Monitoring of Insecticide Resistance in Field-Collected Populations of Culex pipiens pallens (Diptera: Culicidae)

A susceptible strain and six field populations of the northern house mosquito, Culex pipiens pallens Coquillett, were collected in Seoul, Paju, Daejeon, Heongseong, Jinhae, and Buan, and tested for their susceptibility to seven insecticides by topical application. In tests with female Cx. pipiens pallens, marked regional variations of insecticide susceptibility were observed. Five field-collected populations of Cx. pipiens pallens, except for the Seoul colony, exhibitedhightoveryhighlevelsofresistanceto cyfluthrin [resistance ratio (RR), 96–554] and deltamethrin (RR, 192–959). Low to moderate levels of resistance were observed in bendiocarb (RR, 14–46), chlorpyrifos [RR, 7–38; exceptions: Buan (RR, 63) and Jinhae (RR, 80) colonies], λ-cyhalothrin (RR, 5–23), S-bioallethrin (RR, 1–47), and permethrin [RR, 2–27; exception: Jinhae colony (RR, 247)]. These results indicate that careful selection and rotational use of these insecticides should result in continued satisfactory control against field populations of Cx. pipiens pallens.     

Differential susceptibilities of Brontispa longissima (Coleoptera: Hispidae) to insecticides in Southeast Asia

To present the susceptibility of Brontispa longissima (Gestro) (Coleoptera: Hispidae) to insecticides, 22 populations were collected in Southeast Asia from 2007 to 2010. Moreover, the laboratory susceptible strain was maintained. The results demonstrated that the lethal concentration50 value of the susceptible strain to avermectin, beta-cypermethrin, and acetamiprid was 0.034, 0.024, and 1.843 mg liter(-1), respectively. Compared with the susceptible strain, 10 populations developed a medium level of resistance to avermectin (10.8-fold < resistance ratio [RR] < 34.7-fold) and 11 populations had low or minor resistance (3.6-fold < RR < 9.6-fold), only one population from Qionghai still remained susceptible (RR = 2.04-fold). The Zhangjiang population expressed high resistance (RR = 46.1-fold), five populations expressed medium resistance (10.6-fold < RR < 18.1-fold), and 12 populations had low resistance (5.36-fold < RR < 9.66-fold) for beta-cypermethrin, only four populations maintained low resistance or susceptibility (2.08-fold < RR< 3.49-fold). All populations were susceptible to acetamiprid (1.65-fold < RR < 2.89-fold), except for the Jakarta population that developed minor resistance (4.33-fold). This study shows that the resistance to beta-cypermethrin and avermectin in B. longissima is widespread. However, most populations remain susceptible to acetamiprid. Acetamiprid may be considered an available option for the control of B. longissima and should be considered when designing pest management programs.     

Monitoring pyrethroid resistance in field collected Blattella germanica Linn. (Dictyoptera: Blattellidae) in Indonesia

The German cockroach, Blattella germanica , is a major and the most common pest in public areas in Indonesia. Although intensive control measures have been carried out to control the populations of this pest, results have been far from successful, which is believed to be because of its resistance to insecticides. A standard World Health Organization (WHO) glass jar test was carried out to determine the resistance level of this insect to pyrethroid insecticides, the most commonly used insecticides for cockroach control in Indonesia. A susceptible S1 strain collected from Tembagapura Papua was compared with four strains collected from Bandung, West Java: strain S2, from a local restaurant; strain S3, from the Bandung train station; and strains S4 and S5, from two different hotels. All strains showed low resistance to the pyrethroid, except the S5 strain, which had a Resistance Ratio (RR)₅₀ of 95 for permethrin. The addition of piperonyl butoxide (PBO) suggests that the detoxifying enzyme mixed function oxidases (MFO) played an important role in the development of resistance to permethrin in the S5 strain, suggested by the high Synergist Ratio (SR) of 70.4. However, the low level of resistance to cypermethrin was not affected by PBO, suggesting that other mechanisms of pyrethroid resistance are involved. Our study is the first report of German cockroach resistance to permethrin in Indonesia, and the findings can be used in formulating potential strategies for cockroach resistance management.     

Effects of hydroprene exposure on the physiology and insecticide susceptibility of German cockroaches (Orthoptera: Blattellidae).

We determined nonlethal physiological effects of hydroprene on the German cockroach, Blattella germanica L., and the effects of hydroprene exposure on German cockroach susceptibility to traditional insecticides. Live and dry weights of German cockroach adults exposed as nymphs to the juvenile hormone analog hydroprene increased significantly (6-35%) in both sexes. The change in weight was affected by hydroprene dose and time after adult emergence. Hydroprene treatment did not affect total body lipids and carbohydrates consistently. However, total body urates of males and females increased significantly. The susceptible and multiresistant German cockroach strains were significantly more susceptible to propoxur at the lower hydroprene application rate (0.697 mg per tub) than the untreated cockroaches. Hydroprene application also significantly lowered the LD50 for chlorpyrifos of the multiresistant strain at the lower hydroprene application rate. However, hydroprene failed to affect chlorpyrifos susceptibility of the susceptible strain.     

Insecticide Susceptibility of Field‐collected Blattella germanica (Blattaria: Blattalidae) in Busan,Republic of Korea during 2014

The susceptibility values for field‐collected (BG‐BG) and laboratory (KS‐BG) strains of German cockroaches (BS‐BG), Blattella germanica (L.) to five synthetic pyrethroid and three organophosphate insecticides were assessed by topical application. Both KS‐BG and BS‐BG strains demonstrated the highest levels of susceptibility to deltamethrin with LD₅₀ values of 0.0035 μg/female and 1.5744 μg/female, respectively. The KS‐BG strain demonstrated the lowest susceptibility level to fenthion with an LD₅₀ value of 0.0698 μg/female, while the BS‐BG field population showed the lowest susceptibility to bifenthrin with 4.6811 μg/female. The BS‐BG field population showed resistance levels of 50 to 640 fold higher against the eight pesticides evaluated compared to the KS‐BG strain. The resistance ratios (RR) of the eight pesticides were compared using populations of B. germanica that were collected from the same locality during 2007 and 2014. The field population of B. germanica collected during 2014 demonstrated significantly high resistance to an organophosphates and all pyrethroids when compared with those collected during 2007, but RR values were <10 except for chlorpyrifos‐methyl and chlorpyrifos with RR values of 74 and 104, respectively.     

Insecticide susceptibility and resistance of larvae of the Anopheles sinensis Group (Diptera: Culicidae) from Paju, Republic of Korea

The susceptibility of members of the Anopheles sinensis Group in Korea to insecticides was evaluated under laboratory conditions using 15 insecticides currently used by local public health centers in Korea. The insecticides included eight pyrethroids, six organophosphates and a pyrazol analogue. Based on their LC₅₀ values, the order of susceptibility of An. sinensis larvae to the insecticides was bifenthrin, chlorfenapyr, α-cypermethrin and λ-cyhalothrin, with values of 0.009, 0.04, 0.06 and 0.08 p.p.m., respectively. The least susceptibility was obtained with fenitrothion, with an LC₅₀ of 7.7 p.p.m. In the comparative resistance test, the resistance ratios (RR) of 14 insecticides were compared to each other using two strains of members of the An. sinensis Group collected in the locality in 2001 and 2008. Anopheles spp. demonstrated higher RR to organophosphates such as fenthion, and low RR for the pyrethroids. Among the organophosphates, fenthion had the highest RR of 33.3 and 270.0 fold differences for LC₅₀ and LC₉₀ values, respectively. Among the pyrethroids, permethrin was observed to have the highest RR of 3.8 and 1.8 fold differences for LC₅₀ and LC₉₀ values, respectively. However, there were no significant differences in susceptibility to chlorfenapyr, chlorpyrifos, deltamethrin and fenitrothion. An. sinensis s. l. was more susceptible to the six insecticides bifenthrin, λ-cyhalothrin, α-cypermethrin, cypermethrin, cyfluthrin and pyridafenthion, showing 0.03, 0.06, 0.3, 0.3, 0.4 and 0.4 fold differences in resistance rates (RR LC₅₀), respectively.     

Effect of rotational use of insecticides on pyrethroids resistance in Helicoverpa armigera (Lep.: Noctuidae)

Abstract:  To investigate fluctuation in susceptibility to insecticides in the field, natural populations of Helicoverpa armigera were collected from the same field in the region of Multan, Pakistan in 2002 and 2003. The populations were examined against pyrethroids (viz. cypermethrin, esfenvalerate and fenpropathrin) and new insecticides (viz. spinosad, abamectin and indoxacarb). In 2002, the resistance ratio (RR) of cypermethrin and esfenvalerate was significantly higher than fenpropahrin compared with susceptible population. The susceptibility to cypermethrin, esfenvalerate and fenpropathrin increased significantly in 2003; however, the RR for cypermethrin was about half the RR (38) of esfenvalerate (101) and fenpropathrin (89). The toxicity of spinosad, abamectin and indoxacarb was identical in both years. In the field experiments, abamectin was more effective than other compounds tested whereas fenpropathrin, cypermethrin and esfenvalerate had similar toxicity. These results might have important implications in resistance management and suggest that the rotational use of spinosad, abamectin and indoxacarb could help to avoid the development of multiple resistant in H. armigera .     

Survey of susceptibilities to monosultap, triazophos, fipronil, and abamectin in Chilo suppressalis (Lepidoptera: Crambidae)

To provide a foundation for national resistance management of the Asiatic rice borer, Chilo suppressalis (Walker) (Lepidoptera: Crambidae), a study was carried out to determine dose-response and susceptibility changes over a 5-yr period in the insect from representative rice, Oryza sativa L., production regions. In total, 11 populations were collected from 2002 to 2006 in seven rice-growing provinces in China, and they were used to examine their susceptibility levels to monosultap, triazophos, fipronil, and abamectin. Results indicated that most populations had increased tolerance to monosultap. Several field populations, especially those in the southeastern Zhejiang Province, were highly or extremely highly resistant to triazophos (resistance ratio [RR] = 52.57-899.93-fold), and some populations in Anhui, Jiangsu, Shanghai, and the northern rice regions were susceptible or had a low level of resistance to triazophos (RR = 1.00-10.69). Results also showed that most field populations were susceptible to fipronil (RR < 3), but the populations from Ruian and Cangnan, Zhejiang, in 2006 showed moderate levels of resistance to fipronil (RR = 20.99-25.35). All 11 field populations collected in 2002-2006 were susceptible to abamectin (RR < 5). The tolerance levels in the rice stem borer exhibited an increasing trend (or with fluctuation) over a 5-yr period for different insecticides, and they reached a maximal level in 2006 for all four insecticides. Analysis of regional resistance ratios indicated that the history and intensity of insecticide application are the major driving forces for the resistance evolution in C. suppressalis. Strategic development of insecticide resistance management also is proposed.     

Pyrethroid resistance and synergism in a field strain of the German cockroach (Dictyoptera: Blattellidae).

A field-collected strain of the German cockroach, Blattella germanica (L.), was highly resistant to 10 pyrethroid insecticides (cyfluthrin, cyhalothrin, cypermethrin, fenvalerate, esfenvalerate, fluvalinate, permethrin, resmethrin, sumithrin, tralomethrin) based on topical applications and comparison with a known susceptible strain. Resistance ratios ranged from 29 to 337. In general, pyrethroid compounds with an alpha-cyano functional group were more toxic than those lacking this moiety, but resistance ratios were similar for both classes of compound. The metabolic inhibitors DEF and PBO were tested for synergism in conjunction with cypermethrin (alpha-cyano) and permethrin (non alpha-cyano). Application of synergists resulted in partial elimination of resistance, suggesting that the basis of resistance involves enhanced metabolism as well as target site insensitivity. These results suggest that pyrethroid insecticides may have a very short functional life in German cockroach control unless they are used judiciously.     

Ovicidal activity of noviflumuron when fed to adult German cockroaches (Dictyoptera: Blattellidae)

Ovicidal activity of the benzoylphenylurea noviflumuron was evaluated in the laboratory on three adult groups (virgin females, virgin males, and fertilized, nongravid females) of the German cockroach, Blattella germanica (L.), through ingestion of treated bait. Novifumuron caused significant ovicidal effects at concentrations ranging from 10 to 5000 ppm after 5-d feeding exposure to virgin and fertilized females. Untreated females produced little or no viable oothecae when mated with virgin males that had previously ingested bait (5-d exposure) with 1000 ppm or 5000 ppm noviflumuron. The highest tested concentration of noviflumuron (5000 ppm) caused 100% ovicidal activity through two ovarian cycles for all three adult groups. Noviflumuron seems to have broader ovicidal activity against B. germanica than reported for other benzoylphenylurea insecticides and can potentially impact cockroach populations through a combination of nymphal mortality and ovicidal activity.     

Susceptibility of Cypriot Tuta absoluta populations to four targeted insecticides and control failure likelihood

Tuta absoluta, known as the South American tomato pinworm, is one of the most disastrous pests of tomato cultivations, presently menacing tomato cultivations worldwide. In 2006, T. absoluta invaded Spain from South America. Since then, it was rapidly spread to most European, African and Asian countries. Such alien invasive species can minimize crop production, whereas the increasing use of insecticides raises various environmental concerns as well as on control costs, control failure and the toxicity to non‐target organisms. The S. American tomato pinworm is mostly controlled by chemical insecticides, and failure to control it is not a rare phenomenon. Resistance to numerous insecticides has been reported and is mainly due to the fact that farmers do not follow a sustainable resistance management scheme. Several examples have been reported from several countries where the tomato pinworm is present. In order to develop a successful insecticide resistance management (IRM) strategy for any major pest, one needs to identify the baseline toxicity to insecticides and then monitor susceptibility levels . In Cyprus, the current status of susceptibility levels to the main insecticides that are used to control T. absoluta has never been studied before. Herein, nine Cypriot populations of the pest were subjected to laboratory bioassays between 2016 and 2018 using the main insecticides applied against it. We found that the insecticides chlorantraniliprole and indoxacarb could not control the Cypriot T. absoluta populations anymore, with a resistance ratio (RR) >28 and 3–23, respectively. Furthermore, mortality achieved by those two insecticides was 20.6%–72% for chlorantraniliprole and 27.5%–78% for indoxacarb. However, the insecticides emamectin benzoate and spinosad are very effective, since mortality to both of them ranged between 99.5% and 100%.     

Monitoring and mechanisms of organophosphate resistance in San Jose scale,Diaspidiotus perniciosus (Hemiptera: Diaspididae)

San Jose scale, Diaspidiotus perniciosus (Comstock), is a serious pest in Chilean apple tree orchards, and a number of organophosphate insecticides were used to control them for decades. Recently, control failures with these insecticides were reported and linked to insecticide resistance development. In this study, 40 San Jose scale field populations were collected and their susceptibility to two commonly used organophosphate insecticides, that is chlorpyrifos and methidathion, was assessed. The obtained bioassay data suggest moderate levels of resistance to both insecticides when compared to a reference susceptible strain. The highest resistance ratio (RR) detected for chlorpyrifos and methidathion was 31‐fold and 11‐fold, respectively. The bioassay results suggest the occurrence of a significant cross‐resistance between both compounds. Biochemical measurements revealed a role for esterases in conferring resistance to organophosphates, but not modified acetylcholinesterase. The spatial spread and extend of insecticide resistance were also evaluated. Our result shows that no autocorrelation can be assumed, and then, insecticide resistance is caused by random factors.     

Cytochrome P450 MA expression in insecticide-resistant German cockroaches (Dictyoptera: Blattellidae).

Cytochrome P450 monooxygenases are detoxification enzymes commonly involved in insecticide resistance by insects. Recently, an overexpressed form of this enzyme, P450 MA, was purified from an insecticide-resistant strain of German cockroach, Blattella germanica (L.), and polyclonal antisera (anti-P450 MA) was produced. To test hypotheses that the overexpressed condition of P450 MA has evolved in > 1 geographic location and that P450 MA might be involved in insecticide resistance to specific insecticides, investigations were conducted using 4 insecticide-resistant and 1 susceptible German cockroach strains. In western blots that used anti-P450 MA antiserum as a probe, substantial differences in expression of P450 MA were observed. Strains showing the highest P450 MA expression had both the highest tolerance to the organophosphate insecticide chlorpyrifos and cytochrome P450-mediated demethylation activity. Results support the hypothesis that cytochrome P450 MA is potentially overexpressed in insecticide-resistant populations on a global scale.     

Establishing the role of detoxifying enzymes in field‐evolved resistance to various insecticides in the brown planthopper (Nilaparvata lugens) in South India

The brown planthopper (BPH), Nilaparvata lugens (Stål), is one of the major pests of rice throughout Asia. Extensive use of insecticides for suppressing N. lugens has resulted in the development of insecticide resistance leading to frequent control failures in the field. The aim of the present study was to evaluate resistance in the field populations of N. lugens from major rice growing states of South India to various insecticides. We also determined the activity of detoxifying enzymes (esterases [ESTs], glutathione S‐transferases [GSTs], and mixed‐function oxidases [MFOs]). Moderate levels of resistance were detected in the field populations to acephate, thiamethoxam and buprofezin (resistance factors 1.05–20.92 fold, 4.52–14.99 fold, and 1.00–18.09 fold, respectively) as compared with susceptible strain while there were low levels of resistance to imidacloprid (resistance factor 1.23–6.70 fold) and complete sensitivity to etofenoprox (resistance factor 1.05–1.66 fold). EST activities in the field populations were 1.06 to 3.09 times higher than the susceptible strain while for GST and MFO the ratios varied from 1.29 to 3.41 and 1.03 to 1.76, respectively. The EST activity was found to be correlated to acephate resistance (r = 0.999, P ≥ 0.001). The high selection pressure of organophosphate, neonicotinoid, and insect growth regulator (IGR) in the field is likely to be contributing for resistance in BPH to multiple insecticides, leading to control failures. The results obtained will be beneficial to IPM recommendations for the use of effective insecticides against BPH.     

Novel point mutations in the German cockroach para sodium channel gene are associated with knockdown resistance (kdr) to pyrethroid insecticides

Knockdown resistance (kdr) to pyrethroid insecticides has been attributed to point mutations in the para sodium channel gene in more than a half dozen insect pest species. In this study, we identified two novel para mutations in five highly resistant kdr-type German cockroach strains. The two mutations, from glutamic acid (E434) to lysine (K434) and from cysteine (C764) to arginine (R764), respectively, are located in the first intracellular linker connecting domains I and II. E434K is located near the beginning of the linker (closest to domain I), whereas C764R is found toward the end of the linker (closest to domain II). Two additional mutations from aspartic acid (D58) to glycine (G58), and from proline (P1880) to leucine (L1888), respectively, were found in one of the resistant strains. The four mutations coexist with the previously identified leucine to phenylalanine (L993F) kdr mutation in IIS6, and are present only in the highly resistant individuals of a given strain. These findings suggest that these mutations might be responsible for high levels of knockdown resistance toward pyrethroid insecticides in the German cockroach.     

Relationship between the para-homologous sodium channel point mutation (g --> c at nucleotide 2979) and knockdown resistance in the German cockroach using multiplex polymerase chain reaction to discern genotype

Extensive use of pyrethroid insecticides for urban pest control has led to widespread pyrethroid resistance in the German cockroach. A mutation at nucleotide position 2979 (G to C, causing a leucine to phenylalanine change) in the S6 transmembrane segment of domain II of the para-homologous voltage-gated sodium channel has been previously identified in knockdown-resistant cockroaches and demonstrated by site-directed mutagenesis to reduce channel sensitivity to pyrethroids. In a recent survey, 83% of pyrethroid-resistant German cockroach populations were found to possess this mutation. A German cockroach strain with a low incidence of the L993F mutation was subjected to selection pressure with cypermethrin and subsequently evaluated over several generations for the knockdown resistance phenotype. Correspondingly, we determined the genotype of individual cockroaches of each population at the 2979 position of the para-homologous gene. Genotype was discerned by development of a polymerase chain reaction method that employed a mismatched primer-template set. A direct relationship was observed between mean knockdown time and the presence of the kdr mutation. Furthermore, individuals homozygous for the kdr mutation exhibited a significantly higher mean knockdown time than heterozygotes or wildtype cockroaches. This is the first report demonstrating the progressive expression of the kdr allele in response to insecticide selection pressure.     

Insecticide toxicity, synergism and resistance in the German cockroach (Dictyoptera: Blattellidae)

The toxicity of synergism of and resistance to insecticides in four strains of German cockroach, Blattella germanica (L.), were investigated. Toxicity of nine insecticides by topical application to the susceptible strain varied greater than 2,000-fold, with deltamethrin (LD50 = 0.004 micrograms per cockroach) and malathion (LD50 = 8.4 micrograms per cockroach) being the most and least toxic, respectively. Resistance to pyrethrins (9.5-fold) in the Kenly strain was unaffected by the synergists piperonyl butoxide (PBO) or S,S,S-tributylphosphorotrithioate (DEF), suggesting that the metabolism is not involved in this case. Malathion resistance in the Rutgers strain was suppressible with PBO, implicating oxidative metabolism as a resistance mechanism. The Ectiban-R strain was resistant to all the pyrethroids tested, and cypermethrin resistance was not suppressible with PBO or DEF. These findings support results of previous studies that indicated this train has a kdr-like mechanism. Bendiocarb resistance in both the Kenly and Rutgers strains was partially suppressed by either PBO or DEF, suggesting that oxidative and hydrolytic metabolism are involved in the resistance. Trends between the effects of the synergists on the susceptible versus resistant strains are discussed.     

Insecticide resistance in dengue vectors from hotspots in Selangor,Malaysia

BackgroundIn Malaysia, dengue remains a top priority disease and usage of insecticides is the main method for dengue vector control. Limited baseline insecticide resistance data in dengue hotspots has prompted us to conduct this study. The present study reports the use of a map on the insecticide susceptibility status of Aedes aegypti and Aedes albopictus to provide a quick visualization and overview of the distribution of insecticide resistance.Method and resultsThe insecticide resistance status of Aedes populations collected from 24 dengue hotspot areas from the period of December 2018 until June 2019 was proactively monitored using the World Health Organization standard protocol for adult and larval susceptibility testing was conducted, together with elucidation of the mechanisms involved in observed resistance. For resistance monitoring, susceptibility to three adulticides (permethrin, deltamethrin, and malathion) was tested, as well as susceptibility to the larvicide, temephos. Data showed significant resistance to both deltamethrin and permethrin (pyrethroid insecticides), and to malathion (organophosphate insecticide) in all sampled Aedes aegypti populations, while variable resistance patterns were found in the sampled Aedes albopictus populations. Temephos resistance was observed when larvae were tested using the diagnostic dosage of 0.012mg/L but not at the operational dosage of 1mg/L for both species.ConclusionThe present study highlights evidence of a potential threat to the effectiveness of insecticides currently used in dengue vector control, and the urgent requirement for insecticide resistance management to be integrated into the National Dengue Control Program.     

Insecticide resistance in Bemisia tabaci from Cyprus

Abstract A comprehensive study on the Bemisia tabaci (biotype B) resistance to neonicotinoid insecticides imidacloprid, acetamiprid and thiamethoxam, and pyrethroid bifenthrin was conducted in Cyprus. The resistance level to eight field‐collected B. tabaci populations was investigated. The activities of enzymes involved in metabolic detoxification and the frequencies of pyrethroid and organophosphates target site resistance mutations were determined. Moderate to high levels of resistance were detected for imidacloprid (resistance factor [RF] 77–392) and thiamethoxam (RF 50–164) while low resistance levels were observed for acetamiprid (RF 7–12). Uniform responses by the Cypriot whiteflies could be observed against all neonicotinoid insecticides. No cross‐resistance between the neonicotinoids was detected as well as no association with the activity of the P450 microsomal oxidases. Only imidacloprid resistance correlated with carboxylesterase activity. Low to extremely high resistance was observed for insecticide bifenthrin (RF 49–1 243) which was associated with the frequency of the resistant allele in the sodium channel gene but not with the activity of the detoxification enzymes. Finally, the F331W mutation in the acetylcholinesterase enzyme ace1 gene was fixed in all B. tabaci populations from Cyprus.     

Development of neonicotinoid resistance in the cotton aphid Aphis gossypii (Hemiptera: Aphididae) in Japan

A neonicotinoid-resistant Aphis gossypii Glover population was first detected from cucumber and sweet pepper crops in Miyazaki Prefecture, Japan, in April 2012. In this study, we determined the insecticide susceptibility of five field-collected populations of A. gossypii to seven neonicotinoids. Insecticide susceptibility was determined by performing bioassays and using the seedling-treatment method. Insecticide susceptibility of a susceptible population of A. gossypii, originally collected from a cucumber field in 2008, was also determined. High mortality (96.4–100 %) was observed in the susceptible population for the seven insecticides. In contrast, the mortality rate of the five field-collected populations collected in 2012 was low for five of the seven neonicotinoid insecticides tested: imidacloprid (26.7–65.5 %), dinotefuran (0–27.3 %), clothianidin (20.0–35.7 %), thiamethoxam (7.1–42.3 %), and nitenpyram (6.7–32.1 %). Mortality was 86.2–100 % for acetamiprid and 90.2–100 % for thiacloprid. The resistance ratio in comparison with the susceptible population was the highest for clothianidin (687-fold), lowest for thiacloprid (17-fold), and ranged from 43- to 253-fold for the other five neonicotinoids. Thus, the frequency of insecticide use needs to be reduced and other insecticides need to be employed in order to prevent the development of resistance to these two insecticides in the future.