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Antibiotic resistance and the mode of transmission were investigated in bacteria isolated from poultry litter. Total aerobic heterotrophic bacteria were screened and identified for their resistance to different antibiotics such as ampicillin, streptomycin, erythromycin, tetracycline, chloramphenicol, kanamycin, tobramycin, and rifampicin. The distribution of bacteria found in the litter was Staphylococcus (29.1%), which was the predominant group, followed by Streptococcus (25%), Micrococcus (20.8%), Escherichia coli (12.5%), Salmonella (8.3%), and Aeromonas (4.1%). Fifty percent of these isolates were susceptible to ampicillin, 57% to erythromycin, 25% to tetracycline, 4% to chloramphenicol, 40% to kanamycin, 75% to streptomycin, 54% to tobramycin, and 4% to rifampicin. Three randomly selected isolates representing Staphylococcus, Streptococcus, and Micrococcus were examined for plasmids, and plasmid-curing and plasmid-induced transformation studies were conducted. Streptococcus and Micrococcus harbored a plasmid of 4.2 and 5.1 kb, respectively, whereas Staphylococcus did not harbor any plasmids. Plasmids were cured in Streptococcus and Micrococcus at a concentration of 75 and 100 microg/ mL of acridine orange, respectively, and transformation of 4.2- and 5.1-kb plasmids isolated from the Streptococcus and Micrococcus to plasmid-free E. coli DH5alpha strain was possible. In conjugation experiments, the antibiotic resistance profiles of transconjugant cells were found to be the same as the donors with the exception of Staphylococcus. The results of this study suggest that transformation and conjugation could be an important mechanism for horizontal gene transfer between bacteria in poultry litter. An understanding of the mechanism and magnitude of resistance gene transfer may provide a strategy to reduce the potential for dissemination of these genes.

Due to the environmental problems associated with disposal of livestock sludge, many stabilization studies emphasizing on the sludge volume reduction were performed. However, little is known about the microbial risk present in sludge and its stabilized products. This study microbiologically explored the effects of anaerobic lagoon fermentation (ALF) and autothermal thermophilic aerobic digestion (ATAD) on pathogen-related risk of raw swine manure by using culture-independent 16S rDNA cloning and sequencing methods. In raw swine manure, clones closely related to pathogens such as Dialister pneumosintes, Erysipelothrix rhusiopathiae, Succinivibrioan dextrinosolvens, and Schineria sp. were detected. Meanwhile, in the mesophilic ALF-treated swine manure, bacterial community clones closely related to pathogens such as Schineria sp. and Succinivibrio dextrinosolvens were still detected. Interestingly, the ATAD treatment resulted in no detection of clones closely related to pathogens in the stabilized thermophilic bacterial community, with the predominance of novel Clostridia class populations. These findings support the superiority of ATAD in selectively reducing potential human and animal pathogens compared to ALF, which is a typical manure stabilization method used in livestock farms.

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In this study, we microbiologically evaluated antibiotic resistance and pathogenicity in livestock (swine) manure as well as its biologically stabilized products. One of new livestock manure stabilization techniques is ATAD (Autothermal Thermophilic Aerobic Digestion). Because of its high operation temperature (60-65 degrees C), it has been speculated to have effective microbial risk control in livestock manure. This hypothesis was tested by evaluating microbial risk in ATAD-treated swine manure. Antibiotic resistance, multiple antibiotic resistance (MAR), and pathogenicity were microbiologically examined for swine manure as well as its conventionally stabilized (anaerobically fermented) and ATAD-stabilized products. In the swine manure and its conventionally stabilized product, antibiotic resistant (tetracycline-, kanamycine-, ampicillin-, and rifampicin-resistant) bacteria and the pathogen indicator bacteria were detected. Furthermore, approximately 2-5% of the Staphylococcus and Salmonella colonies from their selective culture media were found to exhibit a MAR-phenotypes, suggesting a serious level of microbe induced health risk. In contrast, after the swine manure was stabilized with a pilot-scale ATAD treatment for 3 days at 60-65 degrees C, antibiotic resistant bacteria, pathogen indicator bacteria, and MAR-exhibiting pathogens were all undetected. These findings support the improved control of microbial risk in livestock wastes by ATAD treatment.

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