Department of Veterinary Microbiology and Pathology, Washington State University, 402 Bustad Hall, Pullman, WA 99164-7040, USA.
Appl Environ Microbiol. 2011 Oct;77(20):7255-60. doi: 10.1128/AEM.05352-11. Epub 2011 Aug 19.
It is generally assumed that antibiotic residues in soils select for antibiotic-resistant bacteria. This assumption was tested by separately adding 10 different antibiotics (≥200 ppm) to three soil-water slurries (silt-loam, sand-loam, and sand; 20% soil [wt/vol]) and incubating mixtures for 24 h at room temperature. The antibiotic activity of the resultant supernatant was assessed by culturing a sensitive Escherichia coli strain in the filter-sterilized supernatant augmented with Luria-Bertani broth. We found striking differences in the abilities of supernatants to suppress growth of the indicator E. coli. Ampicillin, cephalothin, cefoxitin, ceftiofur, and florfenicol supernatants completely inhibited growth while bacterial growth was uninhibited in the presence of neomycin, tetracycline, and ciprofloxacin supernatants. High-performance liquid chromatography (HPLC) analysis demonstrated that cefoxitin and florfenicol were almost completely retained in the supernatants, whereas tetracycline and ciprofloxacin were mostly removed. Antibiotic dissipation in soil, presumably dominated by adsorption mechanisms, was sufficient to neutralize 200 ppm of tetracycline; this concentration is considerably higher than reported contamination levels. Soil pellets from the tetracycline slurries were resuspended in a minimal volume of medium to maximize the interaction between bacteria and soil particles, but sensitive bacteria were still unaffected by tetracycline (P = 0.6). Thus, residual antibiotics in soil do not necessarily exert a selective pressure, and the degree to which the pharmaceutical remains bioactive depends on the antibiotic. Efforts to control antibiotic contamination would be better directed toward compounds that retain biological activity in soils (e.g., cephalosporins and florfenicol) because these are the antibiotics that could exert a selective pressure in the environment.
人们通常认为土壤中的抗生素残留会选择出抗药性细菌。为了验证这一假设,我们分别向三种水土混合物(粉砂壤土、砂壤土和砂土;土壤占 20%[重量/体积])中添加了 10 种不同的抗生素(≥200ppm),并在室温下孵育 24 小时。然后通过在过滤灭菌的上清液中添加 Luria-Bertani 肉汤来培养敏感的大肠杆菌菌株,评估所得上清液的抗生素活性。我们发现上清液抑制指示大肠杆菌生长的能力存在显著差异。氨苄青霉素、头孢噻吩、头孢西丁、头孢噻呋和氟苯尼考上清液完全抑制了细菌的生长,而新霉素、四环素和环丙沙星上清液则没有抑制细菌的生长。高效液相色谱(HPLC)分析表明,头孢西丁和氟苯尼考几乎完全保留在上清液中,而四环素和环丙沙星则大部分被去除。土壤中抗生素的降解(推测主要由吸附机制主导)足以中和 200ppm 的四环素;这个浓度远高于报道的污染水平。四环素悬浮液的土壤颗粒重新悬浮在最小体积的培养基中,以最大限度地增加细菌与土壤颗粒之间的相互作用,但敏感细菌仍然不受四环素的影响(P=0.6)。因此,土壤中的残留抗生素不一定会产生选择压力,而药物的生物活性程度取决于抗生素。控制抗生素污染的努力应该更好地针对那些在土壤中保持生物活性的化合物(例如头孢菌素和氟苯尼考),因为这些抗生素可能会对环境产生选择压力。