Susceptible Laboratory Strain Research Articles

Analysis of seasonal and annual field-evolved insecticide resistance in populations of Thrips hawaiiensis in banana orchards

Every season over the past decade, efficacious thrips insecticides have been commonly applied for control of Thrips hawaiiensis on banana crops in China. For effective thrips control, some insecticides have been used extensively against the thrips in numerous banana plantations. This study aims to examine the current resistance status and the dynamics regarding the changing frequencies of resistance in the field T. hawaiiensis populations. A total of 13 field samples from banana orchards (Chengmai, Lingao, Dongfang and Ledong located in Hainan Province, China) were collected monthly in 2014 to 2017, and selected individuals were used in bioassays assessing insecticide resistance and metabolic enzyme activity compared to a laboratory-susceptible strain. In 2014, the initial resistance survey found that all sampled populations were susceptible to spirotetramat and cyantraniliprole, while Lingao and Chengmai population showed a moderate resistance to imidacloprid, acetamiprid, abamectin and spinetoram. Further, seasonal and annual analysis of resistance dynamics with imidacloprid, abamectin and spinetoram demonstrated that resistance in field populations of T. hawaiiensis developed quickly within a season (from April to August 2016), but resistance was unstable over subsequent seasons (April 2015, 2016 and 2017). Moreover, synergism experiments using enzyme inhibitors together with metabolic enzyme assays revealed that field-evolved insecticide resistance in T. hawaiiensis was closely associated with the elevated metabolic enzymes. The observed resistance increase within a season was attributed mainly to the intense use of these insecticides and short generation time of thrips. These findings suggest that a rotation system using efficacious thrips insecticides should be implemented for sustainable thrips control to reduce the potential of resistance development.

ATP-Binding Cassette (ABC) Transporter Genes Involved in Pyrethroid Resistance in the Malaria Vector Anopheles sinensis: Genome-Wide Identification, Characteristics, Phylogenetics, and Expression Profile.

background: The ATP-binding cassette (ABC) transporters family is one of the largest families of membrane proteins existing in all living organisms. Pyrethroid resistance has become the largest unique obstacle for mosquito control worldwide. ABC transporters are thought to be associated with pyrethroid resistance in some agricultural pests, but little information is known for mosquitoes. Herein, we investigated the diversity, location, characteristics, phylogenetics, and evolution of ABC transporter family of genes in the Anopheles sinensis genome, and identified the ABC transporter genes associated with pyrethroid resistance through expression profiles using RNA-seq and qPCR. Results: 61 ABC transporter genes are identified and divided into eight subfamilies (ABCA-H), located on 22 different scaffolds. Phylogenetic and evolution analyses with ABC transporters of A. gambiae, Drosophila melanogaster, and Homo sapiens suggest that the ABCD, ABCG, and ABCH subfamilies are monophyly, and that the ABCC and ABCG subfamilies have experienced a gene duplication event. Both RNA-seq and qPCR analyses show that the AsABCG28 gene is uniquely significantly upregulated gene in all three field pyrethroid-resistant populations (Anhui, Chongqing, and Yunnan provinces) in comparison with a laboratory-susceptible strain from Jiangsu province. The AsABCG28 is significantly upregulated at 12-h and 24-h after deltamethrin exposure in three-day-old female adults. Conclusion: This study provides the information frame for ABC transporter subfamily of genes, and lays an important basis for the better understanding and further research of ABC transporter function in insecticide toxification. The AsABCG28 gene is associated with pyrethroid detoxification, and it functions at later period in the detoxification process for xenobiotics transportation.

Characterization of nitenpyram resistance in Nilaparvata lugens (Stål)

Nitenpyram is very effective in controlling Nilaparvata lugens (brown planthopper, BPH), and its resistance has been reported in field populations; however, the resistance mechanism remains unclear. In the present study, cross-resistance and resistance mechanisms in nitenpyram-resistant BPH were investigated. A resistant strain (NR) with a high resistance level (164.18-fold) to nitenpyram was evolved through successive selection for 42 generations from a laboratory susceptible strain (NS). The bioassay results showed that the NR exhibited cross-resistance to imidacloprid (37.46-fold), thiamethoxam (71.66-fold), clothianidin (149.17-fold), dinotefuran (98.13-fold), sulfoxaflor (47.24-fold), cycloxaprid (9.33-fold), etofenprox (10.51-fold) and isoprocarb (9.97-fold) but not to triflumezopyrim, chlorpyrifos and buprofezin. The NR showed a 3.21-fold increase in cytochrome P450 monooxygenase (P450) activity compared to that in the NS, while resistance was also synergized (4.03-fold) with the inhibitor piperonyl butoxide (PBO), suggesting a role of P450. Furthermore, the mRNA expression levels of cytochrome P450 (CYP) genes by quantitative real-time PCR results indicated that twelve P450 genes were significantly overexpressed in the NR strain, especially CYP6ER1 (203.22-fold). RNA interference (RNAi) suppression of CYP6ER1 through injection of dsCYP6ER1 led to significant susceptibility in the NR strain. The current study expands our understanding of the nitenpyram resistance mechanism in N. lugens, provides an important reference for integrated pest management (IPM), and enriches the theoretical system of insect toxicology.

Evaluation of toxicity of clothianidin (neonicotinoid) and chlorfenapyr (pyrrole) insecticides and cross-resistance to other public health insecticides in Anopheles arabiensis from Ethiopia

BackgroundInsecticide-based interventions play an integral role in malaria vector control. However, the continued spread of insecticide resistance threatens to undermine progress made thus far and may ultimately lead to operational failure of current control measures. Clothianidin and chlorfenapyr both have unique modes of action and have expanded the number of insecticide classes available to vector control programmes. Prior to field use, it is imperative to establish their toxicity against local mosquito populations and evaluate potential cross-resistance with other chemicals used contemporarily or historically. The aim of this study was to determine the diagnostic doses of clothianidin and chlorfenapyr and their efficacies against Anopheles arabiensis, the predominant Ethiopian malaria vector species.MethodsA range of doses of clothianidin and chlorfenapyr were tested, using modified WHO susceptibility tests and CDC bottle bioassays, respectively, against an Ethiopian susceptible laboratory strain and a wild population of An. arabiensis collected from Oromia Region, Ethiopia. Cross-resistance to other public health insecticides: carbamates (bendiocarb and propoxur), organophosphate (malathion) and pyrethroids (deltamethrin and permethrin), was assessed in the same mosquito populations using CDC bottle bioassays.ResultsComplete mosquito mortality was observed with the laboratory strain using the recommended diagnostic doses for clothianidin (2%/filter paper) and chlorfenapyr (100 µg/bottle). The field population was resistant to malathion (83% mortality), capable of surviving 2×, 5× and 10× the diagnostic dose of both deltamethrin and permethrin, but susceptible to bendiocarb and propoxur. The field population of An. arabiensis was significantly more susceptible to clothianidin, reaching 100% mortality by day 2 compared to the laboratory strain (100% mortality by day 3). In contrast, the wild population was less susceptible to chlorfenapyr, with the highest mortality of 99% at 72 h using 200 µg/bottle compared to the laboratory colony, which reached complete mortality at 50 µg/bottle by 24 h.ConclusionsThe putative diagnostic doses of clothianidin and chlorfenapyr are appropriate for monitoring resistance in An. arabiensis from Ethiopia. The unique modes of action and an absence of cross-resistance render clothianidin and chlorfenapyr potential candidates for inclusion in the National Malaria Control Programme vector control efforts, particularly in areas with high pre-existing or emergent resistance to other insecticide classes.

Effects of bistrifluron resistance on the biological traits of Spodoptera litura (Fab.) (Noctuidae: Lepidoptera).

Spodoptera litura is one of the major insect pests of vegetables in Sichuan, China, and it has developed serious resistance to many traditional chemical insecticides. In this present study, S. litura individuals collected from five regions in Sichuan in 2017, which were detected the resistance to bistrifluron. We found that the resistance ratios (RRs) bistrifluron were in the ranges of 15.9- to 77.7-fold, respectively, compared with a laboratory susceptible (Lab-HN) strain. To elucidate the effect of bistrifluron on the life history parameters of S. litura, we established the bistrifluron-susceptible (Bis-UNSEL) and bistrifluron-resistant (Bis-SEL) strains based on the DY field population with the highest RR of 77.7-fold among the all field populations to bistrifluron compared with a laboratory Lab-HN strain. The results show that the Bis-SEL strain developed a 113.8-fold RR while that of the Bis-UNSEL strain was almost equal to that of the Lab-HN strain (with overlapping confidence intervals (CI) of their LC50 values, P < 0.05). Meanwhile, the life span of the egg and pupa stage of the Bis-UNSEL strain was prolonged, and the strain also showed shorter adult duration and lower fecundity. Demographic characteristics, such as the intrinsic rate of increase (r), finite rate of increase (λ) and net reproductive rate (R0) in the Bis-SEL strain were significantly higher than the Bis-UNSEL strain, whereas the mean generation time (T) and gross reproduction rate (GRR) were lower. The results clearly showed that the developing resistance of bistriflueon on S. litura promoted its population growth. Therefore, the present study could provide useful informations for determining potential management methods to prevent the development of bistrifluron resistance in S. litura.

Insecticide Resistance Status and Mechanisms of Anopheles sinensis (Diptera: Culicidae) in Wenzhou, an Important Coastal Port City in China.

Although scaled-up interventions and effective control efforts have drastically reduced malaria morbidity and mortality, malaria remains a serious threat to public health worldwide. Anopheles sinensis Wiedemann 1828 is a historically important vector of Plasmodium vivax (Haemosporida: Plasmodiidae) malaria in China. Insecticide resistance has become a major obstacle to vector-borne disease control. However, little is known about the insecticide resistance of An. sinensis in Wenzhou, an important coastal port city in Zhejiang province, China. The aim of this study was to examine insecticide resistance and mechanisms in An. sinensis field mosquito populations. Evidence of multiple insecticide resistance was found in An. sinensis adult female populations. Medium to high frequencies of target site kdr together with fixed ace-1 mutations was detected in both the Ruian and Yongjia populations. Both populations showed an association between kdr L1014 mutation and resistance phenotype when tested against deltamethrin and DDT. Significantly different metabolic enzyme activities were found between the susceptible laboratory strain and field-collected mosquitoes from both Ruian and Yongjia. Both field collected An. sinensis populations exhibited significantly higher P450 enzyme activity compared with the laboratory strain, while the field-collected resistant mosquitoes exhibited various GST and COE enzyme activities. These results indicate multiple resistance mechanisms in An. sinensis field populations. Effective implementation of insecticide resistance management strategies is urgently needed. The data collected in this study will be valuable for modeling insecticide resistance spread and vector-control interventions.

Contrasting patterns of gene expression indicate differing pyrethroid resistance mechanisms across the range of the New World malaria vector Anopheles albimanus.

Decades of unmanaged insecticide use and routine exposure to agrochemicals have left many populations of malaria vectors in the Americas resistant to multiple classes of insecticides, including pyrethroids. The molecular basis of pyrethroid resistance is relatively uncharacterised in American malaria vectors, preventing the design of suitable resistance management strategies. Using whole transcriptome sequencing, we characterized the mechanisms of pyrethroid resistance in Anopheles albimanus from Peru and Guatemala. An. albimanus were phenotyped as either deltamethrin or alpha-cypermethrin resistant. RNA from 1) resistant, 2) unexposed, and 3) a susceptible laboratory strain of An. albimanus was sequenced and analyzed using RNA-Seq. Expression profiles of the three groups were compared based on the current annotation of the An. albimanus reference genome. Several candidate genes associated with pyrethroid resistance in other malaria vectors were found to be overexpressed in resistant An. albimanus. In addition, gene ontology terms related to serine-type endopeptidase activity, extracellular activity and chitin metabolic process were also commonly overexpressed in the field caught resistant and unexposed samples from both Peru and Guatemala when compared to the susceptible strain. The cytochrome P450 CYP9K1 was overexpressed 14x in deltamethrin and 8x in alpha-cypermethrin-resistant samples from Peru and 2x in deltamethrin-resistant samples from Guatemala, relative to the susceptible laboratory strain. CYP6P5 was overexpressed 68x in deltamethrin-resistant samples from Peru but not in deltamethrin-resistant samples from Guatemala. When comparing overexpressed genes between deltamethrin-resistant and alpha-cypermethrin-resistant samples from Peru, a single P450 gene, CYP4C26, was overexpressed 9.8x (p<0.05) in alpha-cypermethrin-resistant samples. In Peruvian deltamethrin-resistant samples, the knockdown resistance mutation (kdr) variant alleles at position 1014 were rare, with approximately 5% frequency, but in the alpha-cypermethrin-resistant samples, the frequency of these alleles was approximately 15–30%. Functional validation of the candidate genes and the kdr mutation as a resistance marker for alpha-cypermethrin will confirm the role of these mechanisms in conferring pyrethroid resistance.

Status and preliminary mechanism of resistance to insecticides in a field strain of housefly (Musca domestica, L)

Resistance profiles of houseflies (Gol-RR) collected from a field in Golmud city, Qinghai province, China, were determined for seven insecticides using topical bioassays. Resistance ratios of >1219.51, 153.17, >35.43, 6.12, 3.24, 1.73, and 0.86-fold were obtained for propoxur, cypermethrin, imidacloprid, indoxacarb, chlorpyrifos, fipronil, and chlorfenapyr, respectively, relative to a laboratory susceptible strain (SS). Synergism experiments showed that piperonyl butoxide (PBO), triphenylphosphate (TPP), and diethyl maleate (DEM) increased propoxur toxicity by >105.71, >7.88, and >5.15-fold in the Gol-RR strain, compared with 5.25, 2.00, and 1.39-fold in the SS strain, indicating the involvement of P450 monooxygenases, esterases, and glutathione-S-transferase in conferring resistance. Although cypermethrin resistance was significantly suppressed with PBO, TPP, and DEM in the Gol-RR strain, the synergistic potential of these agents to cypermethrin was similar in the SS strain, demonstrating that metabolism-mediated detoxification was not important for conferring resistance to cypermethrin in the Gol-RR strain. However, the three agents did not act synergistically with imidacloprid, indicating that other mechanisms may be responsible for the development of resistance to this insecticide. Acetylcholinesterase (AChE) activity was 13.70-fold higher in the Gol-RR than in the SS strain, suggesting the properties of the AChE enzyme were altered in the Gol-RR strain. Thus, rotation of chlorfenapyr insecticide with other agents acting through a different mode with minimal/no resistance could be an effective resistance management strategy for housefly.

Resistance to Permethrin, β-cyfluthrin, and Diazinon in Florida Horn Fly Populations.

Horn flies, Haematobia irritans, a major cattle pest in the USA, cause substantial economic losses and current control methods rely heavily on insecticides. Three horn fly populations were evaluated for insecticide susceptibility to permethrin, β-cyfluthrin, and diazinon. Susceptibility was variable by population, with the greatest resistance exhibited by a 66-fold resistance ratio (RR) to permethrin and >14-fold RR to diazinon. Mechanisms of resistance were determined using molecular techniques and enzymatic assays. The knockdown resistance (kdr) genotype (L150F) associated with pyrethroid resistance, and a G262A mutation in acetylcholinesterase, previously associated with organophosphate resistance, were found in all field populations evaluated. Insensitivity of diazoxon at the acetylcholinesterase (AChE) target site was significantly different in horn flies from one of the field sites. For metabolic detoxifying enzymes, cytochrome P450 nor general esterases showed a significant difference between field strains and a laboratory susceptible strain. Pyrethroid resistance was likely due to the presence of the L150F mutation in the population. In vitro studies targeting the AChE enzyme did not support the notion that the G262A mutation was the sole cause of resistance to organophosphates, and, therefore, the exact resistance mechanism to diazinon was not able to be confirmed.

Susceptibility Status and Resistance Mechanisms in Permethrin-Selected, Laboratory Susceptible and Field-Collected Aedes aegypti from Malaysia.

This study is intended to provide a comprehensive characterization of the resistance mechanisms in the permethrin-selected (IMR-PSS) and laboratory susceptible (IMR-LS) Aedes aegypti strain from Malaysia. Both IMR-PSS and IMR-LS provide a standard model for use in assessing the pyrethroid resistance in field-collected strains collected from three dengue hotspots: the Taman Seri Bayu (TSB), the Flat Camar (FC), and the Taman Dahlia (TD). Two established methods for determining the resistance mechanisms of the pyrethroid are the quantification of detoxification enzymes via enzyme microassay and the nucleotide sequencing of the domain 2 region from segment 1 to 6 via classical polymerase chain reaction (PCR) amplification—were employed. Enzyme activities in IMR-LS served as the resistance threshold reference, providing a significant standard for comparison with IMR-PSS and other field-collected strains. The amino acids in the domain 2 region of voltage-gated sodium channel (Vgsc) of IMR-LS were served as the reference for detection of any changes of the knockdown resistance (kdr) alleles in IMR-PSS and field-collected strains. Studies clearly indicated that the IMR-LS was highly susceptible to insecticides, whilst the IMR-PSS was highly resistant to pyrethroids and conferred with two resistance mechanisms: the elevated oxidase enzyme activity and the altered target-site mutations. Mutations of V1023G alone, and the combination mutations of V1023G with S996P in IMR-PSS, as well as the in field-collected Aedes aegypti strain, indicate the spread of the (kdr) gene in Aedes aegypti, particularly in dengue-endemic areas in Malaysia.

Molecular identification of four novel cytochrome P450 genes related to the development of resistance of Spodoptera exigua (Lepidoptera: Noctuidae) to chlorantraniliprole.

The beet armyworm, Spodoptera exigua, is an omnivorous insect that damages a variety of crops worldwide. Chlorantraniliprole is a new diamide insecticide that acts on the ryanodine receptors in insects. The aim of this study was to explore key genes related to the development of chlorantraniliprole resistance in S. exigua. Transcriptomes were compared between beet armyworms from a susceptible laboratory strain (Sus-Lab) and Sus-Lab screened with LC25 sublethal doses of chlorantraniliprole for six generations (SE-Sel). Ten of 11 cytochrome P450 genes with upregulated expression verified by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in SE-Sel strains were detected in SE-PZ16 and SE-ZY16, two extremely resistant field populations. In addition, expression of four new cytochrome P450 genes, CYP9A21v1, CYP9A21v2, CYP9A21v3 and CYP9A21v4, was shown in the two field populations and was significantly higher in the SE-Sel strain than in the Sus-Lab strain (P < 0.05). Their full-length and protein tertiary structures were also cloned and predicted. The function of CYP9A21v3 was analysed by RNA interference, and the relative expression of CYP9A21v3 in the SE-ZY16 population after feeding on dsRNA was lower than in the control group. Moreover, mortality rates in insects treated at the LC50 of chlorantraniliprole after dsRNA feeding were significantly higher than in the control group 24 h after treatment (P < 0.05). Overexpression of CYP9A21v3 may be a primary factor in the development of chlorantraniliprole resistance in beet armyworms. © 2018 Society of Chemical Industry.

Target-site mutations (AChE and kdr), and PSMO activity in codling moth (Cydia pomonella (L.) (Lepidoptera: Tortricidae)) populations from Spain

Codling moth, Cydia pomonella (L.) is a key pest of global importance that affects apple fruit production and whose populations have developed resistance to insecticides in many apple production areas. In Spain, enhanced cytochrome P450 polysubstrate monooxygenase (PSMO) activity is the main mechanism involved in insecticide detoxification by codling moth, although acetylcholinesterase (AChE) target site mutations have been described in two populations. However, the extent of AChE and knockdown resistance (kdr) mutations in Spain is unknown. To assess the actual occurrence of AChE and kdr mutations concurrently with the frequency of moths with PSMO enhanced activity (R-PSMO), 32 Spanish field populations from four apple-growing areas of Spain and two susceptible laboratory strains were evaluated. R-PSMO was significantly higher in 23 chemically treated field populations from Extremadura, Catalonia and Aragon, with proportions that varied between 25% and 90%, but no significant differences among strains and the non-chemically treated orchards (organic or abandoned) were observed. The AChE mutation (F290 V) was detected in all field populations from Catalonia (n = 21) and in three field populations from Aragon (n = 5), with resistant phenotype proportions varying from 34.2% to 97.5% and from 7.2% to 65% in Catalonia and Aragon, respectively. In addition, the kdr mutation (L1014F) was detected in twelve Catalonian field populations, at rates of incidence ranging between 2.6% and 56.8%. A positive correlation between R-PSMO and AChE mutation was found. The origin of the mutations and their ability to persist and spread in field populations with different management systems is discussed.

Pyrethroid Resistance Status of Aedes (Stegomyia) Aegypti (Linneaus) from Dengue Endemic Areas in Peninsular Malaysia

Vector control is still the principal method to control dengue and chemical insecticides, especially the pyrethroids such as permethrin are the forerunners of mosquito control agent. Intensive and extensive use of pyrethroids often result in resistance, thereby hampering control efforts. The present study was conducted to evaluate the susceptible status of Aedes aegypti, the primary vector of dengue against permethrin. A nationwide mosquito sampling via ovitrapping was conducted in 12 dengue hotspots across 5 states in Peninsular Malaysia. Field collected Aedes eggs were hatched and reared until L3 larval and further identified it species. Adult F0 Aedes aegypti were reared until F1 progeny and the female were used in adult assay, performed according to World Health Organization (WHO) protocol as to determine the resistance level. The laboratory strain maintained for more than 1000 generations that were susceptible to permethrin served as the control strain. Evaluation of resistance ratio was assessed by comparing the knockdown rate with laboratory susceptible strain. In this present study, 70% ofAe. aegypti population from dengue hotspots was highly resistance to permethrin. The study clearly demonstrated that widespread of permethrin resistant Ae. aegypti in Malaysian mosquito’s population, indicating the need of implementing an efficient pyrethroid resistance management.

Geographical variation in susceptibility of tea red spider mite, Oligonychus coffeae (Nietner) (Acari: Tetranychidae) to commonly used acaricides in tea plantations of Assam, north east India

ABSTRACTThe tea red spider mite, Oligonychus coffeae, is one of the most destructive pests of tea in the world which is mainly managed by chemical acaricides. In the present study, concentration-mortality responses of O. coffeae from eight tea geographical populations along with a laboratory susceptible strain against five acaricides, viz., ethion, dicofol, propargite, fenazaquin and fenpropathrin were assayed based on standard leaf-dipped method. Although observed levels of susceptibility varied between populations and acaricides, population from Cachar region registered a high LC50 value to most of the acaricides evaluated. Almost all of the tested field populations had developed high to very high level of resistance to ethion and dicofol, with resistance ratios ranging from 13.58 to 134.27-fold. Fenazaquin and propargite were very effective against O. coffeae in all tested populations, while fenpropathrin showed minor variation in their susceptibility. Significant positive correlation was only observed between the LC50 values of ethion and dicofol (r = 0.95, P = 0.005) suggesting the potential for cross-resistance. These results indicated that control failure was associated with resistance to ethion and dicofol therefore reinforcing the need of resistance management program and offered useful information about alternative acaricides to control O. coffeae.

Phosphine Resistance in North American Field Populations of the Lesser Grain Borer, Rhyzopertha dominica (Coleoptera: Bostrichidae).

Phosphine is the most widely used fumigant for stored grain insect pests, and resistance to phosphine has evolved in several species worldwide. This study was designed to determine the presence of phosphine resistance in 34 populations of Rhyzopertha dominica (F.) collected from the United States and Canada. Adult R. dominica were sampled and subjected to a discriminatory dose toxicity assay of exposure to 20 ppm of phosphine for 20 h of exposure to distinguish a susceptible R. dominica adult by death from a resistant beetle that survives the treatment. All but two of the 34 geographic populations surveyed had some beetles that were resistant to phosphine, and the frequency of resistance varied from 97% in a population from Parlier, California to 0% in beetles from both Carnduff, Saskatchewan and Starbuck, Manitoba. Probit analyses of dose-mortality bioassays with beetles from a laboratory-susceptible strain and those from five of the populations sampled were used to calculate resistance ratio factors (RRs) based on the ratio of LC50 (estimate for the concentration to kill 50% of a test group) in the sampled population to the LC50 for the susceptible strain. The highest RR for the five resistant populations was nearly 596-fold in beetles from Belle Glade, Florida, whereas the lowest RR in that group was 9-fold in Wamego, Kansas. This study revealed that phosphine resistance in R. dominica is common across North America and some populations have levels of resistance that may pose challenges for continued use of phosphine for their management.

Field cage insecticide resistance tests against Culex quinquefasciatus Say (Diptera: Culicidae) in Harris County, Texas, U.S.A.

The ground adulticiding program in densely populated Harris County has been employed on the basis of virus-positive mosquitoes almost exclusively using the 'rotation of chemicals best practices' stratagem. To evaluate its effectiveness, 15 comparative field cage testing events were conducted from 2011-2015 using seven wild population samples from repeatedly collected locations of Culex quinquefasciatus Say and a laboratory susceptible Sebring strain colony reared to adults. A 3 × 3 plot design was employed for exposure to ultra-low volume applications of malathion and synergized permethrin. No significant differences were found in mortality rate among testing dates, year, row placement, or relative humidity. Mortality was significantly different between adulticides (p<0.001) with mean mortality rates for malathion 96.42% (±7.95%) and permethrin 92.38% (±14.04). There was a significant temperature difference for permethrin (p<0.001) but none for malathion (p=0.644). Mosquito population mortality was statistically different by study operational area (p<0.011) and chemical (p<0.001). Susceptible colony adults used as positive controls downwind strongly aided determination of efficacy and resistance to each adulticide, providing evidence of individual application coverage, though comparative analysis was done with overall mortalities by normal methodology.

Monitoring insecticide resistance and diagnostics of resistance mechanisms in the green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae) in China

Myzus persicae (Sulzer) is one of the most serious agricultural pests in China, and management strategies mainly rely on insecticidal treatment. To evaluate the resistance of field populations of M. persicae to seven insecticides, we assessed the susceptibility of 11 field populations collected from eight provinces in China using leaf-dip bioassays. Toxicity assays showed that M. persicae field populations have developed several levels of resistance to each tested insecticide. For pyrethroids, the field populations have developed a high level of resistance to β-cypermethrin and cypermethrin, while the resistance to bifenthrin is still low. The resistance ratios of field populations to imidacloprid ranged from 1.48 to 52.36, and eight populations have developed moderate to high resistance. Resistance to acetamiprid is low, and only two populations have a moderate level of resistance. Most of the field populations of M. persicae developed moderate to high resistance to methomyl and omethoate. To investigate potential resistance mechanisms, we analyzed the enzyme activity of carboxylesterases, the type of amplified esterase genes, as well as the kdr (L1014F) mutation. All of the field populations exhibited a higher esterase activity compared to the laboratory susceptible strain. An amplified FE4, as well as the L1014F mutation, were also found in all of our experimental field populations. These results provide valuable insight into the current status of insecticide resistance and will prove to be a valuable resource in designing appropriate resistance management strategies for M. persicae in China.

Resistance Management for Asian Citrus Psyllid, Diaphorina citri Kuwayama, in Florida.

The Asian citrus psyllid, Diaphorina citri Kuwayma, is one of the most important pests in citrus production. The objective of this study was to evaluate D. citri resistance management with three insecticide rotations and compare them with no rotation and an untreated check. The different insecticides (modes of action) tested were: dimethoate, imidacloprid, diflubenzuron, abamectin 3% + thiamethoxam 13.9%, and fenpropathrin. Eggs, nymph, and adult psyllids were counted weekly. Five insecticide applications were made in 2016. Insecticide susceptibility was determined by direct comparison with a laboratory susceptible population and field populations before and after all treatments were applied. Rankings of eggs, nymphs, and adults counted in treated plots were significantly lower than in the untreated control plots after each application. Initially, the resistance ratio (RR50) for each rotation model, as compared with laboratory susceptible strain and the field population before application, was less than 5.76 and 4.31, respectively. However, after five applications with dimethoate, the RR50 using the laboratory and pre-treatment field populations was 42.34 and 34.74, respectively. Our results indicate that effectively rotating modes of action can delay and/or prevent development of insecticide resistance in populations of D. citri.

First Data on Resistance to Pyrethroids in Wild Populations of Aedes albopictus from Spain.

The invasive mosquito, Aedes albopictus, found in Spain since 2004, is a competent vector of yellow fever, Zika, dengue, and chikungunya viruses among other diseases. Although controversial, the use of adulticiding is a relevant tool for vector control and could be crucial for the management of any possible outbreak of imported diseases. We present the 1st study in Spain on the susceptibility of field populations from Barcelona, Peñíscola, Castellón, and Mallorca of Ae. albopictus to several pyrethroids using either bioassays under the World Health Organization methodology and biochemical tests. In the bioassays, the discriminating concentrations were calculated using a local, susceptible laboratory strain. Different susceptibility levels were found for some combinations of populations and products. The biochemical tests carried out by enzymatic analysis supported these results, showing an overexpression of glutathione S-transferase activity in 1 population.

A Comparative Analysis of Resistance Testing Methods inAedes albopictus(Diptera: Culicidae) from St. Johns County, Florida

Abstract Aedes albopictus Skuse (Diptera: Culicidae) was tested for resistance to permethrin, bifenthrin, and malathion using Centers for Disease Control and Prevention (CDC) bottle bioassays and topical toxicology assays on adults and larval bioassays. Eggs were collected from 3 locations across St. Johns County, Florida, raised to the F3 generation and compared with an insecticide susceptible laboratory strain. Results from CDC bottle bioassays with permethrin indicate no significant differences between the 3 wild-type strains and the laboratory strain but suggest the possibility of resistance in 1 strain. Bottle bioassay results for malathion were inconclusive. Topical toxicological results for adults and bioassays for larvae showed a significant difference in permethrin resistance between the control strain and 1 of the wild-type strains. Results from this project indicate that insecticide susceptibility testing should be a regular part of mosquito surveillance programs. Upon detection of resistance, ...