Time-Course Changes in Urine Metabolic Profiles of Rats Following 90-Day Exposure to Propoxur.

As a major kind of carbamate insecticide, propoxur plays an important role in agriculture, veterinary medicine, and public health. The acute toxicity of propoxur is mainly neurotoxicity due to the inhibition of cholinesterase. However, little is known regarding the toxicity of propoxur upon long-term exposure at low dose. In this study, Wistar rats were orally administrated with low dose (4.25 mg/kg body weight/day) for consecutive 90 days. And the urine samples in rats treated with propoxur for 30, 60, and 90 days were collected and analyzed by employing 1H NMR-based metabolomics approach. We found that propoxur caused significant changes in the urine metabolites, including taurine, creatinine, citrate, succinate, dimethylamine, and trimethylamine-N-oxide. And the alteration of the metabolites was getting more difference compared with that of the control as the exposure time extending. The present study not only indicated that the changed metabolites could be used as biomarkers of propoxur-induced toxicity but also suggested that the time-course alteration of the urine metabolomic profiles could reflect the progressive development of the toxicity following propoxur exposure.

Wall-type and indoor residual spraying application quality affect the residual efficacy of indoor residual spray against wild malaria vector in southwest Ethiopia.

BACKGROUND: Residual efficacy of indoor residual spray may vary with different spray quality and wall surfaces types. This study evaluated the impact of spray quality and wall surface types on residual efficacy of propoxur against wild Anopheles gambiae sensu lato (s.l.) in southwest Ethiopia. METHODS: Thirty houses of different mud wall surfaces (10 smooth, 10 rough, 10 painted) were selected and randomly allocated into routine and standard spray. The routine spray was conducted by district health office as usual, while the standard spray was done by strictly following guidelines. Three control houses were selected from unsprayed nearby semi-urban. Wild An. gambiae s.l. were used for wall bioassay tests. Two-way mixed model analysis of variance was used to analyse the data. The mean variation between wall and spray types was compared by post hoc analysis of IBM SPSS version 20. RESULTS: On standard spray, knockdown rate was 95.3% on painted, 82% on smooth and 72.5% on rough surface at week 17 of post-spray, whereas on routine spray it was 82.7% on painted, 48.7% on smooth and 60% on rough surface. On standard spray, mortality rate of An. gambiae s.l. was 99.3% on painted surface, 90% on smooth and 80% on rough surface. On routine spray, it was 89.3% on painted, 61.3% on smooth and 65% on rough surface at week 17 of post-spray. The painted wall surface showed the highest knockdown rate (86.4-100%) on standard and (73.8-91.5%) routine spray; mortality rate was more than 80% on both spray types during the 17 weeks of follow-up regardless of spray types. The lower mortality rate and residual effect was observed on routine smooth and rough wall surfaces. The residual efficacy of propoxur was > 80% at week 17 on standard spray regardless of the wall types and it was < 80% on routine spray except painted wall surface. CONCLUSION: The painted wall surface and standard spray showed better residual efficacy. Therefore, it is recommended to consider the wall surface available in the community to estimate the residual lifespan of the insecticide, and strictly to follow the spray guideline to improve the effectiveness of indoor residual spray.

Evaluation of cytotoxicity, genotoxicity and embryotoxicity of insecticide propoxur using flounder gill (FG) cells and zebrafish embryos.

Cytotoxicity, genotoxicity and embryotoxicity of carbamate insecticide propoxur were evaluated using flounder gill (FG) cells and zebrafish embryos. The cytotoxicity of propoxur in FG cells was analyzed by MTT, neutral red uptake (NRU), lactate dehydrogenase (LDH) release and Hoechst 33342 and propidium iodide double staining, and acute cytotoxic effects were observed in a concentration-dependent manner. The 24h-IC50 values of 89.96 ± 1.04, 103.4 ± 1.14 and 86.59 ± 1.13 µg/ml propoxur were obtained by MTT, NRU and LDH assays, respectively. The lethal effects were induced in FG cells mainly through necrosis but not apoptosis as evidenced by double fluorescence staining. Comet assay showed weak genotoxic effects and statistically significant DNA damages were recorded in the cells exposed to highest tested concentration of 75 µg/ml propoxur (p<0.05). Propoxur exerted obvious acute toxic effects on the survival, spontaneous movement, hatching and heart rate, and development (yolk and pericardial sac edema) of zebrafish embryos in both time- and concentration-dependent manner only at ⩾ 100 µg/ml. The corresponding 24h-, 48 h- and 96 h-LC50 values of propoxur in zebrafish embryos were 166.4 ± 1.06, 146.3 ± 1.07 and 134.8 ± 1.06 µg/ml, respectively. The above data obtained suggest a low acute toxicity of propoxur to the in vitro cultured FG cells and zebrafish embryos.

Philornis sp. bot fly larvae in free living scarlet macaw nestlings and a new technique for their extraction.

Bot fly larvae (Philornis genus) are obligate subcutaneous blood-feeding parasites of Neotropical birds including psittacines. We analyze twelve years of data on scarlet macaw (Ara macao) nestlings in natural and artificial nests in the lowland forests of southeastern Peru and report prevalence and intensity of Philornis parasitism. Bot fly prevalence was 28.9% while mean intensity was 5.0 larvae per infected chick. Prevalence in natural nests (11%, N=90 nestlings) was lower than in wooden nest-boxes (39%, N=57) and PVC boxes (39%, N=109). We describe a new technique of removing Philornis larvae using a reverse syringe design snake bite extractor. We compare this new technique to two other methods for removing bots from macaw chicks and find the new method the most suitable.

Effects of propoxur on the reproductive system of male rats.

The reproductive toxicity of propoxur (2-isopropoxy-phenyl-N-methylcarbamate), a carbamate pesticide, was investigated in adult male Wistar rats exposed to 0, 1.73, 2.6, and 5.2 mg/kg body weight/day for 90 successive days. Results obtained from this study showed a significant (p < 0.05) concentration-dependent increase in food consumption, relative weights of testis, epididymis and kidneys with maximum effects observed at the highest dose of propoxur (5.2 mg/kg) compared to the control. On the contrary, there was a significant (p < 0.05) decrease in the body weight gain, sperm density, serum and intratesticular total cholesterol concentrations, and intratesticular total proteins in rats treated with propoxur at the dose of 5.2 mg/kg body weight. Propoxur had no significant effect on gestation, fertility and parturition indices, average birth weight, litter size and pups sex ratio of untreated female rats mated with treated males rats. These results suggested that propoxur increased food consumption, relative weight of reproductive and detoxication organs, decreased body weight gain and sperm density per gram of cauda epididymis without affecting the reproductive performances.

Penetration studies of propoxur and phoxim from eggshell into whole egg after experimental exposure and application in henhouses.

The penetration of propoxur and phoxim from eggshell into whole egg was investigated in vitro by spraying eggs directly and in vivo after application of the compounds in henhouses. Although mean concentrations of the compounds on eggshells were up to 23000 microg kg(-1), mean residue concentrations in whole eggs were far below the current maximum residue levels (50 microg kg(-1) for propoxur and 60 microg kg(-1) for phoxim). These results provide the first evidence that propoxur and phoxim do not penetrate from eggshell into whole egg under experimental and field conditions. Subsequently, residue carry-over after egg cracking in households and during a worst-case situation in an egg-cracking plant was investigated. However, when eggs were cracked manually, a negligible contamination of whole egg values occurred. If, in an automated process, eggshells accidentally come into close contact with whole egg, very high residue levels of propoxur and phoxim may be generated time dependently. These results suggest that eggshell contact with whole egg during egg cracking must be avoided to prevent pesticide carry-over.

[Determination of insecticide-resistance and resistance mechanisms of Blattella germanica (Dictyoptera: Blattellidae)]. / Determinación de la resistencia a insecticidas y mecanismos de resistencia en cepas de Blattella germanica (Dictyoptera: Blattellidae).

In this paper, the level of resistance to four insecticides of 3 Blatella germanica strains collected from various places in the City of Havana province was evaluated. These strains were resistant to two pyrethroids (cypermethrin and lambda-cyalothrine) and to organophosphorate malathion but susceptible to carbamate propoxur. The values of alpha and beta esterases, acetylcholinesterase and gluthatione-S-transferase were estimated in three strains involved in the study. The results of the study showed high esterase activity in all the strains, mainly beta esterases and two of the three strains presented with high gluthation-S-transferase enzyme. No changes in acetylcholinesterase were demonstrated in relation to the reference strain. The association of levels of resistance to insecticides, the possible resistance mechanisms in each strain and the results of the enzymatic activity were also analyzed.

Interactions of skin thickness and physicochemical properties of test compounds in percutaneous penetration studies.

OBJECTIVES: To determine the effect of skin thickness on the percutaneous penetration and distribution of test compounds with varying physicochemical properties using in vitro systems. Studies were carried out in accordance with OECD guidelines on skin absorption tests. METHODS: Percutaneous penetration of caffeine (log P -0.01), testosterone (log P 3.32), propoxur (log P 1.52) (finite dose in ethanol to water vehicle ratio) and butoxyethanol (log P 0.83) (undiluted finite dose or as an infinite dose 50% [v/v] aqueous solution) through skin of varying thicknesses under occluded conditions was measured using flow through cells for 8-24 h. Saline (adjusted to pH 7.4) was used as receptor fluid, with BSA added for studies with testosterone and propoxur. Following exposure, the remaining surface dose was removed by swabbing and the skin digested prior to scintillation counting. RESULTS: The maximum flux of caffeine was increased with decreasing skin thickness, although these differences were found to be non-significant. The presence of caffeine in the skin membrane was not altered by skin thickness. Maximum flux and cumulative dose absorbed of testosterone and butoxyethanol (in both finite and infinite doses) were markedly reduced with full thickness (about 1 mm thick) skin compared with split thickness skin (about 0.5 mm). Maximum flux of propoxur (dissolved in 60% ethanol) was clearly higher through skin of 0.71 mm than through skin of 1.36 mm, but no difference was found between 0.56 and 0.71 mm. The proportion of propoxur present in the membrane after 24 h increased significantly over the complete range of thicknesses tested (0.56-1.36 mm). CONCLUSIONS: A complex relationship exists between skin thickness, lipophilicity and percutaneous penetration and distribution. This has implications for risk assessment studies and for the validation of models with data from different sources.

Stimulus frequency dependence of the central and peripheral somatosensory evoked activity in rats treated with various pesticides.

Rats were treated with a combination of insecticide agents in different timing schemes. In acute administration, 1/5 LD50 of the three insecticides: dimethoate, propoxur and cypermethrin, or their combination, was given once by gavage. In the developmental model, female rats received oral doses of 1/25 LD50 of the above insecticides in combination in three timing schemes including pregnancy and lactation. Responses in the somatosensory cortex and in the tail nerve, evoked by peripheral electric stimulation, were recorded in acute preparation under urethane anesthesia. It was tested whether the parameters of the cortical and peripheral evoked response are dependent on the frequency and whether this dependence is different in control and treated animals. The latency increase of the cortical responses with increasing stimulation frequency was significantly stronger in rats treated acutely with cypermethrin and the combination, and in rats receiving the combination during both intra- and extrauterine development. On the duration, the effects were less clear. Frequency dependent increase of the tail nerve action potential latency was significantly intensified by cypermethrin, and the amplitude decrease, by cypermethrin and dimethoate. Fatigue of this response during a stimulation series was also altered by the insecticides. Frequency dependence and fatigue possibly reflect the actual state of the nervous system and may have the potency to be developed to functional biomarkers.

Effective treatment and control of biting lice, Felicola subrostratus (Nitzsch in Burmeister, 1838), on cats using fipronil formulations.

The efficacy of 0.25% fipronil spray (Frontline Spray, Merial), 10% fipronil spot-on (Frontline Spot-on for Cats, Merial) and 10% fipronil/12% (S)-methoprene (Frontline Plus for Cats, Merial) against the biting louse Felicola subrostratus on cats was assessed and confirmed under laboratory conditions. A field study evaluated the efficacy of a single topical application of Frontline Spray, and spot-on against the parasite on cats. In the laboratory studies, animals were allocated based on pre-treatment louse counts to the treatment groups: (1) untreated control and (2) 10% fipronil spot-on in the exploratory study or (1) untreated control, (2) 0.25% fipronil spray, at 6 ml/kg; (3) 10% fipronil spot-on as per label and (4) 10% fipronil/12% (S)-methoprene as per label in the confirmation study. Cats in treatment groups 2-4 were treated twice topically on Days 0 and 28. No live F. subrostratus were found on cats treated with fipronil formulations at any post-treatment examination. The difference from controls was significant (P < 0.01) for each product at each examination. Based on whole body counts at Day 42, the efficacy of each product was determined to be 100%. In the field study, cats were allocated in strict order of presentation. Cats were randomly allocated to one of the three treatment groups: (1) propoxur collar (Bolfo, Bayer); (2) 0.25% fipronil spray, at 6 ml/kg and (3) 10% fipronil spot-on as per label. Cats were treated once topically on Day 0. Louse counts of cats treated with fipronil formulations were not different than those of cats receiving the propoxur collar. The efficacy was determined to be > 98% on Day 2 and 100% on Days 28 and 42 in all treatment groups. The results of these studies demonstrate that fipronil in topical formulations is effective for treatment and control of biting lice (F. subrostratus) infestations on cats.

Comparative investigation of behavioral, neurotoxicological, and immunotoxicological indices in detection of subacute combined exposure with methyl parathion and propoxur in rats.

The effects of 6 weeks of oral exposure to propoxur (PR; at doses of 0.851 and 8.51 mg/kg body wt.), methylparathion (MP; at doses of 0.218 and 0.872 mg/kg body wt.), and their combinations were investigated in male Wistar rats. Measurement endpoints of the investigation were certain general toxicological parameters (body weight gain, organ weights), plaque-forming cell (PFC) count from the spleen, open field (OF) behavior, auditory startle response (ASR), prepulse inhibition (PPI), rotarod performance, somatosensory and auditory cortical evoked potentials, and peripheral nerve conduction velocity. The treated rats did not show any sign of acute intoxication during the 6 weeks of exposure. The higher dose of PR, but not of MP, significantly decreased the relative liver weight. Both agents produced a significant dose-dependent increase of OF activity, with larger expression after 2 weeks than after 6 weeks. The number of ASR responses and the ASR amplitude increased. The amplitude after PPI was increased by MP but only minimally altered by PR and the combinations. There was a small, but with high-dose PR significant, increase in the latency of the somatosensory evoked potentials. Neither of the two substances alone had any effect on the PFC response. The effect of the combination of high-dose PR and low-dose MP was significantly different from that of high-dose PR alone on the liver weight, on the ASR amplitude, and on the PFC/10(6) cell and PFC/spleen counts. With high-dose MP and low-dose PR, no such interaction was observed. According to the results, the noneffective dose of MP can influence the toxicity of the effective dose of PR in a combined exposure situation.

A study on geno- and immunotoxicological effects of subacute propoxur and pirimicarb exposure in rats.

A 28-day oral exposure with 8.51, 3.40, and 0.851 mg/kg propoxur (PR) and 4.67, 1.87, and 0.467 mg/kg pirimicarb (PI) was performed in male Wistar rats, and the occurrence of numerical and structural chromosome aberrations and the changes in certain immune function parameters (plaque-forming cell (PFC) assay, delayed-type hypersensitivity reaction) and in some basic toxicological (body weight gain and weights of brain, thymus, lung, heart, liver, spleen, kidneys, adrenals, and popliteal lymph node) and hematological (white blood cells, red blood cells, hematocrit (Ht), mean cell volume of red blood cells (MCV) cell content of the femoral bone marrow) parameters were investigated. The high dose of PR increased the relative liver weight and the cell content of femoral bone marrow, and all three doses increased Ht and MCV. The applied doses of PI decreased the relative adrenal weight in a dose-dependent manner, and its highest dose increased the relative liver weight. Among the immune function parameters tested, PFC content of the spleen was decreased by high-dose PR and elevated by high-dose PI, whereas the maximum and the time course of the delayed-type hypersensitivity reaction showed no changes in this dose range. In the genotoxicological investigations only the high PR dose increased the number of numerical, but not the structural, chromosome aberrations. In addition to the changes in relative adrenal weight following PI treatment, the PFC assay showed the highest sensitivity for detection of the 4-week exposure with these carbamates. On the basis of our results, the immunotoxicological approach seems to have the same (PR) or higher (PI) sensitivity in early detection of the repeated low-dose exposure by these carbamates compared to the genotoxicological approach.

Detection of the effects of repeated dose combined propoxur and heavy metal exposure by measurement of certain toxicological, haematological and immune function parameters in rats.

In the present study, an immunotoxicity test system, containing general toxicological (body weight gain, organ weights), haematological (WBC,RBC, Ht, mean cell volume of the RBCs, cell content of the femoral bone marrow), and immune function (PFC assay, DTH reaction) investigations, was used for detection the effects of a 4 weeks repeated low dose combined oral exposure of male Wistar rats with propoxur and the heavy metals arsenic or mercury. Two doses of the compounds were used: a higher one (the lowest dose which resulted in significant change of at least one parameter examined in previous dose-effect experiments), and a lower one (the highest dose which proved to be non-effective). The applied doses were: 8.51 and 0.851 mg kg(-1) of propoxur, 13.3 and 3.33 mg kg(-1) of NaAsO(2), and 3.20 and 0.40 mg kg(-1) of HgCl(2). In the combination treatment, the high dose of propoxur was combined with the low dose of arsenic or mercury, and the high doses of each heavy metals were combined with the low dose of propoxur. The main finding of this study was that some of the combinations significantly altered the relative weight of liver, adrenals and kidneys, related to both the untreated and the high dose internal control. Among the immune functions examined, only the PFC content of the spleen showed a trend of changes in certain combinations versus the corresponding high dose control. According to the present results, combined exposure with propoxur and the heavy metals examined can modify the detection limit of the single compounds and/or may alter their toxic effects.

Resistance to the insecticides lufenuron and propoxur in natural populations of Drosophila melanogaster (Diptera: Drosophilidae).

Lufenuron is a newly marketed benzoylphenyl urea chitin-synthesis inhibitor insecticide that is effective against certain insects, including Drosophila melanogaster (Meigen). Resistance to this class of insecticides is not widespread in pest insect populations and, for the resistance that has been reported, the genetic basis is not understood. In previous work, natural population strains of D. melanogaster from 2 widely separated locations in the United States were found to be as much as 100 times more resistant to lufenuron when compared with laboratory strains. It was postulated that this resistance is the result of cross-resistance that evolved to an earlier, widely used insecticide. In the current study we examined cross-resistance of selected D. melanogaster strains to propoxur, a likely candidate carbamate insecticide that has been extensively used during the past 3 decades. However, no correlation between resistance to lufenuron and propoxur was found. Strains were selected to represent a range of dates of establishment (1936-1996) from natural populations to laboratory culture. Examination of these strains showed susceptibility to propoxur in long-established laboratory strains, but resistance in recently established strains. Susceptibility to lufenuron was also high in long-established strains and apparently slowly decreased in natural populations until approximately equal to 5 yr ago, when it decreased more rapidly. These results suggest that if this loss in susceptibility results from agricultural chemical usage, then these chemicals can significantly affect a non-target insect.

Assessment of the ability of propoxur, methomyl, and aldicarb, three carbamate insecticides, to induce micronuclei in vitro in cultured Chinese hamster ovary cells and in vivo in BALB/c mice.

Three carbamate insecticides (propoxur, methomyl, and aldicarb) were evaluated for their ability to induce micronuclei (MN) in vitro using cultured Chinese hamster ovary (CHO) cells, and in vivo in mouse bone marrow erythrocytes. In vitro, all three insecticides induced a significant increase in micronucleated binucleate cells, which was generally both dose and sample time dependent. The in vivo studies involved treating male BALB/c mice by different routes, either once or on 3 consecutive days, followed by multiple or single sampling. Treatment by intraperitoneal injection or oral gavage induced a significant increase in micronucleated reticulocytes (MNRETs) in peripheral blood. For all three chemicals, the MN response depended on sample time and the number of treatments, while for aldicarb, the response depended also on the route of exposure. These positive results demonstrate that propoxur, methomyl, and aldicarb are capable of inducing structural and/or numerical chromosomal aberrations in mammalian cells either in vitro or in vivo. Furthermore, based on the results obtained, on optimal in vivo MN protocol for carbamate insecticides is a single treatment followed by blood sampling at 24 and 48 hr after treatment.

Effects of single or repeated administration of a carbamate, propoxur, and an organophosphate, DDVP, on jejunal cholinergic activities and contractile responses in rats.

Wistar rats were injected once or repeatedly for 10 days with dichlorvos (DDVP, 5 mg kg-1), propoxur (10 mg kg-1), oxotremorine (0.1 mg kg-1) or atropine (5 mg kg-1). Animals were killed 20 min or 24 h after single or consecutive injections, respectively, for determinations of cholinergic activities and contractile responses to acetylcholine (ACh) of the jejunum. Single treatments: while DDVP and propoxur decreased acetylcholinesterase (AChE) activity, oxotremorine and atropine did not. Although DDVP, propoxur and oxotremorine increased levels of ACh, atropine decreased them. Contractile responses to ACh were enhanced by DDVP and reduced by oxotremorine and atropine. The Bmax value of binding of [3H]quinuclidinyl benzylate (QNB) to muscarinic ACh receptors was decreased by atropine. Consecutive treatments: DDVP and oxotremorine decreased AChE activity markedly and slightly, respectively. Although DDVP and oxotremorine increased levels of ACh, propoxur decreased them. Without affecting the contractile responses, DDVP caused a reduction and propoxur and atropine caused an increase in the Bmax value for binding of [3H]QNB. Both the contractile responses and the value of Bmax for binding of [3H]-QNB were decreased by oxotremorine. In summary, propoxur and DDVP showed similar effects mainly through their anticholinesterase properties in the case of single injection, but DDVP had similar effects to those of oxotremorine and propoxur had similar effects to those of atropine in the case of repeated injection.

Studies to evaluate the effectiveness of sex pheromone-impregnated formulations for control of populations of the American dog tick, Dermacentor variabilis (Say) (Acari: Ixodidae).

The sex pheromone, 2,6-dichlorophenol, was combined with a pesticide to control populations of the American dog tick, Dermacentor variabilis (Say). This pheromone persisted in the fur of treated dogs for at least 18 days. The mixture of pheromone and pesticide was much more effective in reducing mating among the surviving ticks than the treatments without pheromone. The pheromone-pesticide mixture also killed significantly more ticks than the treatment without pheromone. This increased effectiveness was due almost entirely to the significantly greater kill of male ticks. Combining the sex pheromone with pesticide treatments offers a means of suppressing tick populations by curtailing their mating and subsequent reproductive success. The potential applications of this technique and the benefits to be expected are discussed.