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.     

Book reviews

Marilyn Holly is an Associate Professor of Philosophy at the University of Florida. Her special interests include political philosophy, ethics and ecology, Jungian thought, and Native American spirituality in its relation to the environment. She is currently at work on a monograph-length piece on the latter topic. She has taught an undergraduate course in ethics and ecology, and plans to do so again. The following essay emerged from her search for a suitable text for such a course.     

High HEV presence in four different wild boar populations in East and West Germany

Swine Hepatitis E virus (HEV) can be transmitted from pigs to humans causing hepatitis. A high prevalence of HEV in wild boar populations is reported for several European countries, but actual data for Germany are missing. During the hunting season from October to December 2007 liver, bile and blood samples were collected from wild boars in four different German regions. The samples were tested for HEV RNA by quantitative PCR (qPCR) and anti-HEV IgG antibodies by two different ELISAs and a Line immunoassay. A seroprevalence of 29.9% using ELISA and 26.2% in the Line immunoassay was determined. The seroprevalence rate varied greatly within the analyzed regions. However, qPCR analysis revealed a higher prevalence of 68.2% positive animals with regional differences. Surprisingly, also adult wild sows and wild boars were highly HEV positive by qPCR. Compared to liver and serum samples, bile samples showed a higher rate of positive qPCR results. Sequencing and phylogenetic analysis of a 969 nt fragment within ORF 2 revealed that all isolates clustered within genotype 3 but differed in the subtype depending on the hunting spot. Isolates clustered within genotypes 3i, 3h, 3f and 3e. Within one population HEV isolates were closely related, but social groups of animals in close proximity might be infected with different subtypes. Two full-length genomes of subtypes 3i and 3e from two different geographic regions were generated. The wild boar is discussed as one of the main sources of human autochthonous infections in Germany.     

Mixotrophy in a Local Strain of Nannochloropsis granulata for Renewable High-Value Biomass Production on the West Coast of Sweden

A local strain of Nannochloropsis granulata (Ng) has been reported as the most productive microalgal strain in terms of both biomass yield and lipid content when cultivated in photobioreactors that simulate the light and temperature conditions during the summer on the west coast of Sweden. To further increase the biomass and the biotechnological potential of this strain in these conditions, mixotrophic growth (i.e., the simultaneous use of photosynthesis and respiration) with glycerol as an external carbon source was investigated in this study and compared with phototrophic growth that made use of air enriched with 1–2% CO₂. The addition of either glycerol or CO₂-enriched air stimulated the growth of Ng and theproduction of high-value long-chain polyunsaturated fatty acids (EPA) as well as the carotenoid canthaxanthin. Bioassays in human prostate cell lines indicated the highest antitumoral activity for Ng extracts and fractions from mixotrophic conditions. Metabolomics detected betaine lipids specifically in the bioactive fractions, suggesting their involvement in the observed antitumoral effect. Genes related to autophagy were found to be upregulated by the most bioactive fraction, suggesting a possible therapeutic target against prostate cancer progression. Taken together, our results suggest that the local Ng strain can be cultivated mixotrophically in summer conditions on the west coast of Sweden for the production of high-value biomass containing antiproliferative compounds, carotenoids, and EPA.     

Flowering and true seed production in potato (Solanum tuberosum L.). 1. Effects of inflorescence position,nitrogen treatment,and harvest date of berries

Summary Flowering and true seed production from different order inflorescences in potato (Solanum tuberosum L.) were evaluated in two experiments. The number of flowers per inflorescence, berry set, berry weight, number of seeds per berry and seed weight generally decreased from primary inflorescences to tertiary inflorescences and inflorescences on lateral stems. The possible relation with carbohydrate distribution is discussed. Quality of the seeds produced from the different orders of inflorescences was a function of seed size; larger seeds showed better germination, emergence and seedling growth. Late harvest of berries increased berry weight and number of seeds per berry from primary inflorescences. Application of additional nitrogen during the flowering period did not affect flowering or differences between inflorescence positions in seed production, but it significantly decreased the number of seeds per berry. The practical implications of these results for true seed production are discussed.     

Rainbow trout (Oncorhynchus mykiss,Walbaum ) develop a more robust body shape under organic rearing

Prolonged exposure to captive conditions has led to the development of a rainbow trout ‘farmed’ phenotype, which is different from that of wild trout. Selection for desirable productive traits in hatcheries has resulted in the development of some morphological traits that are maladaptive in nature. The recent development of organic aquaculture, guided by the well‐being of the fish, could potentially produce a new farmed phenotype that would be more adaptive in nature. In this study, rainbow trout reared in intensive and organic farms were compared by means of shape analysis, to detect patterns of shape variation associated with rearing environment. The results of this study highlight a significant effect of the rearing method on rainbow trout shape: organically reared trout showed a higher body profile, in particular in the head and trunk regions, shorter median fins and a deeper caudal peduncle. A combined effect of density and habitat complexity could have contributed to the observed shape differences: in organic rearing systems, lower densities and steady water could increase territoriality and aggressive interactions, promoting body designs more functional in rapid attacks and escapes.     

Articular / epiphyseal osteochondrosis in Thoroughbred foals at 5 months of age: influences of growth of the foal and prenatal copper supplementation of the dam

AIMS: To determine the influence of copper (Cu) supplementation by injection of mares in late gestation on the frequency and severity of osteochondrosis (OC) lesions in their foals at around 160 days of age. To determine if there was any influence of the concentration of Cu in the liver, growth rate, birthweight, weight at 160 days of age, fatness, sex, or year of birth of the foal on the frequency and severity of OC lesions. To determine the influence of dam's age, and sex and birthweight of the foal on the growth rate from birth to 160 days of age, and weight at 160 days of age. METHODS: Thirty-three Thoroughbred foals, born in two consecutive years, were weighed every 2 weeks from birth. The dams had been supplemented with parenteral Cu or saline during late gestation, and the supplementation regimens were different in each year. Foals had liver biopsies harvested at birth for determination of Cu concentration. Pasture samples were collected every 4-8 weeks for analysis of concentration of Cu and zinc (Zn). At 160 days of age, articular cartilage of long bones was examined. Gross lesions were counted and scored, then sawn and radiographed, and processed for histopathology. Lesions were given radiographic scores and histopathological scores. Maximum scores for each lesion were combined to give a total OC score for each joint and each foal. The fatness of 20 foals (10 each from Years 1 and 2) at 160 days of age was determined chemically. RESULTS: Supplementation of dams with Cu had no significant effect on the concentration of Cu in the liver of foals at birth, or on the frequency or severity of lesions in articular cartilage at 160 days of age. The Cu and Zn concentrations of pasture were similar in Years 1 and 2, and were lower than current recommendations. All foals in Year 2, and 9/10 foals in Year 1 had irregularities in cartilage that was confirmed histologically to be indicative of OC. The average number of lesions per foal was 4.7 (SD 1.1) and 5.7 (SD 1.1) in Years 1 and 2, respectively. However, the severity of the lesions was considered mild, and no foals showed any clinical evidence of OC while alive. The number of lesions in the tarsocrural (TC) joint and the TC OC score at 160 days were positively associated with average daily weight gain (ADG) in the previous 4 weeks (p=0.005 and p=0.001, respectively). There was no significant effect of sex, fatness, birthweight, weight at 160 days of age, or year of birth of the foal on the frequency and severity of OC lesions. CONCLUSIONS: Many of the lesions classified as OC, using classification systems described by other authors, were likely to be normal variations of the process of endochondral ossification. Despite the high frequency of such lesions, they were considered to be of minor significance and none were clinically evident. The distribution of lesions was not typical, and most probably reflected the subtlety of the lesions. These results support the hypothesis that Cu is an over-emphasised factor in the aetiopathogenesis of OC. The relationship between subtle macroscopic lesions and lesions resulting in clinical signs of disease requires further investigation.     

Pain management in horses and farm animals

Objective: This review discusses the different analgesic drugs and routes of administration used in large animals for acute pain management. General guidelines and doses are given to assist in choosing techniques that provide effective analgesia. Etiology: Noxious stimuli are perceived, recognized, and localized by specialized sensory systems located at spinal and supraspinal levels. Diagnosis: Localizing the source of the noxious stimulus as well as understanding the behavioral aspects and physiological changes that result from such insult is important to adequately diagnose and treat pain. Pain assessment is far from being definite and objective; not only are there species differences, but also individual variation. In addition, the behavioral and physiological manifestations vary with the acute or chronic nature of pain. Therapy: Pain management should include (1) selecting drugs that better control the type of pain elicited by the insult; (2) selecting techniques of analgesic drug administration that act on pathways or anatomical locations where the nociceptive information is being processed or originating from; (3) combining analgesic drugs that act on different pain pathways; and (4) provide the best possible comfort for the animal. Prognosis: Providing pain relief improves the animal's well being and outcome; however, interpreting and diagnosing pain remains difficult. Continuing research in pain management will contribute to the evaluation of the pathophysiology of pain, pain assessment, and newer analgesic drugs and techniques.     

Placing the pieces: Reconstructing the original property mosaic in a warrant and patent watershed

Recent research shows that land use history is an important determinant of current ecosystem function. In the United States, characterization of land use change following European settlement requires reconstruction of the original property mosaic. However, this task is difficult in unsystematically surveyed areas east of the Appalachian Mountains. The Gwynns Falls watershed (Baltimore, MD) was originally surveyed in the 1600-1700s under a system of warrants and patents (commonly known as ‘metes and bounds’). A method for the reconstruction and mapping of warrant and patent properties is presented and used to map the original property mosaic in the Gwynns Falls watershed. Using the mapped mosaic, the persistence of properties and property lines in the current Gwynns Falls landscape is considered. The results of this research indicate that as in agricultural areas, the original property lines in the Gwynns Falls watershed are persistent. At the same time, the results suggest that the property mosaic in heavily urbanized/suburbanized areas is generally ‘reset.’ Further, trends in surveying technique, parcel size, and settlement patterns cause property line density and property shape complexity to increase in the less urbanized upper watershed. The persistence of original patterns may be damping expression of heterogeneity gradients in this urban landscape. This spatial pattern of complexity in the original mosaic is directly opposite of hypothesized patterns of landscape heterogeneity arising from urbanization. The technique reported here and the resulting observations are important for landscape pattern studies in areas settled under unsystematic survey systems, especially the heavily urbanized areas of the eastern United States. This revised version was published online in July 2006 with corrections to the Cover Date.