Performance assessment and validation of a paramagnetic particle-based enzyme-linked immunosorbent assay for chlorpyrifos in agricultural runoff waters

A commercial magnetic particle-based enzyme-linked immunosorbent assay (ELISA) kit for the insecticide chlorpyrifos [O,O-diethyl-O-(3,5,6-trichloro-2-pyridyl) phosphorothioate] was evaluated for its specificity, precision, and accuracy, its susceptibility to matrix interferences in agricultural and environmental surface waters, and its comparability to a gas chromatographic/flame photometric (GC/FPD) method for the determination of organophosphorus pesticides in natural waters. Repeatability, reproducibility, and accuracy studies show that the kit satisfies current U.S. Environmental Protection Agency criteria for the assessment of analytical methods. Observable matrix effects were found to be present in all of the environmental test waters, with the slopes of calibration curves generated in each of the test matrices deviating from that of the control matrix by as much as 16%. Specificity studies indicate that the chlorpyrifos polyclonal antibody adequately differentiates the target compound from other structurally similar organophosphorus pesticides, with the exception of its methyl analogue. Cross-reactivity with chlorpyrifos-methyl was approximately 37%, while reactivity with diazinon, pyridaphenthion, diclofenthion, bromiphos-ethyl, bromiphos-methyl, pirimiphos-ethyl, and chlorpyrifos oxon ranged from 1.6 to 10.7%. Cross-reactivity with pirimiphos-methyl, 3,5,6-trichloro-2-pyridinol, diethyl phosphate, and diethyl thiophosphate was negligible (<1%). Validation of the paramagnetic particle ELISA format was accomplished using water samples from two monitoring studies that were collected, split, and analyzed directly by ELISA and by GC/FPD. Results of the two analytical methods were then compared using standard t tests, regression analysis, and differences against mean measurement (bias) plots. While the agreement between the two methods was determined to be satisfactory, ELISA exhibits consistent positive bias in environmental matrices. Several preanalysis mitigation steps were suggested that may help moderate bias, but additional study is recommended to explicate the exact factors responsible for its consistent overestimation of results.     

Replicative and cytopathic potential of HTLV-III/LAV with sor gene deletions

The genome of the human T-lymphotropic virus type III (HTLV-III/LAV) has the potential to encode at least three polypeptides in addition to those encoded by the gag, pol, and env genes. In this study, the product of the sor (short open reading frame) region, which overlaps the 3' end of the pol gene, was found to be a protein with a molecular weight of 23,000. An assay was developed for testing the ability of cloned HTLV-III proviruses to produce viruses cytopathic for T4+ lymphocytes. In the cell line used, C8166, neither the HTLV-III sor gene product nor the complete 3'-orf gene product were necessary for the replication or cytopathic effects of the HTLV-III.     

Feeding deterrents from Dictamnus dasycarpus Turcz against two stored-product insects

The screening for insecticidal principles from several Chinese medicinal herbs showed that the root bark of Dictamnus dasycarpus possessed significant feeding deterrence against two stored-product insects (Tribolium castaneum and Sitophilus zeamais). From the methanol extract, two feeding deterrents were isolated by bioassay-guided fractionation. The compounds were identified as fraxinellone and dictamnine from their spectroscopic data. Fraxinellone was demonstrated to possess feeding deterrent activity against adults and larvae of T. castaneum as well as S. zeamais adults with EC50 values of 36.4, 29.1, and 71.2 ppm, respectively. Dictamnine was shown to have feeding deterrent activity against adults and larvae of T. castaneum as well as S. zeamais adults with EC50 values of 57.6, 47.9, and 91.7 ppm, respectively.     

棕仁粕基础营养分析及其在尼罗罗非鱼养殖中的应用研究

本文对比研究了棕仁粕、豆柏、花生粕的基础营养及体外胃蛋白酶消化性,通过在实验饲料中添加2％、6％、10％的棕仁粕制备了四份等氮等能的饲料,初步评价了添加棕仁粕对罗非鱼生长、体组分以及血清学参数的影响.结果表明:棕仁柏作为蛋白水平相对略低,纤维含量较高的植物蛋白替代源,其体外胃蛋白酶消化率与豆粕和花生粕无明显差异,可以作为膳食饲料组分适量的添加到水产动物饲料中,而不会影响罗非鱼的正常生长;在罗非鱼养殖实验中,添加2％棕仁粕的饲料组,罗非鱼的增重率,饲料系数,蛋白利用率、鱼体的蛋白组分以及血清学参数与对照组相比无显著差异(P＞0.05),添加至6％～10％后,罗飞鱼的生长表现开始下降,饲料系数开始上升,鱼体蛋白组分下降(P＜0.05).作为棕仁粕在水产养殖领域的初步探索性应用研究,研究认为棕仁粕是可以用于水产饲料的成分使用的,添加比例不宜超过6％.     

Distribution and partial characterisation of acid hydrolysable organic nitrogen in six New Zealand soils

Six New Zealand topsoils of widely different origins and properties were subjected to 6m HC1 hydrolysis and the distribution of N fractions and amino acids were determined qualitatively and quantitatively.Of the total-N in the soils studied 83–91%, was hydrolysable with 6m HCl. The largest proportion of the hydrolysable N was α-amino acid N (38– 42%). followed by hydrolysable-unknown N (HUN) (14–24%), and NH₄⁺-N (14–22%). A significant proportion (25–50%) of the HUN fraction was accounted for by the non α-amino acid-N. Oxidative (3% H₂O₂) hydrolysis released N-phenoxy amino acid-N and possibly N-compounds which were complexed with phenols and sugars. All soils had a similar amino-acid composition with a predominance of acidic amino-acids.     

Psoas compartment and sacral plexus block via electrostimulation for pelvic limb amputation in dogs

Objective

To assess the efficacy of psoas compartment and sacral plexus block for pelvic limb amputation in dogs.

Molecular cloning and physiological effects of brushtail possum interleukin-1beta

Interleukin-1beta (IL-1beta) was isolated from LPS-stimulated brushtail possum alveolar macrophages using PCR primers based on conserved regions of mammalian IL-1beta. The complete cDNA was cloned by 5' and 3' rapid amplification of cDNA ends (RACE). The predicted protein of 269 amino acids shared 4346% identity with several mammalian IL-1beta proteins. Constructs were made to express the mature IL-1beta in Escherichia coli and two recombinant IL-1beta proteins, rpIL-1beta1 and rpIL-1beta2, which differed in length by four amino acids at the N-terminus, were produced. Both proteins induced a weak proliferative response in a possum thymocyte assay. Possums injected intravenously with 100 microg of rpIL-1beta1 or rpIL-1beta2 showed profound changes in body temperature and numbers of circulating leukocytes. A sharp decrease in temperature occurred within 2 h of administration followed by an elevation of temperature peaking at 24 h. The smaller rpIL-1beta1 protein had a greater effect on temperature than rpIL-1beta2. Both rpIL-1beta proteins caused a marked decrease in number of neutrophils and lymphocytes at 2-6 h after injection. At 24 h after injection, neutrophil and lymphocyte numbers were elevated 6.0-fold and 2.6-fold, respectively in the possums injected with rpIL-1beta1 and 3.9-fold and 1.5-fold, respectively in the possums injected with rpIL-1beta2. Fibrinogen levels were elevated at 24 and 72 h after injection with both proteins. In comparison, neither recombinant bovine IL-1beta (rbIL-1beta) nor PBS had significant effects on body temperature or blood haematology. The studies have shown that the two recombinant forms of IL-1beta were biologically active in possums and that the IL-1beta with four fewer amino acids at the N-terminus was the more active.     

Biological nitrogen fixing capacity and biomass production of different understorey pastures in a Pinus radiata-pasture agroforestry system in New Zealand

Quantitative field measurements of biological nitrogen fixation (BNF) and biomass production by four different understorey pastures in a Pinus radiata-pasture agroforestry system were determined over a period of one year. The trees were two years old at the beginning of this study and the understorey pastures were being cut and removed for silage. The BNF was determined using the ¹⁵N dilution technique. Pastures of ryegrass+clover, cocksfoot+clover, phalaris+clover and lucerne were used. Substantial amounts of BNF were found (71 to 230 kg N ha^–1 year^–1) with lucerne showing the highest N fixation. However, lucerne derived only 71 to 72% of its N from the atmosphere (%Ndfa) during the spring/summer period compared to 83–97% with clovers, thus the net N demand from the soil was substantially higher with lucerne. This caused increased N stress to the trees. Clover in ryegrass+clover pasture fixed more N than the other grass+clover pastures. Although pasture position in relation to trees did not affect annual pasture total DMY and %Ndfa, pastures north of tree row grew better than those in other positions. Trees significantly affected the BNF of legumes and the botanical composition of pastures with highest BNF and legume production occurring in pastures midway between two rows of trees. These results suggest that it would be advantageous to evaluate different legumes and grasses for tolerance of shade and moisture stress in future studies. As the trees studied were only 1.5 to 3 m in height, their effects on BNF, seasonal pasture biomass production and botanical composition are expected to increase with tree dominance in the ecosystem with time. Amounts of N fixed were related to the productivity (i.e. dry matter and N yield) and seasonal persistence of the legumes. The productivity was high in spring and summer and low in autumn and winter.     

Duration and method of fixation affects the sensitivity of a digoxygenin-labelled DNA probe in detecting Kudoa thyrsites in Atlantic salmon skeletal muscle

The effect of fixative and duration of fixation on the sensitivity of a non-radioactive in situ hybridisation (ISH) protocol to detect Kudoa thyrsites small subunit ribosomal deoxyribonucleic acid (DNA) was investigated. Strong ISH reactions were detected in 5-μm sections of paraffin-embedded Atlantic salmon muscle after fixation for 1 day in Davidson's solution (DS). Reactions were weak following 3 or 5 days fixation and absent after 17 or 28 days fixation. Strong ISH reactions were observed after 1, 3 or 5 days fixation in neutral buffered formalin (NBF). The reactions were weak after 17 days and weak to nonexistent after 28 days of fixation. Reactions were consistently strong after fixation in 95% ethanol for up to 28 days. Some mature spores reacted weakly or not at all by ISH. Parasite DNA was weakly amplified by polymerase chain reaction (PCR) from paraffin-embedded muscle after 1 day of fixation in DS but not after fixation for 3, 5, 17 or 28 days. Amplified DNA was detected after fixation in NBF for 1, 3 and 5 days, but not after 17 or 28 days. In contrast, PCR consistently amplified DNA from paraffin-embedded, ethanol-fixed muscle. Caution should be used in the choice of fixative and duration of fixation when preserving Atlantic salmon tissues for molecular diagnosis of K. thyrsites.     

Differences in structure of organic matter in two soils as demonstrated by 13C cross polarisation nuclear magnetic resonance spectroscopy with magic angle spinning

Spectra of two whole soils have been recorded by cross polarisation n.m.r. spectroscopy with magic angle spinning. Magic angle spinning allows detailed structural comparisons between the types of organic carbon in whole soils to be made. Sufficient resolution is achieved to show that the two soils differ considerably in polysaccharide content.