Measurement of acephate and its principal metabolite in leaves by gas-liquid chromatography

A semi-micro method is described for monitoring levels of acephate (O, S-dimethyl acetylphosphoramidothioate) and its principal metabolite, methamidophos (O, S-dimethyl phosphoramidothioate) in leaf material derived from treated, hydroponically grown gerberas (Gerbera jamesonii H. Bollus, ex Hook. f.) To minimise injury to the living leaf, analyses were performed on 13-mm diameter discs punched from the interveinal regions of the lamina. The leaf disc samples were then extracted with ethyl acetate. Clean-up of extracts was accomplished by passing the filtered leaf extracts through glass mini-columns containing carbon + filter aid (2 + 5 by mass). Acephate and methamidophos were eluted from the column with methanol and measured simultaneously by programmed temperature gas chromatography using a thermionic specific detector.     

Insecticide resistance patterns among colorado potato beetle,leptinotarsa decemlineata (Say) (Coleoptera: chrysomelidae), populations in north carolina

A residual bioassay was used to determine insecticide susceptibility in populations of Colarado potato beetle, Leptinotarsa decemlineata (Say), in eastern North Carolina. Mean bioassay mortalities were subjected to correlation analysis to assess patterns of resistance among tested insecticides. Within insecticide classes, cross-resistance patterns were observed between carbaryl and carbofuran (carbamates), fenvalerate and permethrin (pyrethroids), and among azinphosmethyl, methamidophos, and phosmet (organophosphates). Significant associations between insecticides from different insecticide classes were concluded to be due to multiple resistance. Knowledge of multiple and cross-resistance patterns are discussed in relation to the bioassay as a tool for monitoring insecticide resistance.     

Synthesis and biological activity of furanyl anti-juvenile hormonal compounds

Twenty-one synthetic compounds, containing one or more furan rings, were demonstrated to possess anti-juvenile hormone (AJH) activity as evidenced by their induction of premature metamorphosis in the milkweed bug, Oncopeltus fasciatus (Dallas) by contact, topical application or fumigation. The ED₅₀ of the four most active analogs required to induce precocious metamorphosis from 3rd-instar nymphs by residue contact in a Petri dish compared favorably with that of precocene II (6,7-dimethoxy-2,2-dimethyl 2H-chromene) a naturally occurring phytochemical AJH. Precocious metamorphosis was fully reversible by co-treatment with juvenile hormone (JH III) or JH analogs, demonstrating that the observed AJH activity resulted from an induced deficiency of juvenile hormone.     

Nonendoscopic Antegrade Percutaneous Gastrostomy: The Effect of Preplacement Gastric Insufflation on Tube Position and Intra-Abdominal Anatomy

Nonendoscopic tube gastrostomy was performed on 41 anesthetized dogs using the technique of Fulton and Dennis with or without gastric insufflation prior to tube placement. Immediately after tube placement, dogs were euthanized and postmortem examinations performed. When gastric insufflation was not performed (group I), gastrostomy tubes penetrated the visceral surface of the stomach in 25% of dogs. The deep leaf of the omentum was interposed between stomach and body wall in the majority of these dogs, exposing other intra-abdominal organs to potential injury. Additionally, displacement and tethering of the spleen cranial to the gastrostomy site were observed in 33% of dogs in group I. Similar results were obtained when preplacement gastric insufflation was performed after the orogastric tube was inserted sufficiently far to displace the stomach laterally against the body wall (group II). In contrast, consistent positioning of gastrostomy tubes through the parietal surface of the stomach was achieved when the stomach was insufflated prior to lateralizing the left abdominal wall with the gastric end of the orogastric tube (group III). It was concluded that the blind percutaneous gastrostomy technique is made safer by insufflating the stomach immediately prior to pushing the gastric wall laterally into contact with the parietal peritoneum. J Vet Intern Med 1996;10:15–20. Copyright © 1996 by the American College of Veterinary Internal Medicine .     

A controlled evaluation of albendazole against natural infections with Fasciola hepatica and Fascioloides magna in cattle

Albendazole administered to cattle at doses of 45, 35, 25, and 15 mg/kg of body weight reduced the number of viable Fasciola hepatica adults by 99.8%, 97.6%, 97.6%, and 95.7%, F hepatica immatures by 100%, 98.6%, 98.6% and 91.9% and Fascioloides magna by 96.5%, 99.1%, 94.0%, and 94.0%, respectively. Statistical evaluation by one-way analysis of variance, showed significant (P less than 0.05) differences in burdens of each fluke species between treated and untreated cattle, but differences in efficacy between any two of the four drug doses were not significant. Signs of toxicosis were not observed between the time of drug administration and slaughter 7 to 10 days later.     

Biphenyl hydroxylation and its induction differ between montane voles and Swiss Webster mice

Hepatic microsomes from female montane voles, Microtus montanus, metabolized biphenyl to 4-hydroxybiphenyl (359 ng/mg microsomal protein/min), 3-hydroxybiphenyl (49 ng/mg/min), and 2-hydroxybiphenyl (29 ng/mg/min). Phenobarbital treatment of the voles (ip) induced in vitro biphenyl hydroxylations by about 324, 865, and 445% for 4-, 3- and 2-hydroxylation, respectively. Comparable studies were made of female, white Swiss Webster mice. The major microsomal metabolite was 4-hydroxybiphenyl (1010 ng/mg/min) but more 2-hydroxybiphenyl (118 ng/mg/min) was formed than 3-hydroxybiphenyl (92 ng/mg/min). Phenobarbital treatment of the mice barely changed biphenyl 2-hydroxylation (?1.3%) but did raise 3- and 4-hydroxylation (55 and 211%, respectively). Treatment of voles with 3-methylcholanthrene raised biphenyl 2-, 3-, and 4-hydroxylation by about 81, 50, and 47%, respectively, whereas in mice the increases were 176, 41, and 31%, respectively. β-Naphthoflavone treatment had similar effects. The vole liver microsomes were dependent upon NADPH for biphenyl hydroxylation. The carbon monoxide difference spectra of reduced cytochrome in the microsomes show a peak at 450.4 nm. This was unchanged by phenobarbital and only slightly shifted (?0.8 nm) by 3-methylcholanthrene or β-naphthoflavone treatment. Voles and mice represent different families of the order Rodentia. The results obtained with voles more closely resemble those reported for hamsters which are in the same family but a different subfamily than voles. Comparative consideration of the taxonomic relations of the rodents, therefore, may be useful in interpreting their comparative toxicology.     

Effects of growth factors (EGF,PDGF-BB and TGF-beta 1) on cultured equine epithelial cells and keratocytes: implications for wound healing

Objective The physiologic mechanisms involving growth factors, including PDGF‐BB, EGF, and TGF‐β1, as potent mediators of fibroblasts and epithelial cells in corneal wound healing remain unknown. The goal of this study was to determine culture methods for equine epithelial cells and keratocytes and to investigate how exogenous growth factors influence proliferation of both cell types. Procedures Cell cultures were established from healthy corneas harvested from horses immediately following euthanasia and maintained using standard tissue culture protocols. To determine the effects of PDGF‐BB, EGF, TGF‐β1, keratocytes (1 × 10⁵/well) and epithelial cells (2 × 10⁵/well) were each cultured in 12 well plates and exposed separately to the growth factors. The cells were exposed to concentrations of EGF between 0 and 50 ng/mL; PDGF‐BB between 0 and 75 ng/mL; and TGF‐β1 between 0 and 10 ng/mL. Cell proliferation was measured using ³H‐thymidine assay and differences in growth determined using anova and Tukey's HSD test (P < 0.05). Results Epithelial cell and keratocyte cultures were successfully established. EGF maximally stimulated keratocyte and epithelial cells at 25 ng/mL and 5 ng/mL, respectively. PDGF‐BB maximally stimulated keratocytes and epithelial cells at 50 ng/mL and 5 ng/mL, respectively. TGF‐β1 inhibited keratocytes at 5 ng/mL and 10 ng/mL, and epithelial cells at 1 ng/mL and 2 ng/mL. Conclusions Methods were established to maintain epithelial cells and keratocytes in vitro. PDGF‐BB and EGF stimulate, while TGF‐β1 inhibits the proliferation of epithelial cells and keratocytes. These growth factors may play a role in maintenance and repair of the equine cornea.     

Flash electroretinography in standing horses using the DTL microfiber electrode

Purpose The goal of our study was the evaluation of a practical method for the recording of flash electroretinograms (ERGs) in sedated, standing horses with the DTL? microfiber electrode. Methods The horses were sedated intravenously with detomidine hydrochloride (0.015 mg/kg). The pupil was dilated and the auriculopalpebral nerve was blocked. The ERGs were recorded with the active electrode on the cornea (DTL?), the reference electrode near the lateral canthus, and the ground electrode over the occipital bone. The light intensities of the white strobe light were 0.03 cd·s/m² (scotopic) and 3 cd·s/m² (scotopic and photopic). Photopic and scotopic single flash and flicker responses to Ganzfeld stimulation were recorded. During the 20‐min dark adaptation period the retina was stimulated every 5 min with the 0.03 cd·s/m² single flash. Results The median b‐wave amplitudes and implicit times were 38 µV and 33 ms (photopic cone‐dominated response), 43 µV and 63 ms (5‐min dark adaptation), 72 µV and 89 ms (10 min), 147 µV and 103 ms (15 min), 188 µV and 109 ms (20 min, 0.03 cd·s/m², rod response), and 186 µV and 77 ms (20 min, 3 cd·s/m², maximal combined rod‐cone response). A steady increase in amplitude and implicit time was noted during dark adaptation. No oscillatory potentials could be isolated. Conclusions The use of detomidine hydrochloride sedation and the DTL? microfiber electrode allowed the recording of good quality ERGs. This protocol should permit the detection of functional problems in the retina without the risk involved with general anesthesia.