Vitamin K requirement in Danish anticoagulant-resistant Norway rats (Rattus norvegicus)

Resistance to warfarin has been connected to an increase in dietary requirement for vitamin K in British strains of the Norway rat, Rattus norvegicus (Berk). This study examines vitamin K requirement of Danish anticoagulant-resistant Norway rats using a vitamin K deficient feeding test. Wild bromadiolone-resistant rats sampled from different localities in Denmark and rats from bromadiolone-resistant and susceptible laboratory strains were fed on a vitamin K deficient diet over a maximum period of 15 days. Development of vitamin K deficiency, measured as reduced blood-clotting capacity, took place in 43% of the Danish resistant rats and was independent of sex, treatment with supplementary vitamin K3 and sampling locality. Development of deficiency was slower for resistant rats that were supplemented with vitamin K3 prior to the feeding test, suggesting storage of the vitamin K in a vitamin body pool. Intraperitoneal administration of vitamin K1 revealed that 80 microg vitamin K1 kg(-1) bodyweight was sufficient to restore normal blood clotting activity in deficient rats, while 60 microg vitamin K1 kg(-1) bodyweight was insufficient. We conclude that vitamin K requirement is moderately increased in Danish homozygous resistant rats whereas heterozygous resistant rats only have a minor increase in vitamin K requirement compared with susceptible rats. We found no indication of different resistance types being present in our test material since vitamin K requirement was not different between rats from separate sampling localities.     

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 tests and field trials with bromadiolone for the control of Norway rats (Rattus norvegicus) on farms in Westphalia, Germany

BACKGROUND: The aim of this study was to determine the incidence and the level of resistance to bromadiolone among rats on farms suspected of being foci of resistance by using the international normalised ratio (INR)‐based blood clotting response (BCR) test. Whether the level of reduced susceptibility constitutes ‘practical resistance’ was subsequently determined in field trials. RESULTS: The 2.5 multiple of the ED₅₀ baseline was used to test for the incidence of resistance, and higher multiples in the range of the suspected resistance factor were used to investigate the degree of resistance. The ED₅₀ values of bromadiolone in resistant rats were confirmed in the range 4.70–7.05 mg kg^?1 for males and 4.62–6.61 mg kg^?1 for females. Variations within these ranges appeared between farms. According to the BCR resistance tests, 50–100% of rats were classified as resistant prior to the field trials; 29–100% of rats survived the treatments. CONCLUSION: BCR tests based on the use of the INR and baselines are suitable for determining the incidence and for assessing the level of resistance in populations of Norway rats. The majority of rats of the Westphalian resistant strain, characterised by the Y139C marker in VKOR, are resistant to bromadiolone under practical control conditions. Copyright © 2011 Society of Chemical Industry     

Rodenticide grain bait ingredient acceptance by Norway rats (Rattus norvegicus), California ground squirrels (Spermophilus beecheyi) and pocket gophers (Thomomys bottae)

Vertebrate pest control in California is often accomplished through the use of rodenticide grain baits. These grain baits are composed of steam-rolled oats (SRO), a toxicant, an indicator dye and an oil combination. A series of tests were performed to determine the effects of various dye and oil formulations on acceptance of grain bait by Norway rats [Rattus norvegicus (Berk)], California ground squirrels [Spermophilus beecheyi (Richardson)] and pocket gophers (Thomomys bottae Eyd & Gerv). Seven different dyes, four oil formulations and clean (untreated) oats were tested for acceptance. The addition of the selected oils and dyes to grain resulted in no significant differences in consumption. This indicates that there is a wide variety of dyes that could be used in the formulation of rodenticides. These alternatives could aid in proper pesticide use, the deterrence of bait consumption by birds and possibly in ingredient adhesion to the finished bait.     

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     

Laboratory and field trials of the rodenticide brodifacoum against warfarin-resistant Norway rats

Norway rats (Rattus norvegicus) trapped in a low-income housing area in Raleigh, North Carolina, and on a poultry farm in Garner, North Carolina, manifested high levels of resistance to warfarin in laboratory tests. Of the rats tested, 59.1 and 91.7% from the Raleigh and Garner trap sites, respectively, survived a 6-day no-choice test of a diet containing 0.005 % warfarin. The warfarin-resistant rats were subsequently given a choice of a 0.005% brodifacoum bait and a placebo diet. Of 27 rats, 19 died after a 3-day test. Survivors had consumed significantly less brodifacoum (0.1 < P > 0.05) than those that died, but the survivors also died when subjected to an additional 1-day no-choice test of the brodifacoum bait. Field trials, in which warfarin-resistant rats in four poultry houses were fed with poison bait for 14–16 days with 0.005% brodifacoum in a pelletised bait (‘Talon’), resulted in 87–99 % control being achieved.     

Toxicity of calciferol,warfarin and their combinations to Rattus norvegicus (albino) and R. rattus

The response of Rattus norvegicus and R. rattus to calciferol, warfarin and a combination of these compounds was tested. Both species succumbed to the recommended bait concentrations of calciferol (1000 mg/kg) and warfarin (250 mg/kg) when tested individually. However, R. rattus was more susceptible to lower concentrations of calciferol (500 and 750 mg/kg) as compared to R. norvegicus, which was sensitive to lower concentrations of warfarin (12.5 and 25 mg/kg bait). Addition of calciferol at different concentrations did not affect the toxicity of warfarin against R. norvegicus. However, with R. rattus calciferol (250–350 mg/kg) or warfarin (50 mg/kg bait), which individually produced partial mortality, in combination produced a complete kill besides accelerating death. Combination of higher dosages of calciferol with warfarin (1000 + 250 mg/kg bait) tested against both species showed no advantages, resulting in neither lower bait consumption nor speedier death, though individually the components gave complete kills.     

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.     

Potentiation of warfarin toxicity to roof rats (Rattus rattus) by L-histidine and by vitamin K adsorbers

The efficacy of warfarin alone or with various additives was tested against roof rats (Rattus rattus). Warfarin alone at 50 mg kg^?1 of bait killed 37% of the roof rats where-as the additives, L -histidine (40 mg kg^?1 of bait), activated-clay, charcoal and carbon (10 g kg^?1 of bait), tested individually, produced no mortality. However, when warfarin wascombined with each of these additives in turn, only the L -histidine combination resulted in 100% mortality while the other combinations killed 88, 75 and 63% of the test rats respectively. Further trials of paired comparison tests were carried out in order to evaluate the efficacy of the warfarin/L -histidine combination under different environmental conditions. All the rats succumbed to this combination in both cage and rattery conditions when there was no alternative food. Mortality decreased to 60% in cage and 70% in rattery when alternative food was provided alongside the poison bait. Rats tended to die more quickly with the L -histidine combination than with warfarin alone. Field trials also confirmed that this combination was effective in reducing roof rat infestations as shown by the yield of corpses, the post-poison bait intake and post-operational surveillance.     

A methodology for preparing a list of recommended regulated non‐quarantine pests (RNQPs)

The concept of ‘regulated non‐quarantine pest’ (RNQP) was introduced in the revised text of the FAO International Plant Protection Convention (IPPC) approved in 1997. Measures against quarantine pests (exclusion, eradication, containment) aim to prevent unacceptable economic, environmental and social impacts resulting from the introduction and/or spread of these pests. On the contrary, the concept of RNQPs is intended to prevent an unacceptable economic impact on the intended use of plants for planting by pests that are already present in the area. RNQPs have been introduced in the new EU plant health regulation, in line with available international standards. This regulation entered into force in December 2016 and will be implemented in the following 3 years. In this context, EPPO agreed to undertake a 2‐year project on RNQPs: the EU Quality Pest Project. The objective of this project was to develop a methodology and then to apply this methodology to a list of about 1400 pest–host combinations to identify which should be recommended as RNQPs. This methodology is presented in this paper, as well as the main issues discussed during its development.     

水稻抗条纹叶枯病鉴定方法的研究

鉴于国内至今尚无水稻条纹叶枯病品种抗性鉴定标准的现状,本文分别研究了接种时间、接种强度与水稻接种苗龄3因素对抗性鉴定的影响。结果表明水稻条纹叶枯病的适合抗性鉴定方法应为：接种时间48～72 h, 接种强度2～6头/株和水稻接种龄期0.5～1.5叶龄。以本研究结果为基础制定了水稻条纹叶枯病品种抗性鉴定的江苏省地方标准。     

谷斑皮蠹与黑斑皮蠹的分子鉴定

本文首次报道了可用于谷斑皮蠹(Trogoderma granarium)和黑斑皮蠹(Trogoderma glabrum)种间鉴定的线粒体CO Ⅰ基因部分标记序列(883bp),两条序列在GenBank数据库的登录号分别为FJ589736和FJ589737.     

The identification of a gene for race-specific resistance to Peronospora parasitica (downy mildew) in Brassica napus var. oleifera (oilseed rape)

The oilseed rape cultivar Cresor was resistant to 14 isolates of Peronospora parasitica derived from crops of Brassica napus in the UK. Segregation for resistance to one isolate among F₂ plants and F₃ progeny of crosses between Cresor and the susceptible cultivars Victor and Jet Neuf indicated that resistance was controlled by a single gene. There was evidence that genetic background and environment could influence the phenotypic expression of this resistance. Two sexual progeny isolates derived from a homothallic isolate of P. parasitica avirulent on Cresor were completely virulent on this cultivar. This suggested that the parental isolate was heterozygous at a matching locus or loci for avirulence and demonstrated the race-specific nature of the resistance.     

Induction of disease resistance in Norway spruce (Picea abies) by necrotizing fungi

Norway spruce trees ( Picea abies ) preinoculated with the root rot fungus Heterobasidion annosum , Nectria fuckeliana or a pathogenic strain of the blue-stain fungus Ceratocystis polonica were more efficiently protected against a subsequent massive inoculation with pathogenic C. polonica than trees pretreated with nonpathogenic C. polonica or sterile malt agar. Control trees that received no pretreatment were extensively colonized by the mass inoculation. There was a strong negative correlation between the length of the phloem necroses induced by the pretreatment inoculations and the extent of host symptoms caused by mass inoculation with pathogenic C. polonica . The degree of induced resistance in Norway spruce thus depended on the amount of host tissue destroyed by the pretreatment.     

PM 7/129 (1) DNA barcoding as an identification tool for a number of regulated pests

Specific scope

This Standard describes the use of DNA barcoding protocols in support of the identification of a number of regulated pests and invasive plant species comparing DNA barcode regions with those deposited in publically available sequence databases. 1 It should be used in conjunction with PM 7/76 Use of EPPO diagnostic protocols.

Specific approval and amendment

2016‐09     

Differential expression of cytochrome P450 genes between bromadiolone-resistant and anticoagulant-susceptible Norway rats: a possible role for pharmacokinetics in bromadiolone resistance

BACKGROUND: Anticoagulant resistance in Norway rats, Rattus norvegicus (Berk.), has been suggested to be conferred by mutations in the VKORC1 gene, encoding the target protein of anticoagulant rodenticides. Other factors, e.g. pharmacokinetics, may also contribute to resistance, however. To examine the involvement of pharmacokinetics in bromadiolone resistance in male and female rats, liver expression profiles of seven cytochrome P450 genes from a Danish bromadiolone-resistant rat strain (with an Y139C-VKORC1 mutation) were compared with profiles from an anticoagulant-susceptible strain. RESULTS: In the presence of bromadiolone, the Cyp2e1, Cyp2c13, Cyp3a2 and Cyp3a3 genes were significantly overexpressed, while Cyp2c12 expression was suppressed in resistant female rats compared with susceptible females. Relative to susceptible males, resistant males showed significant overexpression of the Cyp2a1, Cyp2e1, Cyp3a2 and Cyp3a3 genes. On exposure to bromadiolone, females had higher Cyp2e1 expression than males, which possibly explains why female rats are generally more tolerant to anticoagulants than male rats. CONCLUSION: Results suggest that bromadiolone resistance in a Danish strain of Norway rats involves enhanced anticoagulant metabolism catalysed by cytochrome P450-2e1, -3a2 and -3a3. This pharmacokinetically based bromadiolone resistance is to some extent sex differentiated, as female resistance furthermore seems to involve overexpression of cytochrome P450-2c13 and suppression of P450-2c12, whereas male resistance appears to involve P450-2a1 overexpression.     

Effect of phenology on susceptibility of Norway spruce (Picea abies) to fungal pathogens

Ceratocystis polonica and Heterobasidion parviporum are important fungal pathogens in Norway spruce (Picea abies). Tree susceptibility to these pathogens with respect to phenology was studied using artificial fungal inoculations at six stages of bud development, and assessed by measuring phloem necroses in the stems of 2‐ and 8‐year‐old trees. Tree capacity for resistance was assessed by measuring phloem nonstructural carbohydrates at each stage. Phloem necroses were significantly larger in trees with fungal versus control inoculations and increased significantly over time. Changes in nonstructural carbohydrates occurred in the trees; a significant decline in starch and a slight but significant increase in total sugars occurred over time. These results suggest that susceptibility to fungal pathogens and carbohydrate levels in the stems of the trees were related to fine‐scale changes in bud development. A trade‐off may occur between allocation of starch (the major fraction of the stem carbohydrate pool) to bud development/shoot growth versus defence of the stem. Previous tests of plant defence hypotheses have focused on herbivory on plants growing under different environmental conditions, but the role of phenology and the effect of pathogens are also important to consider in understanding plant resource allocation patterns.