Large-scale identification of rodenticide resistance in Rattus norvegicus and Mus musculus in the Netherlands based on Vkorc1 codon 139 mutations

BACKGROUND

Resistance to rodenticides has been reported globally and poses a considerable problem for efficacy in pest control. The most-documented resistance to rodenticides in commensal rodents is associated with mutations in the Vkorc1 gene, in particular in codon 139. Resistance to anticoagulant rodenticides has been reported in the Netherlands since 1989. A study from 2013 showed that 25% of 169 Norway rats (Rattus norvegicus) had a mutation at codon 139 of the Vkorc1 gene. To gain insight in the current status of rodenticide resistance amongst R. norvegicus and house mice Mus musculus in the Netherlands, we tested these rodents for mutations in codon 139 of the Vkorc1 gene. In addition, we collected data from pest controllers on their use of rodenticides and experience with rodenticide resistance.

RESULTS

A total of 1801 rodent samples were collected throughout the country consisting of 1404 R. norvegicus and 397 M. musculus. In total, 15% of R. norvegicus [95% confidence interval (CI): 13–17%] and 38% of M. musculus (95% CI: 33–43%) carried a genetic mutation at codon 139 of the Vkorc1 gene.

CONCLUSION

This study demonstrates genetic mutations at codon 139 of the Vkorc1 gene in M. musculus in the Netherlands. Resistance to anticoagulant rodenticides is present in R. norvegicus and M. musculus in multiple regions in the Netherlands. The results of this comprehensive study provide a baseline and facilitate trend analyses of Vkorc1 codon 139 mutations and evaluation of integrated pest management (IPM) strategies as these are enrolled in the Netherlands. © 2022 The Dutch Pest and Wildlife. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Resistance to anticoagulant rodenticides in Germany and future strategies to control Rattus norvegicus

Widespread anticoagulant resistance was discovered in populations of the brown rat (Rattus norvegicus Berk.) in an area of roughly 8000 km² in north-west Germany. Resistance testing was performed by feeding tests and/or by measuring blood clotting response after intraperitoneal injection of a sublethal testing solution. A hierarchical resistance system was found with warfarin resistance at the base followed by bromadiolone/coumatetralyl resistance to difenacoum resistance at the top. Warfarin resistance was spread over the whole area with reduced incidence towards the edges. Difenacoum resistance represented the highest level found so far and was restricted to the inner zone of the resistance area where it occurred with a frequency of 6% of all individuals tested. Breeding experiments with bromadiolone-resistant rats showed that expression of the bromadiolone resistance gene differed in the two sexes, suggesting additional sex-linked modifying effects to the resistance gene. Control strategy within the resistance area with respect to prevention of further selection for resistance is discussed.     

Exploring drivers of the usage of anticoagulant rodenticides on UK farms

Anticoagulant rodenticides (ARs) are considered inhumane, show increasingly limited efficacy due to acquired resistance, and carry environmental consequences associated with non-target species uptake. In a questionnaire study of 499 UK farms that all deployed chemical rodenticide we found a high mean reliance (79%), on second generation ARs with just over half of the respondents using no other rodent control methods. Additional methods where deployed, alone or in combination, included predation (41%), kill-trap deployment (16%) and shooting (1%). Nearly 40% of all respondents deployed rodenticides year-round. There was no evidence to suggest that “tidy-farm” measures, such as clearing food spills and minimising on-farm rodent harbourage sites aimed at minimising rodent-associated problems, were associated with a lower likelihood of year-round deployment; in fact trends in our analyses suggested the opposite. We therefore encourage operators to fully evaluate the true necessity of rodenticide deployment before AR use.     

Vitamin K requirement and reproduction in bromadiolone-resistant Norway rats

BACKGROUND: Nucleotide polymorphisms in the VKORC1 gene can be linked to anticoagulant rodenticide resistance in Norway rats (Rattus norvegicus Berkenhout). This provides a fitness advantage to rats exposed to anticoagulant actives, but may also cause fitness costs. The vitamin K requirement and reproductive parameters of bromadiolone‐resistant rats (Westphalian resistant strain; VKOR variant Tyr139Cys) and bromadiolone‐susceptible Norway rats were compared. RESULTS: At vitamin K deficiency, blood clotting times increased in all homozygous resistant males within 8 days and in 80% of homozygous resistant females within 15 days. There was little effect on blood clotting in heterozygous males and no effect in heterozygous females and VKOR wild‐type individuals. Litter size was about 20% higher in sensitive pairs compared with resistant pairs. Testes growth, male gonad weight, sperm motility and testis cell concentration were unaffected by the mutation. CONCLUSIONS: The VKOR variant Tyr139Cys causes considerable physiological cost in Norway rats in terms of vitamin K requirement and reproduction. This may affect the distribution and spread of resistant individuals in the wild. Decreased litter size of resistant parents seems to be due to lowered female reproductive performance, as there was no significant effect of the mutation on any aspects of male reproduction considered, but this requires further study. Copyright © 2011 Society of Chemical Industry     

Distribution of VKORC1 single nucleotide polymorphism in wild Rattus norvegicus in France

BACKGROUND: Anticoagulant rodenticides are commonly used to control rodent pests all over the world. These pesticides inhibit one enzyme of the vitamin K cycle, Vkorc1, and thus prevent blood clotting and cause death by haemorrhage. Resistance to anticoagulants was first observed in Scotland in 1958, and more potent anticoagulants have been developed to overcome this obstacle. Unfortunately, these chemicals are very toxic and cannot be used everywhere. Some authors have shown that resistance to anticoagulants seems closely linked with single nucleotide polymorphism (SNP) in the Vkorc1 gene. RESULTS: This study draws a map of SNP and haplotypes found in Vkorc1 in rats from different areas of France. Some of them had never been described before. Moreover, the Y139F mutation, described previously in France and Belgium, is the most frequent in France. This mutation is known to be associated with a strong resistance to anticoagulants, and it was found in 28% of the samples. CONCLUSION: This biomolecular approach to studying and detecting resistance is easier to carry out than the phenotypic approach measuring blood coagulation time because it can be conducted on biological samples from dead animals, and it is less dangerous for the operator. Copyright © 2009 Society of Chemical Industry     

3种抗凝血类杀鼠剂在喀什农区防治害鼠效果及对家禽的安全性评价

为确定适于防治南疆农区鼠害的抗凝血类杀鼠剂, 在喀什地区疏勒县采用夹捕法、食饵法、粉迹法评估了杀鼠醚、溴敌隆、溴鼠灵的防治效果, 并采用灌胃法给药测试3种杀鼠剂对家鸡Gallus domestiaus和鸽子Columba livia的安全性。结果表明, 连续投放毒饵10 d后, 与溴敌隆防治区相比, 杀鼠醚防治区的阳性粉块和无毒小麦取食量的下降幅度均无显著差异, 但无毒小麦取食量下降的农户比例更高;且防治后褐家鼠比例更低, 防治期间毒饵消耗量更高。连续单独投放溴敌隆30 d的灭鼠效果可达90.9%, 但10 d溴敌隆与20 d溴鼠灵联合防治防效仅为65.9%。灌胃法测试表明, 对当地家禽类的毒性从低到高依次为杀鼠醚、溴敌隆和溴鼠灵。因此, 建议优选毒性较低的杀鼠醚或溴敌隆用于南疆农区鼠害防治。     

Brodifacoum is effective against Norway rats (Rattus norvegicus) in a tyrosine139cysteine focus of anticoagulant resistance in Westphalia,Germany

BACKGROUND: The tyrosine to cysteine amino acid substitution at location 139 of the vkorc1 protein (i.e. tyrosine139cysteine or Y139C) is the most widespread anticoagulant resistance mutation in Norway rats (Rattus norvegicus Berk.) in Europe. Field trials were conducted to determine the incidence of the Y139C mutation at two rat‐infested farms in Westphalia, Germany, and to estimate the practical efficacy against them of applications, using a pulsed baiting treatment regime, of a proprietary bait (Klerat^?) containing 0.005% brodifacoum. RESULTS: DNA analysis for the Y139C mutation showed that resistant rats were prevalent at the two farms, with an incidence of 80.0 and 78.6% respectively. Applications of brodifacoum bait achieved results of 99.2 and 100.0% control at the two farms, when measured by census baiting, although the treatment was somewhat prolonged at one site, possibly owing to the abundance of attractive alternative food. CONCLUSION: The study showed that 0.005% brodifacoum bait is fully effective against Norway rats possessing the Y139C mutation at the Münsterland focus and is likely to be so elsewhere in Europe where this mutation is found. The pulsed baiting regime reduced to relatively low levels the quantity of bait required to control these two substantial resistant Norway rat infestations. Previous studies had shown much larger quantities of bromadiolone and difenacoum baits used in largely ineffective treatments against Y139C resistant rats in the Münsterland. These results should be considered when making decisions about the use of anticoagulants against resistant Norway rats and their potential environmental impacts. Copyright © 2012 Society of Chemical Industry     

Resistance testing and the effectiveness of difenacoum against Norway rats (Rattus norvegicus) in a tyrosine139cysteine focus of anticoagulant resistance,Westphalia, Germany

BACKGROUND: Anticoagulant resistance in Norway rats at foci in Belgium, Denmark, France, Germany, the Netherlands and the United Kingdom is genetically characterised by the same single nucleotide polymorphism (SNP) and consequent amino acid exchange from tyrosine to cysteine at location 139 of the vkorc1 gene (i.e. tyrosine139cysteine or Y139C). The purpose of this study was to assess the degree of resistance among rats at two infested farm sites in the Y139C focus in Westphalia, Germany, using blood clotting response (BCR) tests, and to determine the practical efficacy of applications of a commercial 50 ppm difenacoum bait (Neokil^?) against them. RESULTS: BCR tests showed that the difenacoum resistance factor (RF) among the Y139C rats was about 2.5. DNA analysis for the Y139C mutation revealed that it was present among rats at the two sites with a prevalence of 75 and 93%. Applications of difenacoum bait at the two sites achieved 86.8 and 59.9% control. The different outcomes did not appear to be due to differences either in the degree and prevalence of resistance or in the quantities of poisoned bait consumed. CONCLUSION: The study showed that, although the RF for difenacoum among rats carrying the Y139C SNP was apparently low, an acceptable level of control of resistant Norway rat infestations was not achieved using difenacoum. Continued use of anticoagulants against rats that are resistant to them will exacerbate resistance problems in terms of both increased severity and prevalence. These conclusions are likely to apply elsewhere in Europe where the Y139C SNP occurs. Copyright © 2012 Society of Chemical Industry     

长江中下游稻区毒饵灭鼠后害鼠种群数量的变化

长江中下游稻作区大面积毒饵灭鼠后,褐家鼠种群密度下降很快.相对而言,小型鼠小家鼠(农舍区)与黑线姬鼠(农田区)及栖息在房屋上层的黄胸鼠,容易漏灭.在多种鼠并存而灭鼠效果较差的地区,甚至会出现灭后小型鼠捕获率上升的情况,则是先前受种间竞争抑制其活动之故,所以制定灭鼠技术方案应注意当地鼠种组成.灭鼠质量的好坏和灭鼠面积的大小,对灭鼠后的数量回升速度有较大的影响.灭鼠率越高,灭鼠覆盖面积越大,害鼠种群保持在较低水平的时间越长,害鼠种群回升的速度越慢.但无论如何,若生态环境条件不变,毒饵杀灭后害鼠种群密度总会回升,即仅单纯依靠毒饵灭鼠手段,难以持久控制鼠害.所以既要因地制宜提高化防质量以抑制害鼠数量,更应大力提倡生态防治为主的综合措施.     

Effects of mutations in Drosophila nicotinic acetylcholine receptor subunits on sensitivity to insecticides targeting nicotinic acetylcholine receptors

Several strains of Drosophila melanogaster possess mutant alleles in nicotinic acetylcholine receptor (nAChR) subunits, Dα1 and Dβ2 that confer resistance to neonicotinoids such as imidacloprid and nitenpyram, and Dα6, that confers resistance to spinosyns. These mutant strains were bioassayed with a selected set of nAChR active insecticides including neonicotinoids, spinosad, and sulfoxaflor, a new sulfoximine insecticide. All of the neonicotinoids examined, except dinotefuran showed reduced insecticidal efficacy on larvae of the Dα1 mutant, suggesting that this subunit may be important in the action of these insecticides. All of the neonicotinoids, including dinotefuran, showed reduced insecticidal efficacy on larvae possessing the Dβ2 mutation. A similar pattern of broad neonicotinoid resistance to that of Dβ2 alone was also observed for larvae with both the mutations (Dα1 + Dβ2). The Dβ2 mutation exhibited a lower level of cross-resistance to sulfoxaflor (<3-fold) than to any of the neonicotinoids (>13-fold). In contrast, there was no cross-resistance for any of the neonicotinoids or sulfoxaflor in adult flies with the Dα6 mutation, which confers high levels of resistance to spinosad. Thus in the D. melanogaster strains studied, target site resistance observed for the neonicotinoids and the spinosyns does not translate directly to resistance towards sulfoxaflor.     

Distribution and frequency of VKORC1 sequence variants conferring resistance to anticoagulants in Mus musculus

BACKGROUND: Emerging resistance to anticoagulant rodenticides may significantly impair house mouse (Mus musculus L.) control. As in humans and rats, sequence variants in the gene vitamin K epoxide reductase complex subunit 1 (VKORC1) of house mice are strongly implicated in the responses of mice to anticoagulants. This study gives a first overview of the distribution and frequency of such potentially resistance‐conferring sequence variants in house mice, based on tissue samples from 30 populations in Germany, Switzerland and the Azores. RESULTS: Except for one population from south Germany, sequence variants were found in individuals from all locations sampled (29 out of 30 sites surveyed), with less than 10% of the individuals matching the wild‐type genotype. The most frequent and widespread amino acid substitutions were Leu128Ser, Tyr139Cys and a group of linked sequence changes (Arg12Trp/Ala26Ser/Ala48Thr/Arg61Leu). Where these substitutions occurred as the sole variant, the proportion of homozygous individuals was 72–83%. CONCLUSIONS: An evaluation of published data revealed that the three most frequently found sequence variants are associated with a substantial loss of rodenticide efficacy of first‐generation anticoagulants (e.g. warfarin, coumatetralyl), as well as the second‐generation compound bromadiolone and most probably also difenacoum. Knowledge of the distribution and frequency of resistance‐conferring sequence variants will stimulate their further functional characterisation and facilitate the choice of effective active substances for house mouse control. Copyright © 2011 Society of Chemical Industry     

Common oils and insecticidal control and their resistance to Aleuroclava jasmini (Hem.: Aleyrodidae) on paper mulberry in Iran

Abstract

Aleuroclava jasmini (Hemiptera: Aleyrodidae) is a major insect pest of paper mulberry (Broussonetia papyrifera) in Iran, negatively affecting its production. Considering the importance of oils in the integrated management programs of such pests, the present study examined the possibility of whitefly control on paper mulberry plant to assess mortality rate (MR), synergistic rate (SR), resistance rate (RR), and lethal concentration for 50% of the population (LC₅₀) of oils and common insecticide in populations from four areas of Tehran, Iran (one susceptible and three non-susceptible). The best chemical treatments against A. jasmini adults and nymphs in paper mulberry plants were neem oil (1?ml L^?1) mixed with deltamethrin (0.5?ml L^?1) or with buprofezin (1?ml L^?1). The neem, akylarylpolyglyglycol ether and volk oils mixed with deltamethrin or buprofezin also had synergistic effects on adults and nymphs of A. jasmini, respectively, in Azadi, Shahrake Gharb, and Vanak areas (non-susceptible populations), but with higher concentrations (> LC₅₀) and lower SR than in Garm Dareh area (susceptible population). We observed that A. jasmini adults showed the greatest resistance to deltamethrin in Vanak area and nymphs of this pest to buprofezin in Shahrake Gharb area.     

Detection of virulence to septoria tritici blotch (STB) resistance conferred by the winter wheat cultivar Cougar in the Irish Zymoseptoria tritici population and potential implications for STB control

Septoria tritici blotch (STB) caused by the fungal pathogen Zymoseptoria tritici continues to be the most economically destructive disease of winter wheat throughout Ireland. Due to the widespread development of fungicide resistance in the Irish Z. tritici population, integrated strategies to control STB are increasingly necessary. A key component of such strategies will be the deployment of winter wheat cultivars with improved levels of STB resistance. Unfortunately, due to the nature of Z. tritici, such resistances are at risk of being overcome by the pathogen. In late summer 2020, foci of STB were observed across a range of winter wheat cultivars under evaluation for recommendation in Ireland. Common amongst these was the cultivar Cougar in each of their pedigree. To determine if the foci observed in 2020 resulted from strains virulent to Cougar, isolate collections were established and virulence screens conducted on Cougar and a range of the cultivars currently under evaluation. These confirmed the presence of Cougar-virulent strains in the Irish Z. tritici population, and that this virulence affects not just Cougar, but also cultivars derived from it. Although the foci observed in 2020 were in both fungicide-untreated and -treated plots, there was no evidence that these strains are more sensitive or resistant to fungicides compared to the wider Irish Z. tritici population, with moderate resistance to the SDHIs and azoles dominating. Combined, the present study confirms the need to ensure a diversity of control measures for STB, including ensuring a range of STB resistances are used.