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The applicability of dolomite particlesto control acidificationin a Hyphomicrobium MS3inoculated biofilter removingdimethyl sulphide (Me2S) wasstudied. While direct inoculationof the dolomite particles with theliquid microbial culture was notsuccessful, start-up ofMe2S-degradation in thebiofilter was observed when thedolomite particles were mixed with33% (wt/wt) of Hyphomicrobium MS3-inoculatedcompost or wood bark material.Under optimal conditions, anelimination capacity (EC) of 1680~g Me2S m-3 d-1 wasobtained for the compost/dolomitebiofilter. Contrary to a wood barkor compost biofilter, no reductionin activity due to acidificationwas observed in these biofiltersover a 235 day period because ofthe micro environmentneutralisation of the microbialmetabolite H2SO4 with thecarbonate in the dolomite material.However, performance of thebiofilter decreased when themoisture content of the mixedcompost/dolomite material droppedbelow 15%. Next to this, nutrientlimitation resulted in a gradualdecrease of the EC andsupplementation of a nitrogensource was a prerequisite to obtaina long-term high EC (> 250 gMe2S m-3 d-1) forMe2S. In relation to thisnitrogen supplementation, it wasobserved that stable ECs forMe2S were obtained when thisnutrient was dosed to the biofilterat a Me2S-C/NH4Cl-Nratio of about 10.Abbreviations:DW – dry weight,EC – elimination capacity,Me2S – dimethyl sulphide,OL – organic loading rate,VS - volatile solids

Upon inoculation with Hyphomicrobium MS3, theelimination capacity of a lab-scale biofilter for theodorant dimethyl sulfide (Me2S) can be stronglyincreased from less than 10 to more than 35 and 1000 gm-3d-1 using wood bark and compost as acarrier material, respectively. However, uponsupplementation of isobutyraldehyde (IBA) as a secondgaseous substrate, sequential degradation profiles of IBAand Me2S in physically separated sections wereobserved in the Hyphomicrobium MS3-inoculatedwood bark and compost biofilters. Contrary to this, thebiofiltration efficiency for Me2S remainedunaffected upon the supplementation of toluene as asecond gaseous substrate. Batch experiments with theliquid Hyphomicrobium MS3 culture confirmed thecompetitive effect of IBA on the Me2S degradingactivity: in the presence of both compounds, Hyphomicrobium MS3 preferred degradation of thecarbonyl compound. In technical terms, this means thatthe complete purification of a waste gas stream containingboth IBA and Me2S should be performed usingsufficiently high or bistage HyphomicrobiumMS3-inoculated biofilters. Design criteria have to beconceived in this respect.

Hyphomicrobium sp. strain DM2 was found to grow anaerobically in the presence of nitrate with methanol, formaldehyde, formate or dichloromethane. The estimated growth rate constants with methanol and dichloromethane under denitrification conditions were 0.04 h−1 and 0.015 h−1, respectively, which is twofold and fourfold lower than the rates of aerobic growth with these substrates. Slight accumulation of nitrite was observed in all cultures grown anaerobically with nitrate. Dichloromethane dehalogenase, the key enzyme in the utilization of this carbon source, was induced under denitrification conditions to the same specific activity level as under aerobic conditions. In a fed batch culture under denitrification conditionsHyphomicrobium sp. DM2 cumulatively degraded 35 mM dichloromethane within 24 days. This corresponds to a volumetric degradation rate of 5 mg dichloromethane/l·h and demonstrates that denitrificative degradation offers an attractive possibility for the development of anaerobic treatment systems to remove dichloromethane from contaminated groundwater.

Methamidophos is one of the most widely used organophosphorus insecticides usually detectable in the environment. A facultative methylotroph, Hyphomicrobium sp. MAP-1, capable of high efficiently degrading methamidophos, was isolated from methamidophos-contaminated soil in China. It was found that the addition of methanol significantly promoted the growth of strain MAP-1 and enhanced its degradation of methamidophos. Further, this strain could utilize methamidophos as its sole carbon, nitrogen and phosphorus source for growth and could completely degrade 3,000 mg l(-1) methamidophos in 84 h under optimal conditions (pH 7.0, 30 degrees C). The enzyme responsible for methamidophos degradation was mainly located on the cell inner membrane (90.4%). During methamidophos degradation, three metabolites were detected and identified based on tandem mass spectrometry (MS/MS) and gas chromatography-mass spectrometry (GC-MS) analysis. Using this information, a biochemical degradation pathway of methamidophos by Hyphomicrobium sp. MAP-1 was proposed for the first time. Methamidophos is first cleaved at the P-N bond to form O,S-dimethyl hydrogen thiophosphate and NH(3). Subsequently, O,S-dimethyl hydrogen thiophosphate is hydrolyzed at the P-O bond to release -OCH(3) and form S-methyl dihydrogen thiophosphate. O,S-dimethyl hydrogen thiophosphate can also be hydrolyzed at the P-S bond to release -SCH(3) and form methyl dihydrogen phosphate. Finally, S-methyl dihydrogen thiophosphate and methyl dihydrogen phosphate are likely transformed into phosphoric acid.