Isolation, characterization, and experimental infection of Streptococcus gallolyticus subspecies pasteurianus from commercial turkeys with acute septicemia: a pilot study.

Streptococcus gallolyticus (SG) is a Gram-positive cocci found as commensal gut flora in animals and humans. SG has emerged as a cause of disease in young poults between 1 and 3 wk of age. SG is associated with septicemia resulting in acute mortality with no premonitory signs in turkeys. Three SG isolates were obtained from clinical field cases of acute septicemia of commercial turkeys and used in three independent experiments. In Experiment 1, embryos were inoculated 25 d of embryogenesis with varying concentrations of SG1, SG2, or SG3. In Experiment 2, day of hatch, poults were inoculated with varying concentrations using different routes of administration of SG1, SG2, or SG3. In Experiment 3, day of hatch, poults were inoculated with only isolate SG1 using different paths. Poults were randomly selected for necropsy on d 8 and d 15 and sampled to collect spleen, heart, and liver for SG on d 21, the remaining poults were necropsied and cultured. Samples were plated on Columbia nalidixic acid and colistin agar (CNA) (40°C, 18-24 h). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) confirmed suspect colonies. Data were analyzed using the chi-square test of independence, testing all possible combinations to determine significance (P < 0.05). Weight data were subjected to ANOVA using JMP with significance (P < 0.05). No differences were found in BW or BWG on d 0, 8, 15, or 22. Splenomegaly, focal heart necrosis, and pericarditis were observed in all groups in experiments 1 through 3. In Experiment 3, only airsacculitis was observed in a negative control in separate isolation (P > 0.05). On d 21 of Experiment 3, increased (P < 0.05) recovery of SG from spleens were observed in co-housed negative controls, as well as poults challenged by oral gavage (P > 0.05 for d 7 and d 14). These results confirm numerous previous studies indicating that SG subsp. pasteurianus is a primary infectious microorganism that causes septicemia in young poults.

Intestinal cytokine response of commercial source broiler chicks to Salmonella typhimurium infection.

Development of molecular-based immunotherapeutic strategies for controlling Salmonella Typhimurium (ST) infection in poultry requires a better understanding of intestinal and cecal cytokine responses. Accordingly, an experiment was conducted to measure changes in intestinal cytokine expression when commercial source broiler chickens were challenged with a nalidixic acid-resistant ST. Ross broiler chicks were nonchallenged with ST (control treatment) or challenged by orally giving 7.8 x 10(6) cfu at 4 d of age (STC treatment). Each treatment consisted of 4 replicate pens with 14 chicks per pen. Expression levels of proinflammatory cytokines, interferon-gamma, and antiinflammatory interleukin (IL)-10 were determined at 5 and 10 d postchallenge (PC). Intestinal flushes were also collected from each treatment at 7 d PC to estimate IgA and IgG. Results showed an upregulation in IL-1beta mRNA in STC chicks at 5 d PC. By 10 d PC, the expression of IL-1beta was further increased and accompanied by an upregulation of IL-6 and interferon-gamma mRNA, whereas IL-10 mRNA expression decreased. It was concluded that ST induced an intestinal mucosal inflammatory response in commercial source broiler chicks less than 2 wk of age.

Factors influencing women's quality of life in the later half of life.

BACKGROUND: Among older women in East Asia, and Taiwan in particular, there is little research on quality of life and the health care they receive to address the symptoms of menopause. This study evaluated factors which influence quality of life among post middle-age women in Taiwan. METHODS: This cross-sectional study recruited 1250 women between 43 and 77 years of age during the year 2002. The factors investigated were demographics, menstruation status, menopausal symptoms, osteoporosis status, and use of hormone replacement therapy (HRT). SF-36 was used to assess the health-related quality of life of these women. Correlation, multiple regression and path analysis were used to test for direct and indirect relationships among the variables. RESULTS: There are statistical significances between menopause symptoms and quality of life across different age groups. Path analysis shows a direct positive effect of HRT and a direct negative effect of climacteric symptoms on both physical and mental components of quality of life. Age, marital status, education and osteoporosis also have direct and indirect effects, some positive and others negative, on the components of quality of life. CONCLUSIONS: When developing programs to enhance health in post middle-age women, consideration should be given to symptom relief as well as quality of life.

Crop immune response post-Salmonella enteritidis challenge in eight commercial egg-layer strains and specific-pathogen-free White Leghorn chickens.

The crop immune response against Salmonella Enteritidis (SE) challenge in eight commercial egg-layer strains (five white-egg layer and three brown-egg layer) and specific-pathogen-free (SPF) White Leghorn (WL) hens was investigated. Pre- and post-SE challenge mucosal immune responses within the crops were evaluated. Commercial layers and SPF WL hens were orally challenged with 10(8) CFU/ml SE PT13a and SE nalR PT13, respectively. Crop lavage samples were collected at weekly intervals from day 0 (pre-challenge) to day 25-27 postinfection (PI), and bacteriological examination was performed to monitor progression of SE infection. Crop lavage samples were analyzed for SE-lipopolysaccharide (LPS)-specific IgA using enzyme-linked immunosorbent assay (ELISA). H&E-stained slides of crop sections from day 34 PI and uninfected controls were assessed for lymphoid tissue via light microscopy. Lymphoid areas were graded based on morphology, size, and cellularity using a score 0 to 5 scale. The 0 to 5 (low to high) numerical values represented progressive increases in size and cellular density of lymphoid tissue. Bacterial culture results showed the highest percentage of SE-positive crop lavage samples from all hen groups at day 5-6 PI and day 11-12 PI. A progressive decline in percentage of SE-positive crop lavage samples did occur as time PI lengthened; however, at day 25-27 PI SE persisted in crop lavage samples from SPF WL hens and three commercial white-egg layer strains. A marked increase in SE-LPS-specific IgA was measured in crop lavage samples between day 0 and day 11-12 PI for all hen groups. Crop SE-LPS-specific IgA response remained elevated above day 0 baseline for the duration of the experiment. Well-defined score 3 to 5 lymphoid tissue aggregates were observed in crop tissue sections harvested at day 34 PI. Comparison of crop sections determined a 1.2-4.0 times increase in ratio of lymphoid tissue in day 34 PI SE-challenged hens vs. uninfected control hens.

Organ weight and serum triglyceride responses of older (80 week) commercial laying hens fed an alfalfa meal molt diet.

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.

Immunological cell and serum metabolite response of 60-week-old commercial laying hens to an alfalfa meal molt diet.

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.

The effect of an experimental chlorate product on Salmonella recovery of turkeys when administered prior to feed and water withdrawal.

Previously, an experimental chlorate product (ECP) has been observed to reduce Escherichia coli and Salmonella infections in swine, cattle, and broilers. The following studies were performed to investigate the effects of different concentrations and durations of administering ECP on crop and ceca Salmonella typhimurium (ST) colonization of turkeys. In 2 separate trials, each conducted with 2 replicates, 15-wk-old turkey toms were challenged with 10(7) to 10(9) cfu of ST. In Experiment 1, toms were administered 0, 0.5, 1.0, 2.0, or 4.0x of ECP (a 1.0x concentration is equivalent to a 15 mM chlorate ion concentration) in the drinking water for 38 h. In Experiment 2, toms were administered a 2x concentration of ECP in the drinking water for 0, 14, 26, or 38 h prior to water withdrawal. All treatments were followed by a 10-h water withdrawal and an 8-h feed withdrawal prior to organ sampling. In Experiment 1, turkeys provided ECP had significantly (P < 0.05) lower populations and incidences of crop (>1.4 log reduction) and ceca (>0.6 log reduction) ST as compared with control birds (2.1 and 0.94 log ST average for all trials, respectively), with little or no additional benefit from administration of higher ECP concentrations. In Experiment 2, toms provided ECP had lower populations of crop (>2.2 log reduction) and ceca (>1.5 log reduction) ST when compared with controls (3.1 and 1.8 log ST, respectively). Again, there appeared to be little benefit in longer administration intervals on quantitative reduction of ST. These experiments suggest that the ECP significantly reduces Salmonella colonization in commercial turkeys when administered prior to feed and water withdrawal.

The effect of feed deprivation on tissue invasion by Salmonella enteritidis.

A tissue culture procedure was utilized to compare tissue cell invasion by Salmonella enteritidis from molted and full feed hens. Three identical trials were performed in which 80-wk-old active laying hens were divided into 2 groups of 6 birds each. The molted hen group was subjected to a 14-d feed withdrawal, and the full-fed hen group was administered a standard layer ration. After feed treatment, crop, ileum, cecum, and ovary (small and large yellow follicles removed) were collected, rinsed in PBS, and placed into 50 mL of RPMI medium. The ends of intestine and crop tissues were tied to allow attachment of Salmonella only to the lumen surface. The RPMI medium containing 10(7) to 10(8) cfu of novobiocin and nalidixic acid-resistant phage type 13 Salmonella enteritidis was injected into the lumen of the intestine and crop tissues. Additionally, ovaries were incubated in 50 mL of RPMI medium containing 10(6) to 10(7) cfu of the Salmonella enteritidis. Tissues were incubated with Salmonella at 37 degrees C for 2 h, after which tissues were placed in 50 mL of fresh RPMI medium containing 500 microg/mL of gentamicin and incubated for 5 h at 37 degrees C to remove any Salmonella that had not penetrated tissues. Tissues were rinsed, stomached in 10 mL of PBS, serially diluted, and plated onto brilliant green agar containing novobiocin and nalidixic acid for Salmonella enumeration. Salmonella invasion of ovaries was reduced in tissues from molted hens in trials 1 and 2 as compared with full-fed controls (> 1.2 log reduction) but not in trial 3. Salmonella invasion of ceca from molted hens was numerically increased in trials 1 and 2 and significantly increased in trial 3 as compared with controls (> 0.8 log increase). No significant differences in Salmonella invasion were detected for crops and ileum. These data suggest that molting may affect invasion of tissues by Salmonella enteritidis.

Enhanced gross visualization of chicken Peyer's patch: novel staining technique applied to fresh tissue specimens.

The ileal Peyer's patches (Pp), secondary gut-associated lymphoid tissue of the mucosal immune system, may serve as an important site for monitoring inflammatory and immunologic responses of the host against enteric pathogens. Chicken Pp are often difficult to observe grossly, and a simple technique to enhance visualization of the Pp is lacking. Therefore, we designed a novel staining method that is quick, easy, and accurate to aid in gross identification and recovery of the chicken Pp from fresh tissue specimens. Lower alimentary tracts were harvested from White Leghorn hens and commercial broilers. The ileocecocolic region was excised intact, flushed with deionized water to remove ingesta, and a dilute eosin-Y solution was infused. After 1 min, the eosin-Y was gently extruded. Modified-crystal violet (mCV) was then injected into the gastrointestinal segment, where on the lymphoid tissue area became apparent at the serosal surface. The distal ileal Pp was visible as a pale whitish pink ovoid-focalized area with surrounding gut tissue stained light purple. The exact Pp site could be delineated at the serosal and mucosal surface by gross assessment. Light microscopy evaluation of hematoxylin and eosin-stained tissue slides prepared from the excised Pp site revealed lymphoid tissue aggregations with multiple follicular units indicative of Pp. The novel eosin-Y + mCV staining technique promotes rapid identification and accurate recovery of chicken Pp lymphoid tissue from fresh tissue specimens.

Effect of various combinations of alfalfa and standard layer diet on susceptibility of laying hens to Salmonella enteritidis during forced molt.

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.

Ovarian laying hen follicular maturation and in vitro Salmonella internalization.

Transovarian transmission of paratyphoid Salmonella is well documented and occurs at a low incidence in chickens. However, the exact mechanism of follicular invasion is not well understood. The following study investigates the ability of Salmonella to invade ovarian follicles at different stages of follicular maturity in vitro. Ovarian follicles were collected from Leghorn hens and separated into three stages of maturity: (1) large yellow follicles or F follicles (LYF), (2) small yellow follicles (SYF), and (3) small white follicles (SWF). All follicles were incubated at 37 degrees C in RPMI 1640 medium. Follicles were incubated with 1 x 10(6) CFU/mL of Salmonella typhimurium and Salmonella enteritidis sensitive to gentamicin for 2 h. Samples were then removed from the bacterial culture, and placed in medium containing gentamicin sulfate for 5 h to kill any S. typhimurium or S. enteritidis, which had not invaded the follicular membrane. After the 5 h incubation, follicles were stomached in phosphate buffered saline. Serial dilutions were made of each follicle and viable S. typhimurium and S. enteritidis cells were enumerated on brilliant green agar. Two identical trials were conducted. Data suggest that Salmonella may differentially invade ovarian follicles depending on maturity of the follicle, and that SWF may be more susceptible to S. typhimurium and S. enteritidis invasion than either the SYF or the LYF.

Reduction of Salmonella enterica serovar enteritidis colonization and invasion by an alfalfa diet during molt in Leghorn hens.

The standard method for molting to stimulate multiple egg-laying cycles in laying hens is feed deprivation. However, the physiological changes within hens caused by feed deprivation increase susceptibility of the hens to Salmonella enterica serovar Enteritidis (SE) infection. In an effort to develop an alternative method to induce molting without increasing susceptibility to SE, an alfalfa diet was compared with the standard molting method for the level of ovary regression and SE colonization. Hens over 50 wk of age were divided into 3 treatment groups (12 hens/group): nonmolting by normal feeding (NM), molting by feed deprivation (M), and molting by alfalfa diet (A). Individual hens on all treatments were challenged orally with 10(5) cfu of SE on the fourth day after feed changes and were analyzed for ovary weight and SE colonization or invasion in crop contents, cecal contents, liver, spleen, and ovary on the ninth day. In 3 of the 4 trials, there was a significant decrease in SE colonization of the crop between the alfalfa diet (A) and the feed deprived molt (M). In most of the 4 trials, there was a significant reduction in SE infected organs in birds fed the alfalfa diet (A) compared with birds undergoing feed deprived molt (M). Most of the trials showed no significant difference in overall SE between A and NM. Therefore, the results of this study suggest that an alfalfa diet has the potential to be used as an alternative method for forced molting, without increasing the incidence of SE in eggs and internal organs.

Effects of drinking water treatment on susceptibility of laying hens to Salmonella enteritidis during forced molt.

Feed deprivation is used in the layer industry to induce molting and stimulate multiple egg-laying cycles in laying hens. Unfortunately, the stress involved increases susceptibility to Salmonella enteritidis (SE), the risk of SE-positive eggs, and incidence of SE in internal organs. Leghorn hens over 50 wk of age were divided into 4 treatment groups of 12 hens each in experiment 1 and 3 treatment groups of 12 hens in experiments 2 and 3; hens were placed in individual laying hen cages. Treatment groups were 1) nonmolted (NM) and received feed and distilled water for 9 d, 2) force molted by feed removal for 9 d and received distilled water, 3) force molted by feed removal for 9 d and received 0.5% lactic acid (LA) in distilled water. An additional group (4) in experiment 1 only was force molted by feed removal for 9 d and received 0.5% acetic acid in distilled water. Seven days before feed removal hens were exposed to an 8L:16D photoperiod, which was continued throughout the experiment. Individual hens among all treatments were challenged orally with 10(4) SE on d 4 of feed removal. When compared with the NM treatments, weight losses were significantly higher in the M treatments, regardless of water treatments. When compared with NM treatments, crop pH was significantly higher in the M treatment receiving distilled water. Crop pH was reduced to that of the NM controls by 0.5% acetic acid in the drinking water. No consistent significant changes were observed for volatile fatty acids. The number of hens positive for SE in crop and ceca after culture and the number of SE per crop and per gram of cecal contents were higher in the M treatments, when compared with the NM treatments, but there was no effect of addition of either of the acids to the drinking water. Additional research using different acid treatment regimens may provide a tool for reducing the incidence of SE in eggs and internal organs during and following molting of laying hens.

Evaluation of immunosuppressants and dietary mechanisms in an experimental disease model for necrotic enteritis.

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.

Utilization of the nitrate reductase enzymatic pathway to reduce enteric pathogens in chickens.

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.

Comparison of zinc acetate and propionate addition on gastrointestinal tract fermentation and susceptibility of laying hens to Salmonella enteritidis during forced molt.

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.

Effect of experimental chlorate product administration in the drinking water on Salmonella typhimurium contamination of broilers.

The crop is a known source of Salmonella and Campylobacter contamination. Previously, we evaluated lactic acid in the drinking water during a simulated pretransport feed withdrawal (FW) and reported 0.44% lactic acid significantly (P < 0.05) reduced the number of Salmonella recovered in market-age broiler crops. However, total consumption of the organic acid-treated drinking water was reduced. Presently, we evaluated the effect of experimental chlorate product (ECP; 1x ECP is equivalent to a 15 mM chlorate ion concentration) during a 10-h pretransport FW. Market-age broilers were obtained from a commercial processing plant and randomly assigned to ECP-treated or control (nontreated) groups. Broilers were challenged by crop gavage with 10(8) Salmonella Typhimurium (ST) immediately upon arrival and 1 d prior to termination of the experiment. One day later, broilers were killed for ST enumeration (cfu) in the crop and ceca. Broilers provided ECP 24 h prior to slaughter consumed slightly more ECP water than broilers provided distilled water. Treatment with ECP caused a significant decrease (P < 0.05) in the incidence of ST in crop contents (2%) as compared to the controls (36.7%). Similarly, ECP treatment caused a significant decrease (P < 0.05) in number of ST (0.96 log10 ST/g cecal content) detected in the ceca when compared to controls (2.52 log10 ST). This study suggested that incorporation of ECP in the drinking water 24 to 48 h prior to slaughter could reduce Salmonella contamination in broilers.

Poultry digestive microflora biodiversity as indicated by denaturing gradient gel electrophoresis.

Populations of digestive microflora in chickens change with age and are affected by diet, stressors, and performance enhancers. Culturing techniques used to profile a bacterial community inadvertently select for some organisms while excluding others. Several molecular-based techniques have been used to profile mixed microbial populations on the basis of DNA extracted from the entire community. Denaturing gradient gel electrophoresis was used in the present study to examine PCR-amplified fragments (amplicons) of a 16S ribosomal DNA variable region from predominant digestive bacteria. The objective of the study was to examine changes in digestive microbial communities of developing Leghorn chicks and molted Leghorn hens. Dendrograms of amplicon patterns indicated approximately 51% similarity between cecal bacteria composition in Leghorn chicks less than 20 d old and chicks greater than 20 d old. Cecal communities in Leghorn chicks given a competitive exclusion culture exhibited 21% correlation at all ages with those in control chicks. Nonmolted and molted hens had 40% similarity between cecal communities, whereas diets with low calcium (0.8% wt/wt) and excess zinc (2,800 mg/kg) lessened population differences (90% similarity). Results indicated the potential usefulness of the molecular-based denaturing gradient gel electrophoresis to monitor changes in digestive bacterial communities in chickens.

Effect of bursal anti-steroidogenic peptide and immunoglobulin G on neonatal chicken B-lymphocyte proliferation.

In attempts to identify antibodies for Bursal Anti-Steroidogenic Peptide (BASP), rabbit serum was observed to reduce phorbol ester-stimulated chicken B-lymphocyte proliferation comparable to BASP. These experiments investigated the effects of IgG on B-lymphocyte proliferation. In Experiment 1, 3% rabbit serum decreased B-lymphocyte proliferation. In Experiment 2, 2 mg/ml of intact rabbit IgG or 0.65 mg/ml of IgG papain digest products, Fab and Fc, decreased B-lymphocyte proliferation. The combination of BASP and either Fab or Fc was observed to have at least an additive anti-proliferative effect. In Experiment 3, 0.01 mg/ml of either rabbit or chicken IgG, or 1.0 mg/ml of rabbit or 0.01 mg/ml of chicken Fab, Fc, and the pepsin digestion product F(ab')(2) was observed to have an anti-proliferative effect. No combined effects of BASP and IgG or IgG digest products were observed for this experiment. In Experiment 4, 12 mg/ml of chicken egg yolk IgG or 1.2 mg/ml Fab was found to suppress B-lymphocyte proliferation. Additionally, an additive effect of 12 mg/ml of IgG with BASP was again observed. The present studies suggest that IgG and its digestion products reduce phorbol-stimulated B-lymphocyte proliferation in vitro and combined treatment with IgG and BASP may have at least an additive anti-proliferative effect on B-lymphocyte proliferation.

In situ detection and quantification of bursa of fabricius cellular proliferation or apoptosis in normal or steroid-treated neonatal chicks.

Apoptosis, or programmed cell death, is believed to be the mechanism for depletion of lymphocytes recognizing self-antigens following clonal expansion in the bursa of Fabricius. Although bursal apoptosis has previously been shown to increase following in vivo exposure to glucocorticoids, the microanatomical site of induced or normal apoptosis has not been unequivocally established. Presently, we adapted the existing terminal deoxynucleotidal transferase-mediated dUTP nick-end labeling (TUNEL) assay for use with neonatal bursae. Similar to previous reports, TUNEL revealed that normal apoptosis is preferentially, but not exclusively, ongoing in bursal follicular cortical cells. Administration of a single dose of a synthetic glucocorticoid (dexamethasone) or androgen (19-nortestosterone) did not significantly (P < 0.05) alter follicular lymphocyte numbers or apoptosis per unit of area at the time points evaluated post-administration (6 or 24 h). However, administration of 19-Nortestosterone increased the interfollicular epithelial thickness, a change usually associated with edema, within 6 h following treatment. Additionally, administration of the androgen 19-nortestosterone significantly decreased the number of proliferating cells as detected using mouse anti-proliferating cell nuclear antigen (PCNA) as a primary immunohistochemical antibody. In normal (control) bursal sections, occasional follicles consisting of predominantly apoptotic cells were observed (0.26% of follicles). Such follicles were consistently one-tenth the area of normal follicles. This incidental finding may suggest occasional occurrence of a common signal for deletion, such as a common integral or clonal mistake, viral infection, or an aberrant paracrine signal.