Glutathione transferases and herbicide detoxification in suspension-cultured cells of giant foxtail (Setaria faberi)

Glutathione transferases (GSTs) catalysing the conjugation of 1-chloro-2,4-dinitrobenzene, the chloro-s-triazine herbicide atrazine, the chloroacetanilide herbicides metolachlor and alachlor and the diphenyl ether herbicide fluorodifen have been identified in suspension-cultured cells derived from the grass weed giant foxtail (Setaria faberi Herrm.). In contrast to suspension-cultured cells of maize, where atrazine-conjugating GSTs are lost during de-differentiation, the GSTs active toward this herbicide in S. faberi plants were also expressed in cultures, suggesting that these isoenzymes are subject to different regulation in the crop and weed. As a result, glutathione conjugation was the major route of atrazine metabolism in S. faberi cultures. Activities of these GSTs were maximal three days after sub-culturing when the cells were dividing most actively, when they were determined to be in the order CDNB>alachlor>metolachlor= fluorodifen>atrazine. This indicated that GSTs which are enhanced during cell division can metabolise herbicides. On the basis of activity per mg protein, GST activities in the cultures were between 20 and 60-fold higher than those determined in the foliage of S. faberi seedlings. The GSTs with activity towards CDNB were resolved into three peaks following anion-exchange chromatography at pH 7·8 using Q-Sepharose. Peak 1 GSTs were not retained, while peak 2 and peak 3 were sequentially resolved with an increasing concentration of salt. Peak 1 GSTs showed activity toward metolachlor and atrazine but showed little activity toward fluorodifen. Peak 2 and peak 3 GSTs were active toward atrazine and metolachlor, with peak 3 being particularly associated with activity toward fluorodifen. The GSTs in these peaks were then further purified using S-hexyl-glutathione-agarose affinity chromatography. In each case, the affinity-bound fraction of the GSTs consisted of 28 kDa and 26 kDa polypeptides, suggesting that the GST isoenzymes in S. faberi cultures are composed of related subunits. Our results demonstrate that the GST isoenzymes involved in herbicide metabolism in suspension cultures of a grass weed show a similar level of complexity to that determined in maize cell cultures. © 1998 SCI     

Selective capture of blue cod Parapercis colias by potting: behavioural observations and effects of capture method on peri-mortem fatigue

Selective fishing is the successful capture of target species and size classes in a way that minimises bycatch, minimises damage to flesh, and maximises post-mortem (PM) value. We studied selective harvesting of blue cod Parapercis colias (Pinguipedidae) in central New Zealand, where it is captured mainly by commercial pot-fishing and recreational line-fishing. Potting with paua Haliotis iris (=abalone) guts selectively targeted large blue cod from localities that had six or more fish species and many small blue cod. Video observations of pot entries indicated that blue cod entered and left the pot throughout 30 min sets. Blue cod were observed by video to commence swimming when pot-hauling started. Observations of line-hooked fish indicated a characteristic spinning behaviour while hauling.

Fatigue during harvesting is a major factor in reducing the PM quality and shelf-life of fish muscle. Despite the relatively benign nature of capture via pot, it appears that burst exercise during hauling and a brief flurry of flapping as the pot leaves the water is sufficient to compromise flesh quality. Fish that were fatigued during harvesting had a poorer peri-mortem ‘ATP potential’ than fish that had been tank-rested for a year and then harvested using rested harvesting techniques. Modifying the potting method by providing a reservoir of water reduced peri-mortem fatigue during capture but by an insufficient amount to significantly improve PM flesh quality.

Blue cod flesh is compromised by pot capture, which is widely perceived as a benign harvesting method. In order to maximise value of blue cod, low-stress harvesting methods which take advantage of the behaviour of the fish are required.     

Remediation of Pesticide Contaminated Soil by Planting and Compost Addition

Pesticide contaminated material was obtained from an agrichemical retail facility in Illinois. In an effort to improve physical properties and increase microbial activity and plant growth in the contaminated matrix, the material was mixed with uncontaminated soil or with mature yard waste compost to determine the impact of compost compared to soil on plant establishment and growth, development of soil microbial activity, and herbicide inactivation. Plant growth, particularly weed growth, was significantly greater in compost containing mixes than it was in soil mixes. Microbial activity, as measured by dehydrogenase activity, was significantly higher in compost-containing mixes than in soil mixes. The combination of planting and compost addition resulted in significant increases in herbicide inactivation in the contaminated materials. The results strongly suggest that remediation of herbicide contamination at agrichemical retail facilities can be achieved quite rapidly and at moderate cost and also demonstrate that the combination of compost addition and planting can accelerate xenobiotic degradation in contaminated soils.     

Invasion of enterocytes in cultured porcine small intestinal mucosal explants by Salmonella choleraesuis.

Porcine small intestinal explants maintained in vitro were inoculated with Salmonella choleraesuis to study the characteristics of its invasion of enterocytes. The explants were fixed at selected intervals for up to 12 hours after inoculation and examined by conventional light microscopy, immunoperoxidase staining, and transmission electron microscopy. Although there was diffuse loss of villous enterocytes during the first hour of incubation, the villi were reepithelialized by the end of 2 hours of culture, and the mucosal epithelium remained intact and appeared to be viable through 12 hours of culture. Intraepithelial S choleraesuis were not detected before 6 hours after inoculation, but after 12 hours of incubation, bacteria were numerous within enterocytes. Ultrastructurally, penetration of the brush border by S choleraesuis resulted in focal loss of microvilli. Bacteria were endocytosed into membrane-bound vacuoles where most remained, but a few were free within the cytoplasm of enterocytes. Invasion of the explants closely resembled that described for live animal and cell culture models of Salmonella spp invasion.     

Effects of an oil-based substrate (The Water Cleanser™) and bacterial additives on nitrogen and phosphorous dynamics in freshwater crayfish (Cherax cainii,Austin and Ryan 2002) aquaculture

Two experiments were conducted to evaluate the efficacy of an oil-based substrate, The Water Cleanser? (TWC). The first experiment studied the effects two substrates of different oil composition (TWC, TWC?+), and a commercial bacterial additive (Bio-Aid) on concentrations of nitrogen and phosphorous in indoor aquaria. The second experiment studied the effects of TWC, a bacterial additive, and a combination (TWC?+?B) on concentrations of nitrogen and phosphorous, phytoplankton abundance and diversity in outdoor freshwater crayfish (Cherax cainii, Austin and Ryan, Invertebr Syst 16:357–367, 2002) tanks. In the first experiment, the concentration of TAN decreased more rapidly with Bio-Aid, whilst the maximum concentrations of NO₂-N and NO₃-N were reduced with the substrates. The concentration of orthophosphate was reduced in aquaria with TWC?+?. In the second experiment, concentrations of TAN, NO₂-N, NO₃-N and total phosphate were not significantly affected by TWC. After addition of TWC?+?B, there was a significant decrease in the concentrations of nitrate and total phosphate, and a higher abundance of phytoplankton was maintained than with other treatments. Additionally, a population Bacillus sp. was found on the substrate surface. TWC had no adverse effects on phytoplankton abundance or diversity, or C. cainii physiology, weight gain or survival. TWC and TWC?+?were effective bioremediators of eutrophic water, whilst a combination of TWC?+?B was effective in short term bioremediation and in promoting phytoplankton abundance in C. cainii tank culture.