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Microbial degradation of mefenoxam in rhizosphere of Zinnia angustifolia.

The fate of the fungicide mefenoxam was studied in a containerized rhizosphere system. The rhizosphere system used Zinnia angustifolia (Tropic Snow) in a bark/sand potting mix and was compared to bulk potting mix (no plants). Rhizosphere microbial populations were allowed to establish for 3 weeks prior to fungicide addition (20 microg per g mix). Mefenoxam and degradation product concentrations were determined by High HPLC or capillary electrophoresis after extraction. Seventy eight percent of the fungicide originally applied to the rhizosphere was degraded after 21 days compared to 44% in bulk system (no plant). The primary degradation product was the free acid N-(2,6-dimethylphenyl)-N-(methoxyacetyl)-DL-alanine, which accounted for 71% of the applied parent chemical after 30 days. N-(2,6-dimethylphenyl)-acetamide was also detected, but in lesser amounts. Bacterial populations in the rhizosphere increased during the 30-day period, which correlated with an increase in degradation of the parent compound. Pure cultures of Pseudomonas fluorescens and Chrysobacterium indologenes isolated from the rhizosphere system could degrade the applied fungicide (10 microg/ml) almost completely to the free acid within 54 h.

Alanine↗

Microbial degradation of natural rubber vulcanizates.

An actinomycete, Nocardia sp. strain 835A, grows well on unvulcanized natural rubber and synthetic isoprene rubber, but not on other types of synthetic rubber. Not only unvulcanized but also various kinds of vulcanized natural rubber products were more or less utilized by the organism as the sole source of carbon and energy. The thin film from a latex glove was rapidly degraded, and the weight loss reached 75% after a 2-week cultivation period. Oligomers with molecular weights from 10 to 10 were accumulated during microbial growth on the latex glove. The partially purified oligomers were examined by infrared and H nuclear magnetic resonance and C nuclear magnetic resonance spectroscopy, and the spectra were those expected of cis-1, 4-polyisoprene with the structure, OHC-CH(2)-[-CH(2)-C(-CH(3))=CH -CH(2)-](n)-CH(2)-C(=O)- CH(3), with average values of n of about 114 and 19 for the two oligomers.

Journal Article↗

Microbial degradation of phenanthrene and pyrene in a two-liquid phase-partitioning bioreactor.

A study was conducted to determine the potential of two-liquid phase-bioreactors for the treatment of (polycyclic aromatic hydrocarbons) PAHs. Phenanthrene and pyrene were supplied two times at a concentration of 100 mg/l of reactor broth, either as crystals or dissolved in silicone oil. Complete phenanthrene biodegradation was achieved within 3 days after each addition to the biphasic-inoculated reactor. Its concentration in the monophasic reactors dropped by 93% within 4 days, but remained incomplete for the duration of the experiment. Pyrene removal occurred to a limited extent only in the presence of phenanthrene. Significant pollutant losses were recorded in the monophasic reactors, most likely caused by volatilization. Pollutant degradation was improved upon repeated phenanthrene amendment to the biphasic system. Biphasic reactors allow the fast and complete degradation of PAHs and prevent their hazardous disappearance. The use of biphasic reactors for the degradation of poorly soluble pollutants should become more beneficial when the substrate-interface uptake mechanism is operating. Thus, biphasic reactors should be integrated into the microbial enrichment procedure.

Bacteria↗

Microbial degradation of livestock-generated ammonia using biofilters at typical ambient temperatures.

The purpose of this research was to neutralize livestock-generated ammonia by using biofilters packed with inexpensive inorganic and organic packing material combined with multicultural microbial load at typical ambient temperatures. Peat and inorganic supporting materials were used as biofiltration matrix packed in a perfusion column through which gas was transfused. Results show the ammonia removal significantly fell in between 99 and 100% when ammonia concentration of 200 ppmv was used at different gas flow rates ranged from 0.030 to 0.060 m3 h(-1) at a fluctuating room temperature of 27.5 +/- 4.5 C (Mean +/- SD). Under these conditions, the emission concentration of ammonia that is liberated after biofiltration is less than 1 ppmv (0.707 mg m(-3)) over the period of our study, suggesting the usage of low-cost biofiltration systems for long-term function is effective at wider ranges of temperature fluctuations. The maximum (100%) ammonia removal efficiency was obtained in this biofilter was having an elimination capacity of 2.217 g m(-3) h(-1). This biofilter had high nitrification efficiencies and hence controlled ammonia levels with the reduced backpressure. The response of this biofilter to shut down and start up operation showed that the biofilm has a superior stability.

Air Pollutants↗

The microbial degradation of azimsulfuron and its effect on the soil bacterial community.

AIMS: Azimsulfuron is a recently introduced sulfonylurea herbicide useful in controlling weeds in paddy fields. To date very little information is available on the biodegradation of this pesticide and on its effect on the soil microbial community. The aim of this work was to study its biodegradation both in slurry soil microcosms and in batch tests with mixed and pure cultures. METHODS AND RESULTS: Azimsulfuron was applied to forest bulk soil in order to study its effect on the structure of the bacterial soil community, as detectable by denaturant gradient gel electrophoresis (DGGE) analyses. Biodegradation and abiotic processes were investigated by HPLC analyses. In addition, a microbial consortium was selected, that was able to use azimsulfuron as the sole energy and carbon source. One of the metabolites produced by the consortium was isolated and identified through LC-MS analyses. Cultivable bacteria of the consortium were isolated and identified by 16S rDNA sequencing (1400 bp). CONCLUSIONS: Azimsulfuron treatment seems to have the ability to cause changes in the bacterial community structure that are detectable by DGGE analyses. It is easily biodegraded both in microcosms and in batch tests, with the formation of an intermediate that was identified as 2-methyl-4-(2-methyl-2H-tetrazol-5-yl)-2H-pyrazole-3-sulfonamide. SIGNIFICANCE AND IMPACT OF THE STUDY: The study increases the knowledge on the biodegradation of azimsulfuron and its effects on the soil microbiota.

Bacteria↗

Microbial degradation of explosives and related compounds.

The pollution of soil and water with explosives and related compounds caused by military activities has been known for a long time, but progress in understanding the environmental fate of such substances has only been made in the last few years. Microbial processes could be used for the remediation of explosives-contaminated soils and waste waters because it has been shown that a variety of different microorganisms are able to metabolize these chemical compounds. In some cases even a complete mineralization has been found, whereas in others only biotransformation reactions took place, producing more or less toxic and/or recalcitrant metabolites. Studies with pure cultures of bacteria and fungi have given detailed insights into the biodegradation pathways of at least some nitroorganic compounds. Additionally, some of the key enzymes have been isolated and purified or studied in crude extracts. This review summarizes information on the biodegradation and biotransformation pathways of several important explosives. This may be useful in developing microbiological methods for a safe and economic clean-up of soil and water contaminated with such compounds. It also shows the necessity of further investigations concerning the microbial metabolism of these substances.

Bacteria↗

Effects of alkylphosphates and nitrous oxide on microbial degradation of polycyclic aromatic hydrocarbons.

We conducted a series of liquid-culture experiments to begin to evaluate the abilities of gaseous sources of nitrogen and phosphorus to support biodegradation of polycyclic aromatic hydrocarbons (PAHs). Nutrients examined included nitrous oxide, as well as triethylphosphate (TEP) and tributylphosphate (TBP). Cultures were established using the indigenous microbial populations from one manufactured gas plant (MGP) site and one crude oil-contaminated drilling field site. Mineralization of phenanthrene was measured under alternative nutrient regimes and was compared to that seen with ammoniacal nitrogen and PO(4). Parallel cultures were used to assess removal of a suite of three- to five-ring PAHs. In summary, the abilities of the different communities to degrade PAH when supplemented with N(2)O, TEP, and TBP were highly variable. For example, in the MGP soil, organic P sources, especially TBP, supported a considerably higher degree of removal of low-molecular-weight PAHs than did PO(4); however, loss of high-molecular-weight compounds was impaired under these conditions. The disappearance of most PAHs was significantly less in the oil field soil when organophosphates were used. These results indicate that the utility of gaseous nutrients for PAH bioremediation in situ may be limited and will very likely have to be assessed on a case-by-case basis.

Bacteria↗

Photochemical and microbial degradation of 2,4,5-trichloroaniline in a freshwater lake.

Surface waters from a eutrophic lake in northern Georgia were incubated with C-labeled 2,4,5-trichloroaniline to study the disappearance of the parent compound and production of CO(2). There was no degradation of the compound in the dark. Under 12 h of sunlight and 12 h of darkness, 28% of the trichloroaniline was degraded in both poisoned and untreated samples. Mineralization after 24 h in poisoned and untreated lake water was 5.5 and 6.8%, respectively. Thus, 81% of the mineralization was attributable to photochemical processes, and 19% was attributable to microbial processes. Most biological mineralization was due to microbes of bacterial size (<1.0 mum). Approximately 90% of the trichloroaniline bioaccumulated was associated with organisms larger than 1.0 mum, e.g., algae. When algae were removed by filtration, the amount of trichloroaniline mineralized increased to 9.4%, compared with 6.8% in the presence of algae. The excretion of organic compounds by algae may have inhibited bacterial mineralization of photoproducts.

Journal Article↗

Carbon substrate utilisation profile of a high concentration effluent degrading microbial consortium.

This paper presents the carbon substrate utilisation profile of a group of microorganisms responsible for the biodegradation of a highly concentrated industrial effluent. A 1 litre bioreactor was used to study this consortium's biodegradation potential, with the chemical oxygen demand (COD) of the waste being reduced by 90% from 24000 mg l(-1) within 456 hours. This study also demonstrates that the consortium is capable of degrading organic solvents, such as isopropanol, at concentrations of 260 mg l(-1). The population distribution and biochemical behaviour were also characterised using denaturing gradient gel electrophoresis (DGGE) and Biolog Eco Plates at various stages of bioreactor operation. The DGGE results indicated that the dominant bands of the microbial population profile were stable at various operational stages, and that only a few bands varied with time. Moreover, four Biolog Eco plates were inoculated with samples drawn from the bioreactor at 0, 24, 72 and 120 hours after inoculation. Based on this Biolog Eco Plate profiling, a carbon source utilisation analysis was conducted to group the substrates according to their colour development patterns. Patterns were quantified via measurement of well optical density. Subsequently, cross-correlation statistical techniques were used to establish the existence of recurrent behavioural responses from each carbon source to the various wastewater samples. From the cross correlation, an attempt was made to classify the metabolic potential for future biodegradation processes. Carbohydrates, amino acids and carboxylic acids were the most predominant groups of sole carbon substrates showing similar growth behaviour in the consortium.

2-Propanol↗

Phenol- and toluene-degrading microbial populations from an aquifer in which successful trichloroethene cometabolism occurred.

We characterized the bacterial populations that grew in a Moffett Field, Calif., aquifer following three sequential field tests of phenol- or toluene-driven cometabolism of trichloroethene (TCE). Reducing the toluene and phenol concentrations in most-probable-number (MPN) tubes from 50 to 5 ppm increased the population density measured for these degraders by 1.5 and 1 log units, respectively, suggesting that natural populations might be quite sensitive to these substrates. Phenol and toluene degraders were isolated from the terminal MPN dilution tubes; 63 genetically distinct strains were identified among the 273 phenol- and toluene-degrading isolates obtained. TCE was cometabolized by 60% of the genetically distinct strains. Most strains (57%) grew on both phenol and toluene, and 78% of these strains hybridized to the toluene ortho-monooxygenase (TOM) probe. None of the strains hybridized to probes from the four other toluene oxygenase pathways. Gram-positive strains comprised 30% of the collection; all of these grew on phenol, and 47% of them also grew on toluene, but none hybridized to the TOM probe. Among the gram-negative strains, 86% of those that grew on both toluene and phenol hybridized to the TOM probe, while only 5% of those that were TOM-positive grew on toluene alone. A larger proportion of TCE degraders was found among gram-negative than gram-positive strains and among organisms that grew on phenol than those that grew on toluene. Hybridization of strains to the TOM probe was somewhat predictive of their TCE-cometabolizing ability, especially for strains isolated on toluene, but there was also a significant number (20%) of strains that hybridized to the TOM probe but were poor TCE cooxidizers. No Moffett Field isolates were as effective as Burkholderia cepacia G4 in cooxidizing TCE. Most of the aquifer strains ranged from moderately effective to ineffective in TCE cooxidation. Such populations, however, apparently accounted for the successful phenol- and toluene-stimulated TCE removal that occurred during the field assessment of this remediation process. This suggests that naturally occurring communities of only moderate TCE-cooxidizing ability may support successful TCE bioremediation as long as the phenol or toluene present is not limiting. This activity, however, may not be sustainable for the long term, because TCE-inactive populations that consumed toluene at rates equal to that of the best TCE degraders were present and hence would be expected to eventually dominate the community.

Journal Article↗

Fiber-reinforced polymeric composites are susceptible to microbial degradation.

A mixed culture of fungi, enriched from degraded polymeric materials, formed biofilms on coupons of fiber-reinforced polymeric composites (FRPCs). They grew actively in aqueous extracts of the composites under ambient conditions. The data indicate that the fungi utilized the resins or fiber chemical sizing as carbon and energy sources. A progressive decline in impedance from above 10(7) Ohms to below 10(8) Ohms was detected in the inoculated FRPC panels by electrochemical impedance spectroscopy (EIS) after 179 days of incubation, but not on the sterile controls. The degradation proceeds through an initial ingress of water into the resins, followed by degradation of bonding between fiber surfaces and resins and finally separation of fibers from the resins. At the end of EIS study, the extent of disbonding in the inoculated composite was greater than the control observed by scanning electron microscopy. These results suggest that the composite materials are susceptible to microbial attack by providing nutrients for growth.

Biodegradation, Environmental↗

Microbial degradation of taurine in fecal cultures from cats given commercial and purified diets.

Freshly passed feces were collected from cats (n = 5-7) given cooked and uncooked commercial canned-type diets, casein and soy protein containing purified diets and a commercial extruded diet. The feces were anaerobically cultured in medium containing either taurine, taurocholic acid or [2-3H]taurine for 24 h at 37 degrees C. Taurine degradation in cultures was greatest for cultures from cats receiving diets reputed to cause taurine depletion. Diaminopimelic acid in feces indicated that differences in taurine degradation rate among groups was associated with differences in bacterial numbers in feces. After 6 h of incubation, < 10% of taurocholate and > 60% of taurine remained. Nearly all the tritium on the labeled taurine was recovered as water. These results indicate that deconjugation and deamination are the initial steps in microbial catabolism of taurocholic acid and that enteric microbial growth may be a major determinant of dietary taurine requirement of cats.

Animal Feed↗

Microcosm enrichment of biphenyl-degrading microbial communities from soils and sediments.

A microcosm enrichment approach was employed to isolate bacteria which are representative of long-term biphenyl-adapted microbial communities. Growth of microorganisms was stimulated by incubating soil and sediment samples from polluted and nonpolluted sites with biphenyl crystals. After 6 months, stable population densities between 8 x 10(9) and 2 x 10(11) CFU/ml were established in the microcosms, and a large percentage of the organisms were able to grow on biphenyl-containing minimal medium plates. A total of 177 biphenyl-degrading strains were subsequently isolated and characterized by their ability to grow on biphenyl in liquid culture and to accumulate a yellow meta cleavage product when they were sprayed with dihydroxybiphenyl. Isolates were identified by using a polyphasic approach, including fatty acid methyl ester (FAME) analysis, 16S rRNA gene sequence comparison, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins, and genomic fingerprinting based on sequence variability in the 16S-23S ribosomal DNA intergenic spacer region. In all of the microcosms, isolates identified as Rhodococcus opacus dominated the cultivable microbial community, comprising a cluster of 137 isolates with very similar FAME profiles (Euclidean distances, <10) and identical 16S rRNA gene sequences. The R. opacus isolates from the different microcosms studied could not be distinguished from each other by any of the fingerprint methods used. In addition, three other FAME clusters were found in one or two of the microcosms analyzed; these clusters could be assigned to Alcaligenes sp., Terrabacter sp., and Bacillus thuringiensis on the basis of their FAME profiles and/or comparisons of the 16S rRNA gene sequences of representatives. Thus, the microcosm enrichments were strongly dominated by gram-positive bacteria, especially the species R. opacus, independent of the pollution history of the original sample. R. opacus, therefore, is a promising candidate for development of effective long-term inocula for polychlorinated biphenyl bioremediation.

Bacterial Typing Techniques↗

Culture-dependent and -independent approaches establish the complexity of a PAH-degrading microbial consortium.

A microbial consortium (AM) obtained by sequential enrichment in liquid culture with a polycyclic aromatic hydrocarbon (PAH) mixture of three- and four-ringed PAHs as a sole source of carbon and energy was examined using a triple-approach method based on various cultivation strategies, denaturing gradient gel electrophoresis (DGGE), and the screening of 16S and 18S rRNA gene clone libraries. Eleven different sequences by culture-dependent techniques and seven by both DGGE and clone libraries were obtained. The comparison of three variable regions (V3-V5) of the 16S rRNA gene between the sequences obtained yielded 19 different microbial components. Proteobacteria were the dominant group, representing 83% of the total, while the Cytophaga-Flexibacter-Bacteroides group (CFB) was 11% and the Ascomycota fungi 6%. Beta-proteobacteria were predominant in the DGGE and clone library methods, whereas they were a minority in culturable strains. The highest diversity and number of noncoincident sequences were achieved by the cultivation method that showed members of the alpha-, beta-, and gamma-Proteobacteria; CFB bacterial group; and Ascomycota fungi. Only six of the 11 strains isolated showed PAH-degrading capability. The bacterial strain (AMS7) and the fungal strain (AMF1), which were similar to Sphingomonas sp. and Fusarium sp., respectively, achieved the greatest PAH depletion. The results indicate that polyphasic assessment is necessary for a proper understanding of the composition of a microbial consortium.

Ascomycota↗

Microbial degradation of cephalothin by cephalothin-susceptible Escherichia coli.

Cephalothin (CET)-susceptible Escherichia coli, which can degrade CET after prolonged incubation in broth containing a concentration of the drug greater than the minimum inhibitory concentration, was found in a clinical specimen. The substrate specificity of the partially purified enzyme to cephalosporin analogs strongly indicated the occurrence of CET-specific degradation. Nuclear magnetic resonance analysis of the degradation reaction demonstrated the appearance of two new signals attributed to deacetyl CET. This suggests the possibility of the presence of acylesterase.

Cephalothin↗