Ansari M I, Grohmann E, Malik A
Department of Agricultural Microbiology, Faculty of Agricultural Sciences, Aligarh Muslim University, Aligarh, India.
J Appl Microbiol. 2008 Jun;104(6):1774-81. doi: 10.1111/j.1365-2672.2008.03736.x. Epub 2008 Feb 13.
Determination of heavy metal and antibiotic resistance and presence of conjugative plasmids in bacteria isolated from soil irrigated with wastewater.
Composite soil samples were collected from Ghaziabad, Uttar Pradesh, India. Forty different bacteria were selected from nutrient agar and characterized by morphological, cultural and biochemical tests. All the isolates were tested for their resistance to different heavy metals and antibiotics. The DNA derived from multiple metal and antibiotic-resistant bacterial isolates was PCR amplified and plasmid-specific sequences (IncP, IncN, IncW, IncQ and pMV158-type) were analysed by dot blot hybridization. All isolates gave PCR products with trfA2 and oriT primers of the IncP group. These PCR products also hybridized with the RP4-derived probes. However, the samples were negative for all the other investigated plasmids as proved by PCR and dot blots.
The presence of conjugative/mobilizable IncP plasmids in the isolates indicates that these bacteria have gene-mobilizing capacity with implications for potential dissemination of introduced recombinant DNA.
The detection of IncP plasmids in all the bacterial isolates is another proof for the prevalence of these plasmids. We propose that IncP plasmids are mainly responsible for the spread of multi-resistant bacteria in these soils.
测定从用废水灌溉的土壤中分离出的细菌的重金属抗性、抗生素抗性及接合质粒的存在情况。
从印度北方邦加济阿巴德采集复合土壤样本。从营养琼脂平板上挑选出40种不同细菌,并通过形态学、培养特性及生化试验进行鉴定。对所有分离菌株进行不同重金属和抗生素抗性检测。对多种金属及抗生素抗性细菌分离株的DNA进行PCR扩增,并通过斑点杂交分析质粒特异性序列(IncP、IncN、IncW、IncQ和pMV158型)。所有分离株均用IncP组的trfA2和oriT引物扩增出PCR产物。这些PCR产物也与RP4衍生探针杂交。然而,经PCR和斑点杂交证实,所有其他研究的质粒样本均为阴性。
分离株中存在接合/可移动IncP质粒表明这些细菌具有基因移动能力,这对引入的重组DNA的潜在传播具有影响。
在所有细菌分离株中检测到IncP质粒是这些质粒普遍存在的又一证据。我们认为IncP质粒是这些土壤中多重耐药细菌传播的主要原因。