Pyriproxyfen Resistance Research Articles

The inhibition of ecdysone signal pathway was the key of pyriproxyfen poisoning for silkworm, Bombyx mori

Pyriproxyfen is a juvenile hormone-like pesticide. Once intake occurs, it leads to a series of poisoning characters consequences in silkworm, Bombyx mori (ID: 7091, Lepidoptera), such as non- cocooning, non-pupation, production of low-active eggs, and extended stages. However, the poisoning mechanism is still unclear. Here, silkworms were fed mulberry leaves soaked with different pyriproxyfen concentrations, and the heads were dissected for transcriptome analysis, while the hemolymph was used for determinations of ecdysone and juvenile hormone titers. As a result, after conjoint analysis of 3 feeding groups and a control group, 555 differentially expressed genes (DEGs) were obtained, which were mainly involved in hormone metabolism, glycometabolism and protein metabolism. Meanwhile, 119 genes were significantly correlated with the pyriproxyfen concentrations, and they were mainly involved in drug metabolism and glycometabolism. The ecdysone titers in several feeding groups were significantly lower than those of the control group, while juvenile hormone was not detected in all groups, including the control and feeding groups. Correspondingly, due to activation of the juvenile hormone signaling pathway by pyriproxyfen, key genes in the ecdysone synthesis pathway were downregulated, and a large number of downstream genes were up- or downregulated. In addition, nearly all genes in the detoxification pathway were upregulated. These results suggested that, affected by the juvenile hormone signaling pathway, ecdysone titers decreased and further affected a series of downstream processes, and this was the key reason for pyriproxyfen poisoning in silkworm, B. mori, which could lay a foundation for the study of pyriproxyfen resistance in silkworm.

Laboratory induced selection of pyriproxyfen resistance in Oxycarenus hyalinipennis Costa (Hemiptera: Lygaeidae): Cross-resistance potential, realized heritability, and fitness costs determination using age-stage, two-sex life table

Pyriproxyfen is a biorational insecticide from IGR family, used worldwide against several economic pests. To evaluate the risk of pyriproxyfen resistance in dusky cotton bug, Oxycarenus hyalinipennis Costa (Hemiptera: Lygaeidae), a major concern for cotton producers, and to formulate strategies effective to tackle resistance, a field collected population was selected with pyriproxyfen under laboratory conditions using seed-dip method. A resistant strain designated as Pyr-SEL (G18) was developed after repeatedly selecting O. hyalinipennis with pyriproxyfen over eighteen generations. Thereafter, fitness costs, realized heritability (h2) and cross-resistance were investigated. As a result of selection, Pyr-SEL (G18) developed a very high level of resistance (resistance ratio = 464.23-fold) compared with the susceptible strain unselected over twenty generations Un-SEL (G20). The Pyr-SEL (G18) conferred strong cross-resistance to bifenthrin (146.59-fold), lambda-cyhalothrin (132.96-fold) and fenoxycarb (91.06-fold), whereas showed moderate cross-resistance to diafenthiuron (28.86-fold) and fipronil (22.73-fold). The h2 estimate was 0.16 in Pyr-SEL (G18). The developmental duration of O. hyalinipennis pre-adult prolonged, but traits of λ, r and R0 reduced in Pyr-SEL (G18) compared with the Un-SEL (G20). Also, the population projection obtained lower population size for Pyr-SEL (G18) than Un-SEL (G20). Fitness studies revealed that high resistance development to pyriproxyfen lowered the relative fitness of Pyr-SEL (G18) (Rf = 0.38) compared with the Un-SEL (G20). These findings may be practically valuable in tackling O. hyalinipennis resistance for better pest management.

Insecticide resistance status of Bemisia tabaci MEAM1 (Hemiptera: Aleyrodidae) in Australian cotton production valleys

AbstractThe whitefly Bemisia tabaci Middle East‐Asia Minor 1 (MEAM1), previously known as B biotype, is a major agricultural pest with a reputation for developing resistance to insecticides. DNA‐based identification revealed that B. tabaci MEAM1 is the dominant species within Australian cotton cropping. Discriminating doses of 10 mg/L for pyriproxyfen and 300 mg/L for both diafenthiuron and bifenthrin were determined from bioassay data collected in 2010–2015. Resistance to these insecticides was tested in whitefly populations collected in 2017 from cotton crops in Queensland and New South Wales. This study reports on the presence of pyriproxyfen resistance in B. tabaci MEAM1 at seven locations in Queensland and New South Wales. One population from Goondiwindi was resistant to bifenthrin, whereas no populations tested had resistance to diafenthiuron. To determine the presence of resistance alleles to organophosphates and pyrethroids, reference populations of susceptible and resistant whiteflies (pyriproxyfen, bifenthrin and neonicotinoid) were tested. The resistance mutation L925I was found in a pyrethroid‐resistant population and in some individuals from the pyriproxyfen‐resistant population. The mutation F331W that confers organophosphate and carbamate resistance was found in all individuals tested including the susceptible reference population, indicative of a widespread distribution in Australian B. tabaci MEAM1. The cotton industry has revised the insecticide resistance management strategy for B. tabaci, to restrict the usage of pyriproxyfen to a single application within a 30‐day window. Our results argue against the use of organophosphate and carbamate insecticides in Australian cotton, because B. tabaci MEAM1 populations show significant resistance levels. Furthermore, both pesticide groups are highly disruptive to a diverse range of natural enemy populations, and as such, widespread use likely contributes to pest outbreaks.

Mechanism, stability and fitness cost of resistance to pyriproxyfen in the house fly, Musca domestica L. (Diptera: Muscidae)

Pyriproxyfen, a bio-rational insecticide, used worldwide for the management of many insect pests including the house fly, Musca domestica. To devise a retrospective resistance management strategy, biological parameters of pyriproxyfen resistant (Pyri-SEL), unselected (UNSEL), Cross1 and Cross2M. domestica strains were studied in the laboratory. Additionally, the stability and mechanism of resistance was also investigated. After 30 generations of pyriproxyfen selection, a field-collected strain developed 206-fold resistance compared with susceptible strain. Synergists such as piperonyl butoxide and S,S,S-tributylphosphorotrithioate did not alter the LC50 values, suggesting another cause of target site resistance to pyriproxyfen in the Pyri-SEL strain. The resistance to all tested insecticides was unstable in Pyri-SEL strain. The relative fitness of 0.51 with lower fecundity, hatchability, lower number of next generation larvae, reduced mean population growth rate and net reproductive rate were observed in the Pyri-SEL strain compared with the UNSEL strain. The cost of fitness associated with pyriproxyfen resistance was evident in Pyri-SEL strain. The present study provides useful information for making pro-active resistance management strategies to delay resistance development.

Inheritance mode, cross-resistance and realized heritability of pyriproxyfen resistance in a field strain of Musca domestica L. (Diptera: Muscidae)

Pyriproxyfen is a growth regulator used for the control of different insect pests, including Musca domestica. To assess the risk of resistance and to develop a strategy for resistance management, a field strain of M. domestica was exposed to pyriproxyfen in the laboratory for 30 generations. The inheritance mode, realized heritability of pyriproxyfen resistance and cross-resistance to other insecticides were assessed. Prior to the selection process, the field strain exhibited a resistance ratio (RR) of 25.7, 7.31, 7.67, and 27-fold for pyriproxyfen, methoxyfenozide, cyromazine and lufenuron, respectively, when compared to the pyriproxyfen susceptible strain (Pyri-Sus). After continuous selection with pyriproxyfen, the pyriproxyfen-resistant strain (Pyri-Res) became 206-fold more resistant than the Pyri-Sus strain. The overlapping confidence limits of LC50 values of F1 (Pyri-Res ♂×Pyri-Sus ♀) and F1† (Pyri-Res ♀×Pyri-Sus ♂) suggested an autosomal and completely dominant mode of resistance to pyriproxyfen. Monogenic test of inheritance showed that resistance to pyriproxyfen was governed by multiple genes. The Pyri-Res strain showed very low cross resistance to methoxyfenozide, cyromazine, and lufenuron. The estimated realized heritability was 0.02, 0.05, 0.03 and 0.04 for pyriproxyfen, methoxyfenozide, cyromazine, and lufenuron, respectively. It was concluded that pyriproxyfen resistance in M. domestica was autosomally inherited, completely dominant and polygenic. These results would be helpful for the design of an improved control strategy against M. domestica.

Over-Expression of a Cytochrome P450 Is Associated with Resistance to Pyriproxyfen in the Greenhouse Whitefly Trialeurodes vaporariorum

BackgroundThe juvenile hormone mimic, pyriproxyfen is a suppressor of insect embryogenesis and development, and is effective at controlling pests such as the greenhouse whitefly Trialeurodes vaporariorum (Westwood) which are resistant to other chemical classes of insecticides. Although there are reports of insects evolving resistance to pyriproxyfen, the underlying resistance mechanism(s) are poorly understood.ResultsBioassays against eggs of a German (TV8) population of T. vaporariorum revealed a moderate level (21-fold) of resistance to pyriproxyfen. This is the first time that pyriproxyfen resistance has been confirmed in this species. Sequential selection of TV8 rapidly generated a strain (TV8pyrsel) displaying a much higher resistance ratio (>4000-fold). The enzyme inhibitor piperonyl butoxide (PBO) suppressed this increased resistance, indicating that it was primarily mediated via metabolic detoxification. Microarray analysis identified a number of significantly over-expressed genes in TV8pyrsel as candidates for a role in resistance including cytochrome-P450 dependent monooxygenases (P450s). Quantitative PCR highlighted a single P450 gene (CYP4G61) that was highly over-expressed (81.7-fold) in TV8pyrsel.ConclusionOver-expression of a single cytochrome P450 gene (CYP4G61) has emerged as a strong candidate for causing the enhanced resistance phenotype. Further work is needed to confirm the role of the encoded P450 enzyme CYP4G61 in detoxifying pyriproxyfen.

Analyzing haplodiploid inheritance of insecticide resistance in whitefly biotypes

We developed new methods for analyzing inheritance of insecticide resistance in haplodiploid arthropods and applied them to elucidate resistance of the whitefly Bemisia tabaci (Gennadius) to an insect growth regulator, pyriproxyfen. Two invasive biotypes of this devastating crop pest, the B biotype in Arizona and the Q biotype in Israel, have evolved resistance to pyriproxyfen. Here, we incorporated data from laboratory bioassays and crossing procedures exploiting haplodiploidy into statistical and analytical models to estimate the number of loci affecting pyriproxyfen resistance in strains of both biotypes. In tests with models of one to ten loci, the best fit between expected and observed mortality occurred with a two-locus model for the B biotype strain (QC-02) and for one- and two-locus models for the Q biotype strain (Pyri-R). The estimated minimum number of loci affecting resistance was 1.6 for the B biotype strain and 1.0 for the Q biotype strain. The methods used here can be applied to insecticide resistance and other traits in haplodiploid arthropods.

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.

Effects of Pyriproxyfen on California Red Scale (Hemiptera: Diaspididae) Development and Reproduction

Two life stages of a laboratory colony of California red scale, Aonidiella aurantii (Maskell) (Hemiptera: Diaspididae), were exposed to 10 concentrations of pyriproxyfen to evaluate the effect of this insect growth regulator (IGR) on scale development and fecundity. First instars exposed to pyriproxyfen responded with mortality during the first and second molts. Second instars were more tolerant of pyriproxyfen than were first instars, indicating that growers should treat California red scale in the field before the first molt to achieve maximum efficacy. Male scales responded with an LC50 12-fold lower and an LC90 47-fold lower than was observed for female scales. Female scales that survived first instar exposure to pyriproxyfen experienced reduced fecundity with increasing pyriproxyfen concentration. Reduced fecundity was due to reduced survival of the females rather than sterility. Results from this study provide baseline California red scale susceptibility data for pyriproxyfen, and they suggest that 1 ppm pyriproxyfen can be used as a concentration that would discriminate between susceptible and resistant populations of scales. Field monitoring for incipient pyriproxyfen resistance in California red scale should be implemented in the San Joaquin Valley of California, and if resistance is detected, integrated resistance management strategies should be used to slow the progression of resistance.

Effects of Pyriproxyfen on California Red Scale (Hemiptera: Diaspididae) Development and Reproduction

Two life stages of a laboratory colony of California red scale, Aonidiella aurantii (Maskell) (Hemiptera: Diaspididae), were exposed to 10 concentrations of pyriproxyfen to evaluate the effect of this insect growth regulator (IGR) on scale development and fecundity. First instars exposed to pyriproxyfen responded with mortality during the first and second molts. Second instars were more tolerant of pyriproxyfen than were first instars, indicating that growers should treat California red scale in the field before the first molt to achieve maximum efficacy. Male scales responded with an LC50 12-fold lower and an LC90 47-fold lower than was observed for female scales. Female scales that survived first instar exposure to pyriproxyfen experienced reduced fecundity with increasing pyriproxyfen concentration. Reduced fecundity was due to reduced survival of the females rather than sterility. Results from this study provide baseline California red scale susceptibility data for pyriproxyfen, and they suggest that 1 ppm pyriproxyfen can be used as a concentration that would discriminate between susceptible and resistant populations of scales. Field monitoring for incipient pyriproxyfen resistance in California red scale should be implemented in the San Joaquin Valley of California, and if resistance is detected, integrated resistance management strategies should be used to slow the progression of resistance.

Short communication. Mechanisms in competition, under laboratory conditions, between Spanish biotypes B and Q of Bemisia tabaci (Gennadius)

Laboratory studies were carried out on aspects affecting competition between biotypes B and Q of whitefly [Bemisia tabaci (Gennadius)]: mating behaviour, egg viability and pyriproxyfen resistance. Pairs of biotype B females and males spent more time as courting couples than pairs with Q males, regardless of female biotype. Mixed pairs of biotype Q females and biotype B males spent more time as courting couples than biotype Q pairs. Egg viability from the different crosses and from single females was similar. Pyriproxyfen application to a mixture of whiteflies with a biotype ratio B/Q of one gave a reduction in the ratio because of higher resistance of the Q biotype to the insecticide. The data agrees with the observed displacement of biotype Q by biotype B whiteflies under laboratory conditions, and vice versa in the field.

Biotypes B and Q of Bemisia tabaci and their relevance to neonicotinoid and pyriproxyfen resistance

Resistance monitoring for Bemisia tabaci field populations to the juvenile hormone mimic, pyriproxyfen, was conducted from 1996 to 2003 in commercial cotton fields in two areas of Israel: the Ayalon Valley (central Israel) and the Carmel Coast (northwestern Israel). Although the use of pyriproxyfen ceased in these areas in 1996-1997 (because of the resistance), resistance levels to pyriproxyfen declined to some extent in the fields but remained quite stable, and the susceptibility has not been totally restored. Two strains of B. tabaci collected from the Ayalon Valley in the late 1999 and 2002 cotton seasons (AV99L, AV02L) were assayed for their susceptibility to pyriproxyfen at F1, and subsequently a line of each strain was kept under controlled conditions without exposure to insecticides. After maintenance of more than 20 generations under laboratory conditions, the resistance to pyriproxyfen in the untreated strains substantially declined. This decline was concurrent with a replacement of Q biotype by B-type under non-insecticidal regimes; apparently B biotype was more competitive than the pyriproxyfen-resistant Q-type. Selection under controlled conditions with neonicotinoids on these B. tabaci strains resulted in continued pyriproxyfen resistance, predominantly of Q biotype. Based on our data, applications of either pyriproxyfen or neonicotinoids may select for biotype Q, which would survive to a greater degree where these insecticides are applied.

Dynamics of insecticide resistance in Bemisia tabaci: a case study with the insect growth regulator pyriproxyfen.

The dynamics of pyriproxyfen resistance in Bemisia tabaci (Gennadius) have been studied intensively in cotton fields and greenhouses in Israel. High resistance to pyriproxyfen evolved one year after its introduction for use on flowers in greenhouses, after three successive applications. After ten years of pyriproxyfen use in cotton fields, with only one application per season, a high level of resistance was observed, but its rate of development differed among localities. It is hypothesized that these differences reflect temporal and spatial variations in the availability of alternative host plants for B tabaci, and that pyriproxyfen resistance evolved most readily in geographically isolated areas. It is also likely that the occurrence and development of resistance is partially biotype-related. Pyriproxyfen use has ceased in high-resistance localities, but in areas with low frequencies of resistance, cotton growers have continued to achieve acceptable control of B tabaci with pyriproxyfen. Due to the absence of applications of pyriproxyfen in some cotton fields, resistance levels tended to decline between 1998 and 2001. Laboratory experiments support the hypothesis that this decline reflects, in part, fitness costs associated with pyriproxyfen resistance.

Resistance ofBemisia tabaci (Homoptera: Aleyrodidae) to insecticides in southern Spain with special reference to neonicotinoids

The tobacco whitefly, Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) which occurs in various parts of the world, has developed a high degree of resistance against several chemical classes of insecticide, including organophosphates, carbamates, pyrethroids, insect growth regulators and chlorinated hydrocarbons. The present studies were done in order to monitor the susceptibility of whitefly populations in southern Spain to insecticides commonly used there. Systemic bioassays using Spanish field populations of B tabaci collected in 1994, 1996 and 1998 indicated an increase, albeit a slow one, in resistance to imidacloprid over this period. Comparative studies of other neonicotinoids using the same bioassay revealed a high degree of cross-resistance to acetamiprid and thiamethoxam. Leaf-dip bioassays with adult females from these populations revealed a high level of resistance to cyfluthrin, endosulfan, monocrotophos, methamidophos, and pymetrozine, each at 200 mg litre−1. Buprofezin and pyriproxyfen were tested against second-instar nymphs and eggs, respectively. Buprofezin also showed a lower efficacy against ESP-98, a strain of B tabaci received from Almeria in 1998, but pyriproxyfen resistance was not obvious when tested against eggs of strain ESP-98. Field trials in 1998 revealed good efficacy of imidacloprid in one farm in the Almeria region and two greenhouses in Murcia and Sevilla, but a loss of activity by imidacloprid in another farm in the Almeria region. Cross-resistance between imidacloprid and thiamethoxam was also confirmed under field conditions. © 2000 Society of Chemical Industry

The response of pyriproxyfen-resistant and susceptibleBemisia tabaciGenn (Homoptera: Aleyrodidae) to pyriproxyfen and fenoxycarb alone and in combination with piperonyl butoxide

Pyriproxyfen was effective against susceptible Bemisia tabaci eggs at a LC 50 of 0.003 mg litre -1 and against nymphs at 0.02 mg litre -1 . In comparison, eggs of a laboratory selected, pyriproxyfen-resistant B tabaci strain, originating in an Israeli greenhouse, exhibited 6500-fold resistance and nymphs exhibited 1100-fold resistance. Eggs and nymphs of a strain from an Israeli sunflower field exhibited 450 and 210-fold resistance in comparison to the susceptible standard. Fenoxycarb was generally less effective than pyriproxyfen against B tabaci eggs and nymphs but was unaffected by pyriproxyfen resistance. Piperonyl butoxide (PB) was antagonistic to pyriproxyfen, and this increased with increasing pyriproxyfen resistance. PB had no effect on the toxicity of fenoxycarb. Collectively, these data imply that the modes of action of pyriproxyfen and fenoxycarb are distinct, despite the structural similarities of these molecules. Possible reasons for the antagonism of PB against pyriproxyfen are discussed.

In vitro metabolism of pyriproxyfen by microsomes from susceptible and resistant housefly larvae.

Levels of cytochrome P450 and b5 were investigated in microsomal enzymes of houseflies from the gut and fat body of the third instar larvae of a pyriproxyfen-resistant strain (YPPF) and two pyriproxyfen-susceptible strains (YS and SRS). In comparison to the YS and SRS strains, YPPF microsomes had higher levels of total cytochrome P450s in both the gut and fat body. Furthermore, microsomes from the gut and fat body of YPPF larvae were found to have a much greater ability to hydroxylate aniline than YS larvae. In vitro metabolism studies of pyriproxyfen indicated that the metabolic rates were much higher in both the gut and fat body of YPPF larvae than of YS and SRS larvae. The major metabolites of pyriproxyfen in houseflies were identified to be 4'-OH-pyriproxyfen and 5"-OH-pyriproxyfen. Cytochrome P450 inhibitors, piperonyl butoxide (PB) and 2-propynyl 2,3,6-trichlorophenyl ether (PTPE), decreased the metabolic rates significantly in all three strains. This study confirmed that microsomal cytochrome P450 monooxygenases play an important role in the pyriproxyfen resistance of the housefly. Furthermore, it suggests that the fat body must be as important as the gut for the metabolism of pyriproxyfen in resistant housefly larvae.