Search

The aim of this study was to determine if mobile plasmids carrying antibiotic-resistant genes could survive and be transferred in chicken manure maintained under conditions similar to those found in commercial cage layer operations and during composting. Escherichia coli J5 harboring a self-transmissible plasmid (RP4) and E. coli C600 harboring a mobile plasmid (pIE723) were used as plasmid donors; E. coli CV601 was used as a plasmid recipient. At 23 degrees C both plasmids were transferred to E. coli CV601 in chicken manure and in compost microcosms that consisted of a mixture of chicken manure and peat. The transfer frequencies ranged from 8.1 x 10(-5) to 2.4 x 10(-3) per donor cell in manure and from 2.4 x 10(-5) to 5.5 x 10(-4) per donor cell in compost microcosms. After 45 d of incubation at 23 degrees C, RP4, but not pIE723, was recovered by an exogenous isolation method although their E. coli hosts were not cultured from the microcosms. However, when the temperatures of the compost microcosms were elevated to 50 degrees C or above, neither the plasmids nor their E. coli hosts could be detected. The results suggested that composting of chicken manure at high temperatures could help prevent the spread of antibiotic-resistant genes via plasmids in the environment.

Composting has been used for disposal of poultry carcasses and manure following outbreaks caused by avian influenza virus (AIV) and Newcastle disease virus (NDV), but methods are needed to test for survival of the viruses in compost to ensure biosecurity. Methods developed in the present study include extracting viruses from compost and purifying viral RNA. The extracted viruses were detected by virus isolation using embryonated chicken eggs, and the purified RNA was detected by real-time reverse transcription PCR (RRT-PCR). The virus isolation and the RRT-PCR methods were evaluated with 3 compost preparations that were produced from chicken manure mixed with corn silage, wood shavings, or wheat straw. The detection limits of both methods were 1,700 and 1,000 embryo lethal doses of AIV and NDV per gram of compost, respectively. The copy number of viral RNA quantified by RRT-PCR was highly correlated with the amount of virus in compost. The results suggested that the RRT-PCR method may be used as an alternative to the virus isolation method for rapid detection and accurate quantification of AIV and NDV in compost.

Salmonella enteritidis and Salmonella typhimurium definitive type 104 (DT104) have been detected in the chicken oviduct, and their survival in egg albumen at the chicken body temperature of 42 degrees C may be important in oviductal and transovarian contamination of intact shell eggs. Eight S. enteritidis and 24 S. typhimurium DT104 strains were tested for their in vitro survival in egg albumen. The concentration of the organisms declined more rapidly when incubated at 42 degrees C than at 37 degrees C and dropped to nondetectable levels within 96 h at the higher, but not at the lower, temperature. In another experiment, 3 S. enteritidis and 3 S. typhimurium DT104 strains were randomly selected, and dosages of 20 and 200 cells of each strain were inoculated onto the vitelline membranes of egg yolks, which were then submerged in the original albumen and incubated for 24 h at 42 degrees C. Under these conditions, the organisms survived in albumen but did not penetrate the vitelline membrane. However, in a similar experiment, penetration did occur when the specimens were incubated at 30 degrees C for 72 h. The results suggest that low numbers of S. enteritidis and S. typhimurium DT104 can be expected to survive in egg albumen during the 24-h period of egg formation in the oviduct but would be unlikely to invade the yolk.before oviposition. However, depending on storage conditions following oviposition, S. enteritidis, as well as S. typhimurium DT104, could survive longer and may eventually invade the egg yolks.