Approach for selection of individual enteric bacteria for competitive exclusion in turkey poults.

The intentional early colonization of the intestinal tract with beneficial microflora, known as competitive exclusion, has been shown to successfully protect poultry from selected enteric pathogens. Although effective cultures have been produced and are available, an inexpensive, air-tolerant, and completely defined culture is needed. Presently, we developed an in vitro competition assay to select for individual facultative anaerobes of poultry enteric origin that could exclude Salmonella. Using this assay, 24 isolates were selected and stored individually. These 24 isolates were amplified in batch culture (tryptic soy broth, 4 h at 40 degrees C) and administered at final dilutions of 10, 100, or 1,000 cfu to day-of-hatch poults. Forty-eight hours later, poults were challenged with 100 to 1,000 cfu antibiotic-resistance-marked Salmonella enteritidis PT 13A by oral gavage. Five days later, all poults were killed, and cecal tonsils were aseptically removed for tetrathionate enrichment (24 h at 37 degrees C) followed by selective plating with marker antibiotics. Selected lactose-negative, antibiotic-resistant colonies typical of Salmonella were further confirmed by serogrouping. Treatment-related protection ranged from 0 to 100% in three experiments. Greatest protection was related to the lowest concentrations of the protective microflora in each experiment. These data suggest that effective combinations of competitive enteric microflora can be identified by appropriate in vitro selection methods.

The effect of microaerosolized hydrogen peroxide on bacterial and viral poultry pathogens.

The effect of microaerosolized H2O2 on bacterial and viral poultry pathogens was investigated. Bacterial cultures and viruses were dried on sterile glass Petri dishes and subjected to direct and indirect 5% (H2O2) microaerosol mist. In the trials using Escherichia coli and Staphylococcus aureus, there was complete inactivation following exposure to H2O2. Using Salmonella typhimurium, indirect exposure resulted in only partial inactivation whereas direct exposure to H2O2 gave complete inactivation. For the viruses studied, 5% H2O2 microaerosol mist completely inactivated infectious laryngotracheitis virus. Newcastle disease virus, infectious bronchitis virus, and avian influenza virus showed reduced infectivity but were not completely inactivated. Avian reovirus susceptibility varied with the method of exposure and infectious bursal disease virus was highly resistant. The use of 10% H2O2 mist, however, resulted in total inactivation of infectious bursal disease virus. The effect of 10% H2O2 on equipment and selected materials representative of a hatcher or poultry house was investigated. A solar cell calculator, a thermostat containing a microswitch, and samples of uncoated steel, galvanized steel, and uncoated aluminum were subjected to 10 fumigation cycles. No damage was detected in the calculator and the thermostat. Both the uncoated steel and the galvanized steel showed signs of oxidation. The aluminum did not show signs of oxidation.

Detection of antibody to Bordetella avium using a particle concentration fluorescence immunoassay (PCFIA).

A fluorescent assay for detection of antibody to Bordetella avium was developed and evaluated against current serological techniques. Bordetella antigen was coated on submicron polystyrene beads and used in an antibody sandwich technique with commercially available goat anti-turkey fluorescein-labeled IgG. Positive control sera were taken from turkeys from which B. avium was isolated. Negative control sera were from turkeys from which no B. avium was isolated and which tested negative by agglutination and enzyme immunoassay. A commercial fluorescence concentration analyzer was used to quantitate the epifluorescence in a special 96-well plate designed to minimize background fluorescence. The particle concentration fluorescence immunoassay was found to be as sensitive as the enzyme immunoassay, with good reproducibility. It required less time and smaller quantities of sample and reagents.

Use of an enzyme-linked immunosorbent assay to measure antibody levels in turkey breeder hens, eggs, and progeny following natural infection or immunization with a commercial Bordetella avium bacterin.

Twelve large white turkey hens were immunized with a commercially available Bordetella avium bacterin. Hens and eggs were tested using an enzyme-linked immunosorbent assay (ELISA) to determine the response to the bacterin. Three hundred poults were then obtained from two commercial flocks, the hens of one flock having been immunized with the same bacterin used on the group of 12 turkeys. Titers of the poults were monitored for 7 weeks, and poults were challenged by exposure to infected poults at 1, 7, 14, and 21 days post-hatch. Hens produced an antibody response following immunization, with a parallel antibody response being detected in eggs. Maternal antibodies were present in poults from immunized hens. Poult titers declined to near the level of poults from unimmunized hens by 14 days of age. Poults from immunized hens challenged at 1 and 7 days were resistant to development of clinical disease and gross lesions, whereas all poults from unimmunized hens exhibited clinical signs and gross lesions. After 14 days, the resistance of both groups to development of clinical disease, became near equal, neither group being affected as severely as the unimmunized hens challenged at days 1 and 7. Six commercial turkey breeding flocks and their progeny that had not been vaccinated for B. avium and had no history of B. avium infection were evaluated with the B. avium ELISA. There were variations between the flocks, with poult titers reflecting those found in the hens.