D. E. Corrier

Evaluation of Salmonella Serotype Distributions from Commercial Broiler Hatcheries and Grower Houses

By conventional trayliner (hatcheries) and drag swab assembly (broiler houses) culture methods, the isolation distribution of Salmonella serotypes from five commercial broiler hatcheries (three sample times) and 13 broiler farms (eight sample times) was evaluated. A total of 11 different Salmonella serotypes were isolated from hatcheries, with Salmonella heidelberg (9/30) and Salmonella kentucky (6/30) accounting for 50% of the total isolations. Of 700 chick paperpad trayliners sampled, regardless of lot (breeder flock source) or hatchery, 12% were positive for Salmonella. When 10 individual trayliners were cultured from individual lots (same breeder flock source), Salmonella was detected in 24/57 lots (42%). Multiple serotypes were simultaneously isolated from the same lot on three occasions (6%). Of the 21 lots that were serially sampled, the Salmonella serotype detected was different within lots eight times (38%) on at least one occasion of two or more sampling times. Of the 196 individual broiler houses sampled, 44 were positive for Salmonella (42%). Twelve different serotypes were isolated from broiler houses during this study. The serotypes isolated most frequently were S. heidelberg (34/94) and S. kentucky (22/94). These two serotypes accounted for 59.6% (56/94) of the total broiler house isolations. Of the 38 houses that were serially sampled, two or more serotypes were detected in the same broiler house on 20 occasions (53%). Of the 38 serially sampled houses (four or more times), a consistent Salmonella serotype was detected in five houses (13%). In only 5 of the 38 (13%) serially sampled houses did we fail to detect Salmonella on four or more samplings. No significant difference in Salmonella isolation frequency was observed between poultry houses using new or used litter. These data support previous findings indicating that paratyphoid Salmonella serotypes are prevalent in some broiler hatcheries and houses. Further, the observation of multiple serotypes simultaneously and serially isolated from the same breeder hatchery lots suggests that breeder flocks may be infected with more than one serotype, possibly providing a source for multiple serotype infections in progeny grower flocks.

Effect of Feed Withdrawal on Campylobacter in the Crops of Market-Age Broiler Chickens

The presence of Campylobacter and Salmonella on poultry meat products remains a significant public health concern. Previous research has indicated that feed withdrawal may significantly increase Salmonella contamination of broiler crops and that crop contents may serve as an important source of Salmonella carcass contamination at commercial processing. The present study evaluated the effect of preslaughter feed withdrawal on the incidence of Campylobacter isolation in crops of market-age commercial broiler chickens prior to capture and transport to the processing plant. The incidence of Campylobacter isolation from the crop was determined immediately before and after feed withdrawal in 40 7-wk-old broiler chickens obtained from each of nine separate broiler houses. Ceca were collected from broilers in six of the same flocks for comparison with the crop samples. Feed withdrawal caused a significant (P < 0.025) increase in Campylobacter-positive crop samples in seven of the nine houses sampled. Furthermore, the total number of Campylobacter-positive crops increased significantly (P < 0.001) from 90/360 (25%) before feed removal to 224/359 (62.4%) after the feed withdrawal period. Alternatively, feed withdrawal did not significantly alter the Campylobacter isolation frequency from ceca. Similar to our previous studies with Salmonella, the present results suggest that preharvest feed withdrawal increases the frequency of Campylobacter crop contamination and, thus, provides a source of Campylobacter contamination of carcasses at commercial processing.

Effect of immunosuppression with cyclophosphamide, cyclosporin, or dexamethasone on Salmonella colonization of broiler chicks.

One-day-old broiler chicks were divided into four groups: untreated controls and three groups treated with the immunosuppressive drugs cyclophosphamide, cyclosporin, or dexamethasone on days 1-4 of age. On day 5, all groups were challenged orally with 10(4) Salmonella typhimurium. Suppression of immunocompetency was demonstrated in the treatment groups by significant decrease (P less than 0.05) in weights of bursas of Fabricius, and diminished mitogen-induced T and B lymphocyte blastogenesis and cutaneous basophil hypersensitivity responses. There were no significant differences among the four groups in the number of Salmonella in the cecal contents or in the number of cecal-culture-positive chicks on day 10 post-challenge at 15 days of age. Results indicated that impairment of B or T lymphocyte responses had little influence on the resistance of young chicks to Salmonella cecal colonization.

Effectiveness of dietary propionic acid in controlling Salmonella typhimurium colonization in broiler chicks.

Newly hatched broiler chicks were provided a corn/soybean meal-based ration treated with propionic acid at 30 mumol/g of feed ration. At 3 days of age, the chicks were challenged orally with 10(4) Salmonella typhimurium. Crop contents from 4-day-old chicks that were provided dietary propionic acid contained significantly higher concentrations of propionic acid (4.0 to 6.8 mumol/g crop contents) than crops from challenged control chicks provided untreated feed (0.9 to 1.5 mumol/g crop contents). Provision of dietary propionic acid on feed as a dry powder in five trials or a liquid application in three trials had no significant effect on crop or cecal pH. Significant decreases in Salmonella in the crop and ceca were detected in one trial, but the decreases were likely the result of the presence of anti-salmonellae bacteria rather than the dietary propionic acid. Results indicate that propionic acid in the feed was ineffective in reducing Salmonella infection in the crop and ceca.

COMPARISON OF DRAG-SWAB ENVIRONMENTAL PROTOCOLS FOR THE ISOLATION OF SALMONELLA IN POULTRY HOUSES

Three surveys were conducted during November 1995 and March and May 1996 to compare the use of double-strength skim milk (wet) or no transport media (dry) drag swabs for the detection of salmonellae in 10 broiler houses. Salmonellae were isolated from 57 of 120 individual wet drag-swab samples, compared with 21 of 120 dry drag-swab samples. Furthermore, Salmonella was detected at a higher frequency with wet drag swabs (66.7%) than with dry drag swabs (40%) when compared on an individual growout house basis. A total of seven different serotypes were isolated from the 10 broiler houses. Although double-strength skim milk drag swabs are more labor intensive than dry drag swabs, double-strength skim milk drag swabs are more efficient for detecting Salmonella than are dry drag swabs with no transport media.

Survival of Salmonella in the Crop Contents of Market-Age Broilers During Feed Withdrawal

Recent studies have indicated that crop contamination increases during preslaughter feed withdrawal and that contaminated crop contents may serve as an important source of Salmonella entry into poultry processing plants. During the present study, we evaluated the effect of preslaughter feed withdrawal on crop pH and Salmonella crop contamination in broilers from three commercial broiler flocks. The effect of experimental feed withdrawal on crop pH, lactic acid concentration, and Salmonella crop contamination was also evaluated in market-age broilers challenged experimentally with Salmonella typhimurium. Crop pH increased significantly (P < 0.05) from 3.64 +/- 0.25 before feed removal to 5.14 +/- 0.72 after 8 hr of feed withdrawal in broilers from commercial flocks. The incidence of Salmonella crop contamination in the commercial broilers increased (P < 0.05) from 3.3% before feed removal to 12.6% after 8 hr of feed withdrawal. Similarly, crop pH increased (P < 0.05) by a magnitude of approximately 1 unit in broilers after 8 hr of experimental feed withdrawal. The population of S. typhimurium in the crops of the experimentally challenged broilers increased (P < 0.05) by approximately 1 log unit during the 8-hr experimental feed withdrawal. The concentration of lactic acid in the crop of the broilers during experimental feed withdrawal decreased (P < 0.01) from a range of 119-135 mumol/ml before feed removal to a range of 22-32 mumol/ml after 8 hr of feed withdrawal. The results indicated that feed withdrawal resulted in a decrease in lactic acid in the crop, accompanied by an increase in crop pH, and an increase in Salmonella crop contamination.

Modulation of cell-mediated resistance to listeriosis in mice given T-2 toxin

The modulating effect of preinoculation and postinoculation treatment with a single oral 4.0 mg/kg dosage of T-2 toxin on cell-mediated resistance was studied in mice inoculated with Listeria monocytogenes. Toxin treatment caused significant decreases in thymus and spleen weights, bone marrow cellularity, and in the total number of circulating leukocytes, lymphocytes, and neutrophils. Enhancement or suppression of resistance to listeriosis was dependent on the time of toxin administration relative to the time of Listeria challenge. Preinoculation treatment on Day 2 or 4 prior to Listeria challenge significantly enhanced resistance and decreased mortality due to listeriosis by as much as 50%. In contrast, resistance was suppressed and mortality was increased by 50% in mice that were treated with toxin after Listeria challenge. Enhanced resistance to listeriosis was accompanied by a significant increase in the influx of macrophages into Listeria-elicited peritoneal exudates. In addition, in vivo phagocytosis of sheep red blood cells by peritoneal macrophages was significantly increased in toxin-treated mice that were sensitized with sheep erythrocytes. The data indicate that T-2 toxin has a modulating effect on cell-mediated resistance and that enhancement of resistance to listeriosis in mice pretreated with T-2 toxin is associated with increased migration/activation of macrophage effector cells.

Dynamics of avian inflammatory response toSalmonella-immune lymphokines

Investigations in our laboratories have indicated that an increased resistance to SE organ infectivity in chicks was conferred by the immunoprophylactic administration of SE-immune lymphokines (SE-ILK). This resistance was associated with an increase in the lamina propria thickness due to a marked infiltration of inflammatory polymorphonuclear cells (PMNs). In the present study, we determined whether the hematological profile of SE-ILK-treated chicks might reflect changes that are associated with the protection against organ invasion by SE. As protection has been observed in previous studies within 24 h of SE-ILK administration, we evaluated alterations in the circulating leukocyte profile in 1-day-old Leghorn chicks during this time period. We also determined whether the alterations in the peripheral blood leukocytes correlated with the increased protection against SE organ invasion induced by the SE-ILK. Within 4 h after an intraperitoneal injection of SE-ILK and challenge with SE, the number of circulating leukocytes increased significantly (P < 0.05) from all of the other treatment groups. The number of circulating PMNs was found to account for more than 80% of the increase in the number of circulating leukocytes. Using correlation analysis, we found a strong association between the number of circulating PMNs and the protection induced by SE-ILK against SE organ invasion. These studies associate the expansion of the available pool of circulating PMNs and the expression of innate resistance to organ invasion by SE.Investigations in our laboratories have indicated that an increased resistance to SE organ infectivity in chicks was conferred by the immunoprophylactic administration of SE-immune lymphokines (SE-ILK). This resistance was associated with an increase in the lamina propria thickness due to a marked infiltration of inflammatory polymorphonuclear cells (PMNs). In the present study, we determined whether the hematological profile of SE-ILK-treated chicks might reflect changes that are associated with the protection against organ invasion by SE. As protection has been observed in previous studies within 24 h of SE-ILK administration, we evaluated alterations in the circulating leukocyte profile in 1-day-old Leghorn chicks during this time period. We also determined whether the alterations in the peripheral blood leukocytes correlated with the increased protection against SE organ invasion induced by the SE-ILK. Within 4 h after an intraperitoneal injection of SE-ILK and challenge with SE, the number of circulating leukocytes increased significantly (P < 0.05) from all of the other treatment groups. The number of circulating PMNs was found to account for more than 80% of the increase in the number of circulating leukocytes. Using correlation analysis, we found a strong association between the number of circulating PMNs and the protection induced by SE-ILK against SE organ invasion. These studies associate the expansion of the available pool of circulating PMNs and the expression of innate resistance to organ invasion by SE.

Effect of Selected Antibiotics and Anticoccidials on Salmonella enteritidis Cecal Colonization and Organ Invasion in Leghorn Chicks

One-day-old leghorn chicks were placed in floor pens on previously used poultry litter (potentially providing exposure to normal chicken enteric flora) for 7 days and provided feed containing one of several antibiotics or anticoccidials. On day 7, all groups were challenged orally with an isolate of Salmonella enteritidis (10(6) colony-forming units) that was resistant to bacitracin, novobiocin, nalidixic acid, and nitrofurazone. All chicks were killed on day 13, and liver, spleen, and cecal tonsils were cultured. Dietary administration of novobiocin (0.385 g/kg) caused a significant increase (P < 0.05) in positive chick colonization rate (either liver and spleen or cecal tonsils) compared with the unmedicated controls. Similarly, chicks administered dietary nitrofurazone (0.3 g/kg) were infected with S. enteritidis at a significantly greater frequency than the unmedicated controls. A significant decrease in cecal volatile fatty acid concentration, previously shown to influence susceptibility to selected enteric pathogens, was observed in the novobiocin- and nitrofurazone-treated groups. Treatment with chlortetracycline (11.4 g/kg), monensin (0.91 g/kg), or nicarbazin (0.49 g/kg) had no effect on S. enteritidis invasion or colonization. Bacitracin (0.49 g/kg) significantly increased S. enteritidis cecal colonization rate when administered continuously throughout the study. These data support and extend previous investigations involving other salmonellae and indicate that selected antibiotics may increase the severity and frequency of S. enteritidis colonization and invasion rate in leghorn chicks.

Predictive value of multiple drag-swab sampling for the detection of Salmonella from occupied or vacant poultry houses.

The results of four independent complete drag-swab samplings of a 31-farm broiler-production complex are described. Samplings of vacant poultry farms (i.e., between flocks) and occupied farms (i.e., containing chickens) were completed in four 3-day intervals beginning in July 1992 and ending in May 1993. During the first two sampling periods, two drag-swab assemblies were dragged through each individual house and pooled into one sample to evaluate overall incidence of Salmonella detection among vacant and occupied houses. During the second two sampling periods, four drag-swab assemblies were dragged through houses to evaluate frequency of Salmonella detection on individual assemblies within each house. Over the four sampling periods, mean isolation frequencies were 42.2% for vacant poultry farms and 76.2% for occupied poultry farms. When four assemblies were dragged in vacant houses and cultured separately, the probability of detecting salmonellae contamination increased with increasing numbers of assemblies, indicating that multiple drag-swab assemblies should be used for sampling either vacant or occupied broiler houses for the presence of Salmonella. On vacant farms, 89.5% of the total isolations occurred in only one house of two-house farms; the remaining 10.5% of farms were positive in both houses. Salmonella isolation from occupied farms occurred in 54.2% of the samplings in only one house, whereas 45.8% of occupied farms were positive in both houses. These data show a higher frequency of Salmonella isolation on multiple swabs in occupied poultry houses, possibly indicating a quantitatively greater presence of Salmonella in these houses.