Arthur Hinton

Effect of Dietary Lactose on Cecal pH, Bacteriostatic Volatile Fatty Acids, and Salmonella typhimurium Colonization of Broiler Chicks

One-day-old broiler chicks were inoculated with volatile fatty acid producing cecal flora from adult chickens. The chicks were divided into four groups and provided 1) no lactose, 2) 2.5% lactose in water, 3) 5% lactose in feed, or 4) 10% lactose in feed, until 10 days of age. All groups were challenged at 3 days of age with 10(6) or 10(8) S. typhimurium. At 10 days, the number of Salmonella in the ceca of the chicks challenged with 10(6) Salmonella was significantly decreased (P less than 0.01) in the groups provided lactose as compared with the controls. A significant decrease (P less than 0.01) in Salmonella numbers occurred in the chicks challenged with 10(8) Salmonella and provided 10% lactose. Providing 2.5% lactose or 5% lactose failed to inhibit Salmonella growth in chicks challenged with 10(8) Salmonella. The pH of the ceca of the groups provided lactose decreased significantly (P less than 0.05) and was accompanied by significant increases (P less than 0.01) in the concentrations of bacteriostatic acetic and propionic acids. Results showed that providing dietary lactose to broiler chicks and inoculation with normal cecal flora decreased cecal pH, increased the concentrations of bacteriostatic volatile fatty acids, and inhibited Salmonella colonization.

Biological Control of Salmonella typhimurium in Young Chickens

The effect of dietary lactose and anaerobic cultures of cecal microflora of mature chickens on the colonization of young broiler chickens by Salmonella typhimurium was evaluated. Newly hatched chicks were given either no treatment (controls), anaerobic cecal cultures, lactose (2.5%) in the drinking water, or both anaerobic cultures and lactose. Chicks were challenged per os at 3 days of age with either 10(6) or 10(8) S. typhimurium resistant to nalidixic acid and novobiocin. On day 10, the cecal contents of the chicks were examined for S. typhimurium, pH, short-chained volatile fatty acids (VFAs), undissociated VFAs, and lactic acid. Chicks given either lactose alone or cecal anaerobes alone had significantly (P less than 0.05) fewer S. typhimurium recovered from their ceca than the controls. Chicks given the combination of dietary lactose and cecal anaerobes had significantly fewer S. typhimurium recovered from their ceca than the chicks given dietary lactose or cecal anaerobes alone. Chicks given lactose had significant (P less than 0.05) increases in the lactic acid concentration of their cecal contents. Increased lactic acid concentrations were directly correlated to decreased cecal pH values and caused a reduction in the total concentration of VFAs but a significant (P less than 0.05) increase in the undissociated form of some VFAs.

Effect of dietary lactose on salmonella colonization of market-age broiler chickens.

The effect of providing lactose in feed and inoculation with volatile fatty acid-producing anaerobic cultures (AC) of cecal flora on Salmonella typhimurium colonization was evaluated in broilers. One-day-old chicks were divided into four groups and provided 1) no lactose, no AC; 2) AC, no lactose; 3) AC and lactose on days 1-10; or 4) AC and lactose on days 1-40. All groups were challenged per os with 10(6) Salmonella on day 3 and with 10(8) Salmonella on day 33. Salmonella growth in the cecal contents was significantly decreased (P less than 0.01) on day 10 in the chicks provided lactose from day 1-10. However, after the removal of lactose from the diet, the chicks were susceptible to Salmonella colonization. The number of Salmonella in the ceca was significantly reduced (P less than 0.05) in the chicks provided lactose throughout the 40-day growing period. Dietary lactose decreased the pH of the cecal contents and was accompanied by marked increases in the concentrations of undissociated bacteriostatic volatile fatty acids in the cecal contents.

Intracloacal Salmonella typhimurium infection of broiler chickens: reduction of colonization with anaerobic organisms and dietary lactose.

The combined effect of treatments with dietary lactose plus anaerobic organisms on cecal colonization of broiler chicks by Salmonella typhimurium was evaluated. Chickens treated with a combination of anaerobic organisms and 7% dietary lactose were resistant to cecal colonization by S. typhimurium. The number of recoverable S. typhimurium cells per gram of cecal contents taken on days 10 and 15 after infection was significantly reduced. Treatment with anaerobes without the addition of lactose did not effectively control cecal colonization. Intracloacal inoculations with bacterial concentrations that varied by 10,000-fold resulted in roughly similar levels of colonization. The treatments resulted in reduced cecal pH and elevated levels of undissociated volatile fatty acids. Statistically significant correlations (P less than 0.01) were observed between the S. typhimurium concentrations in cecal material and the concentrations of undissociated fatty acids (r = -0.79, and between the bacterial counts and pH (r = 0.72).

Tracking spoilage bacteria in commercial poultry processing and refrigerated storage of poultry carcasses

Four trials were conducted to examine the effect of commercial processing and refrigerated storage on spoilage bacteria in the native microflora of broiler carcasses. Prescalded, picked, eviscerated, and chilled carcasses were obtained from a commercial processing facility, and psychrotrophs in the bacterial flora were enumerated on Iron Agar, Pseudomonas Agar, and STAA Agar. The size of the population of spoilage bacteria on processed carcasses stored at 4 degrees C for 7, 10, or 14 days was also determined. Bacterial isolates were identified and dendrograms of the fatty acid profiles of the isolates were prepared to determine the degree of relatedness of the isolates. Findings indicated that although some processing steps increased the level of carcass contamination by selected bacteria, the number of spoilage bacteria recovered from processed carcasses was significantly (P< or = 0.05) less than the number of bacteria recovered from carcasses entering the processing line. Acinetobacter and Aeromonas spp. were the primary isolates recovered from carcasses taken from the processing line. During refrigerated storage, there was a significant (P < or =0.05) increase in the population of bacteria on the carcasses, and Pseudomonas spp. were the predominant bacteria recovered from these carcasses. Dendrograms of the fatty acid profiles of the isolates indicated that bacterial cross-contamination of carcasses occurs during all stages of processing and that some bacteria can survive processing and proliferate on carcasses during refrigerated storage. Furthermore, cross-contamination was detected between carcasses processed on different days at the same facility. Findings indicate that although poultry processing decreases carcass contamination by psychrotrophic spoilage bacteria, significant levels of bacterial cross-contamination occur during processing, and bacteria that survive processing may multiply on the carcasses during refrigerated storage.

Effect of Anaerobic Cecal Microflora and Dietary Lactose on Colonization Resistance of Layer Chicks to Invasive Salmonella enteritidis

The effect of oral inoculation with anaerobic cultures of cecal microflora and providing lactose in the feed on colonization resistance to invasive Salmonella enteritidis was evaluated in newly hatched leghorn chicks. Salmonella colonization of the ceca, tissue invasion and organ colonization, horizontal transmission, and seroconversion were significantly decreased (P less than 0.01) in chicks inoculated with cecal flora. The addition of lactose to the feed, in the absence of cecal microflora, failed to provide protection. Dietary lactose enhanced colonization resistance in chicks that were inoculated with anaerobic cultures of cecal flora. The results indicated that establishment of normal cecal flora in layer chicks together with the addition of lactose to the diet markedly increases resistance to cecal colonization and organ invasion, and decreases horizontal transmission of S. enteritidis.

Use of Oleic Acid To Reduce the Population of the Bacterial Flora of Poultry Skin

The effect of oleic acid on native bacterial flora of poultry skin was examined. Skin from commercial broiler carcasses was washed once or twice in solutions of 0, 2, 4, 6, 8, or 10% (wt/vol) oleic acid and rinsed in peptone water. Aerobic bacteria, Enterobacteriaceae, Campylobacter, and enterococci in the rinsates were enumerated. Significantly fewer aerobic bacteria, Enterobacteriaceae, Campylobacter, and enterococci were recovered from rinsates of skin washed in oleic acid than from control samples. Additionally, fewer bacteria were recovered from rinsates of skin washed in higher concentrations of oleic acid than from skin washed in lower concentrations of the fatty acid. In most cases, there was no significant difference in the number of bacteria recovered from rinsates of skin washed once or twice in solutions of oleic acid. Washing skin samples twice in 10% solutions of oleic acid significantly reduced the number of aerobic bacteria, Enterobacteriaceae, Campylobacter, and enterococci that remained attached to the skin. Campylobacter sp., Enterococcus faecalis, and Listeria monocytogenes isolates possessed the least resistance to the antibacterial activity of oleic acid in vitro, while Escherichia coli and Pseudomonas aeruginosa showed higher resistance. Enterobacter cloacae, Staphylococcus lentus, and Salmonella Typhimurium had the greatest resistance to the antibacterial activity of oleic acid. Findings indicate that oleic acid reduces the number of bacteria on the skin of processed broilers and that the fatty acid is bactericidal to several spoilage and pathogenic bacteria associated with poultry.

Coliform, Escherichia coli, and Salmonellae Concentrations in a Multiple-Tank, Counterflow Poultry Scalder

Scald water samples from a commercial broiler processing plant were tested for coliforms, Escherichia coli, and salmonellae to evaluate the numbers of suspended bacteria in a multiple-tank, counterflow scalder. Water samples were taken from each of three tanks on 8 different days after 6-week-old broilers had been processed for 8 h. Coliforms and E. coli were counted using Petrifilm, and the most probable number (MPN) of salmonellae was determined both in water samples and in rinses of defeathered carcasses that were removed from the processing line immediately after taking the water samples. Mean coliform concentrations in tanks 1, 2, and 3 (the last tank that carcasses pass through before being defeathered) were 3.4, 2.0, and 1.2 log10(CFU/ml), respectively. E. coli concentrations followed the same pattern with means of 3.2, 1.5, and 0.8 in tanks 1, 2, and 3, respectively, with significant differences (P < 0.02) in the concentrations of both coliforms and E. coli between the tanks. Sixteen of 24 scald-water samples were positive for salmonellae with a geometric mean of 10.9 MPN/100 ml in the positive samples. Salmonellae were isolated from seven of eight water samples from both tanks 1 and 2, but in only two of eight water samples from tank 3, the last tank that carcasses pass through. It appears that most bacteria removed from carcasses during scalding are washed off during the early part of scalding.

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.

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.