Multi-factors on biodegradation kinetics of polycyclic aromatic hydrocarbons (PAHs) by Sphingomonas sp. a bacterial strain isolated from mangrove sediment

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) in contaminated sediment is an attractive remediation technique and its success depends on biodegradation kinetics, and the optimal condition for the PAH-degrading isolates; however, information on this aspect is still scarce. The effects of multi-factors on biodegradation of phenanthrene, a 3-ring model PAH, in contaminated sediment slurry by Sphingomonas sp. a bacterial strain isolated from surface mangrove sediment, were investigated using the orthogonal experimental design (form L₁₆(4⁵)). The most significant factors were salinity and inoculum size, while the effects of phenanthrene concentrations, nutrient addition and temperatures were insignificant. The optimal biodegradation condition in contaminated mangrove sediment slurry was 30 °C, 15 ppt salinity, a carbon/nitrogen ratio of 100:1 (the background ratio in sediment) and an inoculum size of 10⁶ most probable number g⁻¹ sediment. The phenanthrene biodegradation could be best described by the first order rate model, C = C₀e^−kt, where k (the rate constant) is equaled to 0.1185, under the optimal condition. The kinetic model was verified and its validity in predicting biodegradation by Sphingomonas sp. at various phenanthrene concentrations was proved by experimental data.     

Preliminary study on biodegradation of phenanthrene by bacteria isolated from mangrove sediments in Hong Kong

Elevated concentrations of polycyclic aromatic hydrocarbons (PAHs) have been found in mangrove sediments due to anthropogenic pollution, and microbial degradation has been suggested as the best way to remove PAHs from contaminated sediments. The degradation of phenanthrene, a model PAH compound by bacteria, either the enriched mixed culture or individual isolate isolated from surface mangrove sediments was examined. The effects of salinity, initial phenanthrene concentrations and the addition of glucose on biodegradation potential were also investigated. Results show that surface sediments collected from four mangrove swamps in Hong Kong had different degree of PAH contamination and had different indigenous phenanthrene-degrading bacterial consortia. The enriched bacteria could use phenanthrene as the sole carbon source for growth and degrade this PAH compound accordingly. A significant positive relationship was found between bacterial growth and percentages of phenanthrene degradation. The phenanthrene biodegradation ability of the enriched mixed bacterial culture was not related to the degree of PAH contamination in surface sediments. The growth and biodegradation percentages of the enriched mixed culture were not higher than that of the individual isolate especially at low salinity (0 and 10 ppt). High salinity (35 ppt) inhibited growth and biodegradation of phenanthrene of a bacterial isolate but less inhibitory effect was found on the mixed culture. The inhibitory effects of salinity could be reduced with the addition of glucose.     

Bioremediation of Phenanthrene by Mycoplana sp. MVMB2 Isolated from Contaminated Soil

The effect of nutrient and surfactant addition on the biodegradation of phenanthrene was studied in a batch scale soil–slurry system using isolated Mycoplana sp. MVMB2strain. The study was conducted using an artificially phenanthrene spiked and as well as contaminated soil from petrochemical industrial site. Maximum phenanthrene degradation and subsequent high microbial growth were observed at optimum pH (pH 6) and C/N/P ratio (100:20:3). To investigate maximum substrate degradation potential of Mycoplana sp. MVMB2, very high concentrations of phenanthrene (50–200 mg/kg soil) were used. The organism was capable of degrading >60% for a concentration below 20 mg/kg soil and >40% for concentrations up to 200 mg/kg within 8 days. Further the influence of five different surfactants namely Span 80, Tween 20, Triton X‐100, cetyl trimethyl ammonium bromide, and sodium dodecyl sulfate were tested at their critical micelle concentration (CMC) levels for phenanthrene degradation in the soil. The addition of surfactant enhanced the biodegradation and a maximum of 84.49% was obtained for Triton X‐100. Complete phenanthrene degradation by Mycoplana sp. MVMB2 was observed at 3 CMC concentration of Triton X‐100. The optimized parameters obtained were used for the degradation of phenanthrene present in the contaminated soil and 98.6% biodegradation was obtained. Thus, the results obtained in the study suggested that biodegradation of phenanthrene by Mycoplana sp. MVMB2 appeared to be feasible to remediate phenanthrene rich contaminated sites.     

Fate of polycyclic aromatic hydrocarbon (PAH) contamination in a mangrove swamp in Hong Kong following an oil spill

The fate of polycyclic aromatic hydrocarbon (PAH) contamination in a mangrove swamp (Yi O) in Hong Kong after an oil spill accident was investigated. The concentrations and profiles of PAHs in surface sediments collected from five quadrats (each of 10 m×10 m) covering different degrees of oil contamination and the most contaminated mangrove leaves were examined in December 2000 (30 days after the accident) and March 2001 (126 days later). The concentrations of total PAHs in surface sediments ranged from 138 to 2135 ng g⁻¹, and PAHs concentrations decreased with time. In the most contaminated sediments, total PAHs dropped from 2135 (30 days) to 1196 ng g⁻¹ (120 days), and the decrease was smaller in less contaminated sediments. The percentage reduction in sediment PAHs over three months (44%) was less significant than that in contaminated leaves (85%), indicating PAH in or on leaves disappeared more rapidly. The PAH profiles were very similar in sediments collected from quadrats Q1 and Q2 with benzo[a]anthracene and pyrene being the most abundant PAH compounds, but were different in the other three quadrats. The proportion of the light molecular weight PAHs to total PAHs increased after three months, especially phenanthrene. Results suggest that physical and photo-chemical weathering (tidal washing and photo-oxidation) of crude oil in surface sediments and on plant leaves were important processes in the first few months after the oil spill. The PAH contamination in Yi O swamp came from both petrogenic and pyrolytic sources. The petrogenic characteristic in the most contaminated sediment was confirmed with high values of phenanthrene to anthracene ratio (>10) and low values of fluoranthene to pyrene ratio (0.3–0.4).     

Optimization of Minimal Salt Medium for Efficient Phenanthrene Biodegradation by Mycoplana sp. MVMB2 Isolated from Petroleum Contaminated Soil Using Factorial Design Experiments

This study presents the degradation of phenanthrene by Mycoplana sp. MVMB2 isolated from petroleum contaminated soil and the media optimization by factorial design experiments. The Plackett–Burman design was used to evaluate the effects of eight variables (potassium dihydrogen phosphate, disodium hydrogen phosphate, magnesium sulfate, calcium chloride, ferrous sulfate, glucose, inoculum concentration, and phenanthrene concentration) on phenanthrene degradation. Based on the results, the critical medium components having significant influence on the degradation were found to be disodium hydrogen phosphate, magnesium sulfate, ferrous sulfate, and phenanthrene. Furthermore, these four variables were used as central composite design parameters. The optimum minimal salt medium composition obtained by conventional and factorial design experiments for the degradation of phenanthrene by Mycoplana sp. MVMB2 at pH 6.5 and 30°C were found to be, potassium 2.5 g/L dihydrogen phosphate, 0.3505 g/L disodium hydrogen phosphate, 0.5501 g/L magnesium sulfate, 0.02 g/L calcium chloride, 0.0261 g/L ferrous sulfate, 0.6756 g/L phenanthrene, 0.5 g/L glucose, 0.5 g/L ammonium sulfate, and inoculum 5% v/v. The phenanthrene degradation was confirmed by analyzing the metabolites formed.     

Effectiveness of bacterial inoculum and mangrove plants on remediation of sediment contaminated with polycyclic aromatic hydrocarbons

The remediation of mangrove sediment contaminated with mixed polycyclic aromatic hydrocarbons (PAHs) having 3-, 4- and 5-rings by natural attenuation, bioaugmentation, phytoremediation and its combination was compared by greenhouse microcosm studies. At Days 90 and 154, the decreases of PAHs in contaminated mangrove sediment by phytoremediation, planted with one-year old Aegiceras corniculatum, and bioaugmentation, the inoculation of PAH-degrading bacterial strains isolated from mangrove sediment, either SCSH (Mycobacterium parafortuitum) or SAFY (Sphingobium yanoikuyae), were not better than that by natural attenuation (the non-vegetated and un-inoculated microcosms). The populations of SCSH and SAFY in sediment could not be maintained even with repeated inoculation, suggesting that the two isolates were not able to compete with the indigenous microbes and had little enhancement effect. Although some PAHs were accumulated in roots, root uptake only accounted for <15% of the spiked PAHs and the effect of plants on remediation were also insignificant. At the end of the 154-day experiment, the mass balance calculation revealed that the overall losses of PAHs by phytoremediation were comparable to that by bioaugmentation but were lower than that by natural attenuation, especially for the high molecular weight PAHs. Under natural attenuation, around 90% fluorene, 80% phenanthrene, 70% fluoranthene, 68% pyrene and 32% benzo[a]pyrene in contaminated sediment were removed. These results demonstrated that the mangrove sediment itself had sufficient indigenous microorganisms capable of naturally remedying PAH contamination.     

Natural attenuation, biostimulation and bioaugmentation on biodegradation of polycyclic aromatic hydrocarbons (PAHs) in mangrove sediments

The biodegradability of a mixture of PAHs, namely fluorene (Fl), phenanthrene (Phe) and pyrene (Pyr), in mangrove sediment slurry was investigated. At the end of week 4, natural attenuation based on the presence of autochthonous microorganisms degraded more than 99% Fl and Phe but only around 30% of Pyr were degraded. Biostimulation with addition of mineral salt medium degraded over 97% of all three PAHs, showing that nutrient amendment could enhance Pyr degradation. Bioaugmentation with inoculation of a PAH-degrading bacterial consortium enriched from mangrove sediments did not show any promotion effect and the degradation percentages of three PAHs were similar to that by natural attenuation. Some inhibitory effect was observed in bioaugmentation treatment in week 1 with only 50% Fl and 70% Phe degraded. These results indicate that autochthonous microbes may interact and even compete with the enriched consortium during PAH biodegradation. Natural attenuation appeared to be the most appropriate way to remedy Fl- and Phe-contaminated mangrove sediments while biostimulation was more capable to degrade Pyr-contaminated sediments. The study also shows that although a large portion of the added PAHs (more than 95%) was adsorbed onto the sediments at the beginning of the experiment, most PAHs were degraded in 4 weeks, suggesting that the degraders could utilize the adsorbed PAHs efficiently.     

Effects of Salinity and Mangrove Detritus on Desorption of Metals from Brackish Water Desorption of Metals from Brackish Water and Shrimp

Desorption and bioaccumulation of Cd, Zn, and Pb were studied using naturally contaminated sediment from a brackish water pond in the Sunderban Biosphere Reserve in India. Pattern of desorption of the metals from the sediment and bioaccumulation in fingerlings of the teleost Oreochromis mossambicusand postlarvae of the shrimp Penaeus monodon were studied as a function of salinity and loading of detritus of a mangrove plant. Effects of both salinity and loading of detritus on bioaccumulation of the metals were studied under two conditions: either the animals were allowed free access to the sediment or access was denied. Ninety‐six hour experiments showed that desorption of Cd and Pb from sediment into water increased with salinity of the medium while desorption of Zn decreased. Salinity of the medium also had a significant effect on the bioaccumulation of metals by fish; Cd and Pb accumulation decreased in saline medium while the accumulation of Zn increased. Conditions of access to sediment had no effect on the bioaccumulation of metals by fish; effect of interaction between salinity and access condition was also insignificant. The access conditions, however, significantly influenced accumulation of metal by the shrimp postlarvae. The effect of interaction between salinity and access condition was insignificant in influencing the bioaccumulation of all metals except Zn. The accumulation of Zn increased as a function of the salinity of the medium when shrimp postlarvae were allowed access to the sediment. Desorption of metals from sediment to water were below detection limits when detritus of a mangrove plant was added to the medium. Both the level of detritus and the conditions of access influenced accumulation of metals by fish, but the effect of interaction between the two factors were found to be insignificant. Shrimp postlarvae showed net accumulation only of Pb in the presence of detritus and the accumulation of Pb increased when the larvae were separated from the sediment. The results are important in understanding the mobility of metals between solid and aqueous phases in brackish water environments that experience periodic fluctuations in salinity and fluxes of organic load in the form of mangrove detritus.     

Concentrations of PCBs in coastal mangrove sediments of Hong Kong

The concentrations of polychlorinated biphenyls (PCBs) in surface sediments of ten mangrove sites in Hong Kong were determined by Congener-specific and Arcolor methods. Spatial variations were found between mangrove sites and within the same site, indicating that the sediment samples were heterogeneous and total PCBs concentrations in mangrove sediments should be normalised to compensate for natural variability and for identifying serious anthropogenic contamination. In addition to total organic carbon, Al, Fe and Mn were possible normalisers. Based on PCB/normaliser ratio and 95% prediction limits of scatter plots between total PCB and normaliser, all mangrove sites were not seriously contaminated by human activities. The estimated total PCBs concentrations (quantified by Congener-specific method) for most sediment ranged from 0.5 to 5.8 ng g(-1) (dry weight, n = 92). However, very high concentrations of PCB were found in sub-samples from Yi O, Tolo Pond and Lai Chi Wo, suggesting that these samples represented the PCB contaminated "hot spots".     

Effects of varying estuarine conditions on the sorption of phenanthrene to sediment particles of Yangtze Estuary

The sorption of phenanthrene on the Yangtze Estuary sediment was studied under varying conditions of particle size, sediment organic contents, salinity, and dissolved organic matter (DOM) concentrations. Small sediment particles showed higher trapping capacity for phenanthrene due to the higher organic contents associated. The organic carbon-based partition coefficient of phenanthrene to the Yangtze Estuary sediment was obtained as 7120 L/kg, lower than the values for other soils or sediments reported in previous studies. The magnitude and direction of the salt effect were complicated by the specific DOM studied. The sediment sorption capacity was greatly increased in saline water in the absence of DOM but decreased in the presence of DOM. Given the conditions in the Yangtze Estuary, the equilibrium sorption of phenanthrene would be decreased with increasing salinity. Overall, the nature and content of both sediment-bound and dissolved organic matter dominate the sorption of hydrophobic organic contaminants in the estuary.     

Accumulation and biodegradation of phenanthrene and fluoranthene by the algae enriched from a mangrove aquatic ecosystem

This study focused on the accumulation and biodegradation of two typical polycyclic aromatic hydrocarbons (PAHs), phenanthrene (PHE) and fluoranthene (FLA), by the diatoms enriched from a mangrove aquatic ecosystem in the Jiulong River estuary, China. After separation, purification and culture, Skeletonema costatum (Greville) Cleve and Nitzschia sp. were exposed to different concentrations of PHE, FLA, and a mixture of the two. The results showed that the tolerance of S.costatum to PHE and FLA was greater than that of Nitzschia sp., and that the toxic effect of FLA on S. costatum and Nitzschia sp. was higher than that of PHE. The microalgal species S. costatum and Nitzschia sp. were capable of accumulating and degrading the two typical PAHs simultaneously. The accumulation and degradation abilities of Nitzschia sp. were higher than those of S. costatum. Degradation of FLA by the two algal species was slower, indicating that FLA was a more recalcitrant PAH compound. The microalgal species also showed comparable or higher efficiency in the removal of the PHE-FLA mixture than PHE or FLA singly, suggesting that the presence of one PAH stimulated the degradation of the other.     

The effects of biodegradation on the compositions of aromatic hydrocarbons and maturity indicators in biodegraded oils from Liaohe Basin

By the aid of GC-MS technique, a series of sequentially biodegraded oils from Liaohe Basin have been analyzed. The results show that the concentrations and relative compositions of various aromatic compounds in the biodegraded crude oils will change with increasing biodegradation degree. The concentrations of alkyl naphthalenes, alkyl phenanthrenes, alkyl dibenzothiophene are decreased, and the concentration of triaromatic steroids will increase with increasing biodegradation degree in biodegraded oils. Those phenomena indicate that various aromatic compounds are more easily biodegraded by bacteria like other kinds of hydrocarbons such as alkanes, but different series of aromatic compounds have a varied ability to resistant to biodegradation. The ratios of dibenzothiophene to phenenthrene (DBTH/P) and methyl dibenzothiophene to methyl phenanthrene (MDBTH/MP) are related to the features of depositional environment for source rocks such as redox and ancient salinity. However, in biodegraded oils, the two ratios increase quickly with the increase of the biodegradation degree, indicating that they have lost their geochemical significance. In this case, they could not be used to evaluate the features of depositional environment. Methyl phenanthrene index, methyl phenanthrene ratio and methyl dibenzoyhiophene ratio are useful aromatic maturity indicators for the crude oils and the source rocks without vitrinite. But for biodegraded oils, those aromatic maturity indicators will be affected by biodegradation and decrease with the increase of the biodegradation degree. Therefore, those aromatic molecular maturity indicators could not be used for biodegraded oils.     

PAHs contamination and bacterial communities in mangrove surface sediments of the Jiulong River Estuary, China

Sixteen sediment samples collected from eight transects in a mangrove swamp of the Jiulong River Estuary, Fujian, China were investigated for their content of polycyclic aromatic hydrocarbons (PAHs) and the biodegradation potential of the indigenous microorganisms. The bacterial community structures in the mangrove sediments and in enrichment cultures were also investigated. The results showed that the total PAHs concentration of mangrove sediments ranged from 280 to 1074 ng g(-1) dry weight, that the PAHs composition pattern in the mangrove sediments was dominated by high molecular weight PAH components (4-6 rings), and that Benzo[ghi]perylene and Indeno[1,2,3-cd]pyrene were the most dominant at different stations. Abundant PAH-degrading bacteria were found in all the stations, the values of phenanthrene-degrading bacteria ranged from 5.85 x 10(4) to 7.80 x 10(5) CFU g(-1) dry weight, fluoranthene-degrading bacteria ranged from 5.25 x 10(4) to 5.79 x 10(5) CFU g(-1) dry weight, pyrene-degrading bacteria ranged from 3.10 x 10(4) to 6.97 x 10(5) CFU g(-1) dry weight and the benzo(a)pyrene-degrading bacteria ranged from 5.25 x 10(4) to 7.26 x 10(5) CFU g(-1) dry weight. DGGE analysis of PCR-amplified 16S rDNA gene fragments confirmed that there was a remarkable shift in the composition of the bacterial community due to the addition of the different model PAH compound phenanthrene (three ring PAH), fluoranthene(four ring PAH), pyrene(four ring PAH) and benzo(a)pyrene(five ring PAH) during enrichment batch culture. Eleven strains were obtained with different morphology and different degradation ability. The presence of common bands for microbial species in the cultures and in the native mangrove sediment DNA indicated that these strains could be potential in situ PAH-degraders.     

In situ experimental study of the evolution and recolonization of polluted sediments

When perturbing conditions influencing a benthic ecosystem improve, an extremely polluted sediment may very rapidly be colonized by a structured assemblage. This study was made after the implantation of modules containing chemically very contaminated sediment from a polluted biotope, in a lesser contaminated site. The colonization and dynamic processes of the studied mollusc populations do not depend on the concentrations of the solid phase pollutants but are related to the release of these pollutants in the interstitial water of sediments, which in turn is induced by ambiant condition variations and particularly by salinity.     

Depth-related influences on biodegradation rates of phenanthrene in polluted marine sediments of Puget Sound, WA

A whole-core injection method was used to determine depth-related rates of microbial mineralization of (14)C-phenanthrene added to both contaminated and clean marine sediments of Puget Sound, WA. For 26-day incubations under micro-aerobic conditions, conversions of (14)C-phenanthrene to (14)CO(2) in heavily PAH-contaminated sediments from two sites in Eagle Harbor were much higher (up to 30%) than those in clean sediments from nearby Blakely Harbor (<3%). The averaged (14)C-phenanthrene degradation rates in the surface sediment horizons (0-3 cm) were more rapid (2-3 times) than in the deeper sediment horizons examined (>6 cm), especially in the most PAH polluted EH9 site. Differences in mineralization were associated with properties of the sediments as a function of sediment depth, including grain-size distribution, PAH concentration, total organic matter and total bacterial abundance. When strictly anaerobic incubations (in N(2)/H(2)/CO(2) atmosphere) were used, the phenanthrene biodegradation rates at all sediment depths were two times slower than under micro-aerobic conditions, with methanogenesis observed after 24 days. The main rate-limiting factor for phenanthrene degradation under anaerobic conditions appeared to be the availability of suitable electron acceptors. Addition of calcium sulfate enhanced the first order rate coefficient (k(1) increased from 0.003 to 0.006 day(-1)), whereas addition of soluble nitrate, even at very low concentration (<0.5 mM), inhibited mineralization. Long-term storage of heavily polluted Eagle Harbor sediment as intact cores under micro-aerobic conditions also appeared to enhance anaerobic biodegradation rates (k(1) up to 0.11 day(-1)).     

Laboratory analyses of nutrient release processes from Haihe River sediment

Sediment samples were collected from the heavily contaminated Haihe River to conduct static nutrient release experiments with tap water,and to evaluate the effect of dredging depths,salinity and light ...     

Bioremediation perspective of historically contaminated sediment with polycyclic aromatic hydrocarbons

The purpose of the current study is to compare the influence of different aerobic conditions(biostimulation(BS),bioaugmentation(BA),and a combination of biostimulation and bioaugmentation(BB))on polycyclic aromatic hydrocarbons(PAH)degradation and compare the degraded amount with single step XAD-4 extraction as a new tool for bioavailability assessment for chronically contaminated sediment samples obtained from territory of Autonomous Province Vojvodina of Serbia(S1,S2,and S3).A great number of papers dealing with biodegradation of PAHs in spiked sediment or soil have been published,but to the authors’knowledge,a limited number of papers studied aged,historically polluted sediment and a sum of chosen U.S.Environmental Protection Agency(USEPA)PAHs.A significant reduction(up to67%)in PAH concentration was observed,while the percentage of reduction varied depending on the sediment sample and treatment used.BS treatment successfully stimulated growth of indigenous bacteria.Further,PAH-degrading strain Sphingomonas paucimobilis F8 inoculated in BA and BB treatment survived for up to 7 weeks after it was suppressed by unfavorable conditions or native microbes.Degraded amounts generally showed good correlation with results obtained from XAD-4 extraction.Results obtained in the current study represent a good start for standardizing a XAD-4 extraction technique as a simplified,easier,and lower cost method for bioavailability assessment.     

Intrinsic bioremediation of a petroleum-impacted wetland

Following the 1994 San Jacinto River flood and oil spill in southeast Texas, a petroleum-contaminated wetland was reserved for a long-term research program to evaluate bioremediation as a viable spill response tool. The first phase of this program, presented in this paper, evaluated the intrinsic biodegradation of petroleum in the contaminated wetland. Sediment samples from six test plots were collected 11 times over an 11-month period to assess the temporal and spatial petroleum concentrations. Petroleum concentrations were evaluated using gas chromatography-mass spectrometer analyses of specific target compounds normalized to the conservative biological marker, C(30)17alpha,21beta(H)-hopane. The analyses of specific target compounds were able to characterize that significant petroleum biodegradation had occurred at the site over the one-year period. Total resolved saturate and total resolved aromatic hydrocarbon data indicated the petroleum was degraded more than 95%. In addition, first-order biodegradation rate constants were calculated for the hopane-normalized target compounds and supported expected biodegradation patterns. The rapid degradation rates of the petroleum hydrocarbons are attributed to conditions favorable to biodegradation. Elevated nutrient levels from the flood deposition and the unconsolidated nature of the freshly deposited sediment possibly provided a nutrient rich, oxic environment. Additionally, it is suggested that an active and capable microbial community was present due to prior exposure to petroleum. These factors provided an environment conducive for the rapid bioremediation of the petroleum in the contaminated wetland.     

Effects of environmental stresses on the responses of mangrove plants to spent lubricating oil

The influence of different environmental stresses, including salinity (5-35‰), tidal cycle (6/6, 12/12 and 24/24 h of high/low tidal regimes) and nutrient addition (1-6 times background nitrogen and phosphorus content) on Bruguiera gymnorrhiza and Aegiceras corniculatum grown in sediment contaminated with spent lubricating oil (7.5 L m(-2)) were investigated. The oil-treated 1-year-old mangrove seedlings subject to low (5‰) and high (35‰) salinity had significantly more reduction in growth, more release of superoxide radical (O2·-) and higher activity of superoxide dismutase (SOD) than those subject to moderate salinity (15‰). Extended flooding (24/24 h of high/low tidal regime) enhanced O2·- release and malondialdehyde (MDA) content in both oil-treated species but had little negative effects on biomass production (P>0.05) except the stem of A. corniculatum (P=0.012). The addition of nutrients had no beneficial or even posed harmful effects on the growth and cellular responses of the oil-treated seedlings.     

Immobilization of Mycoplana sp. MVMB2 Isolated from Petroleum Contaminated Soil onto Papaya Stem (Carica papaya L.) and Its Application on Degradation of Phenanthrene

This study presents the degradation of phenanthrene using immobilized Mycoplana sp. MVMB2 isolated from contaminated soil. Papaya stem pretreated by two stage processes, treating with acid or alkali and drying, was used for the immobilization of Mycoplana sp. Alkali pretreated papaya stem was found to be most effective in cell uptake compared to acid treated one. The maximum immobilization capacity at various physiochemical conditions for the alkali pretreated papaya stem was found to be at 320 min time, pH 6.5, 30°C temperature, and 18.6 × 10⁶ cells/mL initial concentrations. The adsorption mechanism of Mycoplana sp. MVMB2 on pretreated papaya stem was assessed using various kinetic and isotherm models. The immobilization of Mycoplana sp. MVMB2 on to pretreated papaya stem was corroborated by scanning electron microscopy and Fourier transformed IR spectroscopy analysis. The performance of immobilized cells in batch reactor showed more than 95% phenanthrene degradation within 72 h, whereas, free cells were found to require 120 h. The immobilized cells also showed better degradation performance in the packed column study.