Kinetic and molecular analyses reveal isoprene degradation potential of Methylobacterium sp.

Efforts were made to isolate and characterize bacteria capable of growing on methane and organic compounds, and to achieve the simultaneous degradation of more than one pollutant. Among the methanotrophs, species of Methylobacterium was able to catabolize a variety of hydrocarbons, including the branched-chain alkenes. Therefore, laboratory incubations experiments were carried out in batch mode to assess the potential of Methylobacterium sp. PV1 for degrading isoprene, the low-molecular-weight alkene, the most abundant non-methane volatile hydrocarbon present in the environment. Methylobacterium sp. PV1, isolated from paddy field soil, was characterized by pmoA and 16S rRNA gene sequencing and FAME analysis, and used for isoprene degradation. The kinetics of biodegradation is studied using the Michaelis-Menten model. The optimum degradation (80%) with maximum average relative degradation rate was observed at 150ppm isoprene. The degradation products were also analyzed using FTIR.

Efficacy of wood charcoal and its modified form as packing media for biofiltration of isoprene.

The efficacy of wood charcoal (WC) and nutrient-enriched wood charcoal (NWC) as biofilter packing media were assessed for isoprene biodegradation in a bioreactor comprising bioscrubber and a biofilter connected in series and inoculated with Pseudomonas sp. The bioreactors using WC and NWC exhibited >90% removal efficiency and around 369 g m-3 h-1 elimination capacity at around 404 g m-3 h-1 inlet loading rate. In both the bioreactors, the biofilter component showed better degradation capacity compared to the bioscrubber unit. The kinetic parameters, maximum elimination capacity, ECmax; substrate constant, Ks and ECmax/Ks for Michaelis-Menten model were evaluated. The lower Ks for the WC packed bioreactor indicated that ECmax achieved, was faster compared to others, while higher ECmax and ECmax/Ks for the NWC packed bioreactor suggests its superiority in isoprene abatement in the continuous mode. A comparison of the available published information on biofiltration of isoprene reflected polyurethane foam as the superior packing media.

Degradation kinetics and metabolites in continuous biodegradation of isoprene.

The kinetic parameters of isoprene biodegradation were studied in a bioreactor, comprising of bioscrubber and polyurethane foam packed biofilter in series and inoculated with Pseudomonas sp., using a Michaelis-Menten type model. The maximum elimination capacity, ECmax; substrate constant, Ks and ECmax/Ks values for bioscrubber were found to be 666.7 g m(-3) h(-1), 9.86 g m(-3) and 67.56 h(-1), respectively while those for biofilter were 3333 g m(-3) h(-1), 13.96 g m(-3) and 238.7 h(-1), respectively. The biofilter section exhibited better degradation efficiency compared to the bioscrubber unit. Around 62-75% of the feed isoprene got converted to carbon dioxide, indicating the efficient capability of bacteria to mineralize isoprene. The FTIR and GC-MS analyses of degradation products indicated oxidative cleavage of unsaturated bond of isoprene. These results were used for proposing a plausible degradation pathway for isoprene.

Whole-Genome Sequence of Listeria monocytogenes Strains from Clinical and Environmental Samples from Varanasi, India.

We present here the whole-genome sequences of Listeria monocytogenes from Ganges River water, agricultural soil, and human clinical samples from Varanasi, India, which will be used for a comparative analysis.

Characterization of Listeria monocytogenes isolated from Ganges water, human clinical and milk samples at Varanasi, India.

Listeria monocytogenes isolated from Ganges water, human clinical and milk samples were characterized by antibiotic susceptibility, serotype identification, detection of virulence genes and ERIC- and REP-PCR fingerprint analyses. All isolates were uniformly resistant to ampicillin, except two isolates, and showed variable resistance to gentamicin, cotrimoxazole, ofloxacin, rifampicin and tetracycline. Of the 20 isolates found positive for pathogens, seven (four human and three water isolates) belong to serogroups 4b, 4d and 4e; six (one human and five water isolates) belong to serogroups 1/2c and 3c; four milk isolates belong to serogroups 1/2b and 3b; and three milk isolates belong to serogroups 1/2a and 3a. Two water isolates, all human isolates, except one (Pb1) lacking inlJ gene, and three milk isolates possess inlA, inlC, plcA, prfA, actA, hlyA and iap genes. The remaining water and milk isolates showed variable presence of inlJ, plcA, prfA, and iap genes. ERIC- and REP-PCR based analyses collectively indicated that isolates of human clinical samples belong to identical or similar clone and isolates of water and milk samples belong to different clones. Overall study demonstrates the prevalence of pathogenic L. monocytogenes species in the environmental and clinical samples. Most of the isolates were resistant to commonly used antibiotics.

Degradation kinetics of chlorpyrifos and 3,5,6-trichloro-2-pyridinol (TCP) by fungal communities.

Fungal isolates obtained from soil were used for degrading chlorpyrifos (CP) and TCP. The percentage degradation ranged from 69.4 to 89.8 for CP and 62.2 to 92.6 for TCP after one week. The values of K(s) and V(max) were different for different isolates. The K(s) ranged from 66.66 to 169.5mg/L and V(max) from 6.56 to 40.4 mg/L/d for CP and from 53.19 to 163.9 mg/L and 3.41 to 40.40 mg/L/d, respectively, for TCP. Fungal community showed high affinity for both CP and TCP. The genetic relatedness of isolate F1 to Aspergillus sp., F2 and F3 to Penicillium sp., F4 to Eurotium sp. and F5 to Emericella sp. were confirmed. The degradation potential was in the order: F1>F2=F3>F4>F5.

Community structure of methanogenic archaea and methane production associated with compost-treated tropical rice-field soil.

The diversity and density of methanogenic archaea and methane production were investigated ex situ at different growth stages of rice plant cultivated in compost-treated tropical rice fields. The qPCR analysis revealed variation in methanogens population from 3.40 × 10(6) to 1.11 × 10(7)  copies g(-1)  dws, in the year 2009 and 4.37 × 10(6) to 1.36 × 10(7)  copies g(-1)  dws in the year 2010. Apart from methanogens, a large number of bacterial (9.60 × 10(9) -1.44 × 10(10)  copies g(-1)  dws) and archaeal (7.13 × 10(7) -3.02 × 10(8)  copies g(-1)  dws) communities were also associated with methanogenesis. Methanogen population size varied in the order: flowering > ripening > tillering > postharvest > preplantation stage. The RFLP-based 16S rRNA gene-targeted phylogenetic analysis showed that clones were closely related to diverse group of methanogens comprising members of Methanomicrobiaceae, Methanosarcinaceae, Methanosaetaceae and RC I. Laboratory incubation studies revealed higher amount of cumulative CH(4) at the flowering stage. The integration of methanogenic community structure and CH(4) production potential of soil resulted in a better understanding of the dynamics of CH(4) production in organically treated rice-field soil. The hypothesis that the stages of plant development influence the methanogenic community structure leading to temporal variation in the CH(4) production has been successfully tested.

Substrate inhibition during bio-filtration of TCE using diazotrophic bacterial community.

The kinetics of biodegradation of TCE in the biofilter packed with wood charcoal and inoculated with diazotrophic bacterial community had been investigated. Use of Michaelis-Menten type model showed that substrate inhibition was present in the system. The kinetic model proposed by Edwards (1970) was used to calculate kinetic parameters-maximum elimination capacity (EC(max)), substrate constant (K(s)), and inhibition constant (K(I)). The model fitted well with the experimental data and the EC(max) was found to be in the range of 10.8-6.1 g/m(3) h. The K(s) values depended upon substrate concentration and ranged from 0.024 to 0.043 g/m(3) indicating the high affinity of diazotrophs for TCE. The K(I) values were low and nearly constant (0.011-0.015 g/m(3)) indicating a moderate substrate inhibition.

Kinetics of bio-filtration of trichloroethylene by methanotrophs in presence of methanol.

The biodegradation of TCE was studied in a laboratory scale biofilter packed with wood charcoal and inoculated with mixed culture of methanotrophs isolated from local soil. The removal efficiency was found to be higher than 90% up to an inlet load of 5.1g/m(3)h. The maximum elimination capacity was 6.7g/m(3)h at an inlet loading rate of 11.3g/m(3)h. The reaction constants EC(max,)K(s) and K(i) calculated from the experimental results are also presented. The biodegradation process is found to be inhibited at higher TCE concentration. The carbon dioxide production rate has been found to be a linear function of elimination capacity. The DNA finger printing techniques has indicated the presence of functionally active methanotrophic community including Methylocystis sp. in the biofilter.

Bio-filtration of trichloroethylene using diazotrophic bacterial community.

Biodegradation of TCE was studied in a biofilter packed with wood charcoal and inoculated with diazotrophic bacterial community isolated from local soil. Steady state TCE removal efficiencies higher than 85% were observed up to inlet load of 2.866 g m(-3) h(-1). The maximum elimination capacity of 5.31 g m(-3) h(-1) was observed at an inlet load of more than 7.90 g m(-3) h(-1). The biofilter was sensitive to fluctuations in the process conditions but could easily recover its performance after 10 days shutdown. Almost constant and small pressure drop per unit length and very negligible compaction was observed during the whole experimental period. The molecular analyses such as RT-PCR and gene sequencing revealed the presence of functionally active Azospirillum species in the biofilm.

Diversity of methanotrophs in urea-fertilized tropical rice agroecosystem.

Laboratory experiments were conducted to study the population size, diversity and methane oxidation potential of methanotrophs in tropical rice agroecosystem under the influence of N-fertilizer. Results indicate that the diversity of methane oxidizing bacteria (MOB) is altered in fertilizer treated soils compared to untreated control. Nevertheless, Type I MOB still dominated in the fertilized soils whereas the diversity of Type II methanotrophs decreases. Control soils have higher MOB population and CH(4) oxidation capacity than fertilized soils. Rhizospheric soil is more populated than non-rhizospheric soil in both unfertilized and fertilized conditions. Variation in K(m) and V(max) of methane oxidation in soils appears to be due to variation in methanotrophic community. Experimental results indicate that methanotrophic community differs both quantitatively and qualitatively in unfertilized and fertilized soils.

Biodegradation of trichloroethylene (TCE) by methanotrophic community.

Laboratory incubation experiments were carried out to assess the potential of methanotrophic culture for degrading TCE. Measurements of the growth rate and TCE degradation showed that the methanotrophs not only grew in presence of TCE but also degraded TCE. The rate of TCE degradation was found to be 0.19 ppm h(-1). The reverse transcriptase-PCR test was conducted to quantify expression of pmoA and mmoX genes. RT-PCR revealed expression of pmoA gene only. This observation provides evidence that the pmoA gene was functionally active for pMMO enzyme during the study. The diversity of the methanotrophs involved in TCE degradation was assessed by PCR amplification, cloning, restriction fragment length polymorphism and phylogenetic analysis of pmoA genes. Results suggested the occurrence of nine different phylotypes belonging to Type II methanotrophs in the enriched cultures. Out of the nine, five clustered with, genera Methylocystis and rest got clustered in to a separate group.