scholarly journals Isolation and characterization of hydrocarbon-degrading bacteria from gas station leaking-contaminated groundwater in the Southern Amazon, Brazil

2020 ◽  
Vol 80 (2) ◽  
pp. 354-361 ◽  
Author(s):  
S. D. Lima ◽  
A. F. Oliveira ◽  
R. Golin ◽  
V. C. P. Lopes ◽  
D. S. Caixeta ◽  
...  
Keyword(s):  
Contaminated Groundwater ◽  
Rrna Gene ◽  
Bacillus Sp ◽  
Alkane Hydroxylase ◽  
Gene Group ◽  
Group Iii ◽  
Isolation And Characterization ◽  
Degrading Bacteria ◽  
Benzene Toluene ◽  
Gas Station

Abstract Twenty-three hydrocarbon-degrading bacteria strains were isolated from gas station leaking-contaminated groundwater located in the Southern Amazon, Brazil. Based on hydrocarbon (diesel, hexadecane, benzene, toluene and xylene) degradation ability, two strains were selected for further study. The amplification and sequencing of the 16S rRNA gene showed that these two strains belonged to the genus Bacillus (Bacillus sp. L26 and Bacillus sp. L30). GC-MS analysis showed that strain L30 was the most effective in degrading n-alkane (C10-C27) from diesel after 7 days of cultivation in mineral medium. Both strains produced biosurfactants and showed emulsification activity, specially the strain L30. Alkane hydroxylase gene (group III), which is important for alkane biodegradation, was present in strains. As a result, this study indicated that these bacteria could have promising applications in hydrocarbon bioremediation.

scholarly journals Isolation and characterization of the cellulose-degrading bacteria from Ngoc Linh ginseng soil in Quang Nam province

2016 ◽  
Vol 14 (1) ◽  
pp. 55-61
Author(s):  
Trần Bảo Trâm ◽  
Phạm Hương Sơn ◽  
Ngô Thị Hiền ◽  
Ngô Thị Hoa ◽  
Nguyễn Thu Hiền ◽  
...  
Keyword(s):  
Thick Layer ◽  
Rrna Gene ◽  
Ideal Condition ◽  
Pseudomonas Sp ◽  
Bacillus Sp ◽  
Isolation And Characterization ◽  
Degrading Bacteria ◽  
Ginseng Soil ◽  
The Ideal

Ngoc Linh ginseng (Panax vietnamensis Ha et Grushv.) is an endemic species in Vietnam and was discovered at the Ngoc Linh mountain (Kon Tum/Quảng Nam). Investigations showed that the soil with a thick layer of humus was the ideal condition for growth and development of Ngoc Linh ginseng. Therefore research on microbial flora as well as cellulose-degrading bacteria in ginseng soil may elucidate factors contributing to acclimatized cultivation of this ginseng in Vietnam. From the soil sample with cultivated Ngoc Linh ginseng in Quang Nam, five bacteria strains with cellulose-degrading activities were isolated (QN1, QN2, QN3, QN4, QN5 with respectively hydrolyzed CMC halos diameters of 10, 11, 22, 7, 22 mm) with cellulase activities of 1,31; 1,23; 2,99; 0,99; 2,51 U/ml. The combination of 16S rRNA gene sequences and cultured/biochemical characteristics of the bacteria showed that the five bacteria strains was classified to be Pseudomonas sp. QN1; Pseudomonas sp. QN4; Bacillus sp. QN2; Bacillus sp. QN3; Roseomonas sp. QN5.

scholarly journals Isolation and Characterization of Polycyclic Aromatic Hydrocarbon-Degrading Bacteria Associated with the Rhizosphere of Salt Marsh Plants

2001 ◽  
Vol 67 (6) ◽  
pp. 2683-2691 ◽  
Author(s):  
L. L. Daane ◽  
I. Harjono ◽  
G. J. Zylstra ◽  
M. M. Häggblom
Keyword(s):  
Salt Marsh ◽  
Polycyclic Aromatic Hydrocarbon ◽  
Aromatic Hydrocarbon ◽  
Rrna Gene ◽  
Gram Positive ◽  
Salt Marsh Plants ◽  
Pah Degradation ◽  
Isolation And Characterization ◽  
Degrading Bacteria ◽  
Polycyclic Aromatic

ABSTRACT Polycyclic aromatic hydrocarbon (PAH)-degrading bacteria were isolated from contaminated estuarine sediment and salt marsh rhizosphere by enrichment using either naphthalene, phenanthrene, or biphenyl as the sole source of carbon and energy. Pasteurization of samples prior to enrichment resulted in isolation of gram-positive, spore-forming bacteria. The isolates were characterized using a variety of phenotypic, morphologic, and molecular properties. Identification of the isolates based on their fatty acid profiles and partial 16S rRNA gene sequences assigned them to three main bacterial groups: gram-negative pseudomonads; gram-positive, non-spore-forming nocardioforms; and the gram-positive, spore-forming group,Paenibacillus. Genomic digest patterns of all isolates were used to determine unique isolates, and representatives from each bacterial group were chosen for further investigation. Southern hybridization was performed using genes for PAH degradation fromPseudomonas putida NCIB 9816-4, Comamonas testosteroni GZ42, Sphingomonas yanoikuyae B1, andMycobacterium sp. strain PY01. None of the isolates from the three groups showed homology to the B1 genes, only two nocardioform isolates showed homology to the PY01 genes, and only members of the pseudomonad group showed homology to the NCIB 9816-4 or GZ42 probes. The Paenibacillus isolates showed no homology to any of the tested gene probes, indicating the possibility of novel genes for PAH degradation. Pure culture substrate utilization experiments using several selected isolates from each of the three groups showed that the phenanthrene-enriched isolates are able to utilize a greater number of PAHs than are the naphthalene-enriched isolates. Inoculating two of the gram-positive isolates to a marine sediment slurry spiked with a mixture of PAHs (naphthalene, fluorene, phenanthrene, and pyrene) and biphenyl resulted in rapid transformation of pyrene, in addition to the two- and three-ringed PAHs and biphenyl. This study indicates that the rhizosphere of salt marsh plants contains a diverse population of PAH-degrading bacteria, and the use of plant-associated microorganisms has the potential for bioremediation of contaminated sediments.

scholarly journals Isolation and Characterization of Microcystin Degrading Bacteria from Holy Ponds in India

2017 ◽  
Vol 4 (4) ◽  
pp. 436-447 ◽  
Author(s):  
Vikram Pal Gandhi ◽  
Anil Kumar
Keyword(s):  
Restriction Analysis ◽  
Water Bodies ◽  
Rrna Gene ◽  
Degrading Bacterium ◽  
Isolation And Characterization ◽  
Chronic Poisoning ◽  
Degrading Bacteria ◽  
Health Related ◽  
Water Treatments

Microcystins (MCs) are toxic cyclic heptapeptides produced by few toxic cyanobacteria and generally form blooms in eutrophic surface fresh water bodies. They cause acute to chronic poisoning and other health related problems mainly by irreversible inhibition of protein phosphatases (PP1 and PP2A) and increased formation of reactive oxygen species (ROS).  Due to limitation of non-biological methods of water treatments the exploration of MCs degrading bacteria is emerging at a quite pace to address, through bioremediation, the problems posed by MCs in water and water-bodies. Report and study of MCs biodegrading bacteria from India were lacking. However it was evident, from our previous study, that microcystin degradation can be achieved by indigenous microcystin degrading bacterial population in its natural place where microcystin producing blooms occur. This study has presented isolation and characterization of indigenous microcystin degrading bacteria from holy ponds in Utter Pradesh of India. Overall 20 bacterial isolates were isolated from Microcystis infested different ponds. Out of these 13 isolates were mlrA positive by PCR and were found to be distinct isolates by amplified ribosomal DNA restriction analysis (ARDRA). However, ARDRA analysis revealed overall four bacterial groups. On the basis of 16S-rRNA gene sequence the Gram-positive-rod isolate PM1 was identified, with 99% identity, as Bacillus licheniformis which was shown earlier to cluster with microcystin degrading bacterium B. subtilis. Thus the present study revealed, for the first time, probable microcystin degrading bacteria in water-bodies from India. The potential and the metabolic pathway of PM1 and other mlrA positive isolates need to be further studied and validated to confirm their application in microcystin bioremediation. Int J Appl Sci Biotechnol, Vol 4(4): 436-447

scholarly journals Isolation and characterization of diesel-degrading bacteria from hydrocarbon-contaminated sites, flower farms and soda lakes

2020 ◽  
Author(s):  
Gessesse Kebede Bekele ◽  
Eshetu Mekonne Bogale ◽  
Tekle Tafesse Fida ◽  
Adugna Abdi Woldesemayat ◽  
Mesfin Tafesse Gemeda ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Soda Lakes ◽  
Bacterial Species ◽  
Growth Patterns ◽  
Rrna Gene ◽  
Gravimetric Analysis ◽  
Mineral Salts ◽  
Isolation And Characterization ◽  
Degrading Bacteria

Abstract Background: Hydrocarbon-derived pollutants are becoming one of the most concerning ecological issues. Thus, there is a need to investigate and develop innovative, low-cost, eco-friendly, and fast techniques to reduce and/or eliminate pollutants using biological agents. The current study is conducted to isolate, characterize, and identify potential diesel-degrading bacteria.Results: Samples were collected from flower farms, lakeshores, old aged garages, asphalt, and bitumen soils and spread on selective medium (Bushnell Hass Mineral Salts Agar) containing diesel as the growth substrate. The isolates were characterized based on their growth patterns using OD measurement, biochemical testing and gravimetric analysis and identified using the Biolog database, and 16S rRNA gene sequencing techniques. Subsequently, six diesel degraders were identified and belong to Pseudomonas , Providencia , Roseomonas , Stenotrophomonas , Achromobacter , and Bacillus . Among these, based on gravimetric analysis, the three potent isolates AAUW23, AAUG11 and AAUG36 achieved 84%, 83.4%, and 83% diesel degradation efficiency, respectively, in 15 days. Consequently, the partial 16S rRNA gene sequences revealed that the two most potent bacterial strains (AAUW23 and AAUG11) were Pseudomonas aeruginosa , while AAUG36 was Bacillus subtilis . Conclusion: This study demonstrated that bacterial species isolated from hydrocarbon-contaminated and/or uncontaminated environments could be optimized to be used as potential bioremediation agents for diesel removal.

scholarly journals RNA-Based Stable Isotope Probing and Isolation of Anaerobic Benzene-Degrading Bacteria from Gasoline-Contaminated Groundwater

2006 ◽  
Vol 72 (5) ◽  
pp. 3586-3592 ◽  
Author(s):  
Yuki Kasai ◽  
Yoh Takahata ◽  
Mike Manefield ◽  
Kazuya Watanabe
Keyword(s):  
Stable Isotope ◽  
16S Rrna ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Stable Isotope Probing ◽  
Contaminated Groundwater ◽  
Rrna Gene ◽  
Major Band ◽  
Benzene Degradation ◽  
Dgge Analysis ◽  
Degrading Bacteria

ABSTRACT Stable isotope probing (SIP) of benzene-degrading bacteria in gasoline-contaminated groundwater was coupled to denaturing gradient gel electrophoresis (DGGE) of DNA fragments amplified by reverse transcription-PCR from community 16S rRNA molecules. Supplementation of the groundwater with [13C6]benzene together with an electron acceptor (nitrate, sulfate, or oxygen) showed that a phylotype affiliated with the genus Azoarcus specifically appeared in the 13C-RNA fraction only when nitrate was supplemented. This phylotype was also observed as the major band in DGGE analysis of bacterial 16S rRNA gene fragments amplified by PCR from the gasoline-contaminated groundwater. In order to isolate the Azoarcus strains, the groundwater sample was streaked on agar plates containing nonselective diluted CGY medium, and the DGGE analysis was used to screen colonies formed on the plates. This procedure identified five bacterial isolates (from 60 colonies) that corresponded to the SIP-identified Azoarcus phylotype, among which two strains (designated DN11 and AN9) degraded benzene under denitrifying conditions. Incubation of these strains with [14C]benzene showed that the labeled carbon was mostly incorporated into 14CO2 within 14 days. These results indicate that the Azoarcus population was involved in benzene degradation in the gasoline-contaminated groundwater under denitrifying conditions. We suggest that RNA-based SIP identification coupled to phylogenetic screening of nonselective isolates facilitates the isolation of enrichment/isolation-resistant microorganisms with a specific function.

scholarly journals Valorization Potential of a Novel Bacterial Strain, Bacillus altitudinis RSP75, towards Lignocellulose Bioconversion: An Assessment of Symbiotic Bacteria from the Stored Grain Pest, Tribolium castaneum

2021 ◽  
Vol 9 (9) ◽  
pp. 1952
Author(s):  
Mudasir A. Dar ◽  
Neeraja P. Dhole ◽  
Rongrong Xie ◽  
Kiran D. Pawar ◽  
Kalim Ullah ◽  
...  
Keyword(s):  
Tribolium Castaneum ◽  
16S Rrna Gene Sequencing ◽  
Maximum Activity ◽  
Rrna Gene ◽  
Bacterial Cells ◽  
Cellulolytic Bacteria ◽  
Stored Grain ◽  
Bacillus Altitudinis ◽  
Isolation And Characterization ◽  
Degrading Bacteria

Bioconversion of lignocellulose into renewable energy and commodity products faces a major obstacle of inefficient saccharification due to its recalcitrant structure. In nature, lignocellulose is efficiently degraded by some insects, including termites and beetles, potentially due to the contribution from symbiotic gut bacteria. To this end, the presented investigation reports the isolation and characterization of cellulolytic bacteria from the gut system of red flour beetle, Tribolium castaneum. Out of the 15 isolated bacteria, strain RSP75 showed the highest cellulolytic activities by forming a clearance zone of 28 mm in diameter with a hydrolytic capacity of ~4.7. The MALDI-TOF biotyping and 16S rRNA gene sequencing revealed that the strain RSP75 belongs to Bacillus altitudinis. Among the tested enzymes, B. altitudinis RSP75 showed maximum activity of 63.2 IU/mL extract for xylanase followed by β-glucosidase (47.1 ± 3 IU/mL extract) which were manifold higher than previously reported activities. The highest substrate degradation was achieved with wheat husk and corn cob powder which accounted for 69.2% and 54.5%, respectively. The scanning electron microscopy showed adhesion of the bacterial cells with the substrate which was further substantiated by FTIR analysis that depicted the absence of the characteristic cellulose bands at wave numbers 1247, 1375, and 1735 cm−1 due to hydrolysis by the bacterium. Furthermore, B. altitudinis RSP75 showed co-culturing competence with Saccharomyces cerevisiae for bioethanol production from lignocellulose as revealed by GC-MS analysis. The overall observations signify the gut of T. castaneum as a unique and impressive reservoir to prospect for lignocellulose-degrading bacteria that can have many biotechnological applications, including biofuels and biorefinery.

scholarly journals Isolation and Characterization of Three and Four Ring Pahs Degrading Bacteria from Contaminated Sites, Ankleshwar, Gujarat, India

2015 ◽  
Vol 4 (1) ◽  
pp. 130-140 ◽  
Author(s):  
Jignasha G. Patel ◽  
J.I. Nirmal Kumar ◽  
Rita N Kumar ◽  
Shamiyan R. Khan
Keyword(s):  
Sole Source ◽  
Ex Situ ◽  
Rrna Gene ◽  
Degradation Study ◽  
Isolation And Characterization ◽  
Degrading Bacteria ◽  
Polycyclic Aromatic ◽  
Bacterial Groups

Polycyclic aromatic hydrocarbon (PAH)-degrading bacteria were isolated from prolong contaminated Amalakhadi sediment and crude oil polluted soil Telva, near Ankleshwar Gujarat India. Organisms were treated with two-model PAHs compound Anthracene (ANT), and Pyrene (PYR) as the sole source of carbon and energy. Identification of the isolates was carried out based on their morphological and partial 16S rRNA gene sequences, which revealed that the isolates belong to two main bacterial groups: gram-negative pseudomonas indoxyladons and gram-positive, spore-forming group, Bacillus benzoevorans. GC-MS based degradation study demonstrated that P. indoxyladons efficiently degrade 98% of ANT and PYR by 93.2 % when treated with 250 mg L-1. However, B. benzoevorans could tolerate to 200 mg L-1of PYR. Thus, the findings of the study provide novel bacterial sp. having different capacity to degrade model PAHs compounds and further could be utilized for the standardization of bioremediation protocols for ex situ and in situ studies in aquatic as well as terrestrial ecosystem.DOI: http://dx.doi.org/10.3126/ije.v4i1.12184International Journal of Environment Volume-4, Issue-1, Dec-Feb 2014/15, Page: 130-140  

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