Pharmacokinetic mechanisms associated with synergism by DEF of cypermethrin toxicity in larval and adult Helicoverpa zea, Spodoptera frugiperda, and Agrotis ipsilon (Lepidoptera: Noctuidae)

Penetration, metabolism, and excretion of radiocarbon were observed after topical treatment of Helicoverpa zea (Boddie), Spodoptera frugiperda (J. E. Smith), and Agrotis ipsilon (Hufnagle) larvae and adults with cypermethrin-14C. These pharmacokinetic events usually were higher with trans-cypermethrin-14C than with cis-cypermethrin-14C. They also were generally higher with H. zea and S. frugiperda than with A. ipsilon, and they were higher in larvae than in adults. No marked sex differences in the degradation of trans-cypermethrin were apparent. Pretreatment of H. zea, S. frugiperda, and A. ipsilon larvae and adults with S,S,S-tri-n-butyl phosphorotrithioate (DEF) 30 min before application of cypermethrin resulted in a perturbation of trans-cypermethrin pharmacokinetics manifested primarily by a lower rate of pyrethroid metabolism as compared with that in the absence of DEF. Appreciably higher internal levels of the toxic parent pyrethroid were often observed in the presence of DEF than in the absence of DEF in most cases. Suppression of cypermethrin penetration and elimination also was usually detected. Inhibition by DEF of the enzymatic degradation of cypermethrin may account for the synergy observed between these two compounds.     

DEF sensitive esterases in homogenates of larval and adult Helicoverpa zea, Spodoptera frugiperda, and Agrotis ipsilon (Lepidoptera: Noctuidae)

Homogenates of Helicoverpa zea (Boddie), Agrotis ipsilon (Hufnagle), and Spodoptera frugiperda (J. E. Smith) third instars and adults contained S,S,S-tri-n-butyl phosphorotrithioate (DEF)-sensitive enzymes that hydrolyzed trans-cypermethrin and two known esterase substrates, alpha-naphthyl acetate and beta-naphthyl acetate. Except for H. zea with alpha-naphthyl acetate, larval preparations were more active than those of adults, and no marked sex differences were apparent. The hydrolysis of trans-cypermethrin in noctuid preparations were inhibited by DEF, with pI50 values ranging from 4.5 to 6.7. DEF was a potent inhibitor of the degradation of general carboxylesterase substrates alpha-naphthyl acetate and beta-naphthyl acetate in some cases. Electrophoretic studies confirmed the presence in noctuid gut homogenates of one or more DEF-sensitive esterases that hydrolyzed alpha-naphthyl acetate and beta-naphthyl acetate and that were completely inhibited by dichtorvos.     

Insecticide resistance and cross-resistance in the house fly (Diptera: Muscidae)

A house fly strain, ALHF, was collected from a poultry farm in Alabama after a control failure with permethrin, and further selected in the laboratory with permethrin for five generations. The level of resistance to permethrin in ALHF was increased rapidly from an initial 260-fold to 1,800-fold after selection. Incomplete suppression of permethrin resistance by piperonyl butoxide (PBO) and S,S,S,-tributylphosphorotrithioate (DEF) reveals that P450 monooxygenase- and hydrolase-mediated detoxication, and one or more additional mechanisms are involved in resistance to permethrin. The ALHF strain showed a great ability to develop resistance or cross-resistance to different insecticides within and outside the pyrethroid group including some relatively new insecticides. Resistance to beta-cypermethrin, cypermethrin, deltamethrin, and propoxur (2,400-4,200-, 10,000-, and > 290-fold, respectively, compared with a susceptible strain, aabys) in ALHF house flies was partially or mostly suppressed by PBO and DEF, indicating that P450 monooxygenases and hydrolases are involved in resistance to these insecticides. Partial reduction in resistance with PBO and DEF implies that multiresistance mechanisms are responsible for resistance. Fifteen- and more than fourfold resistance and cross-resistance to chlorpyrifos and imidacloprid, respectively, were not effected by PBO or DEF, indicating that P450 monooxygenases and hydrolases are not involved in resistance to these two insecticides. Forty-nine-fold cross-resistance to fipronil was mostly suppressed by PBO and DEF, revealing that monooxygenases are a major mechanism of cross-resistance to fipronil. Multiresistance mechanisms in the ALHF house fly strain, however, do not confer cross-resistance to spinosad, a novel insecticide derived from the bacterium Saccharopolyspora spinosa. Thus, we propose that spinosad be used as a potential insecticide against house fly pests, especially resistant flies.     

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.     

Relative resistance of transgenic tomato tissues expressing high levels of tobacco anionic peroxidase to different insect species.

Different parts of genetically transformed tomato (Lycopersicon esculentum L.) plants that express the tobacco anionic peroxidase were compared for insect resistance with corresponding wild type plants. Leaf feeding by first instar Helicoverpa zea and Manduca sexta was often significantly reduced on intact transgenic plants and/or leaf disks compared to wild type plants, but the effect could depend on leaf age. Leaves of transgenic plants were generally as susceptible to feeding damage by third instar Helicoverpa zea (Boddie) and Manduca sexta (L.) as wild type plants. Green fruit was equally susceptible to third instar larvae of H. zea in both type plants, but fruit of transgenic plants were more resistant to first instar larvae as indicated by significantly greater mortality. Basal stem sections were more resistant to neonate larvae of H. zea and adults of Carpophilus lugubris Murray compared to wild type plants as indicated by significantly greater mortality and/or reduced feeding damage. Thus, tobacco anionic peroxidase activity can increase plant resistance to insects in tomato, a plant species closely related to the original source plant species, when expressed at sufficiently high levels. However, the degree of resistance is dependent on the size of insect and plant tissue involved.     

Acquisition, persistence, and species susceptibility of the Hz-2V virus

Hz-2V, formerly called gonad-specific virus, is known to infect the reproductive organs of both males and females of the corn earworm Helicoverpa zea, rendering them agonadal or sterile. The primary mode of transmission is through mating by asymptomatic carrier moths. In this report we show that Hz-2V can be acquired by first instar larvae, through feeding on virus laced diet, although the incidence of agonadal condition was significantly lower. In a laboratory study, the virus appeared to persist for no more than three generations, with the incidence of agonadal progeny decreasing with each generation. Although, Hz-2V has been reported only from H. zea, in our tests when nine species of insects were artificially infected, four of the Noctuid species showed some signs of agonadal condition. Out of the remaining five species, the diamondback moth Plutella xylostella and the German cockroach Blatella germanica, showed no evidence of the virus in progeny of adults that were injected with Hz-2V, even after using the very sensitive PCR based assay.     

Identification of six chymotrypsin cDNAs from larval midguts of Helicoverpa zea and Agrotis ipsilon feeding on the soybean (Kunitz) trypsin inhibitor

Lepidopteran insects like Helicoverpa zea and Agrotis ipsilon produce STI-insensitive trypsins in the midgut following ingestion of dietary plant proteinase inhibitors like STI [Broadway, R. M., J. Insect Physiol. 43(9) (1997) 855-874]. In this paper, the effects of dietary STI on a related family of midgut serine proteinases, the chymotrypsins, were investigated. STI-insensitive midgut chymotrypsins were detected in larvae of H. zea and A. ipsilon feeding on diets containing 1% STI while STI-sensitive chymotrypsins were present in larvae feeding on diets containing 0% STI. These chymotrypsins were unaffected by TPCK, a diagnostic inhibitor of mammalian chymotrypsins but were fully inhibited by chymostatin. Four midgut cDNA libraries were constructed from larvae of each species fed either 0% STI or 1% STI diets. Six full-length cDNAs(1) encoding diverse preprochymotrypsins were isolated (three from H. zea and three from A. ipsilon) with certain sequence motifs that set them apart from their mammalian counterparts. Northern blots showed that some chymotrypsin mRNA were detected at higher levels while others were down-regulated when comparing insects reared on 0% STI and 1% STI diets. Southern hybridizations suggested that (like mammals) both species contained several chymotrypsin genes. A full-length chymotrypsin gene(1) from H. zea was sequenced for the first time and the presence of four introns was deduced. A first time comparison of 5' upstream regions(1) from three chymotrypsin genes and two trypsin genes of A. ipsilon indicated the presence of putative TATA boxes and regulatory elements. However a lack of consensus motifs in these upstream regions suggested the likelihood of multiple trans factors for regulation of genes encoding digestive proteinases and a complex response mechanism linked to ingestion of proteinase inhibitors.     

Some aspects of the host:parasite relationships between adult blackflies (Diptera; Simuliidae) and larvae of the water-mite Sperchon setiger(Acari; Hydrachnellae) in a small chalk stream in southern England

SUMMARY. In Bere Stream three species of adult simuliids, S . ornatum, S. equinum and S. angustipes were parasitized by Sperchon setiger larvae. Trapped adult S. ornatum were more often parasitized than corresponding adults of S. equinum. The lower 95% confidence limits were used as an indication of the relative importance of the statistically significant differences and were high (10.4–33.6%). Laboratory experiments on the occupation of host pupae by pre-parasitic Sperchon setiger larvae also indicated that pupae of S. ornatum were more often occupied than S. equinum. This was confirmed by tests on the difference in percentage occupation of more than 4000 held-collected host pupae. In tests that indicated a significant difference in percentage parasitization between the sexes of the three host species, females were more often parasitized. However, the lower 95% confidence limits for the increase in percentage parasitization of females compared to males were low (0.9–10.2%). The majority of adult simuliids carried only one parasite. The mean number of larvae per trapped parasitized host was significantly greater for S. ornatum (2.69) than for S. equinum (1.77). There was no significant difference between the mean number of parasites carried by the sexes of S. ornatum for each year from 1975 through 1978 but females were significantly more heavily parasitized than males during 1979. The ventral neck region of all three host species is the main attachment site for Sperchon setiger larvae.
The morphology of the pupae of S. ornatum and S. equinum is related to differences in levels of percentage parasitization between these two species. It is suggested that pupal morphology may be the most important factor in the selection of host simuliids by Sperchon setiger larvae.     

Insecticide susceptibility in Coptotermes formosanus and Reticulitermes virginicus (Isoptera: Rhinotermitidae).

Lethal time to mortality responses were established for eight insecticides against workers and soldiers of the Formosan subterranean termite, Coptotermes formosanus Shiraki, and workers of Reticulitermes virginicus (Banks). There were significant differences in the tolerance ratios between workers of C. formosanus colonies to all toxicants tested except fipronil. One colony was 16 times more tolerant than another to deltamethin. C. formosanus soldiers had significant differences in tolerance ratios among colonies exposed to all toxicants except chlorpyrifos. Methoxychlor, permethrin, deltamethrin, and fipronil did not kill soldiers from two, one, one, and three colonies, respectively, within 8 h. Seventy-five percent of R. virginicus colonies were significantly less susceptible than the most susceptible colony to chlordane, methoxychlor, chlorpyrifos, cypermethrin, and fipronil, with 50% of the colonies less susceptible to permethrin and bendiocarb. In 50% of C. formosanus colonies the worker lethal time curves displayed substantial flattening in response to permethrin, and deltamethrin. Lethal time curses for C. formosanus soldiers exposed to chlordane, chlorpyrifos, permethrin, cypermethrin, deltamethrin, and bendiocarb showed substantial flattening. R. virginicus workers demonstrated substantial curve flattening when exposed to chlordane, methoxychlor, chlorpyrifos, deltamethrin, and fipronil. These findings indicate substantial intercolony and intra-colony differences in susceptibility to insecticides.     

拟除虫菊酯的杀虫活性和温度的关系

除了戊酸醚酯对苜蓿蚜呈弱的正温度系数外,杀灭菊酯、二氯苯醚菊酯、氯氰菊酯、溴氰菊酯、氟氰菊酯及百树菊酯,都呈负温度系数。杀灭菊酯、戊酸醚酯、二氯苯醚菊酯、氯氰菊酯、溴氰菊酯和氟氰菊酯,对梨网蝽全部呈正温度系数。氯氰菊酯对粘虫呈负温度系数,杀灭菊酯、戊酸醚酯、二氯苯醚菊酯和溴氰菊酯都呈正温度系数。杀灭菊酯和溴氰菊酯对粘虫卵也呈正温度系数。杀灭菊酯对小菜蛾的杀虫活性,受温度的影响不明显,而戊酸醚酯则呈负温度系数。杀灭菊酯和戊酸醚酯对蚊幼呈正温度系数,而二氯苯醚菊酯则呈负温度系数。     

Bioactivities of methyl allyl disulfide and diallyl trisulfide from essential oil of garlic to two species of stored-product pests, Sitophilus zeamais (Coleoptera: Curculionidae) and Tribolium castaneum (Coleoptera: Tenebrionidae)

Two of the major constituents of the essential oil of garlic, Allium sativum L., methyl allyl disulfide and diallyl trisulfide, were tested against Sitophilus zeamais Motschulsky and Tribolium castaneum (Herbst) for contact toxicity, fumigant toxicity, and antifeedant activity. The contact and fumigant toxicities of diallyl trisulfide were greater than that of methyl allyl disulfide to the adults of these two species of insects. These two compounds were also more toxic to T. castaneum adults than to S. zeamais adults. Older T. castaneum larvae were more susceptible to the contact toxicity of the two compound, whereas younger larvae were more susceptible to the fumigant toxicity of these compounds. Both compounds reduced egg hatching of T. castaneum and subsequent emergence of progeny. Diallyl trisulfide totally suppressed egg hatching at 0.32 mg/cm2, and larval and adult emergence at 0.08 mg/cm2. Methyl allyl disulfide significantly decreased the growth rate, food consumption, and food utilization of adults of both insect species, with feeding deterrence indices of 44% at 6.08 mg/g food for S. zeamais and 1.52 mg/g food for T. castaneum. However, it did not affect any nutritional indices of T. castaneum larvae. Diallyl trisulfide significantly reduced all of the nutritional indices in all of the insects tested. Feeding deterrence indices of 27 and 51% were obtained in S. zeamais adults and T. castaneum larvae, respectively, at the concentration of 2.98 mg/g food, whereas feeding deterrence of 85% was achieved in T. castaneum adults at a much lower concentration of 0.75 mg/g food. Hence, diallyl trisulfide is a more potent contact toxicant, fumigant and feeding deterrent than methyl allyl disulfide.     

Development of permethrin resistance in Culex quinquefasciatus Say in Kuala Lumpur,Malaysia

The resistance status towards permethrin among the laboratory strain, the permethrin-selected strain and four field strains of Culex quinquefasciatus collected in Kuala Lumpur, Malaysia was determined using three standard laboratory methods: WHO larval bioassay, WHO adult bioassay and biochemical microplate assay. Cx. quinquefasciatus permethrin-selected strain larvae were the least susceptible to permethrin with a resistance ratio of 47.28-folds, whereas all field strain larvae of the same species were tolerant to permethrin with resistance ratios of more than 3-folds. In contrast, in adult stage, the permethrin exposed permethrin-selected strain (resistance ratio = 1.27) was found to be more susceptible to permethrin than all permethrin-exposed field strains (resistance ratios = 2.23–2.48). Complete mortalities for all strains of Cx. quinquefasciatus adults proved the effectiveness of the synergist; piperonyl butoxide (PBO). For the biochemical microplate assay, the reduction of the mean optical density of elevated oxidase activity of three field strains upon exposure to PBO confirmed the association between oxidase activity and permethrin tolerance. On the other hand, irregular patterns of the mean optical density of elevated oxidase activity in the laboratory strain, permethrin-selected strain and Jalan Fletcher strain illustrated the gene variation within these mosquito colonies as well as the involvement of other enzyme activities in the permethrin resistance occurred.     

Phenology and field biology of black cutworm (Lepidoptera: Noctuidae) in Ontario no-till corn

Black cutworm, Agrotis ipsilon (Hufnagel), is an occasional corn, Zea mays L., pest that is attracted to no-till fields. Understanding the phenology of black cutworm in Ontario no-till corn, particularly the time of arrival of adults in relation to the onset of crop damage and the stages of larvae that coincide with vulnerable corn seedling leaf stages, is important for their effective control. Pheromone and blacklight trap captures of moths first occurred in early April, whereas significant influxes did not occur until mid- to late April. Males and females were often captured simultaneously, in contrast to findings in Iowa and Illinois where males were captured in pheromone traps on average 3 wk ahead of females or males in blacklight traps. This may be a reflection of a more mature source population for the influxes into Ontario because first captures also were later than in the United States. Females arrived mated and corn seedling cutting occurred within 137 degree-days (DD) (base 10.4 degrees C) of first capture in Ontario corn. Cutworms were present in cornfields before planting, and the mean age of larvae increased along with corn leaf stage, suggesting that no new recruitment took place after planting. The apparent synchrony between corn and cutworm phenology in the northern areas of corn production seems more related to the availability and quality of food for young larvae relative to the development of the crop then the time of arrival of moths.     

The joint toxicity of type I, II, and nonester pyrethroid insecticides

Evidence suggests that there are separate binding domains for type I and II pyrethroid insecticides on the voltage gated sodium channel of the nerve cell axon, but there are no studies that have examined the mixture toxicity of nonester pyrethroids and type I and II pyrethroids. Therefore, we examined the effect of nonester pyrethroid (etofenprox), type I (permethrin), and type II (cypermethrin) pyrethroid insecticides alone and in all combinations to Drosophila melanogaster Meigen. The combination of permethrin + etofenprox and permethrin + cypermethrin demonstrated antagonistic toxicity, while the combination of cypermethrin + etofenprox demonstrated synergistic toxicity. The mixture ofpermethrin + cypermethrin + etofenprox demonstrated additive toxicity. The toxicity of permethrin + cypermethrin was significantly lower than the toxicity of cypermethrin alone, but the combination was not significantly different from permethrin alone. The toxicity of permethrin + cypermethrin + etofenprox was significantly greater than the toxicity of both permethrin and etofenprox alone, but it was significantly lower than cypermethrin alone. The mixture of permethrin and etofenprox was significantly less toxic than permethrin. The explanation for the decreased toxicity observed is most likely because of the competitive binding at the voltage-gated sodium channel, which is supported by physiological and biochemical studies of pyrethroids. Our results demonstrate that the assumption that the mixture toxicity of pyrethroids would be additive is not adequate for modeling the mixture toxicity of pyrethroids to insects.     

Phenotypic variation in adult behavioral response and offspring fitness in Plutella xylostella (Lepidoptera: Plutellidae) in response to permethrin

Phenotypic variation in behavioral response and physiological tolerance to permethrin was measured and compared between two populations of diamondback moth, Plutella xylostella (L.). A field population from Celeryville, OH, and a laboratory population originally collected from Wooster, OH, were compared. In laboratory choice bioassays, females from both populations were less likely to oviposit on cabbage leaf disks and seedlings treated with permethrin. The oviposition-deterrent effect was positively related to permethrin concentration. The laboratory population was significantly more behaviorally responsive to the insecticide and showed a significantly greater avoidance than the field population of the highest concentration of permethrin tested (1.50 g [AI] /liter). The physiological response of each population was measured by feeding bioassays, and the laboratory population was physiologically more susceptible to the permethrin. Larvae from the laboratory population that were fed permethrin-treated leaves had significantly lower growth rate, higher mortality, and lower adult fecundity compared with larvae from the field population. These data suggest that female moths that are more behaviorally responsive to permethrin produce offspring that tend to be more susceptible to the same insecticide, at least for the populations tested. The relationship between adult behavioral response and larval physiological tolerance to permethrin is discussed.     

Laboratory evaluation of certain natural compounds against the melon ladybird beetle,Epilachna chrysomelina F. attacking cucurbit plants

Neem Azal-T/S, Neudosan and Spruzit flüssig at different concentrations were evaluated against the second and fourth instar larvae of E. chrysomelina under laboratory conditions. In case of acute effects, the highest mortality was recorded when larvae were offered squash leaves treated with the highest concentration (2%) of Neem Azal-T/S, and the lowest mortality was, however recorded after treating them with the lowest concentration (0.25%). On the other hand, the second instar larvae were more susceptible to any compound at any concentration than the fourth instar one. One per cent Neudosan formulation showed the highest toxicity to both the 2nd and 4th instar larvae. As the concentration of this compound decreased the mortality percentage of treated larvae decreased to reach the minimum at the lowest concentration (0.25%). The same trend could be applied for Spruzit flüssig compound. When Neem Azal-T/S was compared with Neudosan, the later at any concentration was more toxic than the former. Generally, Neudosan and Spruzit flüssig seemed to be active against E. chrysomelina during the first three days after treatment, then, deterioration occurred among this compound. In case of latent effects, the duration of the treated fourth instar larvae of E. chrysomelina with any tested compound, was prolonged. An increase in the concentration caused prolongation of this period to reach the longest at the highest concentration. This prolongation, was more pronounced after Neem Azal-T/S treatment followed by Neudosan and Spruzit flüssig. The same trend could be applied for the duration of the pupal stage as being affected by the different concentrations of the tested compounds which were applied to the fourth instar larvae. However, adults of E. Chrysomelina treated as fourth instar larvae with high concentrations of Neem Azal-T/S showed some adult malformations. The sex ratios of adults produced from fourth instar larvae treated with different concentrations of the tested natural compounds were about 1?:?1, except for two cases; 1% Neudosan where the number of females were double the number of males and 0.5% Spruzit flüssig where the number of males were double the number of females. An increase in the concentration of any tested compound caused an obvious decrease in the percentage of emerged adults. The least emergence percentage was recorded in the case of Spruzit flüssig, followed by Neudosan and Neem Azal-T/S. Treating the fourth instar larvae with the different concentrations of the tested natural compounds significantly shortened the longevity of produced adults and decreased the fecundity.     

Effects of dietary sterols on development and reproduction of southwestern corn borer Diatraea grandiosella Dyar

Southwestern corn borer larvae, Diatraea grandiosella Dyar, were reared on artificial diets containing individual sterols (cholesterol, sitosterol, or stigmasterol) in concentrations ranging from 0.05 to 0.2%. Female larvae developed to pupae more rapidly as sitosterol and stigmasterol were increased in the diets. Increased cholesterol concentrations did not affect the larval period significantly, and development was not as rapid as with the phytosterols. Female larvae developed at significantly slower rates in all diets than did males, except at the highest concentrations of sitosterol and stigmasterol. Female pupae and adults were significantly heavier than the males, and pupal and adult weight increased as sterol concentrations increased. Number of eggs laid per fertilized female and egg hatchability were significantly increased as concentrations of the three sterols were increased in the larval diets. Sitosterol-reared females produced more eggs than did females reared on other sterols but egg hatchability was not significantly different among sterols.     

Development,survival, body weight,longevity, and reproductive potential of Oemena hirta (Coleoptera: Cerambycidae) under different rearing conditions

Oemona hirta (F.) is a New Zealand native longicorn beetle, whose larvae bore into the wood of branches and stems of living trees and vines, causing serious damage. To explore effective methods for maintaining laboratory colonies and biology of immature stages of this species we evaluated four laboratory rearing methods with both natural and artificial diets and compared biological parameters of laboratory colonies with those of field-collected insects. On an artificial diet, approximately 40% of neonate and 70% of autumn- and 11% of winter-collected larvae reached adulthood. Neonate larvae could not complete their development in cut poplar (Populus nigra variety italica Koehne) twigs; however, when twigs were standing in water >46% of neonate larvae survived to adulthood. Mean larval development time ranged from approximately150 to almost 300 d, depending on rearing methods. Mean pupal stage ranged from 15 to 19 d. Adult females were significantly heavier than males. Although adult females from field-collected twigs and reared on the artificial diet had similar mean body weight, which was significantly greater than that of other rearing colonies, the potential fecundity was the highest in the former (83.1 +/- 29.4 eggs) and the lowest in the latter (33.5 +/- 9.1 eggs). In all laboratory-reared and field-collected insects, the total number of eggs produced (eggs laid + eggs that remained in dead female body) by females was positively correlated with their body weight. When larvae fed on the artificial diet, there was no positive correlation between the number of eggs successfully laid and female body weight. However, when larvae lived on natural food, a positive correlation was found. In laboratory colonies, mean longevity of females (36 to 52 d) was slightly greater than that of males (30 to 50 d) without significant difference between sexes but in the adults from field-collected twigs, males (52 d) lived significantly longer than females (33 d). In terms of time, labor, and the number of resulting adults, collecting larvae in the field in autumn and then transferring them onto artificial diet is the most effective method for maintaining a laboratory colony.     

Effects of spirodiclofen on reproduction in a susceptible and resistant strain of <Emphasis Type="Italic">Tetranychus urticae</Emphasis> (Acari: Tetranychidae)

In this study the reproductive capacity of a laboratory-selected spirodiclofen resistant strain was investigated after treatment with spirodiclofen. Firstly, females were exposed to different concentrations of spirodiclofen (200 and 1,000 mg/l) during 6, 12 or 24 h. In contrast to the susceptible parental strain, the fecundity and fertility of resistant mites was not affected by treatment with these concentrations after any time of exposure tested. Secondly, pre-treatment of the resistant females with the synergists PBO or DEF could increase the inhibitory effect of spirodiclofen on reproduction, demonstrating the possible involvement of monooxygenases and esterases in metabolic detoxification of the acaricide. Because spirodiclofen interferes with lipid biosynthesis, total lipid content was measured in female adults. There were no significant differences between treated and non-treated female adults, both in the susceptible and resistant strain. However, the total lipid content in the resistant females was significantly higher than in susceptible females. Our data shows that the detection of spirodiclofen resistance should not be limited to mortality bioassays with eggs or larvae, but should be combined with inhibitory studies on female fertility and fecundity.     

Role of the exoprotease InA in the pathogenicity of Bacillus thuringiensis in pupae of Hyalophora cecropia

Two geographical biotypes of Nomuraea rileyi (from Ecuador and the United States) were topically bioassayed against seven lepidopteran species, i.e., Anticarsia gemmatalis, Heliothis zea, Heliothis virescens, Heliothis subflexa, Pseudoplusia includens, Spodoptera exigua, and Trichoplusia ni. There was an average difference of 1.7-fold in mortality in how cultures of the same insect species from different sources responded to topical applications of either biotype of N. rileyi. Regression equations and LC₅₀ values were obtained for each insect species and fungal biotype combination. Larvae of S. exigua were equally susceptible to both biotypes of N. rileyi. Although larvae of A. gemmatalis were moderately susceptible to the Ecuadoran biotype, they were relatively nonsusceptible to the Mississippian biotype. Species of Heliothis (H. zea, H. virescens, and H. subflexa) were about equally susceptible to the Mississippian biotype. Larvae of H. subflexa and H. virescens, however, were significantly less susceptible than H. zea to the Ecuadoran biotype. When the integumental barrier was breached via intrahemocoelic injections, larvae of H. virescens were as susceptible as H. zea larvae to blastospores of either biotype of N. rileyi.