Isolation, identification and evaluation of oil hydrocarbon decomposing bacteria from contaminated areas of oil fields

Mohsen Kojuri, F. Ardestani
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Abstract

Microbialbiodegradation is known as an effective and harmless method to overcome environmental pollution with oil hydrocarbon. Some bacterial species were isolated from the Sarvestan oilfields (Iran, Fars province), then identified and applied for oil hydrocarbon decomposition. A carbon-free minimum medium (CFMM) containing 1% crude oil was used to isolate bacteria through incubation at 30°C in the dark at 200 rpm for 7 days. Different methods were used to identify the  hydrocarbon oil decomposing bacteria: gram staining, squalene hydrolysis, catalase, production of arginine dihydrolase, gelatin liquefaction, hydrogen sulfide production, levan production, methyl red, oxidase, nitrite reduction, oxidative/fermentative, starch hydrolysis and Tween-80 hydrolysis tests. Nine different oil decomposing bacterial species were isolated. All the species grew well at 28 and 35°C, while four isolates containing of Bacillus sp. SA13, Pantoea sp. SA1112, Pseudomonas aeruginosa sp. SA21, and Bacillus sp. SA23 were capable of growing in a temperature of up to around 42°C. The minimum salt tolerance for isolates, except for Enterobacter sp. SA711, was 8%; Bacillus sp. SA212 had the highest tolerance of 15% sodium chloride. Acinetobacter sp. SA172, Enterobacter sp. SA711, Pseudomonas sp. SA75, Bacillus sp. SA212 and Bacillus sp. SA23 had the most growth rate in the CFMM. The highest percentages of oil removal obtained were 89% for Enterobacter sp. SA711, 86% for Acinetobacter sp. SA172, and 68% for Pseudomonas sp. SA75. The three isolated bacterial strains from the contaminated soil of the Sarvestan area had a good ability to degrade oil hydrocarbon. Therefore, they could be used commercially for the bioremediation of this region.
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油田污染区的油烃分解细菌的分离、鉴定与评价
微生物降解是克服石油烃类污染的一种有效、无害的方法。从Sarvestan油田(伊朗法尔斯省)中分离出一些细菌,对其进行鉴定并应用于石油烃分解。采用含1%原油的无碳最小培养基(CFMM),在30°C暗箱中200 rpm孵育7天。采用革兰氏染色法、角鲨烯水解法、过氧化氢酶法、精氨酸二水解酶法、明胶液化法、硫化氢法、利末法、甲基红法、氧化酶法、亚硝酸盐还原法、氧化/发酵法、淀粉水解法和吐温-80水解法对烃类油分解菌进行鉴定。分离出9种不同的石油分解细菌。所有菌株在28℃和35℃下均生长良好,而含有Bacillus sp. SA13、Pantoea sp. SA1112、Pseudomonas aeruginosa sp. SA21和Bacillus sp. SA23的4株菌株在42℃左右的温度下均能生长。除肠杆菌SA711外,其余菌株的最低耐盐性为8%;芽孢杆菌SA212对15%氯化钠的耐受性最高。不动杆菌sp. SA172、肠杆菌sp. SA711、假单胞菌sp. SA75、芽孢杆菌sp. SA212和芽孢杆菌sp. SA23在CFMM中生长速度最快。除油率最高的是肠杆菌SA711 89%,不动杆菌SA172 86%,假单胞菌SA75 68%。从Sarvestan地区污染土壤中分离出的3株细菌对石油烃有较好的降解能力。因此,它们可用于该地区的商业生物修复。
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