Detection and characterization of twenty-eight isomers of fumonisin B1 (FB1) mycotoxin in a solid rice culture infected with Fusarium verticillioides by reversed-phase high-performance liquid chromatography/electrospray ionization time-of-flight and ion trap mass spectrometry.

Fumonisin mycotoxins which are hazardous to humans and animals were produced in a Fusarium verticillioides-infected solid rice culture. To decrease the possibility of the formation of artifacts, the fumonisins were analysed by reversed-phase high-performance liquid chromatography/electrospray ionization time-of-flight (RP-HPLC/ESI-TOFMS) and ion trap mass spectrometry (RP-HPLC/ESI-ITMS) immediately after the extraction of the culture material, without any further sample clean-up. The fumonisin isomers were separated by using a flat gradient on a special, high-coverage C(18), narrow-bore HPLC column (YMC-Pack J'sphere ODS H80) suggested for the separation of structural isomers by the manufacturer. Exact mass measurements (TOFMS) of the protonated molecules and extraction of the ion chromatogram corresponding to the empirical formula (C(34)H(59)NO(15)) of FB(1) toxins led to the identification of 29 peaks and shoulders, including those of FB(1). The FB(1) toxin and 28 of its isomers were also detected by ITMS after separation with RP-HPLC. The characteristic m/z values of the product ions, including the backbones obtained by ITMS(2), undoubtedly indicated the structures of the FB(1) isomers for 28 peaks and shoulders. In the MS(2) spectra of the protonated molecules of the FB(1) isomers, with some exceptions, 15 characteristic product ions including the hydrocarbon backbone at m/z 299 were observed. The abundance ratio of the cation at m/z 299 ranged up to 5.8%. The relative quantities of the isomers found in the sample extract were expressed as percentages of the FB(1) content (0.001-0.579%). The total amount of the 28 FB(1) isomers was 2.803% of the quantity of FB(1) that is important from the aspect of food and feed safety.

Biomass productivities in wild type and pigment mutant of Cyclotella sp. (Diatom).

Microalgae are expected to play a significant role in greenhouse gas mitigation because they can utilize CO(2) from power plant flue gases directly while producing a variety of renewable carbon-neutral biofuels. In order for such a microalgal climate change mitigation strategy to become economically feasible, it will be necessary to significantly improve biomass productivities. One approach to achieve this objective is to reduce, via mutagenesis, the number of light-harvesting pigments, which, according to theory, should significantly improve the light utilization efficiency, primarily by increasing the light intensity at which photosynthesis saturates (I(s)). Employing chemical (ethylmethylsulfonate) and UV mutagenesis of a wild-type strain of the diatom Cyclotella, approximately 10,000 pigment mutants were generated, and two of the most promising ones (CM1 and CM1-1) were subjected to further testing in both laboratory cultures and outdoor ponds. Measurements of photosynthetic oxygen production rates as a function of light intensity (i.e., P-I curves) of samples taken from laboratory batch cultures during the exponential and linear growth phase indicated that the light intensity at which photosynthesis saturates (I(s)) was two to three times greater in the pigment mutant CM1-1 than in the wild type, i.e., 355-443 versus 116-169 mumol/m(2) s, respectively. While theory, i.e., the Bush equation, predicts that such a significant gain in I(s) should increase light utilization efficiencies and thus biomass productivities, particularly at high light intensities, no improvements in biomass productivities were observed in either semi-continuous laboratory cultures or outdoor ponds. In fact, the maximum biomass productivity in semi-continuous laboratory culture was always greater in the wild type than in the mutant, namely 883 versus 725 mg/L day, respectively, at low light intensity (200 micromol/m(2) s) and 1,229 versus 1,043 mg/L day, respectively, at high light intensity (1,000 micromol/m(2) s). Similarly, the biomass productivities measured in outdoor ponds were significantly lower for the mutant than for the wild type. Given that these mutants have not been completely characterized in these initial studies, the exact reasons for their poor performance are not known. Most likely, it is possible that the mutation procedure affected other photosynthetic or metabolic processes. This hypothesis was partially validated by the observation that the pigment mutant had a longer lag period following inoculation, a lower maximum specific growth rate, and poorer stability than the wild type.

Cometabolic mineralization of benzo[a]pyrene caused by hydrocarbon additions to soil.

The mineralization of [7-14 C]benzo[a]pyrene (BaP) in soil was investigated in response to additions of individual hydrocarbons, defined hydrocarbon mixtures, crude oil, and crude oil fractions. Neither substantial BaP mineralization nor enrichment of BaP degraders occurred in BaP-spiked soil in the absence of a suitable hydrocarbon supplement. Crude oil, the saturated and aromatic class components of crude oil, the distillates heating oil, jet fuel, and diesel fuel supported up to 60% mineralization of 80 µg [7-14 C]BaP per gram of soil in 40 d. Neither single hydrocarbons nor defined hydrocarbon mixtures containing normal and branched alkanes, alicyclics, and aromatics supported comparable BaP mineralization. Evolution of 14 CO2 occurred after lag periods characteristic to specific petroleum products and their concentrations. Time required for microbial proliferation, hydrocarbon toxicity, and competitive inhibition might have contributed to these lag periods, but the complete inhibition of BaP mineralization by dieselfuel vapors pointed to a dominant role of competitive inhibition. A lack of radiocarbon incorporation into soil biomass from [7-14 C]BaP indicated that at least the initial steps of BaP biodegradation in soil were cometabolic in nature. Suitable hydrocarbon mixtures not only supported BaP mineralization by serving as primary substrates, but also enhanced BaP bioavailability by dissolving this hydrophobic solid.