Hydrocarbonoxidizing bacteria have been isolated from a variety of terrestrial and aquatic environments, using both enrichment and direct plating techniques. In oil fields, some but not all effective oildegrading bacteria produce extracellular biosurfactants to facilitate microbial oil uptake and degradation by emulsifying the hydrocarbon morikawa et al. Oildegrading bacterial community composition of few studies. With respect to oil hydrocarbons, the wide spectrum of over 17,000 compounds can be divided into four classes, resins quinolones, sulfoxides, amides and pyridines, asphaltenes ketones, fatty acids, esters and phenols, saturates and aromatics 11. Also, samples amended with cp were found to have more types and higher numbers of heterotrophic and hydrocarbon degrading bacteria than the samples without amendment controls. The hydrocarbon degrading bacterial types and numbers were found to be lesser than their heterotrophic counterparts. For the total count of bacteria, the diluted suspension was plated onto nutrient agar. Molecular and microbiological insights on the enrichment. The present study describes the isolation and identification of total heterotrophic bacterial thb population and hydrocarbon degrading bacterial hdb population, identification of the efficient degrader, its molecular characterization and phylogenetic analysis based upon 16s rna gene sequencing from the samples collected in ennore creek.
Particle stabilized emulsions provide an environmentally friendly alternative to the chemical dispersion of oil in the event of a spill over water. The ecology of hydrocarbon degradation by microbial populations in the natural. Isolation of hydrocarbon degrading bacteria following the strategy designed in our work to isolate hydrocarbondegrading bacteria both with high growth and biological activity, 39 purified bacterial isolates were recorded amongst hundreds of isolated ones at the first step of the program. Isolation and identification of petroleum hydrocarbon. To degrade large amounts of petroleum hydrocarbons, a substantial bacterial population is required which, in.
Interest in possible hydrocarbon oxidation by sulfatereducing bacteria arose from their frequent presence in oil production. Isolation and characterization of hydrocarbondegrading. Screening and isolation of petroleum hydrocarbon degrading bacteria separation of petroleum hydrocarbon degrading microorganisms. Succession of hydrocarbondegrading bacteria in the aftermath. Gutierrez t, singleton dr, berry d, yang t, aitken md, teske a. Isolation and characterization of hydrocarbondegrading bacteria. The deepwater horizon oil spill produced large subsurface plumes of dispersed oil and gas in the gulf of mexico that stimulated growth of psychrophilic, hydrocarbon degrading bacteria. Microbial biodegradation is the use of bioremediation and biotransformation methods to harness the naturally occurring ability of microbial xenobiotic metabolism to degrade, transform or accumulate environmental pollutants, including hydrocarbons e. In order to isolate the desired bacteria, enrichment culture was used.
Mittal and singh 2009, have reported eleven petroleum hydrocarbon degrading bacterial isolates from oil production site of lingala oil field, ongc and nine isolates from oil contaminated soil of local area at haridwar. Hydrocarbondegrading bacteria exhibit a speciesspecific. Isolation of hydrocarbon degrading bacteria from soils. In the present study, altogether 32 morphologically distinct bacteria were isolated from crude oil contaminated soil and 18 different hydrocarbondegrading bacteria were number. Oil hydrocarbon degradation by caspian sea microbial communities. Isolation and molecular identification of hydrocarbon. We describe the carbonization of these clays using the sustainable biopolymer, chitosan, to optimize wetting. Reconstructing metabolic pathways of hydrocarbondegrading bacteria from the deepwater horizon oil spill nina dombrowski1, john a. Anaerobic degradation of oil hydrocarbons by sulfate. In this study, we intend to isolate, screen and select bacterial strains able to degrade petroleum hydrocarbons specifically in recently or. Reconstructing metabolic pathways of hydrocarbon degrading bacteria from the deepwater horizon oil spill nina dombrowski1, john a. Oxygen is considered by many to be the primary growthlimiting factor for hydrocarbondegrading bacteria, but it is normally depleted in zones that have been contaminated with hydrocarbons.
Abstract many hydrocarboncontaminated environments are characterized by low or elevated temperatures, acidic or alkaline ph, high salt concentrations, or high pressure. Department of chemistry, rochester institute of technology, rochester, ny. Bacteria displaying such capabilities are often exploited for the. Bacteria are the most active agents in petroleum degradation, and they work as primary degraders of spilled oil in environment 37, 38. Isolation of hydrocarbon degrading bacteria following the strategy designed in our work to isolate hydrocarbon degrading bacteria both with high growth and biological activity, 39 purified bacterial isolates were recorded amongst hundreds of isolated ones at the first step of the program. Hydrocarbon degrading bacteria enriched by the deepwater horizon oil spill identified by cultivation and dnasip. Adaptation is brought about by selective enrichment of hydrocarbon utilizing microorganisms and amplification of the pool of hydrocarbon catabolizing genes.
Altogen labs has developed active bioremediation technology based on natural oildegrading bacteria for largescale remediation of hydrocarboncontaminated water and soil. Characterization of hydrocarbondegrading microbial populations. One potent gram negative bacillus bacteria was isolated on the basis of petroleum hydrocarbon degradation efficiency and identified as pseudomonas sp. In oil fields, some but not all effective oil degrading bacteria produce extracellular biosurfactants to facilitate microbial oil uptake and degradation by emulsifying the hydrocarbon morikawa et al. Several bacteria are even known to feed exclusively on hydrocarbons 39.
Isolation and characterization of bacteria degrading. Indeed, the proportion of hydrocarbon degrading bacteria at 10,400 m is the highest observed in any natural environment on earth. Results the result of the bacterial count showed 38. Isolation, identification and characterization of oil degrading bacteria isolated 431 biochemical characterization of selected microorganisms. The function of hydrocarbon degrading bacteria mainly depends on their hydrocarbon degrading enzymes, the expression and activity of which are closely related to the physiological activity of bacteria mukherjee et al. Hydrocarbon degrading microorganisms require an environmental habitat that has a sufficient and preferably sustainable slow release source of nutrients mostly n and p, water, air, mild ambient temperature, and a moderate ph. Isolation and identification of effective hydrocarbon. Pdf proliferation of hydrocarbondegrading microbes at. Floodgate 36 listed 25 genera of hydrocarbon degrading bacteria and 25 genera of hydrocarbon degrading fungi which were isolated. Screening of hydrocarbon degrading microorganisms isolated from three habitats in western new york state by selective enrichment technique, resulted in the collection of 20 distinct species. Isolation, screening and activity of hydrocarbondegrading.
The separation of petroleum hydrocarbon degrading bacteria was as follows. Oil hydrocarbon degradation by caspian sea microbial. Educator materials microbial community response to an. These bacteria were mainly oleibacter, thalassolituus, and. Twentythree hydrocarbondegrading bacteria strains were isolated from gas station leakingcontaminated. Role of microbes in oil spills remediation and degradation. Pnp degrading bacteria and this item was submitted in to occur in microform, scholars, gejun. Engineered clays as sustainable oil dispersants in the. Hydrocarbondegrading bacteria enriched by the deepwater horizon oil spill identified by cultivation and dnasip. Pdf hydrocarbon degrading potential of bacteria isolated from oil.
Study on the biodegradation of crude oil by free and. Microbial degradation of petroleum hydrocarbons sciencedirect. Pdf petroleum hydrocarbondegrading bacteria for the. Mexico that stimulated growth of psychrophilic, hydrocarbon degrading bacteria. Frontiers petroleum hydrocarbondegrading bacteria for. These isolates were purified from soil samples on the basis of their colony morphology, texture, growth, temperature and hydrocarbon utilization ability. Isolation and characterization of hydrocarbon degrading.
Sufficient time is needed to synthesize hydrocarbon degrading enzymes because of the requirements of. We tracked succession of plume bacteria before, during and after the 83day spill to determine the microbial response and biodegradation potential throughout the incident. The stoichiometric ratio of oxygen per hydrocarbon is 3 m o 2 per 1 mole of hydrocarbons. Pdf three dieseldegrading bacterial strains, alcaligenes piechaudii ccesb2, gordonia hirsuta ccgh1 and rhodococcus ruber ccrr1. Interestingly the total numbers were higher than have been reported for more temperate locations such as prince william sound. Isolation, identification and characterization of oil. Microorganisms able to utilize hydrocarbons anaerobically usually degrade only a very restricted range of them. Indeed, the proportion of hydrocarbondegrading bacteria at 10,400 m is the highest observed in any natural environment on earth. All strains were cultivated in liquid media with crude oil as a sole carbon and energy source. Petroleum hydrocarbon degrading bacteria are ubiquitous in nature and can utilize these compounds as sources of carbon and energy. Five ml of the reaction solution was added to 1 ml of the diluted sample of the culture filtrate.
Isolation and identification of hydrocarbon degrading. In the same sand samples for which detailed chemical analysis was performed, mpn counts of cultivatable hydrocarbon degrading bacteria in oilcontaminated sands 2. Petroleum products are generally biodegradable regardless of their molecular weight, as long as indigenous microorganisms have an adequate supply of oxygen and nutrients. Source of isolation the source for bacterial isolation consisted of wastewaters contaminated with. It is evident that many oil degrading microbes exist in the shallow waters of the caspian sea, but the structure of the community has not been investigated. Sep 14, 2016 created using powtoon free sign up at youtube create animated videos and animated presentations for free. Dominant bacteria shifted substantially over time and were dependent on relative quantities of. Baker1 1 the deepwater horizon blowout in the gulf of mexico in 2010, 2 one of the largest marine oil spills1, changed bacterial commu. Jan 03, 2017 oil degrading bacterial community composition of few studies. Hydrocarbondegrading microorganisms, adapted to grow and thrive in these environments, play an important role in the biological treatment of polluted extreme habitats. Isolation and characterization of bacteria degrading sumandak.
Isolation and characterization of crude oil degrading. Crude oil and petroleum is a complex mixture of hydrocarbons, which cause widespread environmental contaminants following a spill. Hydrocarbon oxidizing bacteria have been isolated from a variety of terrestrial and aquatic environments, using both enrichment and direct plating techniques. Forty nine oil degrading bacteria were isolated and among them four bacterial isolates alk14, alk16, alk23 and alk35 have shown maximum potential to degrade petroleum hydrocarbon and identified as alcaligenes species, bacillus species, enterobacter species and corynebacterium species respectively. Hydrocarbondegrading bacteria and the bacterial community. Bayoumi and abulhamd 2010, have reported isolation of one hundred and nine 109 pure bacterial cultures in mineral salt medium msm supplemented with toluene and phenol. Screening and optimization of biosurfactant production by the. Hydrocarbons are degraded primarily by bacteria and fungi. The wide spread use of petroleum products leads to contamination of soil and aquatic environments, thereby poses a serious threat to all life forms counting humans. Characterization of hydrocarbondegrading bacteria isolated from oilcontaminated sediments in the sultanate of oman and evaluation of bioaugmentation and biostimulation approaches in microcosm experiments. Pdf proliferation of hydrocarbondegrading microbes at the.
Washing the inoculum did not improve recovery of petroleum degraders. Similarly, in preliminary study 57 oil degrading bacteria were isolated from oil impregnated soil in search of potent hydrocarbon degrader 14. Petroleum hydrocarbondegrading bacteria are ubiquitous in nature and can utilize these compounds as sources of carbon and energy. Although bacteria able to grow on aliphatic and aromatic hydrocarbons are found in many genera, the genera alcanivorax appear to be special because these bacteria are specialized for. Enrichment and isolation of petroleum hydrocarbon degrading bacteria petroleum hydrocarbon degrading bacteria were isolated from oil contaminated soil as described above. Baker1 1 the deepwater horizon blowout in the gulf of mexico in 2010, 2 one of the largest marine oil spills1, changed bacterial commu 3 nities in the water column and sediment as they. Microbial degradation of petroleum hydrocarbon contaminants.
In the present study, altogether 32 morphologically distinct bacteria were isolated from crude oil contaminated soil and 18 different hydrocarbon degrading bacteria were number hydrocarbon degrading bacterial strain from and. Media used in this study were mineral salts medium msm, which contained per liter. After the deepwater horizon oil spill, there has been a push to discover the species of hydrocarbon degrading bacteria naturally present in an area and their precise role in bioremediation in a hydrocarbon degrading bacterial consortium. A realtime polymerase chain introduction reaction method. Role of microbes in oil spills remediation and degradation of. Isolation and characterization of hydrocarbon degrading bacteria from environmental habitats in western new york state, masters thesis in chemistry, 2005. Pdf with the sharp increase in population and modernization of society, environmental pollution resulting from petroleum hydrocarbons has.
Hydrocarbondegrading bacteria use oxygen to metabolize organic material to yield carbon dioxide and water, a process commonly referred to as aerobic respiration. Isolation, screening and activity of hydrocarbondegrading bacteria. The deepwater horizon blowout in april 2010 represented the largest accidental marine oil spill and the largest release of chemical dispersants into the environment to date. Native halotolerant, hydrocarbon degrading bacteria actively degrade hydrocarbons at 5% and 0% salt concentrations. Biosurfactant production and concomitant hydrocarbon. Briefly, 1 ml of soil mixture was inoculated into 50 ml of msm containing 1% vv crude oil. Data point microbial community response to an oil spill educator materials microbe consumesbreaks down oceanospirillales, pseudomonas linearchain hydrocarbons 6 or more carbons colwellia, cycloclasticus, and pseudoalteromonas cyclic hydrocarbons harder to break down than linear bonds. Toourknowledge, this is the first reported molecular method that targets anaerobic, hydrocarbondegrading bacteria based on a catabolic gene. After the deepwater horizon oil spill, there has been a push to discover the species of hydrocarbondegrading bacteria naturally present in an area and their precise role in bioremediation in a hydrocarbondegrading bacterial consortium. Created using powtoon free sign up at youtube create animated videos and animated presentations for free. Petroleum hydrocarbondegrading microbial communities in. Isolation and identification of effective hydrocarbon degrading bacteria from diesel polluted soil doi. These two cities are representatives of polluted environments, either by the effect of city traffic seville or nearby industries mechelen.
Various researchers have isolated oilhydrocarbon degrading organisms from oily sludge joseph and joseph 2009. However, cultures have not been preserved, or attempts to reproduce hydrocarbon oxidation by desulfovibrio strains failed 1, 2. The realtime pcr method for bssa could be a powerful tool for monitored natural attenuation ofbtexinfuelcontaminatedgroundwater. Adaptation by prior exposure of microbial communities to hydrocarbons increases hydrocarbon degradation rates. Mineral clay particles present in abundance in marine environments adsorb at oilwater interfaces forming stable emulsion droplets. How to evaluate alternative cleanup technologies for. Biodegradation and bioremediation of hydrocarbons in.
The level of petroleum hydrocarbon degradation was determined by gravimetric assay at each 5 days interval. In our survey we found high total numbers of microorganisms about 1010 cells g dry wt1 sediment. The hydrocarbo n degrading bacterial counts for the. Biofilm formation by hydrocarbon degrading marine bacteria and its effects on oil dispersion. Protein expression in response to oil in a marine hydrocarbon. The media found to be best for enumerating petroleum degrading microorganisms contained 0. Oxygen is considered by many to be the primary growthlimiting factor for hydrocarbon degrading bacteria, but it is normally depleted in zones that have been contaminated with hydrocarbons. While dispersant application may provide numerous benefits to oil spill response efforts, the impacts of dispersants and potential synergistic effects with crude oil on individual hydrocarbon degrading bacteria are poorly. Different hydrocarbon degrading microorganisms such as bacteria and archaea have been found in hydrocarbons contaminated environments.
Reconstructing metabolic pathways of hydrocarbondegrading. Succession of hydrocarbondegrading bacteria in the. Screening and optimization of biosurfactant production by. The presence of such bacteria was evidenced by the conversion to 14 c02 of freshlyadded 9. Raeid m m abed, jamal alsabahi, fatema almaqrashi, amal al. Source of isolation the source for bacterial isolation consisted of wastewaters contaminated with used motor oil, diesel and lubricating oils.
Bacteria that are capable of oil degradation play a large role in bioremediation of hydrocarbon based pollutants in their environment. All chemicals, solvents and reagents used in the present study were of analytical grade. Jun 27, 2018 gutierrez t, singleton dr, berry d, yang t, aitken md, teske a. Hydrocarbondegrading bacteria enriched by the deepwater horizon oil spill identified by. While dispersant application may provide numerous benefits to oil spill response efforts, the impacts of dispersants and potential synergistic effects with crude oil on individual hydrocarbondegrading bacteria are poorly. Characterization, phylogenetic distribution and evolutionary. Biodegradation of petroleum hydrocarbons in cold environments, including alpine soils, is a result of indigenous coldadapted microorganisms able to degrade. In addition to making use of inherent catabolic ability of. The media found to be best for enumerating petroleumdegrading microorganisms contained 0. Frontiers petroleum hydrocarbondegrading bacteria for the.