Tuta absoluta (Meyrick) is an invasive tomato pest that occurs worldwide, including Iran. This study investigates the occurrence of resistance to indoxacarb, an oxadiazine insecticide, and the underlying mechanisms in Iranian populations of T. absoluta. Bioassays were performed on second-instar larvae using indoxacarb alone and in combination with three synergists: Piperonyl butoxide (PBO), diethyl maleate (DEM), and triphenyl phosphate (TPP). The activities of the main detoxification enzymes, including glutathione S-transferases (GST), general carboxylesterases (CarEs), and P450 monooxygenases (P450s), were evaluated. In addition, the presence of known amino acid substitutions in the IV segment 6 domain of the T. absoluta sodium channel was investigated. The results showed that resistance rates to indoxacarb in Iranian populations ranged from 2.37- to 14.45-fold. However, pretreatment with synergists did not significantly increase the toxicity of indoxacarb. Enzyme assays showed that Ardabil (Ar) and Kerman (Kr) populations had the highest CarEs activity, while Ar population showed the highest P450 activity. However, the observed increases in enzyme activities were <2-fold. Two indoxacarb resistance mutations, F1845Y and V1848I, were detected. Apart from a significant and positive correlation between LC50 values of indoxacarb and thiocyclam hydrogen oxalate, no cross-resistance between indoxacarb and other insecticides was detected. Overall, these results suggest that populations of T. absoluta in Iran have developed resistance to indoxacarb, primarily through changes at the target site.
Title: Hormetic effects of neonicotinoid insecticides on Rhizoglyphus robini (Acari: Acaridae) Alimirzaee S, Khajehali J, Van Leeuwen T Ref: Pestic Biochem Physiol, 192:105396, 2023 : PubMed
The stimulation of biological processes by sublethal doses of insecticides or other stressors is known as hormesis. Here, we have evaluated whether exposure to field-relevant or low concentrations of neonicotinoids induce changes in the reproductive capacity of the bulb mite Rhizoglyphus robini (Acari: Acaridae). Among the tested neonicotinoids imidacloprid, thiamethoxam, and dinotefuran, the highest hormetic effect on the reproduction of R. robini occurred 24 h after the 48 h exposure period to imidacloprid at concentrations of 70 and 140 mg a.i./L. Despite the stimulating effects of imidacloprid on mite reproduction, no significant differences were observed in the offspring (F(1)) for biological aspects including egg hatch rate, embryonic period and sex ratio, while an increase was found in the duration of development time from egg to adult. Evaluation of the detoxification enzyme activities of treated adults showed that the highest activity of carboxyl/cholinesterases, cytochrome P450s, and glutathione S-transferases was obtained when exposed to 70, 140 and 70 mg a.i./L imidacloprid, immediately after the exposure period, respectively. Also, an increase in the activity of the antioxidant enzyme catalase was observed compared to that of the control. After imidacloprid pretreatment (140 mg a.i./L), the tolerance of adult mites to diazinon was increased about two-fold. This study shows that exposure to imidacloprid can induce hormetic effects on R. robini and could severely complicate its control due to a higher reproduction, enhanced detoxification enzyme activities, and increased tolerance against other pesticides.
Title: Biochemical and insecticidal effects of plant essential oils on insecticide resistant and susceptible populations of Musca domestica L. point to a potential cross-resistance risk Ahmadi E, Khajehali J, Jonckheere W, Van Leeuwen T Ref: Pestic Biochem Physiol, 184:105115, 2022 : PubMed
Essential oils (EOs) can provide important alternatives to chemical insecticides in the control of pests. In this study, 12 EOs of native plant species from Iran were evaluated for their adulticidal activity against the house fly. In addition, we examined the insecticidal activity of Zataria multiflora and Rosmarinus officinalis EOs on adult female house flies from pyrethroid and organophosphate resistant and susceptible populations, using both fumigant and topical bioassays. The involvement of detoxification enzymes in susceptibility was investigated with synergism experiments in vivo, while the inhibitory effects of R. officinalis and Zataria multiflora EOs on the activities of cytochrome P450-dependent monooxygenases (P450s), carboxylesterases (CarEs) and glutathione S-transferases (GSTs) were determined by enzymatic inhibition assays in vitro. The EOs of Z. multiflora, Mentha pulegium, R. officinalis and Thymus vulgaris were the most effective against adults in contact topical assays, while oils extracted from Eucalyptus cinerea, Z. multiflora, Citrus sinensis, R. officinalis, Pinus eldarica and Lavandula angustifolia where the most effective in fumigant assays. Rosmarinus officinalis and Z. multiflora EOs were selected for further investigation and showed higher toxicity against a susceptible population, compared to two insecticide-resistant populations. Correlation analysis suggested cross-resistance between these EOs and pyrethroids in the resistant populations. The toxicity of both EOs on the resistant populations was synergized by three detoxification enzyme inhibitors. Further, in vitro inhibition studies showed that R. officinalis and Z. multiflora EOs more effectively inhibited the activities of the detoxification enzymes from flies of the susceptible population compared to those of the pyrethroid resistant populations. Synergistic and enzymatic assays further revealed that increased activities of P450s, GSTs, and CarEs are possibly involved in the cross-resistance between EOs and pyrethroids. Investigating the molecular mechanisms of P450s, GSTs, and CarEs in the resistance to EOs should be subject to further studies.
Title: Dichlorvos Resistance in the House Fly Populations, Musca domestica, of Iranian Cattle Farms Ahmadi E, Khajehali J Ref: J Arthropod Borne Dis, 14:344, 2020 : PubMed
BACKGROUND: Insecticide resistance is one of the most important problems associated with the control of Musca domestica, due to the potential of the rapid development of resistance to different chemical insecticides. The present study was carried out to evaluate dichlorvos resistance in the house fly populations collected from central regions of Iran, Isfahan Province and Chaharmahal and Bakhtiari Province, during 2017 to 2019. METHODS: Bioassays were carried out using a standard topical application method as well as a fumigation method. The Koohrang population (susceptible) with the lowest LD(50) values to dichlorvos was chosen to calculate the resistance ratios (RR). Altered sensitivity of acetylcholinesterase (AChE), a target enzyme for dichlorvos, was investigated. RESULTS: According to the results, very high levels of dichlorvos resistance were observed in the Mobarake population (RR= 80.25-fold by topical application and 33-fold by fumigation bioassay), and Isfahan population (RR= 107.30-fold by topical application and 43-fold by fumigation bioassay) compared to the Koohrang population. Acetylcholinesterase of the Koohrang population was the most sensitive to inhibition by dichlorvos based on the determination of median inhibitory concentration (IC(50)), but AChE of Mobarake and Isfahan populations were 741.93- and 343.94- fold less sensitive to inhibition. CONCLUSION: The insensitivity of AChE was possibly involved in dichlorvos resistance in the house fly populations.
Spirodiclofen is one of the most recently developed acaricides and belongs to the new family of spirocyclic tetronic acids (ketoenols). This new acaricidal family is an important chemical tool in resistance management strategies providing sustainable control of spider mites such as Tetranychus urticae. Spirodiclofen targets lipid biosynthesis mediated by direct inhibition of acetyl coenzyme A carboxylase (ACCase). In this study, we investigated two genetically distant spider mite strains with high resistance to spirodiclofen. Despite the strong resistance levels to spirodiclofen (up to 680-fold), only limited cross-resistance with other members of this group such as spiromesifen and spirotetramat could be detected. Amplification and sequencing of the ACCase gene from resistant and susceptible strains did not reveal common non-synonymous mutations, and expression levels of ACCase were similar in both resistant and susceptible strains, indicating the absence of target-site resistance. Furthermore, we collected genome-wide expression data of susceptible and resistant T. urticae strains using microarray technology. Analysis of differentially expressed genes revealed a broad response, but within the overlap of two resistant strains, several cytochrome P450s were prominent. Quantitative PCR confirmed the constitutive over-expression of CYP392E7 and CYP392E10 in resistant strains, and CYP392E10 expression was highly induced by spirodiclofen. Furthermore, stage specific expression profiling revealed that expression levels were not significantly different between developing stages, but very low in eggs, matching the age-dependent resistance pattern previously observed. Functional expression of CYP392E7 and CYP392E10 confirmed that CYP392E10 (but not CYP392E7) metabolizes spirodiclofen by hydroxylation as identified by LC-MS/MS, and revealed cooperative substrate binding and a Km of 43 muM spirodiclofen. CYP392E10 also metabolizes spiromesifen, but not spirotetramat. Surprisingly, no metabolism of the hydrolyzed spirodiclofen-enol metabolite could be detected. These findings are discussed in the light of a likely resistance mechanism.
BACKGROUND: In Tetranychus urticae Koch, acetylcholinesterase insensitivity is often involved in organophosphate (OP) and carbamate (CARB) resistance. By combining toxicological, biochemical and molecular data from three reference laboratory and three OP selected strains (OP strains), the AChE1 mutations associated with resistance in T. urticae were characterised. RESULTS: The resistance ratios of the OP strains varied from 9 to 43 for pirimiphos-methyl, from 78 to 586 for chlorpyrifos, from 8 to 333 for methomyl and from 137 to 4164 for dimethoate. The insecticide concentration needed to inhibit 50% of the AChE1 activity was, in the OP strains, at least 2.7, 55, 58 and 31 times higher for the OP pirimiphos-methyl, chlorpyrifos oxon, paraoxon and omethoate respectively, and 87 times higher for the CARB carbaryl. By comparing the AChE1 sequence, four amino acid substitutions were detected in the OP strains: (1) F331W (Torpedo numbering) in all the three OP strains; (2) T280A found in the three OP strains but not in all clones; (3) G328A, found in two OP strains; (4) A201S found in only one OP strain. CONCLUSIONS: Four AChE1 mutations were found in resistant strains of T. urticae, and three of them, F331W, G328A and A201S, are possibly involved in resistance to OP and CARB insecticides. Among them, F331W is probably the most important and the most common in T. urticae. It can be easily detected by the diagnostic PCR-RLFP assay developed in this study.
Title: Susceptibility of an organophosphate resistant strain of the two-spotted spider mite (Tetranychus urticae) to mixtures of bifenazate with organophosphate and carbamate insecticides Khajehali J, Van Leeuwen T, Tirry L Ref: Exp Appl Acarol, 49:185, 2009 : PubMed
Bifenazate, a new and frequently used carbazate, is a pro-acaricide which needs to be activated by carboxylesterases. We evaluated the possible antagonism of organophosphate and carbamate insecticides on bifenazate toxicity in Tetranychus urticae applied in mixtures. Two organophosphate resistant strains were used (WI and MR-VL) and several organophosphate (chlorpyrifos, azinphosmethyl and phosmet) and carbamate (carbaryl and methomyl) insecticides were evaluated. Mixing chlorpyrifos with bifenazate decreased bifenazate toxicity in both tested strains. However, in the strain with a higher esterase activity, antagonism decreased after 2 days. Of all other tested chemicals, only methomyl displayed an antagonistic effect 1 day after treatment. These findings indicate that mixing organophosphate and carbamate insecticides with bifenazate may inhibit bifenazate efficacy under field conditions, especially when resistant strains are present.
