Metabolism of furametpyr. 2. (14)C excretion, (14)C concentrations in tissues, and amounts of metabolites in rats

14C-Labeled furametpyr [N-(1,3-dihydro-1,1, 3-trimethylisobenzofuran-4-yl)-5-chloro-1, 3-dimethylpyrazole-4-carboxamide, Limber] was dosed to male and female rats at 1 (low dose) and 200 or 300 mg/kg (high dose). Elimination of furametpyr was rapid, and the dosed (14)C was substantially excreted within 7 days (45.5-53.3% in feces, 44.1-53. 8% in urine, and 0.01% in expired air). However, (14)C excretion rate showed sex- and dose-related differences, more rapid in males at low dose. (14)C concentrations in tissues decreased rapidly to generally low levels at 7 days (<0.004 ppm with the low dose and <1. 1 ppm with the high dose). Forty metabolites were detected, and 13 metabolites and 4 glucuronides were identified. A small amount of unchanged furametpyr was detected in feces (0.1-0.5% of the dose). The major metabolites in tissues were N-demethylated metabolites. In a bile study, 52.5-54.2% of the dosed (14)C was rapidly excreted into bile within 2 days. The absorption ratio was estimated to be >93.7% for the low dose (1 mg/kg). Major metabolites in bile were glucuronic acid conjugates of furametpyr hydroxides. On the basis of the results, furametpyr is substantially absorbed from the gastrointestinal tract after oral administration, rapidly distributed to tissues, extensively metabolized, and excreted into urine and bile or feces.     

In vitro metabolism of trans-permethrin and its major metabolites, PBalc and PBacid, in humans

To estimate the metabolic profile of trans-permethrin in humans, a comparison of the in vitro metabolism of trans-permethrin in humans and rats was conducted using hepatic microsomes, and cytochrome P450 and UDP-glucuronyltransferase isoforms, which catalyze the metabolism of 3-phenoxybenzyl alcohol (PBalc) and 3-phenoxybenzoic acid (PBacid), respectively. In humans and rats, the major metabolic reaction of trans-permethrin in microsomal incubations was the cleavage of ester linkage to give PBalc, followed by oxidation to 4'-OH-PBalc, 4'-OH-PBacid, and PBacid. As to 4'-hydroxylation of PBalc, several CYPs were able to catalyze the reaction, and CYP2E1 was identified as a predominant isoform. PBacid and its conjugates (glucuronide and glycine) are major urinary metabolites of trans-permethrin in mammals. PBacid is also a metabolite of several pyrethroids, and has been used as a biomarker of human exposure to pyrethroids. Our study indicated that there was no difference in glucuronyltransferase activity of PBacid between humans and rats, and that only UGT1A9 can catalyze the glucuronidation of PBacid among human UGTs. Some UGT1A9 variants are known to have poor glucuronidation activity. From these results, it was assumed that deficiency or polymorphism of UGT1A9 might affect the profile of PBacid and its conjugates in urine collected from persons exposed to trans-permethrin or other pyrethroids. These results are helpful for understanding the metabolism of trans-permethrin in humans and determining methods for quantification of target analytes for assessment of human exposure to trans-permethrin and other pyrethroids that give PBacid and its conjugates as urinary metabolites.     

Development of green onion and cabbage certified reference materials for quantification of organophosphorus and pyrethroid pesticides

Green onion and cabbage certified reference materials for the analysis of pesticide residues were issued by the National Metrology Institute of Japan, part of the National Institute of Advanced Industrial Science and Technology. Green onion and cabbage samples were grown so as to contain several kinds of organophosphorus and pyrethroid pesticides, and those were collected from a field in the Kochi Prefecture in Japan. The certification was carried out by using multiple analytical methods to ensure the reliability of analytical results; the values of target pesticides (diazinon, fenitrothion, cypermethrin, etofenprox, and permethrin for green onion and chlorpyrifos, fenitrothion, and permethrin for cabbage) were obtained by isotope dilution mass spectrometry. Certified values of target pesticides were 0.96-13.9 and 2.41-6.9 mg/kg for green onion and cabbage, respectively. These are the first green onion and cabbage powder certified reference materials in which organophosphorus and pyrethroid pesticides are determined.     

Pyrethroids: mammalian metabolism and toxicity

Synthetic pyrethroids, a major insecticide group, are used worldwide to control agricultural and household pests. Mammalian metabolism of pyrethroids was substantially launched in the 1960s and 1970s by the research groups of Professor Casida and Sumitomo Chemical Co., which made great contributions to the elucidation of their metabolic fates. They showed that ester hydrolysis and oxidation play predominant roles in mammalian metabolism of pyrethroids and that rapid metabolism leads to low mammalian toxicity. These metabolic reactions are mediated by carboxylesterases and CYP isoforms, the resultant metabolites then undergoing various conjugation reactions. In general, there are substantially neither significant species differences in metabolic reactions of pyrethoids nor metabolic differences among their chiral isomers except with fenvalerate, one isomer of which yields a lipophilic conjugate causing toxicity.     

Fine-root dynamics in a young hinoki cypress (<Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>) stand for 3 years following thinning

Fine roots play a key role in carbon and nutrient dynamics in forested ecosystems. Fine-root dynamics can be significantly affected by forest management practices such as thinning, but research on this topic is limited. This study examined dynamics of fine roots <1 mm in diameter in a 10-year-old stand of hinoki cypress (Chamaecyparis obtusa) for 3 years following thinning (65% in basal area). Fine-root production and mortality rates were estimated using a minirhizotron technique in combination with soil coring. In both thinned and un-thinned control plots, fine-root elongation occurred from early spring to winter (March to December) and fluctuated seasonally. In the thinned and the control plots, the annual fine-root production rates were estimated to be 101 and 120 g m⁻² year⁻¹, respectively, whereas the estimated annual fine-root mortality rates were 77 and 69 g m⁻² year⁻¹, respectively. At 3 years after thinning, live fine-root biomass was significantly smaller in the thinned plot (143 g m⁻²) than in the control plot (218 g m⁻²), whereas dead fine-root biomass was not (147 and 103 g m⁻², respectively). Morphological and physiological indices of fine roots such as diameter, specific root length, and root tissue density of the live fine roots was similar in both plots. These results suggested that thinning tended to decrease biomass and production of fine roots, but the effects on characteristics of fine roots would be less evident.     

Salivary mucocele in a laboratory beagle

The histologic characteristics of a salivary mucocele in a beagle used in a toxicity study are described in this report. A pale yellowish cyst under the mandibular skin containing frothy mucus was observed at necropsy. Microscopically, numerous villous projections arose from the internal surface of the cyst and were lined by stratified epithelial-like macrophages, which were immunopositive for macrophage scavenger receptor A. A ruptured sublingual interlobar duct connected to the lumen was observed near the cyst. Luminal amorphous material showed a positive reaction with Alcian blue and periodic acid-Schiff staining as did mucin in the sublingual gland. Ultrastructurally, the epithelial-like macrophages had numerous vacuoles containing electron-lucent material, which was presumed to be lysosomal in origin, and had pseudopods on their cell surfaces interdigitating with those on the adjacent cells. This case report helps to understand the diversity of the background findings in beagles used in toxicity studies.     

Prevalence of hemoplasma infection among cattle in the western part of Japan

We have examined for hemoplasma infection among cattle in the Hiroshima and Miyazaki prefectures by using a sensitive real-time PCR, with SYBR Green I and with melting curve analysis, which allow to distinguish the two bovine hemoplasma species, Mycoplasma wenyonii and 'Candidatus M. haemobos'. We found 69.4% of 36 cattle in Hiroshima and 93.8% of 32 cattle in Miyazaki infected with either of these two hemoplasma species. High morbidity in western part of Japan may reflect the activity of arthropod vectors for hemoplasma transmission. We also demonstrated neonatal calves less than three months old affected with hemoplasmas without grazing in summer, suggesting a possibility of vertical transmission.     

Early-stage impacts of sika deer on structure and function of the soil microbial food webs in a temperate forest: A large-scale experiment

Modification of forest vegetation caused by an overabundance of mammalian herbivores has been reported in temperate and subarctic regions all over the world. However, the indirect effects of these herbivores on the structure and functions of soil decomposer systems are not fully understood, especially in temperate forests. We investigated the early effects of sika deer invasion on soil decomposer systems in a Japanese temperate forest using two large-scale experimental enclosures with low and high densities of deer (LD: 25 ha, 4 deer km⁻²; HD: 6.25 ha, 16 deer km⁻²) including control plots without deer (WD). Three years after deer introduction the understory cover of dwarf bamboo (Sasa nipponica) declined due to deer browsing in both enclosures. At the same time, measurements were made of the soil microbial community, soil nematode community, soil nitrogen (N) mineralization rate, and carbon (C) and N content in dwarf bamboo leaves. In LD, soil microbial biomass was lower from WD, probably due to the decrease of fresh aboveground litter from dwarf bamboo. Surprisingly, there were no cascading effects on total abundance of soil nematodes and soil total N mineralization potential which were unaffected by deer in the LD treatment, while soil NH₄⁺-N content was lower and soil nematode community structure was different (abundance of 4 families was higher and that of 3 families was lower, but the functional structure was not different) from WD. Specifically, the responses to deer introduction varied between microbes and nematodes, and the change of balance in the microbial food webs may have altered N mineralization processes. In contrast, in the HD treatment, all the variables measured were not significantly different from those of WD treatment. Intensive browsing by deer may have cancelled out the effects of the decrease in aboveground litter input on the soil decomposer systems through other pathways, such as a transitory increase in belowground litter input caused by induced changes in allocation patterns of bamboo. No changes in total N mineralization potential, leaf N, and composition of understory vegetation in both enclosures indicated that deer introduction did not facilitate nor retard N cycling regardless of deer density. This study showed that sika deer browsing can affect soil decomposer systems at an early stage of invasion even at low density, which contrasts with previous studies on the subject. Linking our findings of early-stage effects of deer on soil decomposer systems to longer-term dynamics of understory vegetation and tree regeneration will be needed to evaluate the adequacy of deer management practices with respect to the sustainability of soil nutrient supplies.     

A Root Box Method to Estimate Virulence of Helicobasidium mompa Using Carrots and Its Comparison with the Conventional Method Using Apple Stocks

A root box method with carrots was developed to estimate virulence of the violet root rot fungus, Helicobasidium mompa, to facilitate short-term screening of many isolates during a year. The root box consisted of two transparent acrylic plates and a plastic bag of vermiculite in which two taproots of carrot were growing and inoculated with the fungus growing on fragments of mulberry twigs. The boxes were kept in a greenhouse at 25°C, and the surface of carrots was observed weekly up to 14 weeks. The virulence of each isolate was determined based on the number of weeks after inoculation required for the fungus to develop infection cushions on the surface of carrots. Results were compared with those from the conventional inoculation method using apple stocks. Two-year-old 456 apple stocks were planted with or without fungal inoculum in 30-cm diam. plastic pots containing commercial soil and placed outdoors in April 1999. Symptoms on plant tops were observed weekly, and the first stocks were killed 14 weeks after inoculation. At the end of trial 1 (6 months) and trial 2 (14 months), apple stocks were dug up to rate disease index (DI) based on hyphal growth and infection cushion formation on the stem base. There was variability in disease severity among replicates as well as isolate variability ; however, the results were similar in both trails. The level of virulence estimated by both methods was almost parallel for a total of 23 isolates from five plant species, except for two isolates from sweet potato that formed no obvious infection cushion on apple roots but on carrot were the most virulent. Received 11 December 2000/ Accepted in revised form 23 March 2001