Resistance to insect growth regulator insecticides in populations of sheep lice as assessed by a moulting disruption assay

Abstract.  Low-volume, backline applications with the benzoylphenyl urea insecticides triflumuron and diflubenzuron represent in excess of 70% of treatments for the control of sheep lice, Bovicola ovis (Schrank) (Phthiraptera: Trichodectidae), in Australia. Reports of reduced effectiveness from 2003 and subsequent controlled treatment trials suggested the emergence of resistance to these compounds in B. ovis populations. A laboratory assay based on the measurement of moulting success in nymphs was developed and used to assess susceptibility to diflubenzuron and triflumuron in louse populations collected from sheep where a control failure had occurred. These tests confirmed the development of resistance to triflumuron and diflubenzuron in at least two instances, with estimated resistance ratios of 67–94X at LC₅₀.     

Pyrethroid resistance in Australian field populations of the sheep body louse, Bovicola (Damalinia) ovis

Abstract. Synthetic pyrethroid (SP) resistance has developed in Australian field populations of the sheep body louse, Bovicola ( Damalinia ) ovis. Laboratory bioassays were used to measure the susceptibility of lice to cypermethrin and the other registered SPs. Results of these bioassays indicated resistance to cypermethrin, deltamethrin, cyhalothrin and alphacypermethrin. So far, high-level resistance has been diagnosed in only a few strains. The toxicological responses of these strains were clearly separated from those of the majority of louse strains tested. Furthermore, these strains had survived immersion in commercial SP dips. The level of resistance described in some strains was sufficient to cause pour-on products to fail despite the fact that the LC50s of these strains fell within the normal range of field responses.     

Speed of action and in vitro efficacy of spinosad against sheep body lice, Bovicola ovis (Schrank) (Phthiraptera: Trichodectidae), resistant to pyrethroid, organophosphate or insect growth regulator insecticides

Abstract  Results of laboratory bioassays indicated that spinosad was equally effective against sheep lice populations that were susceptible to insecticides or resistant to pyrethroid, organophosphorus or insect growth regulator (IGR) insecticides. Spinosad had similar toxicity against susceptible strains of lice to that previously reported for diazinon, but lower toxicity than cypermethrin. Lethal concentrations of spinosad and diazinon caused knock down of lice within 6 h of exposure and death within 24 h. Prior to the current phasing out of diazinon as a sheep dip, most wool producers, needing to control pyrethroid- or IGR-resistant lice infestations in short-wool, would have chosen to use diazinon. Our results suggest that spinosad is an effective alternative for treatment of lice resistant to other chemical groups.     

Heritability of resistance to infestation with the body louse, Bovicola ovis, in Romney sheep bred for differences in resistance or resilience to gastro-intestinal nematode parasites

The inheritance of resistance to louse infestation and the related allergic skin disease, cockle, was examined in Romney lambs. The lambs used in the study were the 2001- and 2004-born progeny of four experimental breeding lines (“Resistant”, “Susceptible”, “Resilient” and “Control”) developed as part of a long-term study of the genetics of host resistance (maintenance of low faecal egg count (FEC) under nematode challenge) or resilience (maintenance of health and productivity under nematode challenge irrespective of FEC) to nematode parasites in sheep. Between 13 and 22 progeny (equally distributed between males and females, where possible) from each of five sires in each line were selected each year for this trial. All lambs (n = 701) were examined for lice (Bovicola ovis) before artificial infestation; in 2001 the lambs were free of natural infestation, whilst in 2004 naturally acquired infestation was evident. In November 2001 and May 2002, approximately 60 B. ovis were transferred to each lamb, followed by monitoring at approximately 2-monthly intervals until August 2002. Similar procedures, but with fewer monitoring times, were repeated on the 2004 lambs. Overall, lambs in the Control line were significantly more susceptible to louse infestation and cockle compared with those in the other three lines (P < 0.001). Least squares-means (SEM) of log-transformed louse score for the control, resistant, susceptible and resilient lines, respectively, were 2.178 (0.045), 1.499 (0.050), 1.618 (0.050) and 1.587 (0.044), and for cockle score were 1.36 (0.05), 0.76 (0.05), 0.95 (0.05) and 0.78 (0.05). From all progeny together, the heritability of log-transformed louse score was 0.22 (Standard Error (SE) 0.06) in autumn and 0.34 (SE 0.08) in winter, with a value of 0.44 (SE 0.09) when these data were combined. These estimates were similar to those obtained for resistance to gastro-intestinal nematodes in these breeding lines, using log-transformed FECs. Heritability estimates for cockle score in autumn, winter and when combined were 0.06 (SE 0.04), 0.45 (SE 0.09) and 0.40 (SE 0.09), respectively. The genetic correlations of mean log-transformed louse score with mean cockle score and levels of two different louse antigens in wool were, respectively, 0.97 (SE 0.04), 0.96 (SE 0.08) and 0.95 (SE 0.09). However, there was no significant genetic correlation between louse scores and FEC. These results suggest that selective breeding would be effective in reducing louse infestation and cockle in sheep, but that differences in louse burdens were not related to differences in nematode burdens as indicated by FECs.     

Evaluation of non-conventional treatments for control of the biting louse (Bovicola ovis) on sheep

variety of non-conventional treatments was applied to biting louse ( Bovicola ovis ) infested sheep in order to evaluate ways in which farmers could control the louse infestations and still maintain Organic Production Standards.
In one trial, louse scores of sheep shorn but kept dry or wetted by water alone or with water plus detergent were compared with unshorn sheep treated similarly. Shearing alone accounted for a 35.7–66.3% reduction in mean louse scores. Wetting alone either with water or with water and added detergent accounted for a 26.9–35.3% reduction in mean louse scores. The combined effects on mean louse scores of shearing and wetting, as opposed to shearing alone, were statistically significant on two of the three farms at 32–35 days post-treatment. The effects persisted for the duration of the trial (between 48 and 52 days), at which point shearing and wetting with detergent provided 95.3–99.6% control of lice. In a second trial, a range of insecticidal substances considered acceptable by Organic Production Standards, azadirachtin (neem), pyrethrum, soap, was applied to louse-infested sheep and their efficacy compared with that of a commercial formulation of cypermethrin. The sheep treated with azadirachtin and pyrethrum had significantly fewer lice than either the control or soap treated sheep over the 48 days of the trial. Neither azadirachtin nor pyrethrum were significantly less effective than cypermethrin. Control (reduction in louse score) of 85.0–100% was achieved over the period of the trial.
It is concluded that most of the non-conventional treatments evaluated had a useful and cost-effective role to play in reducing louse numbers on sheep for at least 40–50 days. The lack of persistence compared with that obtained with conventional insecticides was the only apparent drawback.     

Resistance to two organophosphorus insecticides in New Zealand populations of the Australian sheep blowfly, Lucilia cuprina

Abstract. A treated-surface test was used in a bioassay to detect insecticide resistance to the organophosphorus compounds diazinon and propetamphos, in larvae of field strains of Lucilia cuprina in New Zealand during 1990/91 and 1991/92 (December-April). Mean LC₅₀s for both diazinon and propetamphos showed a significant increase between seasons although this is not necessarily indicative of a trend. Mean LC₅₀s for diazinon increased from 1.16 to 1.60 mg/1 (P< 0.001) and for propetamphos from 0.49 to 0.59 mg/1 (P< 0.005) in pooled populations. Diazinon and propetamphos resistance was significantly correlated in both seasons, suggesting side-resistance between the two insecticides.     

Resistance of Spanish codling moth (Cydia pomonella) populations to insecticides and activity of detoxifying enzymatic systems

Resistance of codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), to insecticides has become a major problem in many apple and pear production areas. Our aim was to determine the level of insecticide resistance in Spanish field populations. Seven field populations collected from apple, Malus domestica Borkhausen (Rosaceae), orchards, and three laboratory susceptible strains of codling moth were studied. Damage at harvest in all the conventional orchards from which codling moth populations were collected was higher than the economic threshold. The efficacy of eight insecticides, with five modes of action, was evaluated by topical application of the diagnostic concentrations on post‐diapausing larvae. The enzymatic activity of mixed‐function oxidases (MFOs), glutathione transferases (GSTs), and esterases (ESTs) was evaluated for each population. The susceptibility to insecticides and the biochemical activity of the three laboratory strains and one organic orchard population were not significantly different. Field populations were less susceptible to the tested insecticides than the susceptible strains, especially for azinphos‐methyl, diflubenzuron, fenoxycarb, and phosalone. The efficacy of all insecticides was significantly dependent on the activity of MFOs. Only the toxicity of the three insecticides most used in Spain when the populations were collected (azinphos‐methyl, fenoxycarb, and phosalone) was also dependent on the activity of ESTs and GSTs activity. We conclude that the control failures were because of the existence of populations resistant to the main insecticides used.     

小菜蛾对九种杀虫剂的抗药性

采用叶片药膜法测定了9种杀虫剂对海南、山东、广东、湖北武汉和襄樊等5个地区小菜蛾Plutella xylostella(L.)田间种群的毒力,和室内相对敏感品系比较。结果显示,5个地区的小菜蛾种群都对氯氰菊酯产生了10倍以上的抗性,广东和山东种群达到30倍以上的抗性。对阿维菌素的抗性山东小菜蛾种群达到135倍,广东种群达到30倍,其他地区均在10倍以下。但是,作用机制与阿维菌素类似的药剂,氟虫腈的抗性5个地区均在5倍或5倍以下。对昆虫生长调节剂定虫隆和氟虫脲的抗性除广东种群分别在10倍和20倍以上外,其他地区均在5倍以下。广东种群对多杀菌素的抗性为7.8倍,其他地区的种群没有产生抗性,多杀菌素和杀虫双均为作用于乙酰胆碱受体的药剂,但是作用的位点不同。杀虫双对5个种群的LC50值均是比较大的(>800 mg/L),尽管没有明显的抗性,不适宜用于小菜蛾田间防治。对呼吸链电子转移抑制剂溴虫腈的抗性均在10倍以下。对辛硫磷的抗性在2~5倍。     

Identification of an abundant allergen from the sheep louse, Bovicola ovis

Infestation of sheep with the louse Bovicola ovis is common worldwide and leads to an allergic dermatitis referred to as ‘scatter cockle’. IgE from an infested lamb was used in immunoaffinity chromatography to purify allergens from crude preparations of whole B. ovis and its faeces. SDS-PAGE of the affinity-purified eluates from both preparations showed a dominant band with M_r of 28.5 kDa. Spleen cells from a mouse immunised with B. ovis faecal antigens were used to produce hybridomas which were screened by ELISA to identify those producing monoclonal antibodies (mAb) to the allergens purified by IgE immunoaffinity chromatography. Western blotting demonstrated that all of the mAbs examined recognised the 28.5 kDa allergen. The allergen, purified using immunoaffinity columns constructed with one of the specific mAbs, was shown to cause immediate and late-phase responses on intradermal skin testing in B. ovis-infested but not in naïve lambs. Levels of serum IgE specific for the purified allergen were significantly higher in infested than in naïve lambs (P ? 0.0025). N-terminal and internal amino acid (aa) sequences obtained from the purified 28.5 kDa allergen were used to design primers to amplify a partial cDNA probe from B. ovis cDNA by PCR. The amplified probe was radiolabeled and used to screen a B. ovis cDNA library. The complete nucleotide sequence of the allergen was determined from the sequences of the positive clones from the library. The full-length cDNA encodes a 255 aa protein including a secretory leader sequence of 26 aas and a mature protein of 229 aas. The encoded protein showed strong homology to several hypothetical proteins of unknown function from diverse species and weak homology with lipid-binding proteins of Xenopus tropicalis and Galleria mellonella. This is the first allergen to be identified from a louse and it has been designated Bov o 1 in accordance with the criteria of the World Health Organization/International Union of Immunological Societies Allergen Nomenclature Subcommittee.     

广东肇庆柑橘木虱田间种群对常用药剂的抗药性

室内采用浸叶法测定了柑橘木虱Diaphorina citri敏感种群与田间种群对广东常用药剂的敏感性,明确了柑橘木虱田间种群对阿维菌素、高效氯氰菊酯、吡虫啉、噻虫嗪、啶虫脒和甲维盐6种药剂的抗药性.柑橘木虱田间种群对70％吡虫啉水分散粒剂、20％啶虫脒可溶液剂、3％甲维盐微乳剂处于低水平抗药性,LC50抗性倍数分别为5...     

昆虫抗药性分子机制研究的新进展

昆虫抗性机制的研究对于抗性监测、治理及新农药的研制具有重要意义。在过去几十年中,人们对与昆虫杀虫剂抗性有关的昆虫行为、生理代谢活动以及作用靶标等进行了广泛的研究。已经证实,昆虫的抗药性与行为改变、生理功能改变、解毒功能增强以及靶标不敏感性有关。近年来,随着分子生物学以及昆虫基因组学的发展,昆虫抗药性的分子机理有了突破性进展,已发现并克隆了一些靶标基因,与抗药性相关的基因突变也得到广泛验证。本文综述了昆虫的抗药性机理在分子生物学上的研究最新进展,重点阐述了与昆虫抗性相关基因的扩增、表达及基因结构的改变等相关内容。     

Kinetic disposition of an aqueous formulation of alphacypermethrin applied to the dorsal mid-line of sheep with long wool and its effect on lice

A group of 5 adult Merino sheep with fleeces about 70 mm long (7-months growth of wool) was treated with a topical formulation of the synthetic pyrethroid insecticide, alphacypermethrin, applied to the dorsal mid-line. Insecticide concentrations at the tip, middle and base of wool staples collected from meridians along the back, upper and lower flanks were measured at intervals from 1 to 98 days after treatment. Some movement of the alphacypermethrin from the back to the lower body occurred within 24h after treatment, but despite careful application of the insecticide there was wide variation in the concentration between and within meridians. The majority of the alphacypermethrin remained close to the dorsal mid-line and near the tip of the staple. There were significant differences in the concentration between the tip, middle and base segments of the staples in the back and lower flank meridians (P < 0.05). Despite exposure of the sheep to normal weathering, there was no significant difference in the concentration of alphacypermethrin between samples collected at day 1 or day 98 after treatment (P > 0.05). Numbers of pyrethroid-susceptible lice surviving exposure in vitro for 20 h differed significantly between samples collected at different times after treatment (P < 0.05). The numbers of lice surviving in samples collected within 28 days after treatment tended to be lower than in those collected from 28 to 98 days but, in some samples, regardless of time after treatment, lice survived for 20 h in wool taken from parts of the fleece that contained high concentrations of alphacypermethrin.     

害虫抗药性的显性水平与抗性进化

对杀虫剂的代谢抗性和主要靶标抗性的显性水平作了理论解释,其中包括昆虫对Bt抗性的显性水平的解释。并对抗性显性具有的多变性作了阐述。分析抗性显性水平与抗药性进化的关系,认为在抗性进化早期抗性表现为显性的基因频率上升快于抗性表现为隐性时;但在抗性等位基因频率较高且出现抗性纯合子个体时,抗性表现为隐性的基因频率上升显著快于抗性表现为显性时。最后论述抗性显性在抗性治理中的应用。     

甜菜夜蛾的抗性及抗性机理研究进展

甜菜夜蛾Spodoptera exigua(Hübner)是世界性农业害虫,近年来在我国由次要害虫上升为主要害虫。甜菜夜蛾对很多种化学杀虫剂和生物杀虫剂产生了抗性。本文分别阐述了甜菜夜蛾对常用杀虫剂如有机磷类、拟除虫菊酯类、氨基甲酸酯类、生长调节类杀虫剂、Bt杀虫剂等的抗性发展现状,并且对抗药性机理进行了总结,提出了抗性治理措施。     

Insect growth regulator cross-resistance studies in field- and laboratory-selected strains of the Australian sheep blowfly, Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae)

Abstract Resistance to diflubenzuron in the Australian sheep blowfly, Lucilia cuprina , has rendered this insecticide incapable of preventing flystrike in sheep from a few districts in eastern Australia. Wool producers affected by this situation must find suitable alternatives to protect their flocks. Results of laboratory bioassays against one population demonstrated that, despite extremely high diflubenzuron resistance (Resistance Factor >791), it had only very low (2x) tolerance of cyromazine and dicyclanil. It is unlikely that this level of tolerance would have any practical impact on field control with either insecticide. Consequently, wool producers in districts where diflubenzuron-resistant flies are common can rotate insecticide treatment to either of these compounds to prevent flystrike in their flocks. However, unlike the highly diflubenzuron-resistant field strain, a laboratory strain selected for resistance to diflubenzuron (Resistance Factor = 617) was 10 times more resistant to dicyclanil than a susceptible strain but, like the field strain, was only two times more tolerant of cyromazine. Conversely, a field-derived strain selected in the laboratory for cyromazine resistance was 20 times more resistant to dicyclanil and 362 times more resistant to diflubenzuron than the reference susceptible strain.     

Ultrastructural changes in the cuticle of the sheep blowfly, Lucilia, induced by certain insecticides and biological inhibitors

The ultrastructural effects on larval cuticle of Lucilia cuprina of two inhibitors of chitin synthesis, diflubenzuron and polyoxin D and an inhibitor of dihydrofolate reductase, aminopterin, are compared with those of the insecticide, cyromazine. Diflubenzuron and polyoxin D both prevent the formation of a normal lamellate appearance in procuticle and interfere with deposition of epicuticle. Aminopterin and cyromazine cause necrotic lesions in the cuticle which, in the case of cyromazine, are contiguous with invasive processes of epidermal cells. There is an accumulation of electron-dense granules in some epidermal cells in larvae poisoned with aminopterin or cyromazine. Aminopterin has a more drastic cytotoxic effect than cyromazine and it also interferes with the formation of epicuticle. The lesions produced by cyromazine treatment are not mimicked precisely by any of the other chemicals. However, there is closer accord between the effects of cyromazine and aminopterin than between cyromazine and the inhibitors of chitin formation.     

Survey of insect growth regulator (IGR) resistance in house flies (Musca domestica L.) from southwestern Turkey

Insect growth regulators (IGRs) are currently the fastest‐growing class of insecticides, and in Turkey these products represent a new approach to pest control. In recent years, several IGRs were also registered for the control of the house fly, Musca domestica L. (Diptera: Muscidae), in Turkey. A field survey was conducted in the summers of 2006 and 2007 to evaluate resistance to some agriculturally and medically used IGRs on house flies from livestock farms and garbage dumps in the greenhouse production areas (Merkez, Kumluca, Manavgat, and Serik) of Antalya province (Southwestern Turkey). The results of larval feeding assay with technical diflubenzuron, methoprene, novaluron, pyripoxyfen, and triflumuron indicate that low levels (RF<10‐fold) of resistance to the IGRs exist in the house fly populations from Antalya province. Exceptions, however, were two populations, Guzoren and Toptas, from the Kumluca area which showed moderate resistance to diflubenzuron with 11.8‐fold in 2006 and 13.2‐fold in 2007, respectively. We found substantial variation in susceptibility of field‐collected house fly populations from year to year and from product to product. We generally observed an increase in resistance at many localities sampled from 2006 to 2007. The implications of these results to the future use of IGRs for house fly control are discussed. It will be critically important to continue monitoring efforts so that appropriate steps can be taken if resistance levels start to increase.     

Resistance of the codling moth Cydia pomonella (L.) (Lep., Tortricidae) to pesticides in Israel

Abstract:  Resistance of the codling moth Cydia pomonella (L.) (Lep., Tortricidae) to the organophosphorus compound (OP) azinphosmethyl was observed in apple orchards in Israel. The level of resistance varied with the pest control strategy. Compared with a sensitive laboratory population, the resistance level was highest in insects from the preventative pest control strategy, intermediate in integrated pest management (IPM) orchards, and relatively low in the organic orchards. The level of azinphosmethyl resistance in larvae (but not in adults) exposed for 17 generations in the laboratory to a pesticide-free diet was reduced by 50%. Codling moth larvae resistant to azinphosmethyl were also resistant to various insect growth regulators (IGRs). The IGRs include three chitin synthesis inhibitors (diflubenzuron, novaluron and teflubenzuron), two juvenile hormone mimics (pyriproxyfen and fenoxycarb) and one ecdysone agonist (methoxyfenozide). Codling moth resistant to azinphosmethyl was tolerant to methoxyfenozide and novaluron without previous history of application in apple orchards, indicating the possibility of cross-resistance. According to this study, managing resistance programs in apple orchards should be based on IPM principles with minimum use of conventional neuroactive pesticides.     

Effects of pyriproxyfen and buprofezin on immature development and reproduction in the stable fly*

The stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae), is one of the most significant biting flies that affect cattle. The use of traditional insecticides for stable fly control has only a limited success owing to the insect's unique feeding behaviours and immature development sites. A laboratory study was conducted to evaluate the effects of two insect growth regulator (IGR) products, pyriproxyfen and buprofezin, on the development of the immature stages of the stable fly and the effects of pyriproxyfen on oviposition and egg hatch. Both pyriproxyfen and buprofezin had significant inhibitory effects on immature development. The LC₅₀s of pyriproxyfen and buprofezin were 0.002 and 18.92 p.p.m., respectively. Topical treatment of adult females with different doses of pyriproxyfen had significant negative effects on both female oviposition and egg hatching when 1‐ and 3‐day‐old females were treated, and the effects were dose dependent. A significant reduction in the mean number of eggs laid was observed only at the highest pyriproxyfen dose (8 µg/fly) and egg hatch was unaffected by pyriproxyfen treatment when 5‐day‐old females were treated. Results from the present study indicate that pyriproxyfen has the potential to be used as part of an integrated stable fly management programme.