Application of the polymerase chain reaction (PCR) and reverse transcriptase / PCR for determining the fate of phenol-degrading Pseudomonas putida ATCC 11172 in a bioaugmented sequencing batch reactor

Selvaratnam, Shivi; Schoedel, B A; McFarland, B. L.; Kulpa, Charles F.

doi:10.1007/s002530050919

“…However, a major problem in microbial ecology is ascertaining the identity of the target bacterial strains used for bioremediation. Application of PCR techniques to monitor augmented microorganisms have been widely reported [7,29]. In a previous study, the efficacy of ERIC-PCR has been reported to monitor the survivability of wild-type strain A. baumannii S30 in the field [1].…”

Section: Discussionmentioning

confidence: 99%

“…There are certain shortcomings in using PCR-based fingerprinting techniques with repeat element primers to monitor survivability of the introduced strains [3,8]. The use of reporter genes for efficient monitoring of these biological agents circumvents the mentioned shortcomings of conventional PCR-based DNA fingerprinting techniques [3,6,29]. Thus, the aim of the present investigation was to evaluate the stability of a reporter gene in wild strain A. baumannii S30 and observe the survival and effect of the foreign insert on the metabolic capability of the wild-type strain.…”

Section: Discussionmentioning

confidence: 99%

Crude oil degradation efficiency of a recombinant Acinetobacter baumannii strain and its survival in crude oil-contaminated soil microcosm

Mishra¹,

Sarma

²

,

Lal

³

2004

FEMS Microbiology Letters

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A hydrocarbon degrading Acinetobacter baumannii S30 strain, isolated from crude oil-contaminated soil, was inserted with the lux gene from the luciferase gene cassette luxCDABE. Soil microcosms were designed to study the degradation efficacy for total petroleum hydrocarbon (TPH) of crude oil by lux-tagged A. baumannii S30 pJES. Bioaugmentation of a TPH-contaminated microcosm with A baumannii S30 pJES showed that TPH levels were reduced from 89.3 to 53.9 g/kg soil in 90 days. Biodegradation of TPH by A baumannii S30 pJES was also monitored in shake flask conditions, which showed a reduction of initial TPH levels by over 50% at the end of 120 h. A lux-PCR-based approach along with the standard dilution plating with selective antibiotics was successfully utilized to monitor the survivability of the lux-tagged strain A. baumannii S30 pJES in soil microcosms and stability of the lux insert in the host strain A. baumannii S30. The selective plating technique indicated the population of A. baumannii S30 pJES to be 6.5+/-0.13 x 10(8) CFU/g at day zero (just after bioaugmentation) and 2.09+/-0.08 x 10(8) CFU/g of soil after 90 days of incubation. lux-PCR confirmed the stability of the insert in all the randomly selected colonies of A. baumannii strains from the antibiotic plates. The lux insert was stable after 50 generations in Luria Bertini broth and storage at -70 degrees C as glycerol stocks for over a year. These results revealed that the lux insert was stable and lux-tagged A. baumannii S30 strain could survive in a TPH-contaminated soil microcosm and could degrade TPH in the soil microcosm conditions. It can be used as an effective marker to monitor the survival of augmented strains at a bioremediation site.

show abstract

“…However, a major problem in microbial ecology is ascertaining the identity of the target bacterial strains used for bioremediation. Application of PCR techniques to monitor augmented microorganisms have been widely reported [7,29]. In a previous study, the efficacy of ERIC‐PCR has been reported to monitor the survivability of wild‐type strain A. baumannii S30 in the field [1].…”

Section: Discussionmentioning

confidence: 99%

Crude oil degradation efficiency of a recombinantAcinetobacter baumanniistrain and its survival in crude oil-contaminated soil microcosm

Mishra¹,

Sarma²,

Lal

³

2004

FEMS Microbiology Letters

View full text Add to dashboard Cite

A hydrocarbon degrading Acinetobacter baumannii S30 strain, isolated from crude oil-contaminated soil, was inserted with the lux gene from the luciferase gene cassette luxCDABE. Soil microcosms were designed to study the degradation efficacy for total petroleum hydrocarbon (TPH) of crude oil by lux-tagged A. baumannii S30 pJES. Bioaugmentation of a TPH-contaminated microcosm with A baumannii S30 pJES showed that TPH levels were reduced from 89.3 to 53.9 g/kg soil in 90 days. Biodegradation of TPH by A baumannii S30 pJES was also monitored in shake flask conditions, which showed a reduction of initial TPH levels by over 50% at the end of 120 h. A lux-PCR-based approach along with the standard dilution plating with selective antibiotics was successfully utilized to monitor the survivability of the lux-tagged strain A. baumannii S30 pJES in soil microcosms and stability of the lux insert in the host strain A. baumannii S30. The selective plating technique indicated the population of A. baumannii S30 pJES to be 6.5+/-0.13 x 10(8) CFU/g at day zero (just after bioaugmentation) and 2.09+/-0.08 x 10(8) CFU/g of soil after 90 days of incubation. lux-PCR confirmed the stability of the insert in all the randomly selected colonies of A. baumannii strains from the antibiotic plates. The lux insert was stable after 50 generations in Luria Bertini broth and storage at -70 degrees C as glycerol stocks for over a year. These results revealed that the lux insert was stable and lux-tagged A. baumannii S30 strain could survive in a TPH-contaminated soil microcosm and could degrade TPH in the soil microcosm conditions. It can be used as an effective marker to monitor the survival of augmented strains at a bioremediation site.

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“…It has been reported that the periodic change of selected environmental conditions is an effective means to control the abundance and activity of the introduced microorganisms in multi-species microbial communities [15,25,26]. Sequencing batch reactor is operated in such a periodic mode.…”

Section: Long Operation Of Bioaugmented and Control Ansbrsmentioning

confidence: 99%

Bioaugmentation on decolorization of C.I. Direct Blue 71 by using genetically engineered strain Escherichia coli JM109 (pGEX-AZR)

Jin

¹

,

Yang

²

,

Zhang

³

et al. 2009

Journal of Hazardous Materials

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Application of the polymerase chain reaction (PCR) and reverse transcriptase / PCR for determining the fate of phenol-degrading Pseudomonas putida ATCC 11172 in a bioaugmented sequencing batch reactor

Cited by 49 publications

References 8 publications

Crude oil degradation efficiency of a recombinant Acinetobacter baumannii strain and its survival in crude oil-contaminated soil microcosm

Crude oil degradation efficiency of a recombinant Acinetobacter baumannii strain and its survival in crude oil-contaminated soil microcosm

Crude oil degradation efficiency of a recombinantAcinetobacter baumanniistrain and its survival in crude oil-contaminated soil microcosm

Bioaugmentation on decolorization of C.I. Direct Blue 71 by using genetically engineered strain Escherichia coli JM109 (pGEX-AZR)

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