Application of spinosad increases the susceptibility of insecticide-resistant Alphitobius diaperinus (Coleoptera: Tenebrionidae) to pyrethroids.

The effect of spinosad exposure on the susceptibility of pyrethroid- and organophosphate-resistant populations of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), to insecticides was investigated in broiler house farm and laboratory studies. A field pyrethroid- and organophosphate-resistant population showed a 3.6-fold increase in susceptibility to gamma-cyhalothrin following spinosad treatment. Overall, cyfluthrin- and fenitrothion-resistant field populations were more susceptible to these insecticides following spinosad treatments, but populations that were not resistant showed no change in susceptibility following spinosad treatment. In a related study, three broiler farm beetle populations with very similar levels of cyfluthrin and gamma-cyhalothrin resistance and similar susceptibilities to spinosad were used to investigate temporal effects of spinosad field treatments on the susceptibility to pyrethroids. Farm insecticide regimes applied at the start of each flock differed: the control broiler house received no insecticide applications, another house was systematically treated with cyfluthrin at the start of each study flock, and the third house was systematically treated with spinosad at the start of five flocks. Afterwards, treatments reverted to cyfluthrin on all farms. At the end of flocks, beetles were tested with cyfluthrin, gamma-cyhalothrin, and spinosad. The control and cyfluthrin house beetles did not change susceptibility to pyrethroids over the period of the study. In the spinosad house, spinosad had no effect on spinosad susceptibility but dramatically increased cyfluthrin and gamma-cyhalothrin susceptibilities. These new susceptibilities were maintained while spinosad applications continued, but pyrethroid susceptibility declined once spinosad applications ceased. This study provides evidence of a synergistic interaction between spinosad and pyrethroids in pyrethroid-resistant beetles. This evidence has significant implications for management of insecticide-resistant populations through an integrated spinosad-pyrethroid strategy that aims to minimize insecticide use while enhancing control.

Field assessments of control agents for lesser mealworm (Coleoptera: Tenebrionidae) using litter sampling.

Spinosad, diatomaceous earth, and cyfluthrin were assessed on two broiler farms at Gleneagle and Gatton in southeastern Queensland, Australia in 2004-2005 and 2007-2009, respectively to determine their effectiveness in controlling lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae). Insecticide treatments were applied mostly to earth or 'hard' cement floors of broiler houses before the placement of new bedding. Efficacy of each agent was assessed by regular sampling of litter and counting of immature stages and adult beetles, and comparing insect counts in treatments to counts in untreated houses. Generally, the lowest numbers of lesser mealworm were recorded in the house with hard floors, these numbers equalling the most effective spinosad applications. The most effective treatment was a strategic application of spinosad under feed supply lines on a hard floor. In compacted earth floor houses, mean numbers of lesser mealworms for two under-feed-line spinosad treatments (i.e., 2-m-wide application at 0.18 g of active insecticide (g [AI]) in 100-ml water/m2, and 1-m-wide application at 0.11 g ([AI] in 33-ml water/m2), and an entire floor spinosad treatment (0.07 g [AI] in 86-ml water/m2) were significantly lower (i.e., better control) than those numbers for cyfluthrin, and no treatment (controls). The 1-m-wide under-feed-line treatment was the most cost-effective dose, providing similar control to the other two most effective spinosad treatments, but using less than half the active component per broiler house. No efficacy was demonstrated when spinosad was applied to the surface of bedding in relatively large volumes of water. All applications of diatomaceous earth, applied with and without spinosad, and cyfluthrin at the label rate of 0.02 g (AI)/100-ml water/m2 showed no effect, with insect counts not significantly different to untreated controls. Overall, the results of this field assessment indicate that cyfluthrin (the Australian industry standard) and diatomaceous earth were ineffective on these two farms and that spinosad can be a viable alternative for broiler house use.

Metabolic mechanisms only partially explain resistance to pyrethroids in Australian broiler house populations of lesser mealworm (Coleoptera: Tenebrionidae).

The susceptibility of six Australian broiler house populations and an insecticide susceptible population of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), to cyfluthrin, beta-cyfluthrin, gamma-cyhalothrin, and deltamethrin was investigated. One broiler house population had equivalent susceptibility to the susceptible to beta-cyfluthrin and beta-cyhalothrin, with higher susceptibility to cyfluthrin and deltamethrin. The remaining five populations demonstrated strong resistance to cyfluthrin (19-37-fold), the insecticide used most widely for management of A. diaperinus in Australia. Each cyfluthrin-resistant population demonstrated reduced susceptibility to beta-cyfluthrin (resistance ratios were 8-17-fold), deltamethrin (2.5-8-fold), and gamma-cyhalothrin (6-12-fold) compared with the laboratory population, but cross-resistance patterns varied considerably between populations. Adding piperonyl butoxide (PBO) had no effect on the susceptibility of the susceptible population to any of the insecticides, but it increased the susceptibility of each of the five cyfluthrin-resistant populations: to cyfluthrin (synergism ratio range, 1.9-5.0-fold), beta-cyfluthrin (1.6-4.1-fold), and y-cyhalothrin (1.7-2.0-fold). PBO had a more variable effect on susceptibility to deltamethrin, with three of the cyfluthrin-resistant populations being more susceptible to deltamethrin in the presence of PBO, but susceptibility of the remaining two populations was unaffected by adding PBO (synergism ratio range, 0.9-2.5-fold). Overall, the addition of PBO to the four pyrethroids had variable effects on their susceptibility. This variability indicated the presence of other resistance mechanisms in beetle populations apart from metabolic resistance. In addition, the relative importance of metabolic resistance in each beetle population varied widely between pyrethroids. Thus, it cannot be assumed that PBO will reliably synergize pyrethroids against cyfluthrin-resistant lesser mealworm populations when using it to mitigate insecticide resistance.

Responses of susceptible and cyfluthrin-resistant broiler house populations of lesser mealworm (Coleoptera: Tenebrionidae) to gamma-cyhalothrin.

Gamma-Cyhalothrin was proposed as an agent for management of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), in Australian broiler houses. From 2007 to 2009, baseline susceptibility of 20 Australian broiler farm beetle populations plus an insecticide-susceptible laboratory population was determined for gamma-cyhalothrin by using topical application. In addition, repeat testing and regression analyses of specific beetle populations to gamma-cyhalothrin showed that topical application was a very reliable and repeatable testing method. The 21 populations were tested with a cyfluthrin discriminating concentration (based on LC(99.9), 0.0007% [AI]) to identify possible cross-resistance. Across all populations, there was a significant linear relationship between the gamma-cyhalothrin LC50 value and mortality induced by the cyfluthrin LC(99.9). Full cyfluthrin baseline studies of seven populations indicated that gamma-cyhalothrin was twice as toxic as cyfluthrin, even against susceptible beetles and resistance ratios at the cyfluthrin LC50 and LC(99.9), varied considerably, with maxima of 56.6 and 83.6 respectively. Corresponding ratios for gamma-cyhalothrin for the same populations were 8.6 (LC50) and 7.9 (LC(99.9)). There were no significant correlations between beetle weights and gamma-cyhalothrin LC50 or LC(99.9), values. A discriminating concentration of 0.005% (AI) gamma-cyhalothrin was chosen to detect any future changes in susceptibility. Results of this study suggest that cyfluthrin can confer cross-resistance to gamma-cyhalothrin in A. diaperinus, but the magnitude of this resistance is unpredictable. Thus, widespread and frequent cyfluthrin use in broiler houses in eastern Australia, which has selected for cyfluthrin resistance, also has resulted in reduced susceptibility to gamma-cyhalothrin. Due to its higher relative toxicity, gamma-cyhalothrin is still potentially useful for management of lesser mealworm, but due to cross-resistance issues, adoption of gamma-cyhalothrin for broiler house use will require a cautious and judicious approach.

Factors affecting localized abundance and distribution of lesser mealworm in earth-floor broiler houses in subtropical Australia.

Factors that influence the localized abundance and distribution of lesser mealworm, Alphitobius diaperinus (Panzer), in litter of two compacted earth-floor broiler houses in subtropical Australia were studied using various experimental manipulations. Numbers of lesser mealworms substantially increased inside caged areas and under uncaged empty feed pans placed in open areas of the houses. These populations were found to be localized and independent of chicken-feed, manure, and high beetle populations that normally occur under existing feed pans. Substantial horizontal movement of larvae to under feed pans was recorded. Placing metal barriers around these pans significantly restricted this movement. In almost all treatments, lesser mealworms typically peaked in numbers during the middle of the flock time. This temporal pattern of abundance also was observed under pans within barriers, where relatively low insect numbers occurred, but it was not observed in uncaged open areas (where chickens had complete access). It is likely that larvae do not establish in open areas, but fluctuate in numbers as they either move to refuges away from chickens or suffer high rates of mortality. In these refuges, larvae peak in numbers and then leave the litter environment to pupate in the earth floor before the end of the flock time. This behavior might be exploited for management of lesser mealworm by targeting applications of control agents.

Distributions of lesser mealworm (Coleoptera: Tenebrionidae) in litter of a compacted earth floor broiler house in subtropical Queensland, Australia.

Distributions of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), in litter of a compacted earth floor broiler house in southeastern Queensland, Australia, were studied over two flocks. Larvae were the predominant stage recorded. Significantly low densities occurred in open locations and under drinker cups where chickens had complete access, whereas high densities were found under feed pans and along house edges where chicken access was restricted. For each flock, lesser mealworm numbers increased at all locations over the first 14 d, especially under feed pans and along house edges, peaking at approximately 26 d and then declining over the final 28 d. A life stage profile per flock was devised that consisted of the following: beetles emerge from the earth floor at the beginning of each flock, and females lay eggs, producing larvae that peak in numbers at approximately 3 wk; after a further 3 to 4 wk, larvae leave litter to pupate in the earth floor, and beetles then emerge by the end of the flock time. Removing old litter from the brooder section at the end of a flock did not greatly reduce mealworm numbers over the subsequent flock, but it seemed to prevent numbers increasing, while an increase in numbers in the grow-out section was recorded after reusing litter. Areas under feed pans and along house edges accounted for 5% of the total house area, but approximately half the estimated total number of lesser mealworms in the broiler house occurred in these locations. The results of this study will be used to determine optimal deployment of site-specific treatments for lesser mealworm control.

Baseline responses of Alphitobius diaperinus (Coleoptera: Tenebrionidae) to spinosad, and susceptibility of broiler populations in Eastern and Southern Australia.

Spinosad was proposed as a potential chemical for control of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), in Australian broiler houses after the detection of strong cyfluthrin resistance in many beetle populations. In 2004-2006, spinosad susceptibility of 13 beetle populations from eastern and southern Australian broiler houses and a cyfluthrin/fenitrothion-resistant reference population was determined using topical application, and was compared with the susceptibility of an insecticide-susceptible reference population. Comparisons of dose-response curves and baseline data showed that all populations, including the insecticide-susceptible population, were roughly equivalent in their response to spinosad, indicating no preexisting spinosad resistance. Two field populations, including the resistant reference population, which had confirmed cyfluthrin/fenitrothion-resistance, showed no cross-resistance to spinosad. There was no significant correlation between beetle weight and LC9.9. A discriminating concentration of 3% spinosad was set to separate resistant and susceptible individuals. Considering the levels of spinosad resistance that have been recorded in other insect pests, the sustained future usefulness of spinosad as a broiler house treatment will rely on effective integrated beetle management programs combined with carefully planned chemical use strategies.

Baseline responses of Alphitobius diaperinus (Coleoptera: Tenebrionidae) to cyfluthrin and detection of strong resistance in field populations in eastern Australia.

Resistance to cyfluthrin in broiler farm populations of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), in eastern Australia was suspected to have contributed to recent control failures. In 2000-2001, beetles from 11 broiler farms were tested for resistance by comparing them to an insecticide-susceptible reference population by using topical application. Resistance was detected in almost all beetle populations (up to 22 times the susceptible at the LC50), especially in southeastern Queensland where more cyfluthrin applications had been made. Two from outside southeastern Queensland were found to be susceptible. Dose-mortality data generated from the reference population over a range of cyfluthrin concentrations showed that 0.0007% cyfluthrin at a LC99.9 level could be used as a convenient dose to discriminate between susceptible and resistant populations. Using this discriminating concentration, from 2001 to 2005, the susceptibilities of 18 field populations were determined. Of these, 11 did not exhibit complete mortality at the discriminating concentration (mortality range 2.8-97.7%), and in general, cyfluthrin resistance was directly related to the numbers of cyfluthrin applications. As in the full study, populations outside of southeastern Queensland were found to have lower levels of resistance or were susceptible. One population from an intensively farmed broiler area in southeastern Queensland exhibited low mortality despite having no known exposure to cyfluthrin. Comparisons of LC50 values of three broiler populations and a susceptible population, collected in 2000 and 2001 and recollected in 2004 and 2005 indicated that values from the three broiler populations had increased over this time for all populations. The continued use of cyfluthrin for control of A. diaperinus in eastern Australia is currently under consideration.

Baseline responses of adult Alphitobius diaperinus (Coleoptera: Tenebrionidae) to fenitrothion and susceptibility status of populations in Queensland and New South Wales, Australia.

Traditionally in Australia, regular applications of insecticide to the floors and lower walls of broiler houses after cleanout periods have been used in an attempt to control lesser mealworm, Alphitobius diaperinus (Panzer). The Australian chicken meat industry has been concerned in recent years with the failure to control A. diaperinus in its broiler houses by using this method and with large beetle populations breaching farm biosecurity. Resistance to fenitrothion was suspected to be responsible for these recent control failures. In response, beetles from 13 poultry facilities were compared with an insecticide-susceptible reference population by using a topical application method. Generally, strong resistance to fenitrothion (up to 79 times that of the susceptible at the LC50) occurred in populations of A. diaperinus in long-established broiler growing areas of southeastern Queensland, where fenitrothion had been used continuously for up to 20 yr. In newly established broiler growing areas, where considerably less fenitrothion had been used (i.e., approximately 5 yr), much weaker or no resistance occurred. In addition, dose-mortality data generated for the susceptible reference beetle population over a range of fenitrothion concentrations showed that 0.15% fenitrothion at a LC(99.9) level could be used as a convenient topical dose concentration to discriminate between susceptible and resistant individuals. Using this method, the susceptibility of 27 field populations of A. diaperinus was determined. Of this total, 23 populations did not exhibit complete mortality against the discriminating concentration (mortality range 0-98.7%). Application of fenitrothion in Australian broiler houses for control of A. diaperinus has now ceased.