Photolysis experiments on phosmet, an organophosphorus insecticide

The organophosphorus insecticide phosmet is used in plant protection as well as against parasites on animals. Phosmet showed numerous photoinduced reaction pathways, which first were studied in the presence of model environments for animal fur lipids (e.g., wool wax). The model solvents for saturated and unsaturated lipids were cyclohexane and cyclohexene, whereas methanol and 2-propanol were used as models for primary and secondary alcohol moieties of lipids. The measured degradation rates over an irradiation period of 7 h in all solvents used were very similar (49-55%). The obtained photoproducts generally included phthalimide, N-hydroxymethylphthalimide, and N-methoxymethylphthalimide. Furthermore, depending on the solvent used, additional degradation products were detectable as N-isopropoxy- and N-methylphthalimide in the presence of 2-propanol and cyclohexene, respectively. However, in the presence of cyclohexene, despite the similar turnover, distinctly lower concentrations of photoproducts were found, indicating further still unknown degradation pathways. Irradiations in methanol with increasing percentages of water led to higher degradation rates; however, the products were found to be the same. Irradiation experiments with pure phosmet on silica TLC plates and glass surfaces resulted in degradation rates of 19 and 32%, respectively, after 6 h. The results obtained clearly demonstrate for the first time that the photoinduced degradation of phosmet is strongly dependent on the chemical environment, affecting less the turnover than the photoproducts formed. The results additionally demonstrate the need to investigate the degradation behavior of phosmet on wool and in the presence of wool wax.     

Development of an enzyme-linked immunosorbent assay for the organophosphorus insecticide bromophos-ethyl

A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of the organophosphorus insecticide bromophos-ethyl. Four bromophos-ethyl derivatives (haptens) were synthesized and were coupled to carrier proteins through the pesticide thiophosphate group to use as an immunogen or as a coating antigen. Rabbits were immunized with either one of two haptens coupled to bovine serum albumin for production of polyclonal antibodies, and the sera were screened against one of the haptens coupled to ovalbumin. Using the serum with highest titer, an antigen-coated ELISA was developed, which showed an IC(50) of 3.9 ng/mL with a detection limit of 0.3 ng/mL (20% inhibition). An antibody-coated ELISA using an enzyme tracer was also developed, which showed an IC(50) of 6.5 ng/mL with a detection limit of 1.0 ng/mL (20% inhibition). The antibodies showed negligible cross-reactivity with other organophosphorus pesticides except with the insecticides bromophos-methyl and chlorpyrifos in the antibody-coated assay only. Recoveries of bromophos-ethyl from fortified crop and water samples ranged from 82 to 128% and from 95 to 127%, respectively.     

Development of an enzyme-linked immunosorbent assay for the detection of the organophosphorus insecticide acephate

A competitive indirect enzyme-linked immunosorbent assay (ciELISA) for the organophosphorus insecticide acephate, O,S-dimethyl acetylphosphoramidothioate, was developed using a polyclonal antibody. Five different haptens mimicking the analyte were synthesized and conjugated with the carrier proteins bovine serum albumin (BSA) and keyhole limpet hemocyanin (KLH) by the N-hydroxysuccinimide active ester and diazotization methods. Polyclonal antibodies raised against hapten-KLH conjugates in rabbits and hapten-BSA conjugates as coating antigens were screened and selected for the assay in the homologous and heterologous ELISA systems. The effects of various assay conditions such as detergent, organic solvents, pH, and preincubation of the mixture of the polyclonal antibody and the analyte on the sensitivity were evaluated. The IC(50) value for acephate was 25 ng/mL in an optimized heterologous system using hapten-4-BSA as a coating antigen and a polyclonal antibody no. 8377 against hapten-1-KLH, showing the detection range of 5-140 ng/mL and the lowest detection limit of 2 ng/mL. The cross-reactivities of the structurally related organophosphorus insecticides, including the major metabolite of the analyte, methamidophos, were less than 1%. Recoveries from the analyte-fortified tap water, mulberry leaves, and lettuce samples in the assay were in the range of 72-121% by simple extraction, concentration, and dilution. These results indicate that the ELISA could be a convenient and supplemental analytical tool for monitoring acephate residues in environmental and agricultural samples.     

Biotransformation of an organochlorine insecticide,endosulfan, by Anabaena species

This study assesses the role of the blue-green algal species present in the soil in the dissipation of endosulfan and its metabolites in the soil environment. Two Anabaena species, Anabaena sp. PCC 7120 and Anabaena flos-aquae, were used in this study. Anabaena sp. PCC 7120 produced three principal biotransformation compounds, chiefly endosulfan diol (endodiol), and minor amounts of endosulfan hydroxyether and endosulfan lactone. Trace amounts of endosulfan sulfate were detected. In comparison, the biotransformation of endosulfan by Anabaena flos-aquae yielded mainly endodiol with minor amounts of endosulfan sulfate. An unknown compound was produced up to 70% from endosulfan spiked in the medium inoculated by A. flos-aquae after 8 days of incubation. Therefore, the endosulfan fate was dependent on the species. Within 1 day of incubation, two Anabaena species produced low amounts of beta-endosulfan after application of alpha-endosulfan. These results suggest the presence of isomerase in the Anabaena species. Further studies using a fermentor to control the medium pH at 7.2 to minimize chemical hydrolysis of endosulfan revealed a major production of endodiol with minor amounts of endosulfan sulfate and the unknown compound. These results showed that the production of the unknown compound might be dependent on the alkaline pH in the medium and that the production of endodiol by A. flos-aquae might be biologically controlled. This study showed that two algal species could contribute in the detoxification pathways of endosulfan in the soil environment.     

Bioavailability of an organophosphorus pesticide,fenamiphos, sorbed on an organo clay

Hydrolysis of an insecticide/nematicide, fenamiphos [ethyl-3-methyl-4-(methylthio)phenyl-(1-methylethyl)phosphoramidate], immobilized through sorption by cetyltrimethylammonium-exchanged montmorillonite (CTMA-clay) by a soil bacterium, Brevibacterium sp., was examined. X-ray diffraction analysis, infrared spectra, and a negative electrophoretic mobility strongly indicated that fenamiphos was intercalated within the bacterially inaccessible interlayer spaces of CTMA-clay. The bacterium hydrolyzed, within 24 h, 82% of the fenamiphos sorbed by the CTMA-clay complex. There was a concomitant accumulation of hydrolysis product, fenamiphos phenol, in nearly stoichiometric amounts. During the same period, in abiotic (uninoculated) controls, 4.6% of the sorbed insecticide was released into the aqueous phase as compared to 6.0% of the sorbed fenamiphos in another abiotic control where activated carbon, a sink for desorbed fenamiphos, was present. Thus, within 24 h, the bacterium hydrolyzed 77% more fenamiphos sorbed by organo clay than the amounts desorbed in abiotic controls. Such rapid degradation of an intercalated pesticide by a bacterium has not been reported before. Evidence indicated that extracellular enzymes produced by the bacterium rapidly hydrolyzed the nondesorbable fenamiphos, even when the enzyme itself was sorbed. Fenamiphos strongly sorbed to an organo clay appears to be readily available for exceptionally rapid degradation by the bacterium.     

Enantiomeric separation and toxicity of an organophosporus insecticide, pyraclofos

Despite the fact that the biological processes of chiral pesticides are enantioselective, knowledge of the toxicities of pyraclofos due to enantiospecificity is scarce. In this study, the optical isomers of pyraclofos were separated and their toxicities to butyrylcholinesterase (BChE) and Daphnia magna were assessed. Baseline resolution of the enantiomers was obtained on both Chiralcel OD and Chiralpak AD columns. The effect of the mobile phase composition and column temperature were then discussed. The resolved enantiomers were characterized by their optical rotation and circular dichroism signs. The anti-BChE tests demonstrated that (-)-pyraclofos was about 15 times more potent than its (+)-form. However, acute aquatic assays suggested that (+)-pyraclofos was about 6 times more toxic than its antipode. Moreover, the joint toxicity of pyraclofos enantiomers to D. magna was found to be an additive effect. These results demonstrated that the overall toxicity of pyraclofos should be assessed using the individual enantiomers.     

The Aragon region of the Pyrenees—A stronghold for birds of prey in Europe

The importance of the Aragon region of the Pyrenees in northern Spain for birds of prey is discussed. During field observations in the summers of 1979 and 1982, 19 species of bird of prey were recorded within 80 km of the village of Berdun. Three additional species have also been seen by other observers, thus over four-fifths of the raptorial species recorded in Spain have occurred in this small area.Simple daily counts showed an average of 38 birds of prey of 6·4 species seen per field day. These compare favourably with similar counts made in other parts of Europe.The observations suggest that a significant proportion of Europe's griffon vultures (see Table 1) and lammergeiers occur, the latter representing 7% of the European population. The future of vultures in the area is briefly discussed.     

Development of a microtiter plate ELISA and a dipstick ELISA for the determination of the organophosphorus insecticide fenthion

In previous studies, polyclonal antibodies against the organophosphorus insecticide fenthion were obtained and an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed for this pesticide. In this study, using these antibodies and an enzyme tracer, direct competitive ELISAs for fenthion in microtiter plate and dipstick formats were developed. The microtiter plate ELISA showed an IC(50) value of 1.2 microg/L with a detection limit of 0.1 microg/L. The antibodies showed negligible cross-reactivity with other organophosphorus pesticides. The use of the dipstick format using Immunodyne as a support membrane allowed the quick visual detection of fenthion in concentrations >10 microg/L. The IC(50) value of the dipstick format using reflectance detection was 15 microg/L with a detection limit of 0.5 microg/L. The recoveries of fenthion from spiked vegetable samples using the two formats without any prior enrichment or cleanup steps were 87-116%.     

Inhibition of soil urease by organophosphorus insecticides

The effect of three organophosphorus insecticides on soil urease was examined. Inhibition of urea hydrolysis, some 60 days after application of 1000 parts/10⁶ of insecticide to a sandy clay loam, approached 40% (accothion) and exceeded 50% in the case of malathion and thimet. Similar inhibitory effects were recorded using a silt loam soil with which 200 parts/10⁶ application also produced inhibition ranging from 14% (accothion) to 23% (thimet) after 10 days. With lower concentrations of insecticide (50 parts/10⁶) inhibition, though again significant, was of a more ephemeral nature.All three insecticides, at a concentration of 1000 parts/10⁺⁶, prevented almost any hydrolysis of urea by jack bean urease. Ureolytic microorganisms, isolated from the soils under investigation, were inhibited by the organophosphates to a greater or lesser extent but the development of tolerance was common.It is suggested that the application of insecticides to control soil-borne insect pests may be a factor in determining the efficiency of urea fertilizer mineralization.     

Changes caused by Norway spruce in an ochreous brown earth, assessed by the isoquartz method

Previous investigations had shown that about 80 years of Norway spruce monoculture on an ochreous brown earth of the Belgian Ardennes were sufficient to accentuate the weak podzolization process already present in the climax beechwood soil and to increase the weathering of several minerals, e.g. 2:2 clay minerals. In order to evaluate more accurately mineral weathering and spruce-induced losses of nutrient elements, an isoquartz assessment method has been applied to a pair of soil profiles, one developed under beech, the other under spruce.
Results show a clear decrease of several weatherable minerals in the soil of the spruce stand which was more marked than in the beechwood, especially for chloritic minerals: losses reach about 30% of the initial chlorite content in the cambic horizon and regularly increase up to 70% in the humic layer. There were severe spruce-induced losses of Mg and Na: 27% of the initial Mg reserve was removed from the whole soil and the losses reached 60% in the upper 20 cm.
It is concluded that 86 years of Norway spruce monoculture have appreciably increased weathering processes and removal of elements.     

Development of an enzyme-linked immunosorbent assay for the insecticide imidacloprid

Enzyme-linked immunosorbent assays (ELISAs) were developed for imidacloprid, a neonicotinoid insecticide. Haptens were designed in such ways that spacer arms were introduced on either the pyridinyl or the imidazolidinyl ring of imidacloprid. Two sets of polyclonal antibodies were raised from rabbits immunized with two different immunogens and were characterized with an indirect ELISA format. Cross-reactivities and effects of organic solvents on the assays were evaluated. One set of antibodies shows approximately equal cross-reactivities to imidacloprid and its major metabolites with half-maximum inhibition concentrations (I(50)) of 73-88 ppb. Another is specific to imidacloprid with an I(50) of 35 ppb. The assay was initially applied to the analysis of imidacloprid in fortified water, coffee cherry, and bean extracts.     

Development of an enzyme-linked immunosorbent assay for the insecticide thiamethoxam

An enzyme-linked immunosorbent assay (ELISA) was developed for the neonicotinoid insecticide thiamethoxam, 3-(2-chlorothiazol-5-ylmethyl)-5-methyl-4-nitroimino-1,3,5-oxadiazinane. Three antisera were raised from rabbits immunized with the hapten-KLH conjugate. On the basis of the computational analysis of hapten candidates, the hapten with a spacer arm on the thiazolyl ring of thiamethoxam was synthesized to elicit thiamethoxam-specific antisera. The hapten was 3-[2-(2-carboxyethylthio)-5-ylmethyl]-5-methyl-4-nitroimino-1,3,5-oxadiazinane. Antisera were characterized with indirect competitive ELISA. Cross-reactivity and effects of organic solvents, pH, and ionic strengths were evaluated. The antiserum was specific for thiamethoxam and tolerant of up to 5% acetonitrile and 5% acetone. Various ionic strengths and pH values in the tested ranges had negligible effect on the assay performance. Under the optimized conditions, the half-maximal inhibition concentration (IC(50)) and the limit of detection were approximately 9.0 and 0.1 microg/L of thiamethoxam, respectively. ELISA analysis of stream and tap water samples showed an excellent correlation with the fortification levels.     

Development of an immunoassay for the pyrethroid insecticide esfenvalerate.

A competitive enzyme-linked immunosorbent assay was developed for the detection of the pyrethroid insecticide esfenvalerate. Two haptens containing amine or propanoic acid groups on the terminal aromatic ring of the fenvalerate molecule were synthesized and coupled to carrier proteins as immunogens. Five antisera were produced and screened against eight different coating antigens. The assay that had the least interference and was the most sensitive for esfenvalerate was optimized and characterized. The I(50) for esfenvalerate was 30 +/- 6.2 microg/L, and the lower detection limit (LDL) was 3.0 +/- 1.8 microg/L. The assay was very selective. Other pyrethroid analogues and esfenvalerate metabolites tested did not cross-react significantly in this assay. To increase the sensitivity of the overall method, a C(18) sorbent-based solid-phase extraction (SPE) was used for water matrix. With this SPE step, the LDL of the overall method for esfenvalerate was 0.1 microg/L in water samples.     

Cereal aphid control by generalist predators in presence of belowground alternative prey: Complementary predation as affected by prey density

Generalist predators are important antagonists of pest species in agroecosystems. Increasing populations of alternative prey through detrital subsidies is one way to foster these predators. However, alternative prey may also distract generalist predators from their prey thereby diminishing the efficiency of biological control. To develop reliable predictions for biological control, it is essential to evaluate the relative importance of generalist predators, pests, alternative prey and their respective interactions. We investigated the effects of an assemblage of generalist predators on the grain aphid Sitobion avenae in winter wheat. Treatments with 10, 100 and 1000 aphids were established inside 2 m² sized caged plots with control and reduced density of predators. Three weeks after the experiment started, samples were taken to estimate the size of aphid populations and those of alternative prey. Three prey taxa were significantly reduced by generalist predators: the grain aphid S. avenae (Aphidina), the click beetle Adrastus pallens (Elateridae) and the springtail Isotoma viridis (Collembola). Springtails were decreased by generalist predators independent of aphid densities, indicating complementary predation of springtails and aphids. At high aphid densities, grain aphid population peaks were decreased to the threshold level of economic damage, demonstrating efficient aphid suppression by the predator community. Click beetle numbers declined only at low and medium aphid densities. The results suggest that generalist predators preferentially fed on click beetles at low and medium aphid densities and switched to aphids at high aphid densities. Early-season predators likely had the greatest influence on aphid suppression. Our results indicate that alternative prey from the belowground system forms a substantial food resource for generalist predators, suggesting that the belowground subsystem modulates predator–prey interactions above the ground.     

Development of an enzyme-linked immunosorbent assay for the pyrethroid insecticide cyhalothrin

A competitive enzyme-linked immunosorbent assay (ELISA) was developed for detection of the pyrethroid insecticide cyhalothrin. Three haptens with an amine or propanoic acid terminus were synthesized and then conjugated with bovine serum albumin to give immunogens. Eight polyclonal antisera produced by rabbits were screened for titers and affinity using three different coating antigens. The antiserum CWB-C had the highest affinity with cyhalothrin and a low affinity with fenvalerate, fenpropathrin, deltamethrin, and fluvalinate. The half-maximum inhibition concentration for cyhalothrin was 37.2 microg/L, and the limit of detection was 4.7 microg/L. The recoveries of different concentrations of cyhalothrin (0.1-2500 microg/L) from fortified tap water, well water, and wastewater samples as determined with the ELISA were 81-114%.     

Causes of mortality at nests of ground-nesting birds in the Upper Waitaki Basin, South Island, New Zealand: a 5-year video study

We used video cameras over 5 years to quantify causes of mortality at 172 nests of three species of ground-nesting birds that nest on braided riverbeds of the Upper Waitaki Basin, South Island, New Zealand. The species were banded dotterels Charadrius bicinctus (n=114), black stilts Himantopus novaezelandiae (n=23), and black-fronted terns Sterna albostriata (n=35). Of 77 recorded lethal events (excluding four desertions caused by us), 66 involved deaths of only eggs, and 11 involved deaths of adults and/or chicks, and/or eggs. The main predators were cats Felis catus, hedgehogs Erinaceus europaeus, and ferrets Mustela furo, which were responsible for 43, 20, and 18% of lethal events, respectively. Cats were the only predator species to take adult birds. We recorded only two avian predations: a harrier Circus approximans took a chick and a hatching egg from one nest, and an Australian magpie Gymnorhina tibicen ate chicks at one nest. Other causes of mortality were incubating adult birds, floods, and sheep Ovis aries. Each accounted for <4% of lethal events. Ninety percent of visits (151 of 168) by predators or potential predators happened between sunset and sunrise. We found no evidence that video cameras or infra-red lighting influenced predation rates during 2 years of testing for such effects.     

Biotransformation of 3-methyl-4-nitrophenol, a main product of the insecticide fenitrothion, by Aspergillus niger

Biotransformation of the environmental pollutant 3-methyl-4-nitrophenol (MNP), a newly characterized estrogenic chemical, and the primary breakdown product of the heavily used insecticide fenitrothion was investigated using a common soil fungus. In 96 h, daily culture sacrifice, extraction, and analysis showed that the filamentous fungus, Aspergillus niger VKM F-1119, removed more than 85% of the MNP present in solution (original concentration = 25 mg/L), mostly through biodegradation. Additionally, in 16-day time-course studies, A. niger was capable of biotransformation of MNP at concentrations as high as 70 mg/L. Gas chromatography mass spectroscopy (MS) analyses of culture fluid extracts indicated the formation of four metabolites: 2-methyl-1,4-benzenediol, 4-amino-3-methylphenol, and two singly hydroxylated derivatives of MNP. Culture scale up and metabolite analysis by liquid chromatography MS resulted in the confirmation of the original metabolites plus the detection of an azo derivative metabolite that has not been previously reported before during MNP biodegradation by any micro-organisms.     

Modelling populations of long-lived birds of prey for conservation: A study of imperial eagles (Aquila heliaca) in Kazakhstan

Eagle populations worldwide are in decline and their demography is generally poorly understood. We use novel sensitivity analysis of stochastic simulation models to analyse the demography of the worlds highest-density and longest-studied population of eastern imperial eagles (Aquila heliaca), at the Naurzum National Nature Reserve in Kazakhstan. Single variable perturbation (a simple elasticity-type analysis) showed that population growth was most sensitive to changes in adult survival but provided no information on how interactions between parameters may influence population growth. Multiple-variable perturbations (a more comprehensive elasticity-type analysis) suggested that population growth is relatively more sensitive to adult survival than is indicated by single-variable perturbation but also that when adult survival is within a biologically reasonable range, other parameters are still highly consequential to model outputs. For Naurzum’s imperial eagles, and for other structured populations of vertebrates, effective conservation and management likely requires an approach that addresses the importance of simultaneous variation in multiple vital rates including both survivorship and reproductive output.     

Agricultural landscape modification increases the abundance of an important food resource: Mistletoes, birds and brigalow

Agricultural landuse alters landscape pattern, with important consequences for native species. Complex responses to changes in landscape pattern may be observed for species with strong inter-specific relationships. We investigate how the abundance of an ecologically important food resource, the grey mistletoe (Amyema quandang), is influenced by the interactive influences of human-modified landscape pattern, bird disperser abundance and host condition. Mistletoe abundance was estimated in brigalow (Acacia harpophylla) dominated remnants in three separate study areas with differing landscape patterns. Negative binomial generalised linear models of mistletoe abundance were tested with combinations of seven explanatory variables: study area, landscape connectivity and edge contrast, patch shape, bird disperser abundance, brigalow foliage cover and basal area of standing dead trees. Ranking of model averaged parameter estimates showed that study area, bird disperser abundance, patch shape, dead trees and connectivity had the strongest influence on A. quandang abundance. Mistletoe abundance apparently increased with landscape modification, particularly in narrow linear patches, but may be limited by the availability of seed dispersers and host condition. Given high landscape modification and ongoing degradation, management should be targeted towards maintaining brigalow remnant condition to ensure bird and mistletoe populations can be supported in the long-term.