Water-soluble metabolites of p-nitrophenol and 1-naphthyl N-methylcarbamate in flies and grass grubs. Formation of glucose phosphate and phosphate conjugates

Metabolites isolated from houseflies dosed with 1-napththol or p-nitrophenol were identified as the phosphate and glucose phosphate conjugates of these phenols by titrations, hydrolysis, ionophoresis, i.r. spectra and mixed melting point. [(3)H]Carbaryl (1-naphthyl N-methylcarbamate) was metabolized by houseflies, blowflies and grass grubs to water-soluble metabolites which had chromatographic and ionophoretic behaviour similar to those of the conjugates of 1-naphthol with glucose, sulphate, phosphate and glucose 6-phosphate.     

The conjugation of phenols with phosphate in grass grubs and flies

1. Houseflies, blowflies and New Zealand grass grubs were dosed with 1-naphthol, 2-naphthol or p-nitrophenol. 2. The corresponding monoaryl phosphates were identified in extracts of insects or excreta along with the beta-glucosides and ethereal sulphates of the phenols. 3. No diaryl phosphates or glucosiduronates were detected but an unidentified metabolite of [(14)C]1-naphthol was present in extracts of flies dosed with [(14)C]1-naphthol.     

On the role of pentachlorocyclohexene in the metabolism and action of hexachlorocyclohexane. I. Synthesis of beta-pentachlorocyclohexene and its identification as the monodehydrochlorination product of alpha-hexachlorocyclohexane (author's transl)]

It is the aim of a series of investigations to test whether or not beta-pentachloro-1-cyclohexene is an intermediate in the biodegradation of alpha-hexachlorocyclohexane. This paper describes attempts to synthesize this intermediate by chemical methods. 1) Pentachlorocyclohexene was synthesized by partial additive chlorination of chlorobenzene. Combined gas chromatography-mass spectrometry revealed that at least five different isomers of pentachlorocyclohexene had been formed. 2) Treatment of alpha-hexachlorocyclohexane with alkaline buffer (pH 8) produced trichlorobenzenes and, in small yield (4%), a pentachlorocyclohexene. This was isolated and identified as the beta-isomer by melting point (71.8 - 72.6 degrees C, uncorr.), IR- and mass spectrum. Dehydrochlorination of beta-pentachlorocyclohexene produced the trichlorobenzene isomers in a pattern which is characteristic of alpha-hexachlorocyclohexane. The position of the chlorine substituents in the beta-pentachlorocyclohexene molecule as judged from NMR studies is e-aeee. This confirms that it is the monodehydrochlorination product of alpha-hexachlorocyclohexane. The configurations of gamma- and delta-pentachlorocyclohexene, determined for comparison, are e-eeaa and e-eeee, respectively. The kinetics of dehydrochlorination of both alpha-hexachlorocyclohexane and beta-pentachlorocyclohexene in alkaline acetone/water (3 + 2) was studied by means of conductometry. Both reactions are of second order: kappa alpha-HCH 0.0495 [1 times mol- minus 1 times s- minus 1[; kappa beta-PCH 0.905 [1 times mol- minus 1 times s- minus 1] (3.6 degrees C). 3) Dehydrochlorination of alpha-hexachlorocyclohexane in pyridine/xylene (3 + 4) was also studied. An earlier report claiming that gamma-pentachlorocyclohexene (and not the beta isomer) is produced in this medium was confirmed, if the reaction was performed at high temperature (120 - 140 degrees C). Moreover, the ratio of trichlorobenzene isomers formed from alpha-hexachlorocyclohexane shifted to a pattern characteristic of the gamma (or gamma) isomer. However, at temperatures of 90 degrees C or less, beta-pentachlorocyclohexene was the main product. The results strongly suggest that in pyridine/xylene, the same isomer is primarily produced from alpha-hexachlorocyclohexane and is isomerized to the gamma, delta and at least two other isomers of pentachlorocyclohexene before further dehydrochlorination ensues. A simple method for the synthesis of beta-pentachlorocyclohexene is presented.     

Degree of plant root digestion by the larva of the beetle, Costelytra zealandica

The degree of plant root digestion by grass grubs (Costelytra zealandica) has been evaluated by scanning electron microscope examination of plant residues in faeces. Faecal pellets were examined from grubs fed on roots of pure plant species (perennial ryegrass, Lolium perenne, and white clover, Trifolium repens) under soil-free conditions, as well as from naturally occurring grubs on mixed pasture. There was considerable variation in the degree of digestion observed with different plant species. Clover root was more extensively digested with formation of partially digested xylem vessels, whereas ryegrass root appeared to undergo very little digestive breakdown. These results indicate that plant structural carbohydrates undergo limited digestion by the grass grub. Thus grass grubs may require a high turnover of dietary root material, which would explain the highly destructive effect of the larvae on pasture plants.     

Comparison of scarab grub populations and associated pathogens and parasitoids in warm- or cool-season grasses used on transitional zone golf courses

Seven different turfgrass species or mixes used on golf courses in the United States' transitional climatic zone were maintained as randomized and replicated plots in separate stands mowed at fairway (1.6 cm) or rough (6.4 cm) cutting heights and sampled in autumn to assess the density and species composition of scarab grubs; incidence of disease and parasitism thereof; and extent of turf damage from foraging insectivorous skunks, Mephitis mephitis. Influence of grass species on parasitism by spring or autumn-active tiphiid wasps was further assessed on implanted grubs in open enclosures. Masked chafers (Cyclocephala spp.) were three-fold more abundant than Japanese beetle, Popillia japonica Newman, grubs in plots of Zoysia and Cynodon sp. mowed at fairway height, and P. japonica were five-fold more abundant than masked chafer grubs in cool-season turf plots mowed at rough height. Phyllophaga spp. accounted for <1% of grubs in the samples. Milky disease bacteria (Paenibacillus sp.) were the predominant pathogens of Cyclocephala spp., followed by Serratia sp. bacteria and gregarines (Stictospora cf. villani). Cyclocephala grub densities, milky disease incidence (25%), and parasitism by the native tiphiid Tiphia pygidialis Alien (10-12%) were especially high in zoysiagrass. Japanese beetle grubs were infected by Paenibacillus, Serratia, Stictospora, and microsporidia (Ovavesicula sp.), but incidence of individual pathogens was relatively low (<6%) and similar among grasses within each stand. Few nematode-infected grubs were found. Skunk damage was mainly in the cool-season fairway-height grasses, probably reflecting difficulty in foraging in the much tougher stolons and rhizomes of the warm season turfgrasses. The degree of natural suppression of scarab grubs provided by endemic pathogens or parasitoids is unlikely to be compromised by the grass species used on a particular site.     

Synergism of imidacloprid and entomopathogenic nematodes against white grubs: the mechanism

Entomopathogenic nematodes and the chloronicotinyl insecticide, imidacloprid, interact synergistically on the mortality of third-instar white grubs (Coleoptera: Scarabaeidae). The degree of interaction, however, varies with nematode species, being synergistic for Steinernema glaseri (Steiner) and Heterorhabditis bacteriophora Poinar, but only additive for Steinernema kushidai Mamiya. The mechanism of the interaction between imidacloprid and these three entomopathogenic nematodes was studied in the laboratory. In vials with soil and grass, mortality, speed of kill, and nematode establishment were negatively affected by imidacloprid with S. kushidai but positively affected with S. glaseri and H. bacteriophora. In all other experiments, imidacloprid had a similar effect for all three nematode species on various factors important for the successful nematode infection in white grubs. Nematode attraction to grubs was not affected by imidacloprid treatment of the grubs. Establishment of intra-hemocoelically injected nematodes was always higher in imidacloprid-treated grubs but the differences were small and in most cases not significant. The major factor responsible for synergistic interactions between imidacloprid and entomopathogenic nematodes appears to be the general disruption of normal nerve function due to imidacloprid resulting in drastically reduced activity of the grubs. This sluggishness facilitates host attachment of infective juvenile nematodes. Grooming and evasive behavior in response to nematode attack was also reduced in imidacloprid-treated grubs. The degree to which different white grub species responded to entomopathogenic nematode attack varied considerably. Untreated Popillia japonica Newman (Coleoptera: Scarabaeidae) grubs were the most responsive to nematode attack among the species tested. Untreated Cyclocephala borealis Arrow (Coleoptera: Scarabaeidae) grubs showed a weaker grooming and no evasion response, and untreated C. hirta LeConte (Coleoptera: Scarabaeidae) grubs showed no significant response. Chewing/biting behavior was significantly increased in the presence of nematodes in untreated P. japonica and C. borealis but not in C. hirta and imidacloprid-treated P. japonica and C. borealis. Our observations, however, did not provide an explanation for the lack of synergism between imidacloprid and S. kushidai.     

Disturbance intensity and above- and belowground herbivory effects on long-term (14 y) recovery of a semiarid grassland

The importance of disturbance intensity and herbivory by cattle and white grubs, or the larvae of June beetles (including Phyllophaga fimbripes), to recovery of shortgrass steppe ecosystems in Colorado, U.S.A. were evaluated over a fourteen year time period. Disturbance intensity was defined by survival of the dominant grass species (Bouteloua gracilis) after an outbreak of root feeding activity by white grubs. Sixteen patches of vegetation consisting of four pairs of adjacent ungrazed-grazed by cattle locations with two replicates that were recently affected by white grubs were selected in 1977. Disturbance intensity was determined in 1977 by the area in each patch that contained live tillers of B. gracilis. Permanent plots were located both within and outside of each patch. Plant basal cover and density by species were estimated at time of peak aboveground biomass in six different years on each plot.Successional dynamics on patches was similar to areas affected by other types of disturbances, however, rate of recovery was faster for patches affected by grubs. Grazing by cattle was infrequently important to plant recovery, a result similar to effects of grazing on other aspects of shortgrass steppe ecosystems. Disturbance intensity was important to recovery of B. gracilis since tiller survival in 1977 was linearly related to cover in each year of sampling. For ungrazed patches, initial conditions were important to recovery of B. gracilis for as many as 14 years. For grazed patches, initial conditions decreased and grazing increased in importance through time. Changes in resource quality and a more uniform distribution of roots due to grazing likely resulted in more complete mortality of plants by grubs under grazed compared to ungrazed conditions.Persistence of shortgrass ecosystems in spite of disturbances with different intensities are determined at least in part by characteristics of disturbances interacting with the ability of plants to respond, and in part by the evolutionary history of the system. Although white grubs affect shortgrass communities infrequently, they have large and important effects on plant community structure through time, and represent an important class of disturbance defined by intensity.     

Long‐term variation in environmental conditions influences host–parasite fitness

Long‐term data on host and parasite fitness are important for predicting how host–parasite interactions will be altered in an era of global change. Here, we use data collected from 1997 to 2013 to explore effects of changing environmental conditions on bird–blowfly interactions in northern New Mexico. The objectives of this study were to examine what climate variables influence blowfly prevalence and intensity and to determine whether blowflies and climate variables affect bird fledging success. We examined how temperature, precipitation, and drought affect two parasitic blowflies and their hosts, Western Bluebirds (Sialia mexicana) and Ash‐throated Flycatchers (Myiarchus cinerascens). We found that blowfly prevalence did not change over time. Blowfly intensity increased over time in bluebird nests, but not in flycatcher nests. More blowflies result in slightly higher fledging success in bluebirds, but not flycatchers. There was a significant interaction between blowflies and precipitation on bluebird fledging success. For flycatchers, there was a significant interaction between blowflies and temperature and between blowflies and drought severity on fledging success. Given that the southwest is projected to be hotter and have more frequent and prolonged droughts, we predict that flycatchers may be negatively impacted by blowflies if these trends continue. Future work should focus on investigating the role of both blowflies and climate on fledging success. Climate patterns may negatively impact host fitness through altered parasite pressure.     

Impact of four insecticides on Japanese beetle (Coleoptera: Scarabaeidae) egg predators and white grubs in turfgrass

Field experiments were conducted to measure the effects of four commonly used turfgrass insecticides (isofenphos, diazinon, imidacloprid, halofenozide) on white grubs (Coleoptera: Scarabaeidae) and ant predators of white grub eggs. Ant populations were measured over time with canned tuna, whereas predation by the ants was measured with artificially placed Japanese beetle, Popillia japonica Newman, eggs. The effectiveness of each insecticide at controlling Japanese beetle grubs, when applied at different times during the growing season, also was measured. Isofenphos and diazinon significantly reduced both ant numbers and white grub egg predation, whereas imidacloprid and one halofenozide treatment did not significantly impact either measurement. A second halofenozide treatment significantly reduced white grub egg predation. Isofenphos and diazinon were ineffective at controlling Japanese beetle grubs when applied in June but were highly efficacious when applied in August. Evidence of enhanced biodegradation was found in plots that received both June and August applications of diazinon. Both June and August applications of imidacloprid and halofenozide provided good control of white grubs.     

Species differences in the inhibition of glutathione S-aryltransferase by phthaleins and dicarboxylic acids

1. Glutathione-S-aryltransferase activity from grass grubs (Costelytra zealandica) was inhibited by phthaleins, sulphonphthaleins and some dicarboxylic acids. 2. These compounds had no detectable action on the enzyme from sheep liver. 3. In insect enzyme the inhibition was competitive with respect to glutathione and non-competitive with respect to the aromatic substrate. 4. Michaelis constants and inhibitor constants were measured for sheep-liver or grass-grub enzyme between pH5 and pH10 and evidence was obtained for the presence of two groups with pK9.2 in the glutathione-binding site of the insect enzyme. 5. Only one such group was detected in the sheep-liver enzyme.     

Recovery of intravenously infused chromium EDTA and lithium sulphate in the urine of cattle and their use as markers to measure urine volume

A series of metabolism experiments investigated the recovery of continuous-, intravenously infused chromium complexed with ethylenediamine tetra-acetic acid (CrEDTA) and lithium sulphate in the urine of cattle with a view to using the markers to estimate urine and metabolite output in grazing cattle. The recovery of Cr in urine from these infusions was similar (90%) in metabolism trials when cattle consumed three very contrasting diets: high-grain formulated pellet, lucerne hay (Medicago sativa) or low-quality native grass hay (predominantly Heteropogon contortus). By contrast, Li recovery in urine averaged 46.3 ± 0.40% and 72.6 ± 0.43% for native pasture and lucerne hays, respectively, but was not constant across days. There was negligible transfer of Cr from CrEDTA in blood serum to the rumen or faeces, whereas appreciable quantities of infused Li were found in both. The ratio of urine volume estimated by spot samples and marker dilution of Cr, to urine volume measured gravimetrically, was 1.05. In grazing studies using rumen-fistulated (RF) steers grazing seven different tropical and temperate grass and legume pastures, the ratio of concentrations of purine derivatives (PD) to Cr in spot samples of urine was shown to vary diurnally in the range of 49% to 157% of the average 24 h value. This finding indicated the need for regular sampling of urine to achieve an accurate average value for the PD : Cr ratio in urine for use in estimating urinary PD excretion and hence microbial protein production in the rumen. It was concluded that continuous, intravenous infusion of CrEDTA resulted in a constant recovery of Cr in the urine of cattle across diets and, provided an intensive sampling regime was followed to account for diurnal variation, it would be suitable as a marker to estimate urine volume and urinary output of PD in grazing cattle.     

Effect of silage type and concentrate level on conjugated linoleic acids, trans-C18:1 isomers and fat content in milk from dairy cows

The objective of the study was to examine how the fatty acid composition of milk especially concentrations of conjugated linoleic acids (CLA) and trans-C18:1 isomers and milk fat percentage were affected by silage type and concentrate level. Forty dairy cows were blocked and randomly assigned to one of four diets in a 2 x 2 factorial arrangement of treatments and a six week experimental period. Treatments were total mixed rations with maize (M) or grass (G) silage differing in polyunsaturated fatty acid (PUFA) profile and starch content, combined with a high (H) or a low (L) level of concentrate (with or without grain). Treatments had no significant effect on milk, protein and lactose yield, but energy corrected milk yield, milk fat percentage and fat yield was lower and protein percentage higher for maize compared with grass silage diets. Overall, maize silage diets resulted in higher concentrations of CLA isomers compared with grass silage diets, but there was a significant interaction between silage type and concentrate level for concentrations of cis9,trans11-CLA; trans10,cis12-CLA; trans11-C18:1 and trans10-C18:1. A high level of concentrate increased trans10,cis12-CLA and trans10-C18:1 and reduced cis9,trans11-CLA and trans11-C18:1 when maize but not grass silage was provided. The results suggest that high levels of concentrate (grain) do not significantly alter the pattern of PUFA biohydrogenation in the rumen, the concentration of CLA and trans-C18:1 isomers in milk or cause milk fat depression unless combined with forage naturally high in starch and C18:2n-6 such as maize silage.     

Reversible activation of the neutrophil superoxide generating system by hexachlorocyclohexane: correlation with effects on a subcellular superoxide-generating fraction

gamma-Hexachlorocyclohexane was found to exert profound effects on the phosphatidylinositol cycle, cytosolic calcium level, and the respiratory burst of human neutrophils. Exposure of neutrophils prelabelled with 32P to 4 X 10(-4) M gamma-hexachlorocyclohexane almost tripled radioactivity in phosphatidic acid and correspondingly decreased radioactivity in phosphatidylinositol 4,5 bisphosphate. Under similar conditions, gamma-hexachlorocyclohexane evoked the generation of superoxide at a rate of over 11 nmol/min/10(6) cells and more than doubled cytosolic-free calcium concentration as monitored by Quin-2 fluorescence. Because intermediates of the phosphatidylinositol cycle, via increases in available calcium levels or activated protein kinase C, are considered potential second messengers for activation of the NADPH-dependent O-2-generating system, we compared neutrophil responses to gamma-hexachlorocyclohexane with responses to phorbol myristate acetate, an activator of protein kinase C with well known effects on neutrophils. Like phorbol myristate acetate, gamma-hexachlorocyclohexane induced neutrophil degranulation but was not an effective chemotactic stimulus. The ability of gamma-hexachlorocyclohexane to induce a pattern of oxidative activation in neutrophil cytoplasts similar to that in intact cells indicated that concurrent degranulation was not required for sustained O-2 generation in response to this agent. When neutrophils or neutrophil cytoplasts exposed to gamma-hexachlorocyclohexane were centrifuged and resuspended in stimulus-free medium, O-2 generation ceased entirely but could be reinitiated by addition of the same stimulus. This finding was in contrast to the continued O-2 production by phorbol myristate acetate-stimulated neutrophils similarly washed and resuspended in stimulus-free medium. Unlike subcellular fractions of phorbol myristate acetate-stimulated neutrophils, corresponding fractions prepared from gamma-hexachlorocyclohexane-stimulated neutrophils contained almost no detectable NADPH-dependent O-2-generating activity. Subcellular oxidase activity was not recovered when cells and membrane fractions were continuously exposed to gamma-hexachlorocyclohexane during disruption and fractionation after cell stimulation, nor could it be induced by the addition of the stimulus to the subcellular fractions. Thus, the stimulus dependence of continuous neutrophil superoxide release evoked by gamma-hexachlorocyclohexane does not merely reflect a physical interaction of the agonist with the enzyme system involved.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effects of hexachlorocyclohexane isomers on concanavalin A 'capping' in bovine lymphocytes.

The effects of alpha, beta, gamma, and delta isomers of hexachlorocyclohexane on concanavalin A 'capping' in bovine lymphocytes were evaluated, gamma and delta hexochlorocyclohexane inhibited 'capping' whereas the alpha and beta isomers were without effect. In addition, gamma-hexachlorocyclohexane has been shown to antagonize the maintenance of preformed 'caps' and cause the rapid dispersal of the concanavalin A-receptor complexes over the surface of cells by a temperature-dependent mechanism. The possible role of a gamma-hexachlorocyclohexane-sensitive process in the organization of microflow patterns in the lectin-activated lymphocyte membrane is discussed.     

The metabolism of γ-2,3,4,5,6-pentachlorocyclohex-1-ene and γ-hexachlorocyclohexane in rats

1. After intraperitoneal administration, gamma-hexachlorocyclohexane (Gammexane) and gamma-2,3,4,5,6-pentachlorocyclohex-1-ene were converted by rats into 2,3,5- and 2,4,5-trichlorophenol, which were excreted as free phenols and as sulphuric acid and glucuronic acid conjugates. 2. Derivatives of 2,4,5-trichlorophenol and 2,4,5-trichlorophenyl glucosiduronic acid and 2,4-dichlorophenylmercapturic acid were isolated from the urine as metabolites of gamma-2,3,4,5,6-pentachlorocyclohex-1-ene. 3. The phenolic metabolites of gamma-hexachlorocyclohexane and gamma-2,3,4,5,6-pentachlorocyclohex-1-ene isolated from urine were similar to those of 1,2,4-trichlorobenzene, which indicates that the two latter compounds are intermediates in gamma-hexachlorocyclohexane metabolism in rats.     

Cross-specificity in some vertebrate and insect glutathione-transferases with methyl parathion (dimethyl p-nitrophenyl phosphorothionate), 1-chloro-2,4-dinitrobenzene and S-crotonyl-N-acetylcysteamine as substrates

1. Enzymes catalysing the reaction between GSH and methylparathion (dimethyl p-nitrophenyl phosphorothionate), 1-chloro-2,4-dinitrobenzene and S-crotonyl-N-acetylcysteamine were separated by (NH(4))(2)SO(4) precipitation from homogenates of sheep, rat and mouse livers and from homogenates of cockroaches, houseflies and grass grubs. 2. Electrofocusing of the preparations from each of these species separated a number of zones, each of which catalysed the reaction of GSH with all three substrates. 3. Ion-exchange chromatography on CM-cellulose also separated a number of fractions in which activity towards the three substrates coincided. 4. In both separation methods patterns of the activities were consistent with the presence in all species of several GSH transferases each having a degree of cross specificity towards the three substrates.     

Sulpiride (stereoisomers, racemic) and dopamine: actions and interactions on tubular reabsorption

In DOCA-pretreated men during hypotonic polyuria Dopamine (DA) infusion in a subpressor dosage (0.1 microgram X kg -1 X min -1) produces renal hyperemia, inhibition of isosmotic sodium reabsorption, increase in sodium distal load (s. d.l.) and finally inhibition of anisosmotic sodium reabsorption as a percentage of s.d.l. Such DA effects are variously modified by sulpiride isomers. Sodium distal load increase is still apparent in the presence of racemic, but not in the presence of either d- or l-Sulpiride isomers. However DA inhibition of anisosmotic sodium reabsorption % s.d.l. is unaffected by either or both Sulpiride isomers. If the DA tubular inhibition was dependent on specific DA receptors these receptors, unlike vascular DA receptors, would bind in a weaker way both d- and l-Sulpiride isomers.     

Influence of dietary hexachlorocyclohexane isomers on lipid metabolism in albino rats

Dietary intake of beta- and gamma-hexachlorocyclohexane (beta- and gamma-HCH) by albino rats for two weeks (at 800 ppm level) resulted in impairment of lipid metabolism, viz. hyperlipemia and fatty metamorphosis of liver. Liver fat content was increased by both beta- and gamma-HCH. Significant increases were observed in triglyceride and phospholipid fractions of blood in these experimental animals. The incorporation of [14C]acetate and palmitate into liver and blood lipids was higher in HCH pretreated animals, suggesting a higher rate of fat synthesis in liver and of secretion as well.     

The metabolism of 3,5-di-tert.-butylphenyl N-methylcarbamate in insects and by mouse liver enzymes

The oxidation of 3,5-di-tert.-butylphenyl N-methylcarbamate (Butacarb) has been studied in the flies Musca domestica and Lucilia sericata, grass grubs Costelytra zealandica and the mouse. In all species eleven oxidation products, which were formed by hydroxylation of the tert.-butyl groups and the N-methyl group, were detected.     

c-fos Expression as a Model for Studying the Action of Hexachlorocyclohexane Isomers in the CNS

The induction of protooncogene c-fos in the CNS after administration of several convulsants has been studied. The organochlorine insecticide gamma-hexachlorocyclohexane (gamma-HCH, lindane) has been shown to induce c-fos expression in different brain areas. Pentylenetetrazole and picrotoxin, a known gamma-aminobutyric acid-receptor antagonist, have also been considered. The administration of two nonconvulsant isomers of gamma-HCH, alpha-HCH, and delta-HCH, before the mentioned toxicants, affects the protooncogene expression in different ways. The differential pattern of expression displayed by c-fos after these treatments suggests the presence of diverse mechanisms of action for the compounds studied.