Enhanced biodegradation of hexachlorocyclohexane (HCH) in contaminated soils via inoculation with Sphingobium indicum B90A.

Soil pollution with hexachlorocyclohexane (HCH) has caused serious environmental problems. Here we describe the targeted degradation of all HCH isomers by applying the aerobic bacterium Sphingobium indicum B90A. In particular, we examined possibilities for large-scale cultivation of strain B90A, tested immobilization, storage and inoculation procedures, and determined the survival and HCH-degradation activity of inoculated cells in soil. Optimal growth of strain B90A was achieved in glucose-containing mineral medium and up to 65% culturability could be maintained after 60 days storage at 30 degrees C by mixing cells with sterile dry corncob powder. B90A biomass produced in water supplemented with sugarcane molasses and immobilized on corncob powder retained 15-20% culturability after 30 days storage at 30 degrees C, whereas full culturability was maintained when cells were stored frozen at -20 degrees C. On the contrary, cells stored on corncob degraded gamma-HCH faster than those that had been stored frozen, with between 15 and 85% of gamma-HCH disappearance in microcosms within 20 h at 30 degrees C. Soil microcosm tests at 25 degrees C confirmed complete mineralization of [(14)C]-gamma-HCH by corncob-immobilized strain B90A. Experiments conducted in small pits and at an HCH-contaminated agricultural site resulted in between 85 and 95% HCH degradation by strain B90A applied via corncob, depending on the type of HCH isomer and even at residual HCH concentrations. Up to 20% of the inoculated B90A cells survived under field conditions after 8 days and could be traced among other soil microorganisms by a combination of natural antibiotic resistance properties, unique pigmentation and PCR amplification of the linA genes. Neither the addition of corncob nor of corncob immobilized B90A did measurably change the microbial community structure as determined by T-RFLP analysis. Overall, these results indicate that on-site aerobic bioremediation of HCH exploiting the biodegradation activity of S. indicum B90A cells stored on corncob powder is a promising technology.

Cyst-based ecotoxicological tests using Anostracans: comparison of two species of Streptocephalus

Cyst-based ecotoxicological tests are simple and low-cost methods for assessing acute toxicity. Nevertheless, only a few comparative studies on their sensitivity are known. In the present study, the suitability of the use of two freshwater Anostracan species, Streptocephalus rubricaudatus and S. texanus, was assessed. The impact of 16 priority pollutants (4 heavy metals, 11 organic, and 1 organometallic compounds) on these two species, as well as on Artemia salina (Artoxkit M), Daphnia magna (International Organization for Standardization 6341), and S. proboscideus (Streptoxkit F) was assessed. For indicative comparison, bioassays using Brachionus calyciflorus (Rotoxkit F) and Photobacterium phosphoreum (Microtox) were also performed. For heavy metals (K2Cr2O7, Cd2+, Zn2+, Cu2+), the sensitivity of the two studied Streptocephalus species was slightly higher than that of D. magna. It was significantly more elevated than for the marine A. salina. For organic and organometallic micropollutants [phenol, 3,5-dichlorophenol, pentachlorophenol (PCP), hydroquinone, linear alkylbenzene sulfonate, sodium dodecyl sulfate, tributylphosphate, dimethylphthalate, atrazine, lindane, malathion, tributyltin chloride (TBT-Cl)], the sensitivity of the 4 anostracan species was of the same order of magnitude as that of D. magna. Artemia salina was slightly less sensitive to some organic compounds (PCP, hydroquinone, TBT-Cl). The sensitivity of S. rubricaudatus to organic solvents was low. On the other hand, this anostracan was quite sensitive to NaCl. Thus, its use is restricted to freshwater samples. The evaluation of global practicability of these two tests confirms that cyst-based freshwater anostracans may be used to perform low-cost tests at a sensitivity comparable to that of D. magna (24 h immobilization test).