Potential estrogenic and antiandrogenic effects of permethrin in rats

Many environmental chemicals including pesticides have been reported to possess hormonal activities, and thus are classified as endocrine disruptors. Permethrin, a synthetic pyrethroid insecticide, is used worldwide, which provides potential environmental exposure. However, relatively few studies have reported on hormonal activities, particularly estrogenic and androgenic activities of permethrin, and the results of these studies are in some respects contradictory. Therefore, this study investigated the potential estrogenic and androgenic activities of permethrin in vitro and in vivo. We conducted an uterine Calbindin-D9k (CaBP-9k) gene expression assay and an uterotrophic assay for estrogenic activity, and a Hershberger assay for androgenic activity. The CaBP-9k gene, one of the intracellular calcium binding proteins, is estrogen-responsive in the uterus. The rat uterotrophic and Hershberger assays are generally used as in vivo short-term screening assays for detecting the estrogenic and androgenic activities of chemicals, although these assays are still being validated by the Organization for Economic Cooperation and Development (OECD). Northern blot analysis showed the induction of uterine CaBP-9k mRNA level in response to permethrin as well as co-administration of permethrin with E2. In the uterotrophic assay using 18-day-old female rats, subcutaneous treatments with permethrin (10 to 800 mg/kg) for three days increased relative uterine wet weights, and E2-induced uterine weights. These effects were statistically significant at 800 and 200 mg/kg, respectively. Moreover, permethrin-induced uterine weights were inhibited by the co-administration of ICI 182,780, an antiestrogen. In the Hershberger assay, the administration of permethrin orally to testosterone propionate-treated castrated male rats led to statistically significant reductions in androgen-dependent sex accessory tissue (ventral prostate, seminal vesicles, levator ani and bulbocavernosus muscles, Cowper's gland and glans penis) weights at all doses tested (10, 50 and 100 mg/kg). These results suggest that permethrin might have estrogen-like effects on female rats, but antiandrogen-like effects on males.     

Differentiation of resistance in cattle to larval Hypoderma lineatum

Cattle were successively infested for 4 years with larval cattle grubs, Hypoderma lineatum, to demonstrate the acquisition of resistance. The effects of host age and dietary-induced state with Vitamin A on susceptibility to infestation and their interactions with effects from previous infestations were also determined.Cattle acquired resistance to infestation after a single infestation. This resistance was not affected by host age for at least 4 years, was not affected by a deficiency of vitamin A, and was effective in the skin and connective tissues of the host. The effects of this acquired resistance were highly variable in degree, were produced primarily during the first 90 days of infestation and increased after repeated provocation to provide nearly complete protection in some animals.Calves and yearlings had innate resistance that decreased with age, was localized in the skin, was impaired by a deficiency of Vitamin A, and resulted in a mean loss of 65–75% of the larvae in an infestation dose of up to 150.The combined effects of the 2 types of resistance reduced the grub-burden in cattle throughout their life.     

A larval development test for the detection of anthelmintic resistance in nematodes of sheep

First stage larvae of a number of species of parasitic nematodes of sheep have been shown to develop to third stage larvae in the presence of a defined medium consisting of Earle's balanced salt solution and yeast extract. A larval development test, based on this culture technique, was used as a screen for detecting the presence of anthelmintic resistance in nematodes of sheep. It was found to be sensitive and simple to use and also appeared capable of detecting resistance to any of the main anthelmintic groups. Available anthelmintic sensitive and resistant strains of Haemonchus contortus and Ostertagia circumcincta showed differences in development when incubated in the presence of either thiabendazole, levamisole and ivermectin. These differences were expressed as the minimum inhibitory concentration required to prevent larval development over the incubation period.     

Single-dose toxicokinetics of permethrin in broiler chickens

1. Single-dose toxicokinetics of permethrin was investigated in broiler chickens.

2. A total of 20 male broiler chickens were assigned at random to two groups of 10 at 30 days of age. A single dose of 10 mg/kg body weight of permethrin was administered intravenously to the first group; in the second group, the same dose was administered into the crop.

3. Serum permethrin was measured using an electron capture detector and gas chromatography equipment. The derived serum permethrin concentration/time curve demonstrated that the distribution kinetics of permethrin was well described by a two-compartment open model.

4. For intravenous permethrin administration, the half-life at λ phase (t_1/2λ), mean residence time (MRT) and area under the concentration–time curve in 0→∞ (AUC_0→∞) values respectively were 4.73 ± 1.00 h, 5.06 ± 1.05 h and 16.45 ± 3.28 mg/h/l. In contrast, the C_max, t_max, t_1/2λ, MRT and AUC_0→∞ values respectively of the group given intra-crop permethrin were 0.60 ± 0.42 μg/ml, 0.55 ± 0.19 h, 5.54 ± 0.78 h, 7.06 ± 0.63 h and 1.95 ± 0.97 mg/h/l. The bioavailability of permethrin was 0.11.

5. For both administration routes, the residence time of permethrin in the body was short and the bioavailability of permethrin was low. These results are relevant for assessing the use and safety of permethrin.

     

High-level ivermectin resistance in a field isolate of Haemonchus contortus associated with a low level of resistance in the larval stage: implications for resistance detection

The IVPro isolate of Haemonchus contortus was isolated in 1999 after significant numbers of the parasite survived an ivermectin capsule treatment of grazing sheep acquiring a natural infection at Prospect, NSW, Australia. The isolate shows high-level resistance to ivermectin (faecal egg count is unaffected by ivermectin oral treatment at 0.2mg kg(-1)). The larval LC(50), as assessed by larval development assays (LDAs), is only approximately two-fold higher than several susceptible isolates, making it difficult to detect the resistance using larval LC(50) as an indicator. However, the isolate shows the presence of a small proportion of the population (<20%) able to develop at significantly higher drug concentrations than the susceptible isolates. Hence, if the IVPro and susceptible isolates are compared at the LC(99) level, the IVPro isolate is readily identifiable as resistant. This degree of distinction at the LC(99) allows the IVPro isolate to be identified as resistant by simply observing the highest drug concentration in the development assay at which some larvae develop relative to the susceptible isolates. Assessing the development assay using this criterion allows a distinction between IVPro and the susceptible isolates equivalent to 10-fold differences in drug concentration, greatly increasing the likelihood of detecting the resistant isolate in routine resistance tests. This study highlights the need to examine this aspect of LDAs in order to detect the type of resistance displayed by IVPro H. contortus.     

Relative importance of larval and adult Mecistocirrus digitatus in inducing resistance to a reinfection in calves

Calves immunized with adult Mecistocirrus digitatus implanted directly into the abomasum did not develop a substantial degree of immunity to a subsequent large oral (challenge) dose of larvae, which developed to maturity. In contrast animals immunized by oral infection developed strong resistance. The calves implanted with adult worms appeared to show a greater degree of susceptibility to maturation of the challenge infection than controls which received a challenge of the same magnitude without any previous immunization. The implanted female adult worms established in the hosts and continued to produce more eggs for a longer time than those which developed to maturity from the oral immunizing infection with third-stage larvae. Passive haemagglutination studies revealed that the implanted adult worms stimulated little or no antibody response in the hosts. In the calves which did not show a response to the adult worm implant the subsequent challenge with an oral infective dose of third-stage larvae also failed to stimulate a response. Likewise the two calves from the group which showed a weak antibody response to the adult worm implant did not show an increased response when challenged. In contrast, calves immunized with an oral infection of third-stage larvae had an antibody response which showed a vigorous rise on challenge in four of the five calves. Thus a direct relationship between resistance to challenge infection and the antibody response determined by the passive haemagglutination and gel-diffusion tests was observed in the calves immunized orally.     

Clinical effects and outcome of feline permethrin spot-on poisonings reported to the Veterinary Poisons Information Service (VPIS), London

Permethrin is a pyrethroid insecticide used in dermally applied spot-on flea treatments for dogs. Permethrin-based spot-on preparations are contraindicated in cats because of the high risk of toxicosis. The Veterinary Poisons Information Service (VPIS) is a 24-h access telephone service that provides veterinary professionals in the United Kingdom with information on the management of poisoned animals. In a review of 286 cases reported to the VPIS regarding inappropriate feline exposure to permethrin spot-on (PSO) preparations, 96.9% were symptomatic. Increased muscular activity (as evidenced by twitching, tremor, muscle fasciculations or convulsions) was common and occurred in 87.8% of cases. The duration of increased muscle activity was long, with convulsions lasting on average 38.9 h and tremors 32 h. Recovery typically occurred within 2 to 3 days but in some cases took 5 to 7 days. Death occurred in 10.5% of cases.     

Culicoides species associated with sheep in the Netherlands and the effect of a permethrin insecticide

Culicoides spp. act as vectors for a number of viral diseases of animals including bluetongue in sheep. The aims of this study were to determine: (1) which Culicoides spp. are associated with sheep in The Netherlands; (2) the time of the day when they are most active; and (3) the effect of treatment of animals with a permethrin insecticide. Two pairs of sheep were each housed within mosquito tents of either one or two layers of netting and all trapped Culicoides spp. were identified microscopically. For the permethrin insecticide study, one of two pairs of sheep was treated with 3.6% permethrin and all animals were housed in tents of similar design. Of the 6210 midges captured, 54.1% were identified as C. chiopterus and 42.7% as C. obsoletus. C. imicola was not identified. The average insect feeding rate was 35-40% and midge activity was greatest around sunset. Permethrin treatment reduced the number of midges captured by 50% and also resulted in a decrease in the percentage of midges that had fed. The findings provide useful information on the behaviour and distribution of Culicoides spp. that will facilitate the development of appropriate control strategies to minimise the risk of insect-vector borne virus diseases such as bluetongue.     

Evaluation of a larval development assay (DrenchRite) for the detection of anthelmintic resistance in cyathostomin nematodes of horses

A larval development assay (LDA, DrenchRite) was evaluated to determine the effectiveness of this method in detecting anthelmintic resistance in cyathostomin nematodes of horses. A total of 15 horse farms from Georgia and South Carolina (USA) and Population S ponies from the University of Kentucky (USA) were included in this study. Nematode eggs were extracted from pooled fecal samples and placed into the wells of a DrenchRite plate for testing against thiabendazole (TBZ), levamisole (LEV) and 2 ivermectin (IVM) analogs (IVM-1, IVM-2). After a 7-day incubation larvae in each well were counted and data were analyzed by logistic regression. Resistance status of each farm for different drugs was determined in a separate study using a fecal egg count reduction test. LDA were performed on the 15 farms once, however, the Population S cyathostomins were assayed on 3 separate occasions to estimate the consistency of results between assays. Mean TBZ LC50 for oxibendazole resistant, suspected resistant and sensitive farms were 0.2015, 0.1625, and 0.1355 microM, respectively. For LEV, mean LC50 for PYR resistant, suspected resistant and sensitive farms were 1.590, 1.8018 and 1.4219 microM, respectively. All 15 farms had worms susceptible to IVM; mean LC50 for IVM-1 and for IVM-2 were 7.5727 and 87.9718 nM, respectively. A linear mixed model was fitted to the data to determine the relationship between LC50 and LC95 and resistance status for each farm. No meaningful relations were found. Consistency of assays varied between drugs, being best for TBZ and worst for LEV and IVM-1. All farms in this study had benzimidazole-resistant nematodes; therefore usefulness of DrenchRite for discriminating susceptibility versus resistance to this drug class could not be accurately assessed. Moreover, since all farms tested were sensitive to IVM and resistance to this drug class has not yet been reported in cyathostomins, it is not possible to assess accurately the usefulness of DrenchRite LDA for detecting IVM resistance at this time. Assay results for LEV suggest that LEV in a LDA does not yield data that is useful in estimating PYR efficacy in vivo. Based on results for PYR/LEV, the current high prevalence of benzimidazole resistance, no known cases of IVM resistance, and the sometimes extreme variation in results seen in many of the assays, DrenchRite LDA cannot be considered a useful tool for the diagnosis of resistance in cyathostomins of horses at present.     

Comparison of two versions of larval development test to detect anthelmintic resistance in Haemonchus contortus

Larval development (LDT) and micro-agar larval development tests (MALDT) were used to compare the reliability and sensitivity of two methods for detecting anthelmintic resistance in Haemonchus contortus. The tests were conducted using three resistant and four susceptible isolates of H. contortus. Both versions of the tests provided comparable results with regard to the characterization of benzimidazole and levamisole susceptibility but neither test was sufficiently sensitive to discrimination between an ivermectin (IVM) susceptible and an IVM resistant isolate. Each test has its own merits with the LDT having the advantage of being less time-consuming.     

家蝇抗菌肽的分离纯化及生物学活性

将诱导家蝇提取的家蝇抗菌肽提取物，经凝胶过滤层析和离子交换层析，得到具有抗菌活性的单一峰，质谱结果显示抗菌肽分离物含有4种小肽，相对分子质量678．3～1017．2。家蝇抗菌肽对革兰氏阴性菌（大肠杆菌、巴氏杆菌）的抗菌活性优于革兰氏阳性菌（金黄葡萄球菌、枯草芽孢杆菌），对耐药性巴氏杆菌有较强的抗菌活性，对大肠杆菌的MIC为4．2～8．4mg／L，对耐药性巴氏杆菌的MIC为3．8～7．8mg／L。扫描电镜观察发现，家蝇抗菌肽通过损伤细胞壁使细胞质外流而达到杀菌的目的。家蝇抗菌肽对家兔、鸡、猪、小鼠、番鸭、鹅和家鸭红细胞无溶血反应和凝集反应。‘     

The effects of ivermectin and moxidectin on egg viability and larval development of ivermectin-resistant Haemonchus contortus

The in vivo effects of ivermectin and moxidectin on egg viability and larval development of ivermectin-resistant Haemonchus contortus were examined over time after anthelmintic treatment of sheep. Twenty merino sheep, (12 months old) were allocated to five treatment groups and infected with ivermectin-resistant H. contortus. Thirty one days later, the sheep were treated with intraruminal ivermectin capsules, oral ivermectin, oral moxidectin or injectable moxidectin at the manufacturer's recommended dosages, or left untreated. At various times up to 112 days after treatment, faecal egg counts (FEC) were determined and development rates of infective larvae (L3) cultured in faeces or on agar were measured. Eggs in faecal cultures from ivermectin capsule treated sheep showed reduced L3 development percentages in comparison to faecal cultures from untreated sheep. Eggs from ivermectin capsule treated sheep, isolated from faeces, and cultured on agar showed similar L3 development to eggs from control sheep. These results demonstrate an inhibitory effect of excreted ivermectin in faeces on larval development of ivermectin-resistant H. contortus. L3 development in faecal culture from animals receiving oral ivermectin were reduced for only 3 days after treatment. Faecal egg counts and development of L3 larvae in both culture systems from moxidectin treated sheep were low, due to the high efficacy of the drug. Egg counts in moxidectin treated sheep were reduced by approximately 90% 24h after treatment, before decreasing to almost 100% at 48h, suggesting that the current quarantine recommendation of holding sheep off pasture for 24h after treatment may still lead to some subsequent pasture contamination with worm eggs.     

Reliability and reproducibility of the larval paralysis test as an in vitro method for the detection of anthelmintic resistance of nematodes against levamisole and morantel tartrate

In order to study the reliability of the larval paralysis test as an in vitro assay for the detection of resistance of nematodes to levamisole and morantel tartrate, the influence of different parameters was evaluated using resistant and susceptible Ostertagia ostertagi strains. The operator, the sample (10% of the larvae present in the suspension), the incubation time (24, 48 or 72 h), the incubation temperature (20 or 25 degrees C) and the observation period of the larvae (5 or 15 s) had no statistically significant influence on the test results. Statistical differences were obtained only when L3 larvae of different ages (1 or 2 months) were used. Reversibility of the paralysis did not occur when concentrations of levamisole of less than or equal to 200 micrograms ml-1 or morantel tartrate of less than 2000 micrograms ml-1 were used. The reproducibility of the test was fairly good, with a mean standard deviation of 21.3% for the LC50 values. Morantel resistance in a strain of O. ostertagi was not confirmed as such by the larval paralysis test. The resistance factor was less than or equal to 1, in spite of an efficacy of morantel of 77.4% as shown in vivo.     

Inhibited development of Obeliscoides cuniculi in rabbits: the effects of active and passive immunization and resumption of larval development

Rabbits were either actively or passively immunized against Obeliscoides cuniculi and challenged with 5 000 infective larvae (IL). Worm development was inhibited in actively immunized animals and worm egg production was impaired by administration of immune serum. Inhibited fourth stage larvae (L₄) from high-level infections (200 000 IL) and normal L₄ from low-level infections (4 000 IL) were transferred to actively immunized and normal rabbits. In both instances, large numbers of L₄ developed to maturity; however, a significant number of inhibited L₄ failed to resume development. Inhibition of larvae resulted in the development of fewer adult male than female worms, and the change in sex ratio was further accentuated in actively immunized recipients. Worm egg production was also reduced in actively immunized rabbits. These data indicate that host immune responses contribute to inhibition of O. cuniculi in rabbits and that worm egg production is also responsive to immunologic control.     

蝇蛆油对实验动物皮肤的刺激性、过敏性及烫伤治疗药效研究

蝇蛆具有良好的药用价值,蝇蛆油对试验动物损伤也有良好的治愈效果.试验证明蝇蛆油对正常皮肤及破损皮肤均无刺激性作用,对皮肤也无过敏反应.药效学试验表明,蝇蛆油溶液对浓盐酸引起的化学致炎及烧伤具有明显的治疗作用.     

Transmission of Campylobacter jejuni by the housefly (Musca domestica)

Houseflies (Musca domestica) were infected with Campylobacter jejuni after being confined for 5 days in a Horsfall isolator containing 25-day-old chickens known to be fecal excretors of the organism. Contaminated flies, when subsequently transferred to a second unit, transmitted C. jejuni to specific-pathogen-free chickens. Allowing a sample of 32 houseflies to ingest C. jejuni in a liquid suspension resulted in recovery rates of 20% from the feet and ventral surface of the body and 70% from the viscera. These experiments demonstrated the potential role of flies in the dissemination of avian campylobacteriosis.     

Residual feed intake of purebred Angus steers: effects on meat quality and palatability

Relationships between residual feed intake (RFI) and other performance variables were determined using 54 purebred Angus steers. Individual feed intake and BW gain were recorded during a 70-d post-weaning period to calculate RFI. After the 70-d post-weaning test, steers were fed a finishing ration to a similar fat thickness (FT), transported to a commercial facility, and slaughtered. A subsample of carcasses (n = 32) was selected to examine the relationships among RFI, meat quality, and palatability. Steers were categorized into high (> 0.5 SD above the mean; n = 16), medium (mid; +/- 0.5 SD from the mean; n = 21), and low (< 0.5 SD below the mean; n = 17) RFI groups. No differences were detected in ADG, initial BW, and d 71 BW among the high, mid, and low RFI steers. Steers from the high RFI group had a greater DMI (P = 0.004) and feed conversion ratio (FCR; DMI:ADG; P = 0.002) compared with the low RFI steers. Residual feed intake was positively correlated with DMI (r = 0.54; P = 0.003) and FCR (r = 0.42; P = 0.002), but not with initial BW, d 71 BW, d 71 ultrasound FT, initial ultrasound LM area, d 71 ultrasound LM area, or ADG. The FCR was positively correlated with initial BW (r = 0.46; P = 0.0005), d 71 BW (r = 0.34; P = 0.01), and DMI (r = 0.40; P = 0.003) and was negatively correlated with ADG (r = -0.65; P = 0.001). There were no differences among RFI groups for HCW, LM area, FT, KPH, USDA yield grade, marbling score, or quality grade. Reflectance color b* scores of steaks from high RFI steers were greater (P = 0.02) than those from low RFI steers. There was no difference between high and low RFI groups for LM calpastatin activity. Warner-Bratzler shear force and sensory panel tenderness and flavor scores of steaks were similar across RFI groups. Steaks from high RFI steers had lower (P = 0.04) off-flavor scores than those from low RFI steers. Cook loss percentages were greater (P = 0.005) for steaks from low RFI steers than for those from mid RFI steers. These data support current views that RFI is independent of ADG, but is correlated with DMI and FCR. Importantly, the data also support the hypothesis that there is no relationship between RFI and beef quality in purebred Angus steers.