Veterinary manure is an important pollution reservoir of antibiotics and antibiotic-resistant bacteria (ARB). However, little is known of the distribution of ARB in plant endophytic bacteria and the number/types of ARB in chicken manure. In this study, 454-pyrosequencing was used to investigate the distribution and composition of ARBs in chicken manure and fertilized vegetables. The prevalence of ARB in the samples of the chicken manure compost recovered from farms on which amoxicillin, kanamycin, gentamicin, and cephalexin were used was 20.91-65.9% for ARBs and 8.24-20.63% simultaneously resistant to two or more antibiotics (multiple antibiotic resistant bacteria (MARB)). Antibiotic-resistant endophytic bacteria were widely detected in celery, pakchoi, and cucumber with the highest rate of resistance to cephalexin. The pyrosequencing indicated that the chicken manure dominantly harbored Firmicutes, Bacteroidetes, Synergistetes, and Proteobacteria and that Bacteroidetes was significantly enhanced in farms utilizing antibiotics. In the total cultivable colonies, 62.58-89.43% ARBs and 95.29% MARB were clustered in Bacteroidetes with the dominant species (Myroides ordoratimimus and Spningobacterium spp., respectively) related to human clinical opportunistic pathogens.
Through livestock manure fertilization, antibiotics, antibiotic-resistant bacteria and genes are transferred to agricultural soils, resulting in a high prevalence of antibiotic-resistant bacteria in the soil. It is not clear, however, whether a correlation exists between resistant bacterial populations in manure and manure-amended soil. In this work, we demonstrate that the prevalence of cephalexin-, amoxicillin-, kanamycin- and gentamicin-resistant bacteria as well as bacteria simultaneously resistant to all four antibiotics was much higher in manure-amended soils than in manure-free soil. 454-pyrosequencing indicated that the ARB and multiple antibiotic-resistant bacteria (MARB) in swine or chicken manure and manure-amended soil were mainly distributed among Sphingobacterium, Myroides, Enterococcus, Comamonas and unclassified Flavobacteriaceae. The genus Sphingobacterium was highly prevalent among ARB from swine manure and manure-amended soil, and was also the most dominant genus among MARB from chicken manure and manure-amended soil. Other dominant genera among ARB or MARB populations in manure samples, including Myroides, Enterococcus and Comamonas, could not be detected or were detected at very low relative abundance in manure-amended soil. The present study suggests the possibility of transfer of ARBs from livestock manures to soils and persistence of ARB in these environments.
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