detected in yolks from hens treated with enrofloxacin. Residue detection occurred on days 2-6 of antibiotic administration, on day 9 of antibiotic administration, on days 1-3 post-antibiotic administration, and also on day 7 post-antibiotic administration. A subset of eggs from each experimental group, corresponding to days 2-6 of antibiotic administration, days 4-6 post-antibiotic administration, and days 14-16 post-antibiotic administration, were pooled for incubation, and chicks hatched from these pools of fertile eggs were treated with PREEMPT at hatch. When 48-h cecal propionate concentrations were used as an index of culture establishment, reduced (P < 0.05) efficacy was observed only in chicks derived from enrofloxacin-treated hens at either collection period. Although several antibiotics do not appear to produce detectable egg residues or interfere with CE culture establishment, these data suggest that chicks derived from enrofloxacin-treated hens may not be candidates for safe and effective CE culture treatment.

Competitive exclusion of intestinal pathogens by administration of beneficial and defined cultures of normal intestinal microflora is a safe and effective means of reducing the incidence and severity of chick infections with Salmonella and other intestinal pathogens. It is important that competitive exclusion cultures not carry genetic material (e.g., plasmids), which could transfer antibiotic resistance to other microflora, including pathogens. As such, safe and effective competitive exclusion cultures must be sensitive to commonly used antimicrobial agents. By necessity, intentional or accidental exposure of these beneficial microflora to antibiotics will reduce or eliminate the protection provided by competitive exclusion culture establishment. As antibiotic residues can be present from embryonic, hatchling, or maternal administration, a rapid and sensitive assay for the nonspecific detection of residues, which could interfere with competitive exclusion culture establishment, is needed. This study was conducted to develop a rapid and inexpensive bioassay to detect multiple antimicrobial residues in egg yolk and neonatal yolk sacs. Aerobic bacterial strains with known sensitivity to several antibiotics used by the poultry industry were selected and individually compared for sensitivity to enrofloxacin, gentamicin, tetracycline, ceftiofur, and tylosin concentrations in egg yolks. This assay was found to be relatively sensitive for the detection of these antimicrobials, and detection of residues was associated with reduced competitive exclusion culture (PREEMPT) establishment in one experiment. Importantly, this assay can be implemented with minimal training or equipment under commercial hatchery practices and could be used to determine embryo groups, in advance of hatch, that are not suitable candidates for competitive exclusion treatment in the hatchery.

The effect of in ovo or day-of-hatch subcutaneous antibiotic administration on the detection of antibiotic residues in yolk sac or blood serum samples and the potential for observed residues to interfere with competitive exclusion (CE) culture establishment was compared in three experiments. The in ovo or subcutaneous administration of gentamicin sulfate or ceftiofur sodium was associated with detectable levels of antibiotic residues in yolk sac or blood serum samples in Experiment 1. Further, the ability to detect antibiotic residues in day-of-hatch chicks was associated with reduced levels of CE culture establishment when cecal propionate level, an indicator of PREEMPT establishment, was determined following PREEMPT application by oral gavage on the day of hatch in Experiments 1 and 2. Restricting chicks from feed, as opposed to providing access ad libitum to a starter ration, for 6 h immediately following administration of PREEMPT improved (P < 0.05) CE culture establishment in Experiment 2 in nonantibiotic injected control chicks, but did not affect (P > 0.05) experimental groups receiving either gentamicin sulfate or ceftiofur sodium by either in ovo or subcutaneous routes. The in ovo administration of 0.1 or 0.2 mg ceftiofur sodium to individual embryos on Day 18 of embryogenesis in Experiment 3 was associated with marked depressions (P < 0.05) in cecal propionate levels compared with uninjected control chicks. When feed was restricted and the time of PREEMPT administration was delayed for 48 or 72 h posthatch, mean cecal propionate levels in in ovo ceftiofur sodium-injected chicks were not significantly different (P > 0.05) from controls, indicating a time- and feed restriction-associated effect on improving CE culture establishment.