Soil microbial populations, community composition, and activity as affected by repeated applications of hog and cattle manure in eastern Saskatchewan.

A field site near Humboldt, Saskatchewan, was annually treated with hog or cattle manure and cropped to canola, spring wheat, barley, and canola from 1997 to 2000. During each growing season, soil was analyzed for microbial populations in terms of activity and community structure, and crops were assessed for root rot and foliar diseases. Microbial activity in soils treated with cattle manure was higher than in soils treated with hog manure or urea. Similarly, nitrous oxide emissions from soil increased with increasing rates of hog and cattle manure. Potential human pathogens, including Rahnella, Serratia, Proteus, Leclercia, and Salmonella species, were identified in soils that received cattle manure, whereas pseudomonads were the dominant species in the hog-manure-treated soil. Fecal coliforms were confirmed in soils that received hog or cattle manure. However, Enterobacteriaceae populations were 10-fold higher in soils receiving cattle manure than in soils receiving the other treatments. Increasing cattle manure rates increased fecal coliform population, but there was no indication that increased hog manure rates increased fecal coliform populations. Addition of urea, hog manure, or cattle manure to the soil did not increase foliar disease in wheat, barley, and canola and had variable effects on root rot incidence in cereals.

Use of anion-exchange membrane extraction for the high-performance liquid chromatographic analysis of mustard seed glucosinolates.

A new one-step extraction using anion-exchange membranes for the HPLC determination of glucosinolates in mustard seeds is reported. The exchange of glucosinolates on the membranes was studied using sinigrin in solutions and sinigrin added as an internal standard to seeds of yellow mustard. By varying time of extraction, membrane size, and sample size, the optimal conditions for maximum glucosinolate recovery were determined and the following procedure was adopted: 0.2 g of ground mustard seeds are heated in 20 mL of boiling water for 5 min. After cooling, samples are transferred to plastic centrifuge tubes, 9-cm(2) membranes are added, and suspensions are shaken on a mechanical shaker for 2.5 h. Glucosinolates are then eluted from the membranes with 25 mL of 1 N KCl by shaking again for 2.5 h. Using this procedure, the sinigrin extraction from solutions and from mustard seeds was linear with 80% recovery. Seeds of yellow, brown, oriental, and Indian mustard were analyzed by this procedure; excellent reproducibility, with coefficients of variation in the range 1.0-4.3% were obtained. This method offers a simple and inexpensive alternative to complicated and tedious procedures for glucosinolate isolation/purification required for chromatographic determinations.

Determination of glucosinolates in canola seeds using anion exchange membrane extraction combined with the high-pressure liquid chromatography detection.

A rapid, simple, and reliable method for the determination of individual glucosinolates in canola seeds was developed using a semiquantitative extraction of glucosinolates with anion exchange membranes and HPLC detection. In this one-step extraction procedure, a membrane (7 cm(2)) is placed in the seed suspension prepared by grinding and boiling 0.8 g of seeds in 20 mL of water. After 10 min of shaking on the mechanical shaker, the membrane is removed from the suspension, washed, and transferred to a vial containing 5 mL of 1 N tetramethylammonium chloride. The glucosinolates are eluted from the membrane by shaking the membrane for 10 min with the eluting solvent. The glucosinolate content in membrane eluates is determined by HPLC using sinigrin standards. A coefficient of variation ranging from 1.9 to 7.6% for aliphatic glucosinolates indicated very good reproducibility of the method. Because of the instability of 4-hydroxyglucobrassicin, the coefficient of variation for the determination of this indolyl glucosinolate was 13.9%. To verify the results of the membrane extraction/HPLC detection, this new method was compared with the existing colorimetric and GC procedures. Very good correlation (R(2) = 0.98) was obtained between the total glucosinolates determined by the membrane extraction/HPLC method and the palladate colorimetric procedure for 17 canola varieties. Concentrations of individual glucosinolates in five canola varieties were compared with the GC data. Very good agreement between these two methods was obtained for aliphatic glucosinolates. However, the membrane extraction/HPLC method yielded slightly higher values for 4-hydroxyglucobrassicin than the GC method, possibly indicating that the decomposition of this glucosinolate was reduced during the sample extraction with the membranes. The simplicity and low cost of the membrane extraction/HPLC method make it an attractive alternative to the existing procedures for glucosinolate analysis in canola seeds.