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.     

Method of insecticide delivery affects horizontal transfer of fipronil in the German cockroach (Dictyoptera: Blattellidae).

Horizontal transmission of insecticide occurs when foragers contact or ingest an insecticide, return to the aggregation or nest, and translocate the insecticide to the shelter and its vicinity. Relatively more sedentary members of the population then contact or eat the translocated insecticide and die. We evaluated three different methods of delivering fipronil to adult male German cockroaches, Blattella germanica (L.), for their potential to cause such secondary mortality in various developmental stages of the cockroach. Adult males topically treated with 5 ng of fipronil (approximately LD99) caused low mortality in untreated nymphs and no mortality in untreated adults within the same aggregation. Males exposed to residual fipronil on a glass surface translocated more insecticide, resulting in higher mortality of cockroaches they contacted, but only early instars were affected and no adult mortality was observed. Ingested fipronil bait, however, was most effectively translocated, and caused high mortality of untreated adults and nymphs. Ingestion of fipronil also caused greater secondary kill compared with a topical application of 25 ng, approximately the same amount recovered from the exterior of males that ingested 1 mg of 0.05% fipronil bait. Secondary mortality in the untreated population was significantly affected by the duration of contact between the treated and untreated cockroaches, the quantity and freshness of excretions from the treated insects, and the accessibility of the secretions to untreated cockroaches. The mechanisms that cause secondary kill may include ingestion of excreted fipronil residues, cannibalism of bait-fed cockroaches, as well as contact with fipronil-contaminated substrates.     

Biochemical characterization and purification of esterases from three strains of German cockroach,Blattella germanica (Dictyoptera: Blattellidae)

Three strains of German cockroach, Blattella germanica (L.) showed varying levels of resistance to chlorpyrifos, methyl parathion, propoxur, bendiocarb, and cypermethrin. The general esterase activity was at least twofold higher than susceptible strain. The subcellular distribution studies revealed that the majority of the esterase activity is present in the 100,000g cytosolic fraction. Only a small portion of the activity was membrane bound. Using non-denaturing gel electrophoresis, ten isozymes were identified in German cockroaches. These isozymes were isolated individually from the gels and analyzed for differences in activity. The isozymes E5, E6, and E7 of resistant strains had significantly higher specific activities when compared with the susceptible strain. The purification process using various column chromatography and preparative gel electrophoresis resulted in 9–11% of total esterase recovery. About double the amount of E6 was recovered from the resistant strains when compared with the susceptible strain. Kinetic analyses of E6 did not indicate differences in K_m and V_max values between the resistant and susceptible strains. Also, inhibition of esterase activity by paraoxon, chlorpyrifos, and propoxur did not suggest any structural differences in esterase E6 between strains. The results suggest that the increased production of E6 esterase contributes to insecticide resistance in German cockroaches. The role of E6 may be sequestration of toxic molecules rather than hydrolysis. © 1996 Wiley-Liss, Inc.     

Extended selections for pyrethroid resistance in the German cockroach (Dictyoptera: Blattellidae).

Selection experiments with a pyrethrins-susceptible and a pyrethrins-resistant strain of German cockroaches, Blattella germanica (L.), were conducted for 17 generations with either permethrin or fenvalerate as the selecting agent. Large nymphs were left on treated glass surfaces for extended periods of time each generation. Mortality was assessed at 24 h. The level of resistance was determined periodically by time-mortality testing. The VPI-susceptible strain served as the basis for comparison. The pyrethrins-susceptible strain developed resistance to pyrethrins early in the selection process; this strain ultimately became resistant to allethrin, phenothrin, permethrin, fenvalerate, cyfluthrin, and cypermethrin. Fenvalerate caused faster development of resistance than did permethrin. The pyrethrins-resistant strain, selected with fenvalerate, quickly became resistant to allethrin, permethrin, phenothrin, and fenvalerate. Ultimately, it developed resistance to all nine pyrethroids tested.     

Trail-following behavior in the German cockroach (Dictyoptera: Blattellidae)

The trail-following behavior of the German cockroach, Blattella germanica (L.), was evaluated by comparing the distance between cockroach movement paths and preapplied "trails" of fecal extract. For each cockroach group tested (adult males, females, gravid females, and late instars), the mean perpendicular distance of the cockroach from the trail was significantly less than the distance from a control trail. The results indicated that the German cockroaches did exhibit trail-following behavior. Trail- following accuracy varied among the cockroach groups. The mean distance from the fecal trail ranged from 18.45 to 110.05 cm with adult males < or = adult females < or = late instars < gravid females. Very dilute fecal extract could still induce trail-following behavior in adult male cockroaches. A 5.6% concentration of fecal extract in methanol was able to induce trail-following behavior in 50% of the cockroaches. Although German cockroaches have demonstrated trail-following behavior, fecal trails are still not thought to be actively deposited. Rather, the passive distribution of fecal material within the home range results in the accumulation of trails along frequently traveled routes (i.e., between resources and the cockroach harborage).     

Detection of insecticide resistance in the German cockroach (Dictyoptera: Blattellidae) with glue-toxin traps.

Glue that contained an insecticide was evaluated for its ability to yield useful toxicological data for German cockroaches, Blattella germanica (L.). Toxicities of three classes of insecticides were evaluated by topical applications (LD50) and exposure to insecticide-impregnated glue (LC50). Cockroaches that were resistant to topical insecticide applications were also resistant to the glue formulation. Reliability was greatest when mortality was scored 40 to 48 h after the cockroaches were placed on the glue. This method should be adaptable for insecticide resistance monitoring of German cockroaches.     

Influence of insecticide treatment on German cockroach (Dictyoptera: Blattellidae) movement and dispersal within apartments

Two similar studies were done to investigate effects of insecticide treatments on German cockroach, Blattella germanica (L.), movement and dispersal within individual units in multifamily housing. In the first study, Whitmire PT-565 Pyrethrum Insect Fogger (pyrethrins), Diazinon 4E (diazinon), Protector (permethrin), and Baygon 1.5 (propoxur) did not induce any significant changes in cockroach distribution, and rates of movement within apartments decreased. In the second study, populations treated with sublethal doses of pyrethrins and resmethrin settled to their original distribution 24 h after treatment. Results from both studies indicate that thorough insecticide applications do not significantly affect German cockroach population dispersal or movement patterns within apartments.     

The occurrence of translocations in insecticide resistant and susceptible field-collected strains of the German cockroach

Insecticide resistant and susceptible field-collected strains of the German cockroach were surveyed for the occurrence of chromosome polymorphisms. Analyses were on meiotic cells of males that hatched from oothecae with unusual numbers (>5) of incompletely developed eggs (aberrant oothecae). Three males were reciprocal translocation heterozygotes. A spontaneous translocation was noted in one cell of a fourth male. The males were from insecticide resistant strains. Partially developed eggs in oothecae from outcrosses of two of the translocation herozygotes ceased development in early stages and were characterized by gross morphologic abnormalities. Chromosome abnormalities other than translocations occurred in a low percentage of males from aberrant oothecae in both resistant and susceptible strains. The only general differences between susceptible and resistant strains were that, in pooled data, the frequencies of unhatched oothecae and of aberrant oothecae were higher in the resistant than susceptible strains.     

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.     

Age-related changes in foraging in the German cockroach (Dictyoptera: Blattellidae)

The dynamics of exploitation of standard experimental food sources by the German cockroach, Blattella germanicaL. (Blattellidae), were analyzed in an urban habitat in relation to developmental stage. The data presented here stress differences in foraging capacities between small (first-and second-instar) larvae and animals of other developmental stages. The first animals to arrive in a food patch presented a developmental-stage distribution significantly different from that of the general population. Adults and large larvae (fifth and sixth instars) were the first to find food sources and, in particular, before small larvae. Significant differences appeared between developmental stages concerning givingup time and the time animals left a patch. Small larvae were significantly underrepresented in a patch just before food exhaustion but they were significantly more numerous than expected just after depletion. Small larvae remained in the vicinity of a depleted food dish longer than animals of other developmental stages. Adults left patches as soon as these were depleted, long before small larvae did. Developmental stage influenced rate of departure. These observations indicate that cockroaches improve their foraging performance as they grow larger.     

Cross-resistance between dieldrin and fipronil in German cockroach (Dictyoptera: Blattellidae)

The toxicity of fipronil and dieldrin was determined in one susceptible laboratory strain and seven insecticide-resistant field-collected strains of Blattella germanica (L). The Zo960302 and Ga021001 strains were 1,270- and 2,030-fold resistant to dieldrin and 15- and 14-fold resistant to fipronil. The Su960304 and Od010803 strains were 15- and 13-fold resistant to dieldrin and two- and four-fold resistant to fipronil. Three strains showed no or a low level of resistance to dieldrin and fipronil. Crosses were performed between the susceptible strain Danish Pest Infestation Laboratory (DPIL)-SUS and the resistant strains Zo960302 and Su960304 and resistance to dieldrin and fipronil were intermediate compared with the susceptible and the resistant strains. Backcrosses to both of the parental strains showed cosegregation of dieldrin and fipronil resistance. The toxicity of dieldrin and fipronil was correlated when compared at LD50, and 93% of the observed variation in LD50 of fipronil can be ascribed to variation among predictions based on the value of LD50 of dieldrin. The frequency of the A302S substitution in the resistance to dieldrin (Rdl) gene in the highly dieldrin- and fipronil-resistant strains Zo960302 and Ga021001 and the moderately resistant Su960304 was 0.97, 1.0, and 0.38, respectively. We consider the connection between the frequency of the Rdl mutation and dieldrin and fipronil resistance a causal connection and not merely a coincidence.     

Effect of temperature on the toxicities of ten pyrethroids to German cockroach (Dictyoptera: Blattellidae).

Of 10 pyrethroids tested by topical application of male German cockroaches, Blattella germanica (L.), d-phenothrin was the least toxic at 19, 26, or 31 degrees C. lambda-cyhalothrin was most toxic. Pyrethroids with the alpha-cyano moiety (fluvalinate, fenvalerate, cypermethrin, esfenvalerate, tralomethrin, cyfluthrin, and lambda-cyhalothrin) were more toxic than those without this functional group (d-phenothrin, resmethrin, and permethrin). At LC50, toxicity was negatively related to temperature. Temperature-toxicity responses of five of seven alpha-cyano pyrethroids were parallel, possibly indicating qualitatively identical but quantitatively different levels of detoxification enzymes.     

Comparative toxicity and repellency of microencapsulated and other liquid insecticide formulations to the German cockroach (Dictyoptera: Blattellidae)

Responses of German cockroaches, Blattella germanica (L.) (Dictyoptera: Blattellidae), to microencapsulated (ME) formulations of six insecticides (bifenthrin, chlorpyrifos, cyfluthrin, deltamethrin, lambda-cyhalothrin, and permethrin) were compared with emulsifiable concentrates (EC) (chlorpyrifos, cyfluthrin, deltamethrin, and permethrin) or ready-to-use (RTU) formulations (bifenthin and lambda-cyhalothrin). Two rates were tested per comparison. Baseline toxicity (LT50 value) was determined by continuous exposure to residual deposits. Repellency, toxicity (LT50), and performance index (PI) values were determined using Ebeling choice boxes. Baseline toxicity of the permethrin formulations was similar, but all other active ingredients had significant toxicity differences at one or both formulation x dose comparisons. Baseline toxicity and repellency were negatively correlated. Choice box LT50 and the time to reach 50% of the maximum PI were positively correlated. The maximum PI was positively correlated (P < 0.06) with baseline LT50 and negatively correlated (P < 0.07) with repellency. Chlorpyrifos had the lowest repellency except for the EC at 0.25%. Bifenthrin ME and lambda-cyhalothrin ME had greater PI values than comparative RTU formulations. Cyfluthrin EC at 0.03% and deltamethrin ME at 0.01% had significantly lower PI values than comparison treatments. Permethrin PI value for the EC at 0.03% exceeded that for the ME, but at 0.05% the ME had a significantly greater PI. These data demonstrate the difficulty in making generalizations about the relative performance of ME compared with EC or RTU formulations. Variable results observed within, and between, formulations may be influenced by application rate, formulation type, other formulation components, and the toxicity-repellency of the active ingredient.     

Secondary transmission of toxic baits in German cockroach (Dictyoptera: Blattellidae)

The death rates of three toxic gel baits (fipronil bait, 0.05%; hydramethylnon, bait 2.15%; and abamectin B1 bait, 0.05%) were estimated in Blattella germanica (L.). After ingestion, all three baits killed similar proportions of cockroaches, but they died more rapidly after ingestion of fipronil bait than after ingestion of abamectin B1 bait or of hydramethylnon bait. Laboratory experiments evaluated the impact of secondary transmission of fipronil bait and hydramethylnon bait. Mortality rates caused by secondary transmission were higher for fipronil bait than for hydramethylnon bait. Under controlled laboratory conditions, secondary transmission occurred mainly through direct contact with, or ingestion of, traces of baits dispersed in the environment by contaminated cockroaches. These traces were either deposited by "trampling" in the environment or on dead contaminated cockroaches. Social interactions and cannibalism played a minor role in secondary transmission of these two baits.     

Topical and oral toxicity of sulfluramid, a delayed-action insecticide, against the German cockroach (Dictyoptera: Blattellidae)

The LD50 of sulfluramid topically applied to 2-d-old, fifth instars of the German cockroach, Blattella germanica (L.), was estimated at 14.5 micrograms/g (95% FL = 13.7-15.4 micrograms/g). Sulfluramid was significantly more toxic than topically applied hydramethylnon (LD50 = 29.2 [19.0-46.5] micrograms/g). Sulfluramid had delayed toxicity but caused mortality significantly faster than hydramethylnon after topical application. The oral LD50 against newly enclosed, fifth instars was estimated to be 4.1 (3.9-4.4) micrograms/g; this toxicity was significantly greater than when sulfluramid was topically applied. Mortality caused by sulfluramid occurred significantly more slowly in the dietary exposures than in the topical applications. Sulfluramid at 1,000 ppm in diets was not a feeding deterrent to nymphal B. germanica.     

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

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

Incorporation of pyriproxyfen in a German cockroach (Dictyoptera: Blattellidae) management program

Pyriproxyfen (Nylar), a juvenile hormone analog (JHA), and acephate were used in a German cockroach, Blattella germanica (L.), management program. Monthly applications of acephate for 6 mo reduced nightly trap catches from 24.68-25.25 to 4.46-5.31 at 2 mo, and trap catches averaged 1.49-8.04 from 2-6 mo. After acephate applications were discontinued, trap catches increased from 6.86 at 7 mo to 28.08 at 12 mo. Pyriproxyfen was applied at months 0, 3, and 6. Monthly pyriproxyfen treatments were applied from 7 to 11 mo because, at month 6, only 21.00-27.30% of adults had wing twisting (one effect of pyriproxyfen exposure). These monthly applications resulted in 74.55-78.53% of adults with twisted wings in the German cockroach population at 12 mo. Trap catches in apartments treated with pyriproxyfen aerosol were significantly lower at 7, 8, 10, and 12 mo after the initial treatment than those that did not receive JHA treatment. At 7-12 mo, the pyriproxyfen residual spray did not significantly reduce German cockroach populations, but trap catches were consistently lower. German cockroach populations were sampled for 1 yr (from 13 to 24 mo) after pyriproxyfen applications were discontinued. Compared with those in apartments treated with acephate, trap catches in apartments treated with pyriproxyfen were significantly reduced from 12 to 18 mo after the initial treatment. At 18-24 mo, trap catches in apartments treated with pyriproxyfen aerosol were consistently lower than those treated with acephate, but these differences were not significant.     

Comparative activity of fenoxycarb and hydroprene in sterilizing the German cockroach (Dictyoptera: Blattellidae)

Last-instar German cockroaches, Blattella germanica (L.), were treated topically with the juvenoids, fenoxycarb and hydroprene, and observed for reproductive capabilities during adulthood. Hydroprene had a significantly lower SD50 (dose required to sterilize 50% of the cockroaches) for males (39.82 micrograms/g body weight) compared with females (86.64 micrograms/g body weight). In contrast, SD50's of fenoxycarb did not differ significantly between the sexes (18.04 and 13.66 micrograms/g body weight for females and males) and were significantly lower when compared with SD50's of hydroprene. A strong positive relationship between wing twisting (a morphogenetic effect caused by juvenoids) and sterility was observed among cockroaches treated with 100 and 10 micrograms/microliters of each juvenoid. However, only 16% of females displaying twisted wings were sterile when treated with 1 microgram/microliter of hydroprene. Cockroaches treated with 100 micrograms/microliters of hydroprene lived 10-14 wk fewer than control adults and often attempted a sixth (supernumerary) molt.     

Differences in the response of German cockroach (Dictyoptera: Blattellidae) field strains to vapors of pyrethroid formulations.

Late-instar German cockroaches from an insecticide-susceptible field strain (Fairbanks) and four pyrethroid-resistant field strains were exposed to vapors of a cyfluthrin flowable concentrate (FC) and the formulation blank (formulation without the active ingredient). Experiments with other pyrethroid formulations were done with the Fairbanks strain and an insecticide-resistant field strain (K-851) because these strains exemplified the maximum interstrain variation in experiments with the FC. In these experiments, I used a cyfluthrin wettable powder (WP) and its blank, a cyfluthrin emulsifiable concentrate (EC), and a permethrin aerosol and its blank. Vapors of the FC and its blank caused the most rapid dispersal. Dispersal induced by vapors of the WP, EC, and aerosol formulations was low and variable. In general, less of a dispersal response occurred in the resistant strains than in the susceptible strain. The strain mostly highly resistant to pyrethroids (K-851) dispersed slowly compared with the other resistant strains in experiments with the FC and its blank. When vapor-induced dispersal was relatively low (less than 50%), a trend was still evident towards less dispersal in the K-851 strain than in the Fairbanks strain. Although dispersal in experiments with the FC did not differ significantly from that in experiments with the FC formulation blank, mean percentage dispersal at the end of the experiments was higher with the formulation blanks than with the complete formulations in experiments with the WP and permethrin aerosol.