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.     

Survival and chill-coma in the adult lesser mealworm,Alphitobius diaperinus (Coleoptera: Tenebrionidae), exposed to low temperatures

The survival of adult Alphitobius diaperinus (Coleoptera: Tenebrionidae) at low temperatures has been investigated under laboratory conditions: LT₅₀ ranges from 9.5 days at 6°C to 47.6 days at 10°C and LT₉₉ ranges from 20 days at 6°C to 102.2 days at 10°C. Mean survival shows no significant differences between sexes, but at 10°C, all the males are dead after 88 days and all the females after 107 days. Temperature of 6°C is progressively lethal for the beetles: activity ceases, all the insects are in chill-coma after 12 days and died after 22 days. At 10°C, after 1 month, 70% of the insects are still active. Chill-coma is a reversible state but insects which entered chill-coma last did not recover in most cases and also died the last. During winter sanitary disinfections, A. diaperinus is submitted to subzero temperatures: the rate of adult survival is a key factor governing the population dynamics in the poultry houses.     

Susceptibility of Alphitobius diaperinus (Coleoptera: Tenebrionidae) from broiler facilities in Texas to four insecticides

Lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), adults were collected from six eastern Texas broiler facilities and examined for susceptibility to four formulated insecticides. Data indicate that A. diaperinus adults exposed to filter papers treated with the label rates of the insecticides exhibit some level of recovery. Approximately 20% or less A. diaperinus adults treated with Tempo SC Ultra (8 ml/92.9 m2, 11.8% beta-cyfluthrin) or Talstar WP Insecticide/Miticide (23.3 ml/92.9 m2, 10% bifenthrin) and recorded as moribund at the 4-h observation period recovered by the 24-h observation period. A. diaperinus adults treated with Tempo SC Ultra and Talstar WP also had the greatest percentage of mortality for both observation periods. A. diaperinus adults treated with Dragnet SFR (49.7 ml/92.9 m2, 36.8% permethrin) had the greatest level of recovery at approximately 50-60% overall, which was similar to that recorded for the water-only control. Additionally, five of the six A. diaperinus populations treated with Dragnet SFR resulted in < 10% mortality for both observation periods. Unlike the other insecticides examined, Talstar Professional Insecticide (10 ml/92.9 m2, "Talstar Pro," 7.9% bifenthrin) resulted in approximately 50% more A. diaperinus mortality at the 24-h than the 4-h observation period due primarily to increased mortality recorded for Farm F. A. diaperinus adults from farm D had > or = 87% knockdown 4 h after treatment to all compounds examined, indicating a high degree of sensitivity to these compounds. However, approximately 90% of the A. diaperinus adults from this population treated with Dragnet SFR and recorded as moribund recovered by the 24-h observation. A. diaperinus adults from this population treated with the other insecticides exhibited limited recovery. Susceptible adult A. diaperinus populations are still present in Texas, based on the populations examined. But, identifying these populations is difficult and time-consuming. Consequently, screening populations before treatment might not be feasible. However, newer generation pyrethroids examined in this study seem to be suitable for suppressing A. diaperinus populations in broiler facilities.     

Changes in free amino acids in Alphitobius diaperinus (Coleoptera: Tenebrionidae) during thermal and food stress

Temporal changes in free amino acid pools are examined in starved and cold-exposed (10 degrees C) beetles (Alphitobius diaperinus) (control beetles; 4, 7, 14, 28 and 56 days of cold-exposure). The range of individual responses and the effect of gender on survival and free amino acid levels are evaluated. Females survived significantly longer (Lt50 and Lt90) than males under stressing conditions. Pro, Gln, Ala, Arg and Thr are the major components of the free amino acid pool in the whole body of adult beetles. Multivariate analysis indicates that the duration of exposure explains most of the observed variation in amino acid levels, while sex effect is not significant. Moreover, amino acid levels fluctuate strongly within each sampled date, revealing a high inter-individual heterogeneity. However, this heterogeneity decreases after four weeks of cold-exposure and starvation. The increase in Ala level and the concomitant decrease of Pro after four weeks suggest that Pro might be an important fuel for metabolism when fat reserves are reduced. We conclude that changes in free amino acid pools are due to a combination of reduced individual heterogeneity, cold-acclimation and amino acid degradation for energetic purposes.     

Comparative study of the metabolic responses during food shortage and subsequent recovery at different temperatures in the adult lesser mealworm, Alphitobius diaperinus (Coleoptera: Tenebrionidae)

Metabolic responses to prolonged food shortage (35 days) and subsequent re‐feeding (14 days) were investigated in adults of an introduced beetle, Alphitobius diaperinus Panzer, as a function of temperature (12, 16, 20 and 24 °C). Various qualitative and quantitative changes that greatly vary according to the temperature experienced occurred in metabolite levels during prolonged starvation. Whereas levels of protein and ATP did not change significantly, triglycerides decreased markedly and glycogen changed little. Metabolite levels were differently affected by temperature, with triglycerides being less rapidly degraded at 20 than at 24 °C and almost completely depleted at 12 and 16 °C; in contrast to higher temperatures, glycerol is accumulated at 12 °C. Physiological adaptation to starvation and low temperatures are highly linked and energy allocation for starvation vs. temperature acclimation must be strictly regulated, both being essential for insect survival. Re‐synthesis rates during recovery are probably highly temperature‐dependent for all metabolites. The proteins retained during starvation and the preferential degradation of lipids allowed a rapid recovery. Above 16 °C, adult A. diaperinus regained locomotory activity rapidly and the triglyceride, glycerol and glycogen reserves were restored. This tropical species may be able to colonize other environments such as natural and/or artificial biotopes where conditions are close to those of its natural habitat.     

Carcinops troglodytes (Erichson) (Coleoptera: Histeridae) preying on Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) in poultry houses

We first record Carcinops troglodytes (Paykull) as a predator of Alphitobius diaperinus (Panzer) larvae in poultry houses in North and Northwest regions of the state of Paraná. Carcinops spp. are commonly recorded as predators of dipterans, and this record in poultry houses indicates the possibility of exploiting such predator for future studies aiming the development of management strategies for A. diaperinus.     

Effects of temperature and humidity on transpiration in adults of the lesser mealworm,Alphitobius diaperinus (Coleoptera: Tenebrionidae)

Measurements of water loss were made on adults of the lesser mealworm, Alphitobius diaperinus, using a recording micro-electrobalance and a programmable heat circulator bath. This species originates in tropical regions and infests poultry houses in temperate countries. Two routes of water loss were examined: the general cuticle and via the spiracles. Temperature and relative humidity of the ambient air substantially affect the cuticular transpiration in adults (fresh body weights from 12 mg to 22 mg). At near 0% R.H., between 20 and 40 degrees C the rate of body water loss gradually increased; on the other hand, the insects gained weight in an atmosphere close to saturation. Above 40 degrees C transpiration flow abruptly increased coinciding with the start of vigorous locomotor activity. This critical point corresponds to the opening of the spiracles from which the water is expelled from the tracheal system.In dead specimens, killed by cyanide or solvent, the water vapour slowly diffused out of the spiracles and, as in atracheate insects, the transpiration curves did not show a peak as the air temperature was increased.The thermostupor point (TSP) occurred as the insects became motionless; the corresponding temperature is significantly affected by atmospheric relative humidity (TSP=47.4+/-0.6 degrees C at c. 0% R.H.; TSP=46.6+/-0.7 degrees C at c. 100% R.H.).The transpiration flow was about four times as fast in specimens treated with solvent as in the individuals (live or cyanide-killed) that had undamaged water-proof cuticle. This species has to cope with a double challenge: (i) to adapt its physiology and ecology to poultry-house conditions which constitutes an extension of its primary habitats, and (ii) to survive over winter; high drought resistance and heat tolerance may constitute a pre-adaptation to conquer anthropogenic air-conditioned sites.     

Transient gut retention and persistence of Salmonella through metamorphosis in the lesser mealworm, Alphitobius diaperinus (Coleoptera: Tenebrionidae)

Aims: This study was undertaken to determine the retention of Salmonella through Alphitobius diaperinus metamorphosis and its contribution, through defecation, to external contamination. Methods and Results: Insects were exposed to a tagged Salmonella enterica and evaluated for external elimination. (i) Each day for 3 weeks, a filter collected frass from a restrained insect for analysis. (ii) Exposed larvae in a closed container were followed through pupation, and newly emerged adults were examined for their retention of marker bacteria. Conclusions: Exposed adults and larvae produced Salmonella‐positive frass for an average of 8 days, ranging from 6 to 11 days and 6 to 12 days, respectively. Nineteen per cent of the larvae carried Salmonella through metamorphosis and eclosion, with 5% of the pupal exuviae being positive as well. Significance and Impact of the Study: Many sources of foodborne pathogens within the poultry production facilities, including reservoir populations, currently go unrecognized. This diminishes the ability of producers to mitigate the transfer of pathogens between animals, humans and the environment. Poultry management standards accept the reutilization of litter. Alphitobius diaperinus survive between flock rotations on the reutilized litter, and it was demonstrated in this study that the Salmonella they carry can survive with them.     

Infectivity of Steinernema spp. (Nematoda: Steinernematidae) to Adult Litter Beetles, Alphitobius diaperinus (Coleoptera: Tenebrionidae) in the Laboratory

Three species of Steinernema, S. carpocapsae, S. feltiae, and S. scapterisci consisting of 12 different strains, were tested for their infectivity towards adults of the litter beetle Alphitobius diaperinus. Of the five most promising nematode strains, LC₅₀ values ranged from 1.5 to 77.0 nematodes per host in the filter paper assays. Assays in poultry litter material revealed LC₅₀ values to be 5.8 and 14.6 nematodes per host for the Mexican S. carpocapsae strain and Pye S. feltiae strain.     

Effect of food shortage and temperature on oxygen consumption in the lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae)

Abstract.  Temperature and food availability are limiting factors for the establishment of tropical insects in temperate countries. In the alien pest beetle, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), starvation and temperature have a significant impact on metabolic rate with oxygen consumption ranging from 0.5 µmol/g fresh mass (FM)/h at 12 °C to 3.4 µmol/g FM/h at 24 °C. At 12 °C, oxygen consumption decreased continuously during an entire period of starvation. However, at 16, 20 and 24 °C, beetles display a marked hyperactivity that leads to an increase in the oxygen consumption level during the first week of starvation, followed by a steep decrease until the end of the starvation period. Oxygen consumption either does not decline in fed beetles (observed at higher temperatures) or declines at a much shallower rate than in starved beetles (observed at cooler temperatures). During the first week of refeeding, Oxygen consumption rose steeply at 16, 20 and 24 °C before levelling off to the initial value ( t ₀). At 12 °C, no compensation process was observed during recovery. This study reveals that an important threshold in the biology of A. diaperinus lies between 12 and 16 °C, leading to the onset of reduced locomotor activity and the promotion of survival to the detriment of reproduction. This 'sit and wait' behaviour is proposed as an adaptive strategy (i.e. inactivity and lower oxygen consumption coupled with low energetic requirements and high recovery abilities). Such behaviour and the observed hyperactivity were rarely described in insects before the present study. Together, the previous and present results suggest that A. diaperinus populations are likely maintained in temperate regions by immigration from warmer situations.     

A sexually dimorphic response in supercooling temperature, enhanced by starvation, in the lesser mealworm Alphitobius diaperinus (Coleoptera: Tenebrionidae)

The effect of starvation on supercooling temperature (SCP) distribution was investigated in adults and larvae of Alphitobius diaperinus (Coleoptera: Tenebrionidae).

The mean values for SCPs of adults fed at 20°C were −14.5±2.4°C (31 males) and −10.3±1.3°C (29 females). The distribution of the SCPs of these control adults was unimodal. No significant differences were observed in either mean wet weight or mean dry weight between males and females.

The mean values for SCPs of adults starved for 1 month at 20°C were found to be bimodal due to sexual dimorphism. The mean SCPs for males was lower (−17±2.6°C; 28) than that for females (−11.2±1.8°C; 26). No significant differences were observed in either mean dry weight or wet weight between males and females.

The SCPs of both fed and starved larvae, kept for 1 month at 20°C were −12.3±2.4°C (fed) and −18.0±2.6°C (starved).     

Populational fluctuation and spatial distribution of Alphitobius diaperinus (Panzer) (Coleoptera; Tenebrionidae) in a poultry house, Cascavel, Parana state, Brazil.

Knowledge of the population fluctuation and spatial distribution of pests is fundamental for establishing an appropriate control method. The population fluctuation and spatial distribution of the Alphitobius diaperinus in a poultry house in Cascavel, in the state of Parana, Brazil, was studied between October, 2001 and October 2002. Larvae and adults of the lesser mealworm were sampled weekly using Arends tube traps (n = 22) for six consecutive flock grow-outs. The temperature of the litter and of the poultry house was measured at the same locations of the tube traps. Beetle numbers increased continuously throughout all the sampling dates (average 5,137 in the first week and 18,494 insects on the sixth week). Significantly greater numbers of larvae were collected than adults (1 to 20 times in 95% of the sampling points). There was no correlation between temperature and the number of larvae and adults collected, therefore no fluctuation was observed during the sampling period. The population growth was correlated to litter re-use. The highest temperatures were observed in deep litter. The spatial distribution of larvae and adults in the poultry house was heterogeneous during the whole period of evaluation. Results suggest that monitoring in poultry houses is necessary prior to adopting and evaluating control measures due to the great variability of the insect distribution in the poultry house.     

The importance of fluctuating thermal regimes for repairing chill injuries in the tropical beetle Alphitobius diaperinus (Coleoptera: Tenebrionidae) during exposure to low temperature

Abstract. In this study, the impact of acclimation (1 month at 15 °C vs. breeding at 30 °C) and fluctuating thermal regimes (daily transfers from low temperatures to various higher temperatures for 2 h) on the cold tolerance of the tropical beetle, Alphitobius diaperinus Panzer (Coleoptera: Tenebrionidae) was examined. Acclimation increased significantly the duration of survival (Lt₅₀) at a constant 5 °C (7.7 ± 0.3 days to 9.7 ± 0.5 days). Survival of acclimated and nonacclimated beetles increased slightly at alternating temperatures of 5 °C/10 °C or 5 °C/15 °C. When daily transfer to 20 °C was applied, survival (Lt₅₀) was improved markedly (nonacclimated: 15.5 ± 0.7 days, acclimated: 19.6 ± 0.6 days). The higher temperatures may allow progressive repair of injuries, and the effects of chilling may be repaired completely at 25 and 30 °C, a phenomenon recorded here for the first time. It is estimated that the theoretical upper threshold of chill injury (Th) of nonacclimated beetles is 15.1 °C whereas it is shifted down to 11.2 °C in acclimated beetles, which might enable this temperature to allow effective repair of injury.     

The genus Alphitobius Stephens (Coleoptera,Tenebrionidae, Alphitobiini) in Africa and adjacent islands

All species of the genus Alphitobius Stephens, 1829 (Alphitobiini Reitter, 1917, subfamily Tenebrioninae Latreille, 1802) from Africa and adjacent islands are revised. New species: Alphitobius capitaneus sp. n. from Kenya. New synonyms: Cryptops ulomoides Solier, 1851, syn. n. of Alphitobius diaperinus (Panzer, 1796); Alphitobius rufus Ardoin, 1976, syn. n. of Alphitobius hobohmi Koch, 1953); Peltoides (Micropeltoides) crypticoides Pic, 1916, syn. n. of Peltoides (Micropeltoides) opacus (Gerstaecker, 1871), comb. n. Homonym: Alphitobius ulomoides Koch, 1953 = Alphitobius arnoldi nom. n. New combinations from Alphitobius: Ulomoides basilewskyi (Ardoin, 1969), comb. n.; Peltoides (Micropeltoides) opacus (Gerstaecker, 1871), comb. n. Figures of all examined species are added and a species key is compiled.     

Selection of Beauveria bassiana (Bals.) Vuill. and Metarhizium anisopliae (Metsch.) Sorok. isolates against Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae)

This study was carried out to evaluate entomopathogenic fungi isolates as microbial control agents ofthe lesser mealworm, Alphitobius diaperinus (Panzer). Larvae and adults were inoculated with conidial suspension of 99 isolates of Beauveria bassiana (Bals.) Vuill. and Metarhizium anisopliae (Metsch.) Sorok. (10(5) to 10(9) conidia/ml). Vegetative growth on culture media and sporulation on culture media, cooked rice and lesser mealworm cadavers were also evaluated. Isolates of B. bassiana were more effective than the M. anisopliae isolates and larvae were more susceptible than adults. The isolates UNIOESTE 04 and UNIOESTE 02 were originally obtained from adults of the lesser mealworm and were the most efficient isolates. In addition, UNIOESTE 04 showed high level of sporulation on different culture media and is considered a potential useful tool to lesser mealworm control.     

Controlling the mealworm Alphitobius diaperinus (Coleoptera: Tenebrionidae) in broiler and turkey houses: field trials with a combined insecticide treatment: insect growth regulator and pyrethroid

Trials were conducted during one year under field conditions to control the lesser mealworm, Alphitobius diaperinus (Panzer), in broiler and turkey houses. The tested combined treatment included an adulticidal compound (pyrethroid: cyfluthrin) and a larvicidal compound (insect growth regulator [IGR]: triflumuron). The combined insecticide treatment greatly reduced the adult and larval stocks throughout the different broiler growing periods, and control of A. diaperinus populations was achieved by the end of the second treatment. Control of the insect population in a turkey house was not similar. A reestablishment of the insect population was observed during the second turkey growing period in summer. Building characteristics and management practices of the breeding system (duration of the breeding period, management of the litter) interact with the combined insecticide treatment and lead to a different efficiency.