the corn earworm, Helicoverpa zea, is a major target pest of the insecticidal Vip3Aa protein used in pyramided transgenic Bt corn and cotton with Cry1 and Cry2 proteins in the U.S. The widespread resistance to Cry1 and Cry2 proteins in H. zea will challenge the long-term efficacy of Vip3Aa technology. Determining the frequency of resistant alleles to Vip3Aa in field populations of H. zea is critically important for resistance management. Here, we provided the first F 2 screen study to estimate the resistance allele frequency for Vip3Aa in H. zea populations in Texas, U.S. In 2019, 128 H. zea neonates per isofamily for a total of 114 F 2 families were screened with a diagnostic concentration of 3.0 μg/cm 2 of Vip3Aa39 protein in diet-overlay bioassays. The F 2 screen detected two families carrying a major Vip3Aa resistance allele. The estimated frequency of major resistance alleles against Vip3Aa39 in H. zea in texas from this study was 0.0065 with a 95% CI of 0.0014-0.0157. A Vip3Aa-resistant strain (RR) derived from the f 2 screen showed a high level of resistance to Vip3Aa39 protein, with a resistance ratio of >588.0fold relative to a susceptible population (SS) based on diet-overlay bioassays. We provide the first documentation of a major resistance allele conferring high levels of Vip3Aa resistance in a field-derived strain of H. zea in the U.S. Data generated from this study contribute to development of management strategies for the sustainable use of the Vip3Aa technology to control H. zea in the U.S. Genetically engineered crops producing insecticidal Cry and Vip proteins from the bacterium Bacillus thuringiensis (Bt) have been planted for control of insect pests for more than two decades 1. Field efficacy of these Bt crops has been outstanding in controlling most target species, resulting in substantial economic, environmental and social gains 2-7. However, with large scale adoption comes intense selection pressure for development of resistance and challenges for long-term sustainability 8,9. To date, field-evolved practical resistance to Bt crops has been globally reported in at least 21 cases 10-15. To delay insect resistance development, an insecticide resistance management (IRM) plan based on a "high-dose refuge" strategy has been implemented in the U.S 16. Monitoring for evolution of resistance in field populations of the target insect species is an essential component of this IRM plan to maintain sustainability of Bt crop technologies. The corn earworm/cotton bollworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), is a major target pest of both Bt cotton and Bt corn in North America. Control of lepidopteran pests is achieved by the adoption of corn hybrids producing combinations of Cry1Ab, Cry1F, Cry1A.105, Cry2Ab2 and Vip3Aa20 insecticidal Bt proteins, and Bt cotton varieties producing combinations of Cry1Ac, Cry1F, Cry1Ab, Cry2Ab, Cry2Ae, and Vip3Aa19 17 .

Synergists were used to diagnose possible mechanisms of permethrin resistance in permethrin-selected strains of the tobacco budworm, Heliothis uirescens (F.). In addition to permethrin, these strains of the tobacco budworm were resistant to a-cyano-pyrethroid insecticides, organophosphorus insecticides and DDT. The monooxygenase-inhibiting prop-2-ynyl aryl ethers were the only effective synergists of permethrin among 16 candidates tested. The most effective synergist was 1,2,4-trichloro-3-(2-propynyloxy)benzene. Piperonyl butoxide, a common monooxygenase-inhibiting synergist in other species and tobacco budworm strains, was inactive. These results suggested the presence and contribution of an unusual monooxygenase in the enzymatic detoxication of permethrin. DDT cross-resistance, which was not synergized, and broad pyrethroid cross-resistance supported previous evidence for target site insensitivity as a second pyrethroid-resistance mechanism in these strains. The actions of S,S,Stributyl phosphorotrithioate (TBPT) and triphenyl phosphate (TPP) suggested that hydrolytic detoxication, important in methyl parathion-resistance tobacco budworm strains, had little or no role in conferring pyrethroid resistance in these strains.

Following the apparent failure of levamisole to control infections of Haemonchus contortus in sheep at Lawes in south eastern Queensland, a strain of this parasite was isolated at the Animal Research Institute, Yeerongpilly. This strain was used to infect sheep at Yeerongpilly and the Merrindale Research Station, Victoria where four experiments to classify the resistance pattern of the parasite were carried out. Resistance to thiabendazole was first suspected in 1969, and these experiments confirmed that resistance to this drug was still present. They also showed that a strong degree of resistance had been developed to both levamisole and morantel tartrate. Other benzimidazole anthelmintics and also the organophosphorus compound naphthalophos were only moderately effective against the original isolate but rafoxanide, nitroxynil and phenothiazine were almost 100% effective. Other highly effective chemicals were disophenol and closantel. After passaging the strain for four generations with both levamisole and albendazole, resistance to both naphthalophos and the newer benzimidazole anthelmintics increased dramatically. This is the first report of a field strain of H. contortus exhibiting resistance to benzimidazole, non-benzimidazole and organophosphorus anthelmintics.

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The tobacco budworm, Heliothis virescens (F.) (Lepidoptera: Noctuidae), is one of the most important pests of cotton, Gossypium hirsutum L, that has become resistant to a wide range of synthetic insecticides. CrylAc-expressing cotton has proven its effectiveness against this insect since its introduction in North America in 1996. However, the constant exposure of tobacco budworm to this protein toxin may result in the development of resistance to it. To estimate the frequency of alleles that confer resistance to a 1.0 microg of Bacillus thuringiensis Cry1Ac diagnostic concentration in field-collected insects, the second generation (F2) of 1,001 single-pair families from seven geographical regions representing 2,202 alleles from natural populations was screened in 2006 and 2007 without finding major resistant alleles. Neonates of 56 single-pair families were able to develop to second instar on the diagnostic concentration in the initial screen, but only seven of these lines did so again in a second confirmatory screen. Minor resistance alleles to Cry1Ac may be quite common in natural populations of H. virescens. Our estimated resistance allele frequencies (0.0036-0.0263) were not significantly different from a previously published estimate from 1993. There is no evidence that H. virescens populations have become more resistant to Cry1Ac.

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