Laboratory-selected Resistant Strain Research Articles

Transcriptomic investigation of the molecular mechanisms underlying resistance to the neonicotinoid thiamethoxam and the pyrethroid lambda-cyhalothrin in Euschistus heros (Hemiptera: Pentatomidae).

Laboratory-selected resistant strains of Euschistus heros to thiamethoxam (NEO) and lambda-cyhalothrin (PYR) were recently reported in Brazil. However, the mechanisms conferring resistance to these insecticides in E. heros remain unresolved. We utilized comparative transcriptome profiling and single nucleotide polymorphism (SNP) calling of susceptible and resistant strains of E. heros to investigate the molecular mechanism(s) underlying resistance. The E. heros transcriptome was assembled, generating 91 673 transcripts with a mean length of 720 bp and N50 of 1795 bp. Comparative gene expression analysis between the susceptible (SUS) and NEO strains identified 215 significantly differentially expressed (DE) transcripts. DE transcripts associated with the xenobiotic metabolism were all up-regulated in the NEO strain. The comparative analysis of the SUS and PYR strains identified 204 DE transcripts, including an esterase (esterase FE4), a glutathione-S-transferase, an ABC transporter (ABCC1) and aquaporins that were up-regulated in the PYR strain. We identified 9588 and 15 043 nonsynonymous SNPs in the PYR and NEO strains. One of the SNPs (D70N) detected in the NEO strain occurs in a subunit (α5) of the nAChRs, the target site of neonicotinoid insecticides. Nevertheless, this residue position in α5 is not conserved among insects. Neonicotinoid and pyrethroid resistance in laboratory-selected E. heros is associated with a potential metabolic resistance mechanism by the overexpression of proteins commonly involved in the three phases of xenobiotic metabolism. Together these findings provide insight into the potential basis of resistance in E. heros and will inform the development and implementation of resistance management strategies against this important pest. © 2023 Society of Chemical Industry.

Biochemical Mechanisms, Cross-resistance and Stability of Resistance to Metaflumizone in Plutella xylostella.

The diamondback moth, Plutella xylostella (L.) is an important pest of cruciferous crops worldwide. It has developed resistance to many conventional and novel insecticide classes. Metaflumizone belongs to the new chemical class of semicarbazone insecticides. To delay the development of metaflumizone resistance in P. xylostella and to guide insecticide use in the field, the biochemical mechanisms, cross-resistance spectrum, and stability of resistance to metaflumizone were studied in a laboratory-selected resistant strain (metaflu-SEL). Synergism tests with the carboxylesterase inhibitor triphenyl phosphate (TPP), the glutathione S-transferase depletor diethyl maleate (DEM), and the P450 inhibitor piperonyl butoxide(PBO) had no obvious effect on metaflumizone in the metaflu-SEL strain and the susceptible strain (SS) of P. xylostella, with synergism ratios that ranged from 1.02 to 1.86. Biochemical studies revealed that the cytochrome P450-dependent monooxygenase increased only 1.13-fold in the metaflu-SEL strain compared with the UNSEL stain; meanwhile, carboxylesterase and glutathione S-transferase activity showed no difference. These results suggest that these detoxification enzymes may be not actively involved in metaflumizone resistance. Furthermore, the metaflu-SEL population showed a moderate level of cross-resistance to indoxacarb (11.63-fold), but only very low cross-resistance to spinosad (1.75-fold), spinetoram (3.52-fold), abamectin (2.81-fold), beta-cypermethrin (0.71-fold), diafenthiuron (0.79-fold), chlorantraniliprole (2.16-fold), BT (WG-001) (3.34-fold), chlorfenapyr (0.49-fold), and chlorfluazuron (0.97-fold). Moreover, metaflumizone resistance decreased from 1087.85- to 1.23-fold in the metaflu-SEL strain after 12 generations without exposure to metaflumizone. These results are useful for formulating insecticide resistance management strategies to control P. xylostella and to delay the development of metaflumizone resistance in the field.

Gut bacteria of field-collected larvae of Spodoptera frugiperda undergo selection and are more diverse and active in metabolizing multiple insecticides than laboratory-selected resistant strains

Bacterial symbiosis affects the physiology and development of insects, facilitating their adaptation to new environmental conditions. Insects and their associated microbiota acquire phenotypic features that guarantee survival in the face of biotic and abiotic stress factors. We used Spodoptera frugiperda (Lepidoptera: Noctuidae) to investigate the effect of selection pressures on the structure, diversity, and capacity of the gut microbiota to metabolize pesticides. We compared the composition of the gut microbiota of susceptible and insecticide-resistant strains of S. frugiperda, as well as the gut microbiota from larvae sampled in cornfields in five Brazilian states. The experiments were conducted through metagenomic analysis of the V3–V4 regions of the 16S ribosomal gene (16S rRNA), isolation, and culture of bacteria in selective minimal medium, and analysis of the growth of isolates using multiple insecticides as the only source of carbon. Field exposure to a range of xenobiotics affected the composition of the gut microbiota of S. frugiperda. Moreover, bacteria from field-collected larvae grew better and showed potential to metabolize more insecticides than the bacteria isolated from laboratory-selected resistant strains. In addition, most of the phylotypes isolated in insecticide-based media were fixed in the gut microbiota of natural populations of S. frugiperda. The higher diversity and capability of gut microbes to metabolize insecticides in field populations of S. frugiperda demonstrate the need for increased investigation of the role of gut microbes in the detoxification of insecticides in the host and their impact on the successful use of pesticides in pest control.

Inheritance and fitness costs of sulfoxaflor resistance in Nilaparvata lugens (Stål).

Sulfoxaflor has been considered as a new tool for Nilaparvata lugens control in the field. In this study, a laboratory-selected resistant strain and a susceptible strain were used to evaluate the inheritance and fitness costs of sulfoxaflor resistance in N. lugens. The resistant strain (SFX-SEL) showed 123.63-fold resistance compared with the susceptible strain (SS). Progenies of reciprocal crosses (F1 RS and F1 SR) showed similar concentration-mortality responses (LC50 ) to sulfoxaflor and also exhibited a similar degree of dominance; -0.16 for F1 RS and -0.26 for F1 SR. Significant differences between the observed and expected mortalities of F2 individuals suggested that sulfoxaflor resistance is associated with multiple genes. The resistant strain had a relative fitness of 0.75 with substantially decreased female adult period, oviposition days, total fecundity, egg hatchability and female adult survival rate, and prolonged pre-adult period and total pre-oviposition period. Sulfoxaflor resistance in N. lugens was inherited as autosomal, incompletely recessive and multigene. The development of resistance may have a significant fitness cost for the resistant population. Current research provides valuable information for researchers to establish management strategies to delay the development of sulfoxaflor resistance and control N. lugens sustainably in the field. © 2019 Society of Chemical Industry.

Selection of influenza virus resistant to the novel camphor-based antiviral camphecene results in loss of pathogenicity

Due to the ability of influenza virus to develop drug resistance, the search for novel antivirals is an important goal of medical science and health care systems. We assessed the ability of the influenza virus to develop resistance to the hemagglutinin inhibitor camphecene and characterized laboratory-selected resistant strains. We showed by electron microscopy that camphecene decreases the number of virions fusing their envelopes with endosomal membranes. A 160-fold decrease in virus susceptibility was observed after six passages in cells. This was associated with the emergence of a V458L mutation in the HA2 subunit of HA and with a decrease in viral pathogenicity. Molecular modeling predicts that this substitution results in a more stable HA molecule compared to wild-type HA; and an altered camphecene-binding site. Therefore, despite the relatively rapid development of resistance, camphecene remains promising as a potential antiviral due to the low pathogenicity of resistant viruses that may arise.

Cross-Resistance and Baseline Susceptibility of Brown Planthopper Nilaparvata lugens (Hemiptera: Delphacidae) From China to Cycloxaprid.

The brown planthopper (BPH), Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), is a serious pest of rice. At present, the application of chemical insecticides is the main control option for this pest. BPH has evolved resistance to various classes of insecticides. Cycloxaprid, a new oxabridged cis-configuration neonicotinoid insecticide, is a (nitromethylene) imidazole analog of imidacloprid. This study focused on the baseline susceptibility to cycloxaprid of 18 field samples of N. lugens collected from nine geographical locations in China, as well as possible cross-resistances between cycloxaprid and other important neonicotinoids in one laboratory-selected resistant strain of N. lugens to imidacloprid. The median lethal concentration (LC50) of cycloxaprid for the 18 samples ranged from 1.26 to 14.90 mg/liter. Furthermore, the cross-resistance studies showed that the imidacloprid-resistant strain exhibited a 27.63-fold resistance to imidacloprid and lower levels of cross-resistance to acetamiprid (16.64-fold), thiacloprid (12.64-fold), and nitenpyram (16.90-fold); however, there was no cross-resistance to cycloxaprid (1.92-fold). These results indicate that cycloxaprid could be an effective alternative insecticide for the management of N. lugens, which is urgently needed to prevent or delay further increases in insecticide resistance in N. lugens.

Biophysical and Pharmacological Characterization of Energy-Dependent Efflux of Sb in Laboratory-Selected Resistant Strains of Leishmania (Viannia) Subgenus.

The growing resistance of leishmaniasis to first-line drugs like antimonials in some regions limits the control of this parasitic disease. The precise mechanisms involved in Leishmania antimony resistance are still subject to debate. The reduction of intracellular SbIII accumulation is a common change observed in both laboratory-selected and field isolated resistant Leishmania strains, but the exact transport pathways involved in antimony resistance have not yet been elucidated. In order to functionally characterize the antimony transport routes responsible for resistance, we performed systematic transport studies of SbIII in wild-type and resistant strains of L. (Viannia) guyanensis and L. (V.) braziliensis. Those include influx and efflux assays and the influence of ABC transporters and metabolism inhibitors: prochlorperazine, probenecid, verapamil, BSO, and sodium azide. The mRNA levels of genes associated with antimony resistance (MRPA, GSH1, ODC, AQP1, ABCI4, and ARM58) were also investigated in addition to intracellular thiol levels. A strong reduction of Sb influx was observed in L. guyanensis resistant mutant (LgSbR), but not in L. braziliensis (LbSbR). Both mutants showed increased energy-dependent efflux of SbIII, when compared to their respective parental strains. In LgSbR, BSO and prochlorperazine inhibited antimony efflux and resistance was associated with increased MRPA and GSH1 mRNA levels, while in LbSbR antimony efflux was inhibited by probenicid and prochlorperazine in absence of resistance-associated gene modulation. Intracellular thiol levels were increased in both Sb-resistant mutants. An energy-dependent SbIII efflux pathway sensitive to prochlorperazine was clearly evidenced in both Sb-resistant mutants. In conclusion, the present study allowed the biophysical and pharmacological characterization of energy-dependent Sb efflux pathway apparently independent of MRPA, ABCI4, and ARM58 upregulation, in Leishmania (Vianna) mutant selected in vitro for resistance to SbIII. Prochlorperazine has also been identified as an effective chemosensitizer in both Sb resistant mutants, which acts through inhibition of the active efflux of Sb.

Baseline susceptibility and cross-resistance of cycloxaprid, a novel cis-nitromethylene neonicotinoid insecticide, in Bemisia tabaci MED from China

The tobacco whitefly Bemisia tabaci (Gennadius) is an important crop pest which threatens agriculture worldwide. It has evolved resistance to various classes of insecticides. Cycloxaprid is a new-generation neonicotinoid which is a (nitromethylene) imidazole analogue of imidacloprid which is highly efficient for the control of various Hemipteran pests in China. Studies were conducted to determine the baseline susceptibility to cycloxaprid of 18 field samples of B. tabaci collected from 9 geographical locations in China and the possibility of cross-resistance between cycloxaprid and other important neonicotinoids of one laboratory-selected resistant strain. The 50% lethal concentrations (LC50) of cycloxaprid to these 18 samples ranged from 0.84 to 12.17 mg/L. Furthermore, compared with the susceptible laboratory strain, the imidacloprid-resistant strain exhibited a 27.9-fold resistance to imidacloprid and lower level of cross-resistance to acetamiprid (16.3-fold), thiacloprid (13.7-fold) and nitenpyram (16.6-fold), but no cross-resistance to cycloxaprid (1.9-fold). These results demonstrate that cycloxaprid could be one effective alternative insecticide for whitefly management that could reduce imidacloprid selection pressure.

Activation of Bt Protoxin Cry1Ac in Resistant and Susceptible Cotton Bollworm.

Crystalline (Cry) proteins from Bacillus thuringiensis (Bt) are used extensively for insect control in sprays and transgenic plants, but their efficacy is reduced by evolution of resistance in pests. Here we evaluated reduced activation of Cry1Ac protoxin as a potential mechanism of resistance in the invasive pest Helicoverpa armigera. Based on the concentration killing 50% of larvae (LC50) for a laboratory-selected resistant strain (LF120) divided by the LC50 for its susceptible parent strain (LF), the resistance ratio was 1600 for Cry1Ac protoxin and 1200 for trypsin-activated Cry1Ac toxin. The high level of resistance to activated toxin as well as to protoxin indicates reduced activation of protoxin is not a major mechanism of resistance to Cry1Ac in LF120. For both insect strains, treatment with either the trypsin inhibitor N-a-tosyl-L-lysine chloromethyl ketone (TLCK) or the chymotrypsin inhibitor N-a-tosyl-L-phenylalanine chloromethyl ketone (TPCK) did not significantly affect the LC50 of Cry1Ac protoxin. Enzyme activity was higher for LF than LF120 for trypsin-like proteases, but did not differ between strains for chymotrypsin-like proteases. The results here are consistent with previous reports indicating that reduced activation of protoxin is generally not a major mechanism of resistance to Bt proteins.

Adult Exposure to Bt Toxin Cry1Ac Reduces Life Span and Reproduction of Resistant and Susceptible Pink Bollworm (Lepidoptera: Gelechiidae).

Insecticidal proteins from Bacillus thuringiensis (Bt) are used widely in sprays and transgenic plants to control insect pests. Although much research has elucidated the effects of Bt toxins on larvae, relatively little is known about their effects on adults. Here, we evaluated the effects of exposing adults to Bt toxin Cry1Ac on the life span and reproduction of two strains of pink bollworm ( Pectinophora gossypiella (Saunders)). In larval diet bioassays, the concentration of Cry1Ac killing 50% of larvae (LC 50 ) was 640 times higher for the laboratory-selected resistant strain (AZP-R) than the susceptible strain (APHIS-S). In experiments with adults, the highest concentrations of Cry1Ac tested (160 and 640 µg Cry1Ac per ml of 5% honey water) reduced life span for both strains. Treatments with 10, 40, and 160 µg Cry1Ac per ml reduced the duration of the oviposition period as well as the number of eggs laid by both strains, but did not affect the percentage of pairs producing eggs, the duration of the preoviposition period, or the percentage of eggs hatching for either strain. Adult life span did not differ between strains at low to moderate concentrations of Cry1Ac, but it was significantly greater for the resistant strain than the susceptible strain at the two highest concentrations of Cry1Ac tested. The reduced susceptibility to high concentrations of Cry1Ac in adults of the AZP-R strain relative to the APHIS-S strain provides the first evidence of expression of resistance to a Bt toxin in adult Lepidoptera.

Inheritance, Realized Heritability, and Biochemical Mechanisms of Malathion Resistance in Bactrocera dorsalis (Diptera: Tephritidae).

To better characterize the resistance development and therefore establish effective pest management strategies, this study was undertaken to investigate the inheritance mode and biochemical mechanisms of malathion resistance in the oriental fruit fly, Bactrocera dorsalis (Hendel), which is one of the most notorious pests in the world. After 22 generations of selection with malathion, the malathion-resistant (MR) strain of B. dorsalis developed a 34-fold resistance compared with a laboratory susceptible strain [malathion-susceptible (MS)]. Bioassay results showed that there was no significant difference between the LD50 values of malathion against the progenies from both reciprocal crosses (F(1)-SR and F(1)-RS). The degree of dominance values (D) was calculated as 0.39 and 0.32 for F(1)-RS and F(1)-SR, respectively. The logarithm dosage-probit mortality lines of the F(2) generation and progeny from the backcross showed no clear plateaus of mortality across a range of doses. In addition, Chi-square analysis revealed significant differences between the mortality data and the theoretical expectations. The realized heritability (h(2)) value was 0.16 in the laboratory-selected resistant strain of B. dorsalis. Enzymatic activities identified significant changes of carboxylesterases, cytochrome P450 (general oxidases), and glutathione S-transferases in MR compared with the MS strain of B. dorsalis. Taken together, this study revealed for the first time that malathion resistance in B. dorsalis follows an autosomal, incompletely dominant, and polygenic mode of inheritance and is closely associated with significantly elevated activities of three major detoxification enzymes.

Resistance to lambda-cyhalothrin in Spanish field populations of Ceratitis capitata and metabolic resistance mediated by P450 in a resistant strain.

The withdrawal of malathion in the European Union in 2009 resulted in a large increase in lambda-cyhalothrin applications for the control of the Mediterranean fruit fly, Ceratitis capitata, in Spanish citrus crops. Spanish field populations of C. capitata have developed resistance to lambda-cyhalothrin (6-14-fold), achieving LC50 values (129-287 ppm) higher than the recommended concentration for field treatments (125 ppm). These results contrast with the high susceptibility to lambda-cyhalothrin found in three Tunisian field populations. We have studied the mechanism of resistance in the laboratory-selected resistant strain W-1Kλ (205-fold resistance). Bioassays with synergists showed that resistance was almost completely suppressed by the P450 inhibitor PBO. The study of the expression of 53 P450 genes belonging to the CYP4, CYP6, CYP9 and CYP12 families in C. capitata revealed that CYP6A51 was overexpressed (13-18-fold) in the resistant strain. The W-1Kλ strain also showed high levels of cross-resistance to etofenprox (240-fold) and deltamethrin (150-fold). Field-evolved resistance to lambda-cyhalothrin has been found in C. capitata. Metabolic resistance mediated by P450 appears to be the main resistance mechanism in the resistant strain W-1Kλ. The levels of cross-resistance found may compromise the effectiveness of other pyrethroids for the control of this species. © 2014 Society of Chemical Industry.

Cross-resistance and baseline susceptibility of spirotetramat in Frankliniella occidentalis (Thysanoptera: Thripidae).

Western flower thrips, Frankliniella occidentalis (Pergande), is an economically important pest all over the world. New products against thrips are necessary, as there are few effective compounds exhibiting cross-resistance among them. Lethal effects, cross-resistance, and baseline susceptibility to spirotetramat were evaluated in this study. A new bioassay method for testing thrips against spirotetramat was developed. Spirotetramat showed a significant mortality effect on larvae after 7 d of exposure, but a low effect was observed on adults. Baseline results for larval instars showed LC50 values ranging from 11.59 to 49.81 mg AI/liter, with a low natural variability (3.2-fold). Cross-resistance studies showed overlapping confidence limits of the LC50 values for laboratory-selected (against acrinathrin, methiocarb, formetanate, and spinosad) resistant and susceptible strains, and low resistance factors, from 0.5 to 1.9, suggesting no cross-resistance to conventional insecticides previously used. A slight ovicidal effect (21-40% reduction) was also detected. Despite presenting low effects on adults, spirotetramat showed high but slow efficacy on F. occidentalis larvae. Field populations in southeast Spain showed a consistent susceptibility to spirotetramat. Given the scarcity of effective products and the lack of cross-resistance to other insecticides, spirotetramat can be considered as a good chemical tool to control F. occidentalis.

Overexpression of Multiple Detoxification Genes in Deltamethrin Resistant Laodelphax striatellus (Hemiptera: Delphacidae) in China

BackgroundThe small brown planthopper (SBPH), Laodelphax striatellus (Fallén), is one of the major rice pests in Asia and has developed resistance to multiple classes of insecticides. Understanding resistance mechanisms is essential to the management of this pest. Biochemical and molecular assays were performed in this study to systematically characterize deltamethrin resistance mechanisms with laboratory-selected resistant and susceptible strains of SBPH.Methodology/Principal FindingsDeltamethrin resistant strains of SBPH (JH-del) were derived from a field population by continuously selections (up to 30 generations) in the laboratory, while a susceptible strain (JHS) was obtained from the same population by removing insecticide pressure for 30 generations. The role of detoxification enzymes in the resistance was investigated using synergism and enzyme activity assays with strains of different resistant levels. Furthermore, 71 cytochrome P450, 93 esterases and 12 glutathione-S-transferases cDNAs were cloned based on transcriptome data of a field collected population. Semi-quantitative RT-PCR screening analysis of 176 identified detoxification genes demonstrated that multiple P450 and esterase genes were overexpressed (>2-fold) in JH-del strains (G4 and G30) when compared to that in JHS, and the results of quantitative PCR coincided with the semi-quantitative RT-PCR results. Target mutation at IIS3–IIS6 regions encoded by the voltage-gated sodium channel gene was ruled out for conferring the observed resistance. Conclusion/SignificanceAs the first attempt to discover genes potentially involved in SBPH pyrethroid resistance, this study putatively identified several candidate genes of detoxification enzymes that were significantly overexpressed in the resistant strain, which matched the synergism and enzyme activity testing. The biochemical and molecular evidences suggest that the high level pyrethroid resistance in L. striatellus could be due to enhanced detoxification rather than target insensitivity. The findings lay a solid ground for further resistance mechanism elucidation studies.

Life-Stage Variation in Insecticide Resistance of the Western Flower Thrips (Thysanoptera: Thripidae)

The life-stage variations in insecticide resistance of western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), to selective insecticides (acrinathrin, formetanate, and methiocarb) were studied using resistant laboratory strains. In each strain, the second-instar larva was less susceptible to the insecticides tested than the adults. The lower the resistance level of the adults, the higher the difference between larva and adult susceptibility: 32-fold to methiocarb, 15.4-fold to formetanate, and 180-fold to acrinathrin in the reference strain. In laboratory-selected resistant strains, these differences were much lower: 5.8-fold to methiocarb, 4.8-fold to formetanate, and 2.0-fold to acrinathrin. In selected strains, higher resistance levels for each insecticide were found, both for larvae and adults, compared with the reference strain. These results show that after insecticide resistance selection in adults, the resistance is carried over to the larvae, but at lower levels.

Lack of fitness costs associated with pyriproxyfen resistance in the B biotype of Bemisia tabaci

The insect growth regulator pyriproxyfen has provided effective control of the whitefly Bemisia tabaci Gennadius in many countries. Here, whether or not fitness costs were associated with pyriproxyfen resistance in a laboratory-selected resistant strain (QC02-R) of the B biotype was determined. Mortality caused by pyriproxyfen and fitness traits over time were measured in unselected and selected hybrid strains, which were created by crossing individuals of the resistant strain with individuals of a susceptible strain. Fitness costs were not associated with resistance in QC02-R, as mortality caused by pyriproxyfen did not increase over time in unselected hybrid strains and fitness traits were similar in unselected and selected hybrid strains. Using a new method to examine the inheritance of resistance, based on data from fitness cost experiments, it was estimated that pyriproxyfen resistance is controlled by two loci in the QC02-R strain. The lack of fitness costs associated with pyriproxyfen resistance could promote the evolution of resistance in field populations with similar traits to QC02-R.

MicroRNAs: An exciting and open field calls for extensive study from initial and established investigators

The tobacco whitefly Bemisia tabaci (Gennadius) is an important crop pest which threatens agriculture worldwide. It has evolved resistance to various classes of insecticides. Cycloxaprid is a new-generation neonicotinoid which is a (nitromethylene) imidazole analogue of imidacloprid which is highly efficient for the control of various Hemipteran pests in China. Studies were conducted to determine the baseline susceptibility to cycloxaprid of 18 field samples of B. tabaci collected from 9 geographical locations in China and the possibility of cross-resistance between cycloxaprid and other important neonicotinoids of one laboratory-selected resistant strain. The 50% lethal concentrations (LC50) of cycloxaprid to these 18 samples ranged from 0.84 to 12.17 mg/L. Furthermore, compared with the susceptible laboratory strain, the imidacloprid-resistant strain exhibited a 27.9-fold resistance to imidacloprid and lower level of cross-resistance to acetamiprid (16.3-fold), thiacloprid (13.7-fold) and nitenpyram (16.6-fold), but no cross-resistance to cycloxaprid (1.9-fold). These results demonstrate that cycloxaprid could be one effective alternative insecticide for whitefly management that could reduce imidacloprid selection pressure.

Inheritance of Resistance to Pyriproxyfen in <I>Bemisia tabaci</I> (Hemiptera: Aleyrodidae) Males and Females (B Biotype)

We evaluated effects of the insect growth regulator pyriproxyfen on Bemisia tabaci (Gennadius) (B biotype) (Hemiptera: Aleyrodidae) males and females in laboratory bioassays. Insects were treated with pyriproxyfen as either eggs or nymphs. In all tests, the LC50 for a laboratory-selected resistant strain was at least 620 times greater than for an unselected susceptible strain. When insects were treated as eggs, survival did not differ between males and females of either strain. When insects were treated as nymphs, survival did not differ between susceptible males and susceptible females, but resistant males had higher mortality than resistant females. The dominance of resistance decreased as pyriproxyfen concentration increased. Resistance was partially or completely dominant at the lowest concentration tested and completely recessive at the highest concentration tested. Hybrid female progeny from reciprocal crosses between the susceptible and resistant strains responded alike in bioassays; thus, maternal effects were not evident. Rapid evolution of resistance to pyriproxyfen could occur if individuals in field populations had resistance with traits similar to those of the laboratory-selected strain examined here.

Inheritance of Resistance to Pyriproxyfen in Bemisia tabaci (Hemiptera: Aleyrodidae) Males and Females (B Biotype)

We evaluated effects of the insect growth regulator pyriproxyfen on Bemisia tabaci (Gennadius) (B biotype) (Hemiptera: Aleyrodidae) males and females in laboratory bioassays. Insects were treated with pyriproxyfen as either eggs or nymphs. In all tests, the LC50 for a laboratory-selected resistant strain was at least 620 times greater than for an unselected susceptible strain. When insects were treated as eggs, survival did not differ between males and females of either strain. When insects were treated as nymphs, survival did not differ between susceptible males and susceptible females, but resistant males had higher mortality than resistant females. The dominance of resistance decreased as pyriproxyfen concentration increased. Resistance was partially or completely dominant at the lowest concentration tested and completely recessive at the highest concentration tested. Hybrid female progeny from reciprocal crosses between the susceptible and resistant strains responded alike in bioassays; thus, maternal effects were not evident. Rapid evolution of resistance to pyriproxyfen could occur if individuals in field populations had resistance with traits similar to those of the laboratory-selected strain examined here.

Comparison of gyrA gene mutations between laboratory-selected ofloxacin-resistant Mycobacterium tuberculosis strains and clinical isolates

To understand the relationship between mutations in the quinolone resistance-determining region (QRDR) of the gyrA gene and drug resistance to ofloxacin, 85 laboratory-selected ofloxacin-resistant Mycobacterium tuberculosis mutant strains and 110 M. tuberculosis clinical isolates, screened by denaturing high-performance liquid chromatography to contain mutations, were analysed for their mutation patterns by sequencing as well as their ofloxacin minimal inhibitory concentrations (MICs). All mutations detected occurred at the codons Ala74, Ala90, Ser91 and Asp94 in all strains. One of the five different forms of missense mutation in Asp94 occurred in 60% of the laboratory-selected strains and 78% of the clinical isolates. However, 53 clinical isolates (48%) and only 2 laboratory-selected strains (2.4%) harboured double point mutations. The mutation Ala74Ser occurred only in the clinical isolates and only in combination with the Asp94Gly mutation. The ofloxacin MIC for the clinical isolates ranged from 0.5 μg/mL to 20 μg/mL, whilst the MICs for the laboratory-selected strains were ≥10μg/mL. The differences in gyrA gene mutation patterns and MICs between the laboratory-selected resistant strains and clinically isolated resistant strains identified here might help to understand the mechanisms involved in fluoroquinolone resistance.