• ISSN 1008-505X
  • CN 11-3996/S
孟应宏, 冯瑶, 黎晓峰, 刘元望, 李兆君. 土霉素降解菌筛选及降解特性研究[J]. 植物营养与肥料学报, 2018, 24(3): 720-727. DOI: 10.11674/zwyf.17161
引用本文: 孟应宏, 冯瑶, 黎晓峰, 刘元望, 李兆君. 土霉素降解菌筛选及降解特性研究[J]. 植物营养与肥料学报, 2018, 24(3): 720-727. DOI: 10.11674/zwyf.17161
MENG Ying-hong, FENG Yao, LI Xiao-feng, LIU Yuan-wang, LI Zhao-jun. Isolation of an oxytetracycline-degrading bacterial strain and its biodegradation characteristics[J]. Journal of Plant Nutrition and Fertilizers, 2018, 24(3): 720-727. DOI: 10.11674/zwyf.17161
Citation: MENG Ying-hong, FENG Yao, LI Xiao-feng, LIU Yuan-wang, LI Zhao-jun. Isolation of an oxytetracycline-degrading bacterial strain and its biodegradation characteristics[J]. Journal of Plant Nutrition and Fertilizers, 2018, 24(3): 720-727. DOI: 10.11674/zwyf.17161

土霉素降解菌筛选及降解特性研究

Isolation of an oxytetracycline-degrading bacterial strain and its biodegradation characteristics

  • 摘要:
    目的 近年来,抗生素在畜禽及水产养殖中的使用量增加,导致固体废弃物和污水中存在大量的抗生素和耐药菌。土霉素作为用于养殖业主要的抗生素之一,在畜禽粪便和污水中的残留含量较高,因此,筛选并鉴定了能降解残留土霉素的微生物。
    方法 采用富集驯化法,以菌肥、药渣和畜禽粪便为原料,采用摇床震荡的方法进行微生物培养,采用高效液相色谱法 (HPLC) 进行土霉素含量测定,筛选出能够高效降解土霉素的微生物。本研究还对降解菌在不同温度、pH、转速和接种量条件下的土霉素降解效果进行优化,并最终利用16S rDNA的方法鉴定菌种。
    结果 筛选出一株能够高效降解土霉素的菌株T4菌,经16S rDNA测序鉴定该菌株为假单胞菌 (Pseudomonas sp.) ,该菌株在30℃时对土霉素的降解率最高,达到了26.75%;不同pH梯度下,T4菌在pH为7时对土霉素的降解率达到最高,为27.03%;转速为150 rpm和170 rpm时,T4菌对土霉素的降解率分别为26.18%和25.59%,考虑到摇床高转速耗能高的因素,因此选择150 rpm为优化的转速;接种量对T4菌降解土霉素的影响较小,而且二者之间呈负相关,接种量1%时降解率最高,为26.88%。优化条件下,T4菌对100 mg/L土霉素的降解率为26.29%;堆肥试验表明,添加了T4菌之后,土霉素去除率更高,为93.21%。
    结论 本研究筛选出的菌株T4对土霉素有较好的降解能力。通过16S rDNA基因序列分析,T4菌属于假单胞菌 (Pseudomonas sp.),其降解土霉素的优化条件为温度30℃、pH 7.0、转速150 rpm、接种量1%。在堆肥中接种T4菌后,提高了对土霉素的去除作用,表明T4菌作为土霉素降解菌具有污染治理的潜力。

     

    Abstract:
    Objectives With the rapid development of intensive livestock breeding, the use of antibiotics has been increased sharply, which results in a large amount of antibiotics residuals and drug-resistant bacteria in solid waste and sewage. The improper discharge of these solid waste and sewage leads to accumulation of antibiotics and drug-resistant bacteria in soil and water. As one of the main antibiotics used in animal husbandry industry, high residual content of oxytetracycline exists in animal manure and sewage. Effective strains need to be developed for the efficient degradation and cleanup of residual oxytetracycline.
    Methods A method of enrichment and acclimation was applied to screen oxytetracycline degradation bacteria from microbial fertilizers, antibiotic fermentation residues and livestock manures. The screened strains were identified using the 16S rDNA sequencing technology. Oxytetracycline content was determined using high performance liquid chromatography (HPLC) method. The biodegradation condition of oxytetracycline by the screened bacteria was optimized under different temperature, pH, rotation speed and inoculum size.
    Results The screened strain of T4 was found capable of efficiently degrading oxytetracycline, and was identified as Pseudomonas sp. The degradation rate of oxytetracycline was the highest (26.75%) at 30℃, and was 27.03% at pH 7. When the rotation speed was 150 rpm and 170 rpm, oxytetracycline showed higher degradation rates of 26.18% and 25.59%, respectively. Considering the high energy consumption of the high-speed shaking, 150 rpm was chosen as the optimal speed. The inoculation was negatively correlated with the degradation of oxytetracycline, and finally 1% of inoculum size was chosen as the optimized one. The highest oxytetracycline degradation rate was 26.29% under the final optimized condition at the medium of 100 mg/L oxytetracycline. In addition, the composting experiment showed that T4 could effectively improve the degradation of oxytetracycline during composting.
    Conclusions The strain of T4, identified as Pseudomonas sp., is proved to be capable of degrading oxytetracycline efficiently. The optimized conditions are 30℃, pH 7.0, 150 rpm, and 1% of inoculum size. The composting test showed that the T4 had a positive effect on the degradation of oxytetracycline, indicating potential of using the T4 as oxytetracycline degrading bacterium in environment pollution prevention.

     

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