Screening and identification of an oxytetracycline degradation strain resistant to Cu2+ and Zn2+ and its characteristics
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摘要:目的
铜、锌离子和土霉素作为养殖业中常用饲料添加剂，在畜禽粪便和污水中大量残留，形成了复合污染。本文在铜、锌离子的胁迫下筛选出降解土霉素的菌株。
方法通过选择培养基，从养殖废水中筛选分离出1株土霉素降解菌DJI，优化其降解条件，测定DJ1菌株对其他四环素类抗生素和氯霉素的耐受性。
结果通过菌株形态学特征及18S rDNA序列比对分析，确定DJ1菌株为Cutaneotrichosporon cutaneum。该菌在pH7、温度30 ℃、装液量50 mL(使用250 mL三角瓶)、底物质量浓度200 mg/L、接种量为1% (φ)的条件下，培养5 d后对土霉素的降解率为78.83%。在低质量浓度(50 mg/L)土霉素的培养基中，添加50 mg/L Zn2+抑制了土霉素的降解；而在高质量浓度(200 mg/L)土霉素培养基中，添加50 mg/L Cu2+抑制了土霉素的降解。菌株DJ1对四环素类抗生素及氯霉素具有高耐受能力，耐受均超过700 mg/L，能在抗生素与重金属铜、锌离子二元交叉培养基平板上生长，具有耐抗生素与重金属的双重抗性。
结论菌株DJI具有高耐受四环素类抗生素、氯霉素、铜和锌离子的能力，能高效降解土霉素，可在环境抗生素污染防治中发挥积极作用。
Abstract:ObjectiveAs the main feed additive in breeding industry, Cu2+, Zn2+ and oxytetracycline contribute to high amounts of residues in livestock and poultry excrement and sewage, forming complex pollution. We are aimed at screening strains to degrade oxytetracycline under the stress of Cu2+ and Zn2+.
MethodThe strain DJ1 of oxytetracycline degrading bacteria was screened and isolated from culture wastewater using selective medium. The biodegrading conditions of oxytetracycline by strain DJ1 were optimized. The tolerances of strain DJ1 to other tetracycline antibiotics and chloramphenicol were determined.
ResultThe strain DJ1 was identified as Cutaneotrichosporon cutaneum based on morphology and 18S rDNA sequence alignment. The oxytetracycline degradation rate of DJ1 reached 78.83% after five days of culturations under the condition of pH7, 30 ℃ temperature, 50 mL liquid in 250 mL flask, 200 mg/L substrate and 1%(φ) inoculum size. In medium with lower concentration (50 mg/L) of oxytetracycline, 50 mg/L Zn2+ inhibited oxytetracycline degradation, while 50 mg/L Cu2+ inhibited oxytetracycline degradation in medium with higher concentration (200 mg/L) of oxytetracycline. DJ1 strain had high tolerance to tetracycline antibiotics and chloramphenicol, the tolerating concentrations to both were above 700 mg/L. DJ1 strain could grow on the plate of binary cross medium with antibiotics and heavy metals of Cu2+ and Zn2+, showing dual resistance to antibiotics and heavy metals.
ConclusionDJ1 strain has high tolerance to tetracycline antibiotics, chloramphenicol, Cu2+ and Zn2+. It can efficiently degrade oxytetracycline and can be used to control antibiotic contamination in the environment.
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Keywords:
- oxytetracycline /
- degradation rate /
- antibiotic /
- heavy metal /
- tolerance
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图 3 pH对DJ1菌株降解土霉素的影响
柱子上方的不同小写字母表示差异显著(P<0.05，Duncan’s法)
Figure 3. Effect of pH on the degradation of oxytetracycline by strain DJ1
Different lowercase letters on bars indicate significant differences (P<0.05, Duncan’s method)
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图 4 接种温度对DJ1菌株降解土霉素的影响
柱子上方的不同小写字母表示差异显著(P<0.05，Duncan’s法)
Figure 4. Effect of incubation temperature on the degradation of oxytetracycline by strain DJ1
Different lowercase letters on bars indicate significant differences (P<0.05, Duncan’s method)
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图 5 接种量对DJ1菌株降解土霉素的影响
柱子上方的不同小写字母表示差异显著(P<0.05，Duncan’s法)
Figure 5. Effect of inoculum size on the degradation of oxytetracycline by strain DJ1
Different lowercase letters on bars indicate significant differences (P<0.05, Duncan’s method)
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图 6 装液量对DJ1菌株降解土霉素的影响
柱子上方的不同小写字母表示差异显著(P<0.05，Duncan’s法)
Figure 6. Effect of substrate volume on the degradation of oxytetracycline by strain DJ1
Different lowercase letters on bars indicate significant differences (P<0.05, Duncan’s method)
![]()
图 7 土霉素浓度对DJ1菌株降解土霉素的影响
柱子上方的不同小写字母差异表示显著(P<0.05，Duncan’s法)
Figure 7. Effect of oxytetracycline concentration on the degradation of oxytetracycline by strain DJ1
Different lowercase letters on bars indicate significant differences (P<0.05, Duncan’s method)
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