Carbaryl-induced histopathologic alterations in the digestive tract of the Levantine frog, Pelophylax bedriagae (Anura: Ranidae).

In this study, histopathologic changes following carbaryl exposure for 96 hr were investigated in the digestive tract of Levantine frog, Pelophylax bedriagae. Adult frogs were exposed to carbaryl once by oral gavage in concentrations of 0.05, 0.1, and 0.2 mg/g. Histopathological changes were more prominent in medium- (0.1 mg/g) and high-dose (0.2 mg/g) groups than in the low-dose (0.05 mg/g) group. Esophageal cells showed vacuolization, cellular swelling, nuclear pyknosis, karyolysis, and necrosis. Additionally, esophageal glandular atrophy and infiltration of inflammatory cells around esophageal glands were observed at medium and high doses. In the stomach, there were prominent histopathologic defects such as cellular swelling and necrosis in gastric glands, necrotic cells within the interstitial spaces, separation of epithelial cell layer, congested vessels, and hemorrhage at medium and high doses. In the intestine, detachment of epithelial layer, epithelial cell disorganization, inflammation, and necrosis were detected at medium and high doses. The results of this study showed that carbaryl caused adverse effects on the digestive tract of the Levantine frog, P. bedriagae.

Evidence for links between feeding inhibition, population characteristics, and sensitivity to acute toxicity for Daphnia magna.

A population experiment with Daphnia magna tested the hypothesis that short-term feeding inhibition provokes a shift in population structure that will vary with conspecific pressure (e.g., pressure occurring from individuals of the same species due to competition for food and space) and increases population sensitivity to a xenobiotic exposure due to size-dependent toxicity (e.g., decreasing sensitivity with increasing body length). Populations were exposed for one week to a feeding inhibitor (imidacloprid, 0.15 or 12.0 mg/L) followed by one week of recovery and one day of exposure to an acute toxin (carbaryl, 0.0098 mg/L). Identical exposure under low and high conspecific pressure was studied by delaying the start of exposure for half of the populations by two weeks; thus populations were in a different stage of population development when exposure occurred. Feeding inhibition of 97% (12.0 mg/L imidacloprid) caused a shift in population structure toward smaller individuals but also reduced population abundance by up to 56 ± 7% with a strong influence of conspecific pressure. Increased population sensitivity to carbaryl was observed after feeding inhibition of 97% as hypothesized. Carbaryl exposure for one day resulted in population decline of up to 23 ± 6% when populations were not previously exposed to imidacloprid. Identical carbaryl exposure provoked a four times stronger decline in population abundance when exposure occurred following feeding inhibition of 97%. In conflict with the hypothesis, this was at least in part due to changes in the reproductive strategy of daphnids following exposure to imidacloprid rather than driven by the shift in population structure. The differences in population sensitivity to additional stress (carbaryl) occurring one week after feeding inhibition caused by exposure to imidacloprid adds a further challenge to understanding potential impacts from multiple stressors as occurring in the field at the population level.

In vitro induction/inhibition of germinal vesicle breakdown (GVBD) in frog (Euphlyctis cyanophlyctis) oocytes by endocrine active compounds.

Oocyte maturation is transformation of oocytes into a fertilizable egg. This study examined the effects of four classes of chemicals: 1) acephate (organophosphate); 2) atrazine (herbicide); 3) cypermethrin and fenvalerate (synthetic pyrethroids); and 4) carbaryl (carbamate) on in vitro germinal vesicle breakdown (GVBD) of Euphlyctis cyanophlyctis oocytes. Follicles were isolated and defolliculated from surgically removed ovaries of E. cyanophlyctis and exposed to either progesterone (1 µM/mL) or graded concentrations (1, 5, 10, 15, and 20 µg/mL) of test chemicals. GVBD was evident by the presence of a white spot in the animal pole as well as the absence of germinal vesicles in sectioned heat-fixed oocytes. Percent GVBD was scored every 4 hours until 24 hours. Progesterone induced 77-84% GVBD, compared to 29-33% in controls, at 24 hours. Acephate induced 46-67% GVBD, whereas atrazine elicited 58-77% of GVBD. In cypermethrin or carbaryl- or fenvalerate-exposed oocytes, GVBD was limited to 22-28, 17-29 and 18-24%, respectively. The study infers that some chemical contaminants in the aquatic system may interfere with GVBD in amphibians. Because oocyte maturation is a prerequisite for the production of fertilizable eggs, any alteration in this process potentially impairs the fecundity of females.

Evaluation of cucurbitacin-based gustatory stimulant to facilitate cucumber beetle (Coleoptera: Chrysomelidae) management with foliar insecticides in melons.

The bitter plant-derived compounds cucurbitacins are known to stimulate feeding of adult cucumber beetles (Coleoptera: Chrysomelidae). A cucurbitacin-based gustatory stimulant applied as a flowable bait combined with either spinosad or carbaryl was compared with foliar sprays of spinosad and carbaryl for controlling two cucumber beetle species (Diabrotica undecimpunctata undecimpunctata Mannerheim and Acalymma trivittatum Mannerheim) in honeydew melons (Cucumis melo L.). Field studies were conducted on the University of California-Davis plant pathology farm in 2008 and 2009. Beetle densities after applications and fruit damage from beetle feeding were compared among treatments. In addition, beetle survival was compared within field cages placed over the treated foliage infested with beetles. Using all three measures of efficacy, we determined that the addition of cucurbitacin bait had no effect on the level of cucumber beetle control with carbaryl in either 2008 or 2009. In both years, spinosad did not significantly reduce cucumber beetle densities in either field cages or field plots and did not reduce fruit damage relative to the untreated control. The addition of the bait to spinosad did not improve its efficacy. A laboratory bioassay of the spinosad formulation used in the field showed it had significant lethal effects on adults of both cucumber beetle species. Results indicated that the bait formulation used did not improve cucumber beetle control but may benefit from the addition of floral attractants or using a different type of cucurbitacin.

Assessment of acetylcholinesterase activity in Clarias gariepinus as a biomarker of organophosphate and carbamate exposure.

The objective of this study was to investigate the response of acetylcholinesterase (AChE) activities in Clarias gariepinus in response to Organophosphates (Ops) and carbamate exposure. The AChE activities were determined in plasma, and eye and brain homogenates of unexposed and exposed fish using Ellman's method and 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) chromophore. The baseline AChE activities in plasma, eyes and brain tissues in unexposed fish were comparable between males and females (P > 0.05). Concentrations of pesticides that inhibited 50% (IC(50)) of AChE activities in brain homogenates following in vitro exposures were 0.003, 0.03, 0.15, 190, 0.2, 0.003 and 0.002 microM for carbaryl, chlorfenvinphos, diazinon, dimethoate, fenitrothion, pirimiphosmethyl and profenofos, respectively. The in vivo dose-effect relationships were assessed using chlorfenvinphos and carbaryl at different concentrations that ranged from 0.0003 to 0.06 microM and 0.0005 to 0.05 microM, respectively. Acetylcholinesterase activities were comparable in plasma, and eye and brain homogenates from control and carbaryl-exposed fish. Following exposure of fish to chlorfenvinphos at concentrations above 0.03 microM, a significant inhibition of AChE activities in plasma (84%) and eye homogenate (50%) was observed. The AChE activities in brain homogenate were comparable between chlorfenvinphos-exposed fish and controls. Because carbaryl cause reversible inhibition of AChE activities was found to be more potent than chlorfenvinphos that cause irreversible inhibition following in vitro exposure. Contrary, carbaryl was less potent than chlorfenvinphos after in vivo exposure possibly due to more rapid biotransformation of carbaryl than chlorfenvinphos. Findings from this study have demonstrated that inhibition of AChE activity in C. gariepinus is a useful biomarker in assessing aquatic environment contaminated by anticholinesterases.

Assessment of the toxicological interaction of sertraline with cholinesterase inhibiting insecticides in aquatic insects using the black fly, Simulium vittatum IS-7.

Sertraline is a selective serotonin reuptake inhibitor (SSRI) prescribed as an antidepressant. Although SSRIs are known to block serotonin reuptake sites on cell membranes, they also have been shown to inhibit acetylcholinesterase (AChE) activity. Thus, the interaction of these chemicals with other AChE inhibitors, namely, organophosphate and carbamate insecticides, is of interest. In addition, these insecticides have been shown to interact with serotonergic neuronal pathways creating questions as to how these chemicals might interact. In this study, the interactive effect of sertraline (SSRI) in binary combinations with carbaryl (carbamate insecticide) and diazinon (organophosphate insecticide) was assessed using a 48-h acute toxicity test with black fly larvae, Simulium vittatum IS-7. Results showed that observed mortality was bracketed by the independent action model and concentration addition model with the independent action model slightly underestimating mortality and the concentration addition model slightly overestimating mortality. Varying the concentration of the chemicals in the mixture did not indicate that sertraline was interacting with the insecticides to make them more toxic or vice versa. These results indicate that sertraline and the insecticides are likely eliciting toxicity at separate neuronal pathways since no interaction was observed.

Efficacy of Spring and Fall Treatments of Carbaryl for Protecting Ponderosa Pine From Mortality Attributed to Mountain Pine Beetle (Coleoptera: Curculionidae).

High-value trees, such as those growing in residential, recreational, or administrative sites, are often susceptible to colonization by bark beetles (Coleoptera: Curculionidae: Scolytinae) as a result of increased amounts of stress associated with off-site plantings, drought, soil compaction, and/or mechanical injury. The value of these trees, cost of removing dead trees, and loss of aesthetics often justify the use of insecticides to protect trees from mortality attributed to bark beetles. Carbaryl (1-naphthyl methylcarbamate) is among the most effective, economically-viable, and ecologically-compatible insecticides available for protecting conifers from several species of bark beetles in the western United States. Treatments are usually applied in spring prior to initiation of flight of the target species. We evaluated the efficacy of spring and fall applications of carbaryl for protecting individual ponderosa pine, Pinus ponderosa Dougl. ex Laws. (Pinales: Pinaceae), from mortality attributed to mountain pine beetle, Dendroctonus ponderosae Hopkins (Coleoptera: Curculionidae), in Idaho. Both spring and fall treatments of 2.0% a.i. carbaryl (maximum label rate; Sevin SL, Bayer Environmental Science, Montvale, NJ 07645) provided one field season of protection, and thus should be applied annually if tree protection is desired for multiple years. Our research also provides some insight on the efficacy of carbaryl treatments after wildfire. We found no evidence that a mixed-severity wildfire negatively affected the efficacy of carbaryl treatments.

Differential sensitivity to anticholinesterase insecticides in the juvenile rat: effects on thermoregulation.

Organophosphate (OP) and carbamate (CB) insecticides inhibit cholinesterase (ChE) activity and induce acute hypothermia in adult rats. Studies showed that juveniles are generally more susceptible to neurotoxic insult than adults. However, little is known concerning the effects of OP and CB pesticides on thermoregulation in developing animals. Thus, alterations in core body temperature (Tc) in juvenile animals exposed to an OP and CB insecticide were investigated. Male rat pups were anesthetized on postnatal day (PND) 15 with metofane and a radio transmitter (Data Sciences) was implanted in the abdominal cavity to monitor Tc and motor activity (MA). Two days were allowed for recovery. The PND 17 pups were then dosed by oral gavage with the OP chlorpyrifos (CHP) (1, 5, 10, or 15 mg/kg) or the CB carbaryl (CAR) (10, 20, 80, 120, or 160 mg/kg) or the corn oil vehicle. Pups were returned to their dams and littermates immediately after dosing and monitored for the next several days. CHP doses of 10 and 15 mg/kg resulted in 1.0 degrees C and 2.4 degrees C reductions in Tc, respectively. Tc recovered to control levels by approximately 16 h after dosing. There was significant mortality in rats dosed with 15 mg/kg CHP (6 of 11). CAR doses of 10 to 80 mg/kg had little effect on Tc. The highest dose of CAR (160 mg/kg) resulted in a 1.3 degrees C reduction in Tc that recovered in 9 h. In contrast, past studies found that adult male rats become hypothermic at CHP doses of >25 mg/kg, whereas a CAR dose of 50 mg/kg is effective at inducing hypothermia. Overall, it appears that during the development from preweanling to adult rat, there is a progressive attenuation in CHP-induced hypothermia. Conversely, CAR-induced hypothermia increases as a function of development.

Thermoregulatory response to an organophosphate and carbamate insecticide mixture: testing the assumption of dose-additivity.

Most toxicity data are based on studies using single compounds. This study assessed if there is an interaction between mixtures of the anticholinesterase insecticides chlorpyrifos (CHP) and carbaryl (CAR) using hypothermia and cholinesterase (ChE) inhibition as toxicological endpoints. Core temperature (T(c)) was continuously monitored by radiotelemetry in adult Long-Evans rats administered CHP at doses ranging from 0 to 50mg/kg and CAR doses of 0-150 mg/kg. The temperature index (TI), an integration of the change in T(c) over a 12h period, was quantified. Effects of mixtures of CHP and CAR in 2:1 and 1:1 ratios on the TI were examined and the data analyzed using a statistical model designed to assess significant departures from additivity for chemical mixtures. CHP and CAR elicited a marked hypothermia and dose-related decrease in the TI. The TI response to a 2:1 ratio of CHP:CAR was significantly less than that predicted by additivity. The TI response to a 1:1 ratio of CHP and CAR was not significantly different from the predicted additivity. Plasma and brain ChE activity were measured 4h after dosing with CHP, CAR, and mixtures in separate groups of rats. There was a dose-additive interaction for the inhibition of brain ChE for the 2:1 ratio, but an antagonistic effect for the 1:1 ratio. The 2:1 and 1:1 mixtures had an antagonistic interaction on plasma ChE. Overall, the departures from additivity for the physiological (i.e., temperature) and biochemical (i.e., ChE inhibition) endpoints for the 2:1 and 1:1 mixtures studies did not coincide as expected. An interaction between CHP and CAR appears to depend on the ratio of compounds in the mixture as well as the biological endpoint.

Carcinogenic and cocarcinogenic studies with carbaryl following topical exposure in mice.

Carbaryl (1-naphthyl methyl carbamate: C12H11NO2) CAS Reg. No. 63-25-2) is a widely used broad spectrum carbamate insecticide known to exert various toxic effects on experimental animals. Along with various other toxicological effects carbaryl is reported to increase the incidence of neoplasm in various tissues in rats after oral or intraperitoneal administration. No study has so far been reported in rodents to assess its carcinogenic/cocarcinogenic potential after topical exposure. In this set of investigations, the complete carcinogenic, tumour initiating and tumour promoting property of carbaryl was tested on the skin of female Swiss albino mice. The animals were exposed to carbaryl through topical painting on the interscapular region at a dose of 100 mg/kg body wt. The results revealed that carbaryl has tumour initiating potential, at the test dose, on mouse skin following two stage, initiation-promotion protocol, but, it failed to induce the tumour(s) when tested for complete carcinogenic and tumour promoting properties.

Carcinogenicity in rats of high oral doses of N-nitroso-carbaryl, a nitrosated pesticide.

N-Nitrosocarbaryl was administered orally to 31 male Sprague-Dawley rats at doses of 130 mg/kg body weight (b.w.) twice weekly. Of the treated animals 29% died with squamous cell carcinomas of the forestomach after an average inductiom time of 167 days. The first carcinoma was observed as early as 63 days after the beginning of the experiment. In addition, 19% of the treated animals died with hyperkeratoses and 6% with papillomas in the forestomach. In contrast to the untreated controls, which had a normal life expectancy, treated animals had a very short life span due to high cumulative toxicity and the early appearance of tumors.

Distribution of radioactivity following oral administration of carcinogenic 14CH3-labelled nitrosocarbaryl in the rat.

After a single intragastric administration of 14C-labelled carcinogenic nitrosocarbaryl, a nitrosated pesticide, the distribution of radioactivity was investigated in the blood and a number of organs in male rats. The animals received 0.25 mg/kg of labelled nitrosamine and were killed following administration at timed intervals between 0.5 h and 24 h. Our results show that the greatest amount of the 14CH3--group was associated with the forestomach, tumor-susceptible tissue; the level of radioactivity is noteworthy but less important in the glandular stomach. There are also sites of radioactivity accumulation mainly in the liver. Moreover, [14C]nitrosocarbaryl was revealed in the blood suggesting that nitrosamine itself rapidly (0.5 h) crosses the intestinal barrier and in a significant quantity (13%). These facts constitute a potential carcinogenic risk.

Carcinogenesis in Sprague-Dawley rats of N-nitroso-N-alkylcarbamate esters.

Nitrosocarbaryl, nitroso-N-methylurethane and nitroso-N-ethylurethane were administered by gavage in olive oil solution to groups of 12 female Sprague-Dawley rats. The dose was 0.2 ml of 0.11 M solution once a week for 10 weeks, a total dose of 0.22 mmol. The rats given nitrosocarbaryl survived longer, but had as high an incidence of tumors (75%) as did rats given nitrosomethylurethane. Most of the tumors induced were invasive squamous carcinomas of the stomach. Nitrosoethylurethane appeared to be a little more potent than nitrosomethylurethane; all 12 animals in this group had squamous stomach tumors at death. A higher total dose of nitrosocarbaryl, 1.3 mmol, given to male rats twice weekly for 20 weeks did not produce a higher incidence of stomach tumors than did thelower dose in females, although the males died earlier with tumors.

Episodic exposures to chemicals: what relevance to chemical intolerance?

Episodic exposures refer to intermittent acute exposures to chemicals that ordinarily have a rapid onset and short duration of effect. There has been a long tradition in preclinical behavioral pharmacology of using episodic-exposure paradigms in order to establish dose-response functions in individual organisms. In these experiments, stable baselines of behavior are first established and then followed by administering varying doses of a drug intermittently, for example, once or twice a week. The power of this approach is well established; the within-subjects design reduces error variance, allows exploration of the entire range of effective doses, and can be used to identify individual differences in drug sensitivity. Of course, the approach is only applicable to reversibly acting compounds, and checks need to be included to insure effects of one dose are not influenced by prior exposure to another dose. We have used baseline approaches to evaluate the effects of pesticides and solvents on the behavior of adult male rats and mice. Moreover, a novel probabilistic dose-tolerance analysis applied to the data suggests substantial individual differences in chemical sensitivity, often spanning orders of magnitude. These results suggest that individual differences in chemical sensitivity may be much greater than previously acknowledged.

A comparison of a single occasion treatment of head louse infestation with phenothrin liquid shampoo or a carbaryl lotion.

Fifty subjects with head louse infestation were recruited into a controlled trial to compare a phenothrin liquid shampoo with a carbaryl lotion. Twenty-seven subjects were treated with phenothrin and 23 with carbaryl, each formulation being applied only on a single occasion. Subjects were inspected for evidence of live lice and eggs at 24 hours and 3 to 4 weeks after application of treatment. The results showed that both the phenothrin liquid shampoo and the carbaryl lotion were effective in killing adult lice and viable eggs. No statistically significant difference in treatment efficacy was observed between the two groups. Fewer side-effects, however, were observed with the phenothrin liquid shampoo than with the carbaryl lotion. These results indicate that, when applied as a single treatment, a phenothrin liquid shampoo was as effective as a carbaryl lotion in eradicating head lice and eggs.

Variations of the digestive absorption kinetics of carbaryl with the nature of the vehicle.

Blood kinetics of 1-naphthyl-N-methyl[14C] carbamate were determined after intravenous injection in DMSO, and after intragastric and intraduodenal administration in DMSO, oil, gum tragacanth and milk. The acetylcholinesterase inhibition and the level of 14C-activity were both determined in the blood over various periods of time. The values of the absorption rate constants after intragastric and intraduodenal administration were found to be: 0.5 h(-1) and 7 h(-1) with DMSO, 0.6 h(-1) and 0.42 h(-1) with oil, 0.13 h(-1) and 0.22 h (-1) with gum tragacanth and 0.10 h(-1) with milk. Appearance of the toxic effect (the inhibition of the acetylcholinesterases) was closely related to the absorption rate constants which themselves depend on the administration vehicle employed.

Histological and ultrastructural studies of rats exposed to carbaryl.

The aim of the study was to assess the general toxic effects of dermally applied carbaryl, based on histological and ultrastructural examinations of internal organs and to relate these effects to earlier own studies where 14C carbaryl was used for determining the pesticide penetration. The pesticide was applied in doses of 1/5 and 1/10 LD50, administered to the tail skin of male Wistar rats 4 hours daily, for 4 weeks except Saturdays and Sundays. After the experiment, the animals were anaesthetized and the following organs were taken for histological study: brain, lung, heart, liver, kidney, skin from the site of exposure and skin from a place at least 2 cm distant from the exposure site. Lung, liver, kidney, heart and skin were used for ultrastructural studies. Dermal application of carbaryl resulted only in slight histological changes in skin, liver, brain and lung. Even in brain and liver, where large amounts of 14C carbaryl, compared to other organs (lung, kidney, heart), where the intensity of histologic changes was earlier stated to below. Ultrastructural changes were observed in skin, liver, lung, heart and kidney.

The relative toxicities of insecticides to earthworms of the Pheretima group (Oligochaeta).

An artificial soil test was used to determine the LC50 values of carbaryl, chlorpyrifos, imidacloprid, cyfluthrin and fipronil against earthworms of the Pheretima group. For a 24-h interval, carbaryl was the most toxic to earthworms (LC50 = 77 mg kg-1), followed by imidacloprid (155 mg kg-1), cyfluthrin (351 mg kg-1), chlorpyrifos (390 mg kg-1) and fipronil (> 8550 mg kg-1) as the least toxic. For the 48-h and 7-day intervals, imidacloprid was the most toxic to earthworms (LC50 = 5 mg kg-1 and 3 mg kg-1 respectively), followed by carbaryl (16 mg kg-1; 9 mg kg-1), cyfluthrin (128 mg kg-1; 110 mg kg-1), chlorpyrifos (330 mg kg-1; 180 mg kg-1) and the least toxic was fipronil (> 8550 mg kg-1 both intervals). The surface application rates required to achieve these values are compared with the rates recommended for the control of turfgrass pests.

Spermatotoxic effects of carbaryl in rats.

The spermatotoxic effect of carbaryl in adult and young male rats has been examined. Carbaryl 50 and 100 mg/kg b.wt. Male fed 5 d/week for 60 days, caused dose and age-dependent decline in epididymal sperm count and sperm motility, an increase in sperm with abnormal morphology. The dose of 25 mg/kg/d was a 'No observed effect level' for the indices studied. Young animals in comparison to adults exhibited pronounced spermatotoxic effects.

Influence of inert ingredients in pesticide formulations on dermal absorption of carbaryl.

OBJECTIVES: To assess the influence of solvent plus various mixtures on percutaneous absorption and disposition of the carbamate insecticide, carbaryl (CA). ANIMALS: Skin was obtained from the dorsum of 14 female weanling specific-pathogen-free Yorkshire pigs. PROCEDURE: In this 8-hour in vitro flow-through diffusion study, porcine skin sections were dosed with 40 micrograms of CA/cm2 of surface area, different amounts of solvents (40 or 80% acetone or dimethyl sulfoxide [DMSO]), different amounts of a surfactant (0, 1, or 5% sodium lauryl sulfate [SLS]), an insect repellent (0 or 15% diethyl-m-toluamide [DEET]), an insecticide synergist (0 or 2% piperonyl butoxide [PB]), and a CA metabolite (40 micrograms/cm2 1-naphthol [1-NA]). RESULTS: In general, CA absorption was greater from acetone than from DMSO mixtures, and CA penetration into skin and stratum corneum was greater from DMSO at 8 hours. This is consistent with the flux-time profiles, which depicted initial peak flux within 2 to 3 hours for most acetone mixtures, but a slow increase in flux for DMSO mixtures. Irrespective of the solvent, increasing water content in pesticide dosing mixtures significantly increased CA absorption from SLS mixtures only. The SLS also enhanced CA absorption, especially at low solvent concentrations. The DEET significantly reduced CA absorption from acetone, but not from DMSO mixtures, and 1-NA enhanced CA absorption from acetone, but not from DMSO mixtures. Piperonyl butoxide significantly enhanced CA absorption from acetone and DMSO mixtures. However, addition of PB or PB plus SLS did not significantly increase CA flux above that observed from solvent plus surfactant mixtures. CONCLUSIONS: Inert ingredients can modulate percutaneous absorption of toxicologically important pesticides and their effect or activity on CA disposition is dependent on solvent specificity and solvent concentration. Whereas SLS, PB, and 1-NA can enhance pesticide absorption, DEET can reduce absorption.