Measuring fitness and intergenic interactions: The evolution of resistance to diazinon inLucilia cuprina

Resistance at the diazinon-resistance locus (Rop-1) is recessive with respect to fitness. Selection initially occurs at concentrations lower than those required to controlLucilia cuprina. The presence of theRop-1 allele initially disrupted development so that in the absence of diazinon, carriers were at a relative selective disadvantage. Continued use of the chemical, subsequent to resistance evolving, selected a modifier to ameliorate this effect. Modified resistant phenotypes show similar developmental stability and relative fitness to susceptible individuals. Frequency-dependent interactions are observed between resistant and susceptible phenotypes of theRop-1 locus. The interactions are determined by the concentration of diazinon and range from competitive to facilitative. The results are discussed in the context of the contribution insecticide resistance systems can make to the study of general evolutionary phenomena.     

Diazinon resistance, fluctuating asymmetry and fitness in the Australian sheep blowfly, lucilia cuprina

Genetic evidence suggests that the evolution of resistance to the insecticide diazinon in Lucilia cuprina initially produced an increase in asymmetry. At that time resistant flies were presumed to be at a selective disadvantage in the absence of diazinon. Subsequent evolution in natural populations selected modifiers to ameliorate these effects. The fitness and fluctuating asymmetry levels of resistant flies are currently similar to those of susceptibles. Previous genetic analyses have shown the fitness modifier to co-segregate with the region of chromosome III marked by the white eyes, w, locus, unlinked to the diazinon resistance locus, Rop-1, on chromosome IV. This study maps the asymmetry modifier to the same region, shows, as in the case of the fitness modifier, its effect to be dominant and presents data consistent with the fitness/asymmetry modifier being the same gene (gene complex). These results suggest changes in fluctuating asymmetry reflect changes in fitness.     

Selection of dieldrin-resistant strains of Lucilia cuprina (Diptera: Calliphoridae) after ethyl methanesulfonate mutagenesis of a susceptible strain.

After ethyl methanesulfonate (EMS) mutagenesis of a susceptible strain (SWT), selective screening of Lucilia cuprina (Wiedemann) resulted in four strains that were resistant to the insecticide dieldrin. Concentrations used for selection were greater than LC99 of susceptible phenotypes. No resistant variants were screened from the standard laboratory strain (SWT) not treated with EMS. The resistance phenotypes of the four resistant strains were similar to each other and to that of a field-selected resistant strain. The genetic basis of resistance is monogenic in all strains and the data are consistent with the same locus, Rdl, determining resistance status in each strain. The Rdl locus maps to chromosome V, approximately 3.5 map units distal to the Sut locus. Dieldrin resistance may be caused by less effective blocking of insect neuronal GABA receptors by the chemical in resistant strains. The data indicate that the evolution of resistance to an insecticide in the field may be constrained by a limited number of genetical and biochemical options if a monogenic response is selected for and that the spontaneous mutation rate to the Rdl allele is less than 1 in 10(6) in the laboratory.     

Selection for resistance to imidacloprid in the house fly (Diptera: Muscidae)

The house fly, Musca domestica L. (Diptera: Muscidae), continues to be a primary pest of livestock facilities worldwide. This pest also has shown a propensity for pesticide resistance development when under high selection pressures. In this study the house fly strain FDm was created by a 20% contribution from each of five colonies collected from dairies in Florida with known imidacloprid resistance. The FDm strain was used to evaluate the level ofimidacloprid resistance after five selections near the LC70 value of each selected generation. Overall, the mean selection mortality was 72.7, with males being considerably more susceptible than females. The unselected (F0) FDm strain showed considerable susceptibility to imidacloprid after its creation, compared with the five parental strains. Between 9500 and 14,000 virgin house flies were used in each selection. After the fifth and final selection, a 331-fold increase in imidacloprid resistance at the LC70 was observed over the parental FDm strain. In parallel studies, the FDm strain showed increasing tolerance of the commercial imidacloprid product QuickBayt. These results suggest that livestock producers should use caution when choosing pesticides and consider rotating fly baits, as is encouraged with other pesticide treatment regimes on farms.     

Organophosphorus resistance in Aphis gossypii (Hemiptera: Aphididae) on chrysanthemum in the UK

The FAO standard dip-test was used to test resistance of glasshouse populations of Aphis gossypii from chrysanthemums to the organophosphorus insecticides heptenophos and diazinon. Compared with a susceptible population collected in 1981, resistance factors to diazinon of up to 538 were found in populations collected after 1984. Resistance to heptenophos was less than that to diazinon but some populations collected after 1984 were 34 times as resistant as the susceptible strain. Possible reasons for the sudden increase in resistance in A. gossypii and current prospects for control of resistant populations are discussed.     

Factors affecting the toxicity of diazinon to Musca domestica L

Processes affecting the toxicity of diazinon to a susceptible and a resistant strain of houseflies were examined. More evidence was obtained to show that slower penetration of diazinon through the integument of resistant flies is a cause of resistance. Small amounts of two decomposition products were found in both strains. The decomposition mechanisms, in these strains were differently distributed and, although detoxication of diazinon in the two strains is quantitatively similar and small, it may contribute to resistance. Traces of diazoxon were detected when diazinon was incubated with tissue extracts of either strain. Tissue extracts of resistant, but not of susceptible, flies decomposed significant amounts of diazinon in 1 hr. and the ability to decompose diazoxon seems to be an important cause of resistance. Tissues of both strains sorbed diazinon from aqueous solution similarly; the quantities sorbed were large and suggest that sorption may increase the amount of poison needed inside the insects to kill, by between five and forty times.     

Genetics of resistance to transgenic Bacillus thuringiensis poplars in Chrysomela tremulae (Coleoptera: Chrysomelidae)

The area under genetically engineered plants producing Bacillus thuringiensis (Bt) toxins is steadily increasing. This increase has magnified the risk of alleles conferring resistance to these toxins being selected in natural populations of target insect pests. The speed at which this selection is likely to occur depends on the genetic characteristics of Bt resistance. We selected a strain of the beetle Chrysomela tremulae Fabricius on a transgenic Bt poplar clone Populus tremula L. x Populus tremuloides Michx producing high levels of B. thuringiensis Cry3Aa toxin. This strain was derived from an isofemale line that generated some F2 offspring that actively fed on this Bt poplar clone. The resistance ratio of the strain was >6400. Susceptibility had decreased to such an extent that the mortality of beetles of the strain fed Bt poplar leaves was similar to that of beetles fed nontransgenic poplar leaves. Genetic crosses between susceptible, resistant, and F1 hybrids showed that resistance to the Cry3Aa toxin was almost completely recessive (D(LC) = 0.07) and conferred by a single autosomal gene. The concentration of Cry3Aa produced in the transgenic Bt poplar used in this study was 6.34 times higher than the LC99 of the F1 hybrids, accounting for the complete recessivity (D(ML) = 0) of survival on Bt poplar leaves. Overall, the genetic characteristics of the resistance of C. tremulae to the Cry3Aa toxin are consistent with the assumptions underlying the high-dose refuge strategy, which aims to decrease the selection of Bt resistance alleles in natural target pest populations.     

Toxicity of organophosphorus insecticides to diazinon-reversely-selected,pesticide-pressure-relaxed and field-resistant strains of bulb miteRhizoglyphus robini (Acari: Acaridae)

Some field strains ofRhizoglyphus robini in Taiwan, developed high multi-resistance to organophosphorus insecticides including diazinon, dimethoate, fenitrothion, malathion, methyl parathion, omethoate, parathion and prothiophos. The organophosphorus resistance of bulb mites remained stable when the pesticide pressure was relaxed. Changes of the susceptibility to several organophosphorus insecticides for a strain after relaxation of selection pressure for 3 and 27 generations were compared. Since susceptible bulb mites could not be found in the field, a reverse selection for susceptibility to diazinon was designed and conducted. Although heterogeneity was observed in the progeny of the mixed populations, the susceptibility of these reversely-selected mites increased slowly. A stable and susceptible strain was obtained after 26 generations of selection.Contribution No. 1412 from Taiwan Agricultural Research Institute.     

Inheritance of Resistance to the Bacillus thuringiensis Toxin Cry1C in the Diamondback Moth

Laboratory selection increased resistance to the Bacillus thuringiensis toxin Cry1C in a strain of diamondback moth (Plutella xylostella). The selected strain was derived from a field population that had evolved high levels of resistance to Bacillus thuringiensis subsp. kurstaki and moderate resistance to Cry1C. Relative to the responses of a susceptible strain of diamondback moth, the resistance to Cry1C of the selected strain increased to 62-fold after six generations of selection. The realized heritability of resistance was 0.10. Analysis of F(inf1) hybrid progeny from reciprocal crosses between the selected strain and a susceptible strain showed that resistance to Cry1C was autosomally inherited. The dominance of resistance to Cry1C depended on the concentration; inheritance was increasingly dominant as the concentration decreased. Responses of progeny from single-pair families showed that resistance to Cry1C and resistance to Cry1Ab were inherited independently, which enhances opportunities for managing resistance. However, compared with projections based on previously reported recessive inheritance of resistance to Cry1A toxins, the potentially dominant inheritance of resistance to Cry1C observed here could accelerate evolution of resistance.     

Dieldrin and diazinon resistance in populations of the Australian sheep blowfly, Lucilia cuprina, from sheep-grazing areas and rubbish tips

Populations of L. cuprina collected from adjacent sheep-grazing areas and rubbish tips in Victoria (Mansfield and Warrnambool) and New South Wales (Lismore) were tested for resistance to the insecticides diazinon and dieldrin. Populations from sheep-grazing areas had a significantly higher diazinon Rop-1 allele frequency than those from adjacent tips with the Victorian populations being more resistant than those from Lismore. Victorian sheep and tip populations had similar gene frequencies at the dieldrin resistance locus, but the Rdl allele frequency was significantly greater in the population at the tip than in the population from the sheep-grazing area at Lismore. The Rdl allele is at a higher frequency in flies from the Lismore area than in Victorian populations. The results at both loci are explained by a balance of selection and gene flow between sheep and tip populations and by selective differences between geographical areas. The exceptionally high frequency of the dieldrin Rdl allele in populations at the Lismore tip may be partially explained by the use of dichlorvos for fly control. Dosage mortality curve and genetic analyses suggest that dichlorvos (an organophosphorus compound) may select at the dieldrin resistance locus. Possible mechanisms for this are discussed. The consequences of genetic differentiation between L. cuprina populations within a region for an autocidal control program are considered.     

Tolerance of Bradyrhizobium sp. (Lupini) strains to salinity, pH, CaCO3 and antibiotics

AIMS: Ten rhizobial isolates obtained from different locations in Egypt were examined for their ability to survive under stress conditions and their growth response to increasing levels of NaCl (1-8% w/v), pH (4-10), CaCO3 (1-10% w/v) and 12 antibiotics. METHODS AND RESULTS: All the rhizobial isolates tolerated a NaCl concentration up to 5% and were divided into two groups with respect to NaCl tolerance. The rhizobial isolates from group two showed significantly (P < 0.05) better survival under high NaCl concentration. All the tested isolates survived acidic (pH 4-5) and alkaline conditions (pH 9-10) and CaCO3 (up to 10% w/v) in liqued YEM medium. CONCLUSION: Antibiotic resistance patterns did not correlate to NaCl, pH or CaCO3 tolerance. Variations among different strains showed that there is potential to improve strain performance under stress conditions. Significance and Impact of the Study: The results suggest that selection of adapted strains under stress conditions is possible and can be used as inoculants for successful lupin growth.     

Genetics of organophosphate resistance in field populations of the house fly (Diptera: Muscidae)

The genetics of resistance to the organophosphate insecticide diazinon were investigated in four populations of the house fly, Musca domestica L., collected in the southern United States. Crosses were made between individual females of lines derived from each population and males of a susceptible strain with three recessive mutants on chromosome II. Individual F1 females were crossed to mutant males, and the progenies were scored for resistance to diazinon and for the presence of mutant phenotypes. A major chromosome II gene for resistance to diazinon was present in all populations at an overall frequency of 83%. Map distances between the resistance gene and the mutant aristapedia and between the mutants aristapedia and stubby wing were highly variable in all populations. Recombination among the visible mutants was usually reduced in resistant progenies relative to susceptible progenies. The data suggest that a single major gene for resistance to diazinon was present on chromosome II in all test populations at variable map positions and is usually associated with a chromosome rearrangement, probably an inversion. The results are similar to those obtained earlier with house fly populations selected for resistance to insecticides in the laboratory; therefore, they seem to be characteristic of field and laboratory populations of the house fly. Overall, the data offer an explanation for previous results suggesting the existence of multiple, closely linked genes for metabolic resistance to insecticides on house fly chromosome II.     

西花蓟马对多杀菌素的抗性汰选和遗传方式

在室内进行了西花蓟马Frankliniella occidentalis(Pergande)种群对多杀菌素的抗性筛选和抗性遗传方式分析。经过2年多的汰选,抗性水平相比较初始种群提高了30倍,与同时测定的敏感品系相比抗性达到80.8倍。抗性遗传方式分析结果表明,正、反交后代显性度分别为0.51和0.43,二者差异不显著,说明西花蓟马对多杀菌素的抗性为常染色体、不完全显性遗传;剂量对数死亡机率值曲线分析结果显示回交后代在死亡率50%处,自交后代在死亡率25%和75%处未出现明显平坡,回交和自交后代实测值卡方检验进一步证实抗性为多基因控制。     

Aspects of insecticide resistance in the melon and cotton aphid, Aphis gossypii (Hemiptera: Aphididae)

Strains of the aphid Aphis gossypii, taken from cucumber in UK glasshouses were tested for resistance to pirimicarb, diazinon and heptenophos. Compared to a susceptible strain, resistance factors to diazinon of up to 659 were found in populations collected between 1987 and 1989. Compared to the same susceptible strain, diazinon-resistant populations also showed a reduced response to heptenophos. All field populations tested against pirimicarb were resistant. Host plant preference tests showed that all strains (including stock cultures) tested could readily breed on cotton. However strains originally collected from chrysanthemum did not transfer to cucumber and likewise strains from cucumber would not colonise chrysanthemum. Polyacrylamide gel electrophoresis of four of the aphid strains showed differences in migration rates and intensity of staining of esterase bands. Intensity of staining was correlated with the degree of diazinon resistance.     

Control of insecticide resistance in a field population of houseflies,Musca domestica,by releasing susceptible flies

Susceptible houseflies, Musca domestica, were released at a waste disposal site to control insecticide resistance in a field housefly population. In the first experiment, a total of 163,000 pupae of the susceptible Takatsuki strain were released in October–November 1977. LD₅₀ values to fenitrothion and diazinon decreased to about one-sixth in April 1978, five months after the releases, of those before the releases. For the second experiment, a susceptible colony was derived by cross and backcross between a white-eyed substrain of the Takatsuki and a field colony. This susceptible colony consisted of whiteeyed flies with low activity and normal-eyed flies bearing no or one white eye gene. The results of large cage experiments suggested that the normal-eyed males of the susceptible colony had half the mating competitiveness of wild males. Approximately 31,000–46,000 susceptible pupae were used in each of five releases from October to November 1980. The population number of each sex, estimated by a mark-release-recapture method, increased from 12,000 in late September to 35,000–43,000 in middle November and then decreased to 5,000–8,000 in early December. The frequency of field-collected males bearing one white eye gene and those bearing one male determining factor, which were characteristics of the susceptible colony released, increased gradually during the period of releases. The susceptibility of the field population to fenitrothion and diazinon was examined five times in the period from September to December 1980. With time, the dosage-mortality regression gradually shifted towards that of the susceptible colony after starting the releases. LD₅₀ values to fenitrothion and diazinon decreased to about one-sixth and one-fifth, respectively, in June 1981, six months after the second series of susceptible fly releases, of those before the releases. Ratios of the wild flies to the released fiies were estimated to be between 4.7∶1 and 9.8∶1 in males and between 3.0∶1 and 3.9∶1 in females by taking the quality of the released colony and the population parameters of the field houseflies into consideration. Under several assumptions, the manner of resistant phenotype reduction was discussed, based on the dosage-mortality regressions and the ratios of released flies. These results showed that the releases of susceptible flies were successful in suppression of insecticide resistance in the field housefly population.     

Genetics of pink bollworm resistance to Bacillus thuringiensis toxin Cry1Ac

Laboratory selection increased resistance of pink bollworm (Pectinophora gossypiella) to the Bacillus thuringiensis toxin Cry1Ac. Three selections with Cry1Ac in artificial diet increased resistance from a low level to >100-fold relative to a susceptible strain. We used artificial diet bioassays to test F1 hybrid progeny from reciprocal crosses between resistant and susceptible strains. The similarity between F1 progeny from the two reciprocal crosses indicates autosomal inheritance of resistance. The dominance of resistance to Cry1Ac depended on the concentration. Resistance was codominant at a low concentration of Cry1Ac, partially recessive at an intermediate concentration, and completely recessive at a high concentration. Comparison of the artificial diet results with previously reported results from greenhouse bioassays shows that the high concentration of Cry1Ac in bolls of transgenic cotton is essential for achieving functionally recessive inheritance of resistance.     

Inheritance of resistance to chlorpyrifos in the Mt Alford strain and to diazinon in the Gracemere strain of the cattle tick (Boophilus microplus)

Reciprocal crossing of the Mt Alford (A) strain of the cattle tick B. microplus with a susceptible (S) strain and phenotype analysis of F1, testcross and F2 progeny showed that high chlorpyrifos resistance in strain A was due to two genes that were complementary and jointly exhibited incomplete dominance. Diazinon resistance in the Gracemere (G) strain appeared to be similarly inherited. The 'average' degree of dominance ('average dominance', Dav) of high chlorpyrifos resistance over susceptibility, exhibited by F1 hybrids from A X S reciprocal crossings, was +0.54 on a -1 to +1 scale and was not significantly different from the parametric value of +0.5 (semi-dominance). The corresponding Dav values revealed by G X S crossings were +0.42 for diazinon resistance (significantly less than +0.5) and -0.031 for chlorpyrifos resistance (not significantly different from zero and therefore exhibiting zero dominance/recessivity). Resistance factors for chlorpyrifos in strains A and G for homozygotes were 74 and 35, respectively, and for F1 hybrids were 25-29 and 5-7, respectively. The resistance factors for diazinon in strain G for homozygotes and F1 hybrids were 174 and 37-41, respectively.     

High-Level Field Resistance to Bacillus sphaericus C3-41 in Culex quinquefasciatus from Southern China

A flowable mosquito-larvicidal formulation of Bacillus sphaericus strain C3-41 has been continuously used for 8 years to control Culex quinquefasciatus larvae in Dongguan, Guangdong Province, China. This formulation had high efficacy against the target larvae during the first 6 years of treatment. However, under this high selection pressure, C. quinquefasciatus showed a significant level of resistance to C3-41 from years seven to eight. The resistance ratio of field-collected larvae at LC 50 was calculated to be 22 672-fold in comparison with the susceptible laboratory colony. Interestingly, no cross-resistance was observed to B. sphaericus strain LP1-G which had the same toxicity against both susceptible and resistant larvae, and B. thuringiensis subsp. israelensis was found to be more active to the latter than the former. After six months treatment with the B. thuringiensis subsp. israelensis formulation in the B. sphaericus resistant population area, the mosquito population recovered its susceptibility to B. sphaericus C3-41, with the resistance ratio of field-collected larvae dropping from 22 672- to 5.67-fold.     

Monitoring for insecticide resistance in field-collected strains of the German cockroach (Dictyoptera: Blattellidae)

Forty-five field-collected strains of German cockroaches, Blattella germanica were tested for resistance to 12 different insecticides by the time-mortality response method in comparison with a known susceptible strain. Only low to moderate resistance to diazinon, chlorpyrifos, and acephate was detected. Resistance to malathion was widespread; about half of the strains tested showed high resistance. High resistance to the carbamates propoxur and bendiocarb also occurred. High resistance was uncommon with propoxur, but about 35 strains were highly resistant to bendiocarb. High resistance to pyrethrins was observed in about of the strains tested. Resistance to the pyrethroids allethrin, permethrin, phenothrin, fenvalerate, and cyfluthrin was detected in some of the strains examined. All of the strains tested were susceptible to one or more of the insecticides used. These results indicate that, although resistance is a serious problem in this species, satisfactory control should be possible by selection of an appropriate insecticide.     

Attempts to induce double resistance to drugs in the flagellate,Chilomonas paramecium

Summary Adaptation to two antipurines, 8-azaguanine and 6-mercaptopurine, has resulted in increased resistance to the inducing agent without modifying an original resistance to sulfanilamide in a strain of Chilomonas paramecium. In the case of isoniazid, a sulfonamide-resistant strain proved to be much more susceptible than the normal strain. Adaptation has reduced this susceptibility but, so far, has not brought sensitivity back to the level of the normal strain. However, this adaptation of a sulfonamide-resistant strain to isoniazid has not appreciably decreased the original resistance to sulfanilamide.The effects of 5-bromouracil and 2,6-diaminopurine were negligible and in each case adaptation caused no apparent change in a previous sulfonamide-resistance. Benzimidazole, although moderately inhibitory for all strains tested, also did not modify a preexisting sulfonamideresistance.To Professor Dr. E. G. Pringsheim on his 80th birthday.