A Combination Sulfuryl Fluoride and Propylene Oxide Treatment for Trogoderma granarium (Coleoptera: Dermestidae).

The khapra beetle, Trogoderma granarium, is an important quarantine pest of stored-products. While it is not established in the North America, frequent interceptions highlight the need for treatments to eradicate isolated populations if they occur. Methyl bromide has long been the standard fumigant used for this purpose; however, increasing restrictions on its use necessitate the development of new treatment options. Here we evaluate a treatment for khapra beetle using a combination of sulfuryl fluoride (SF) and propylene oxide (PPO). Experiments were conducted to determine the most tolerant stages and the effective rates for each compound. Combination treatments using both products were then evaluated at several temperatures to determine effective doses for quarantine level control. The egg stage was by far the most tolerant to SF, requiring concentration × time (CT) products roughly 10-fold greater than the next most tolerant stage, diapausing larvae. Diapausing larvae were significantly more tolerant to SF than non-diapausing larvae in a recently collected 'field' strain, but not in a more than 30-yr-old 'laboratory' culture. PPO treatments were shown to completely control the egg stage at CTs greater than 155 hr-mgL-1 at both 10 and 20°C. Resulting combination treatments with initial dose rates of 96 and 40 mgL-1 at 10°C and 80 and 40 mgL-1 at 20°C are proposed for SF and PPO, respectively. These combination treatments utilize SF's efficacy against the larval stage along with PPO's superior ovicidal properties to provide enhanced treatment efficacy over each product alone against all life stages of khapra beetle.

Occurrence and larval movement of Diatraea saccharalis (Lepidoptera: Crambidae) in seed mixes of non-Bt and Bt pyramid corn.

BACKGROUND: Larval movement of target pest populations among Bt and non-Bt plants is a major concern in the use of a seed mixture refuge strategy for Bt resistance management. In this study, occurrence and larval movement of the sugarcane borer, Diatraea saccharalis (F.), were evaluated in four planting patterns of non-Bt and Bt plants containing Genuity® SmartStax(TM) traits in 2009-2011. The four planting patterns were: (1) a pure stand of 27 Bt plants; (2) one non-Bt plant in the center, surrounded by 26 Bt plants; (3) a pure stand of 27 non-Bt plants; (4) one Bt plant in the center, surrounded by 26 non-Bt plants. Studies were conducted under four conditions: (1) open field with natural infestation; (2) greenhouse with artificial infestations; open field with artificial infestations (3) on the center plants only and (4) on every plant. The major objective of this study was to determine whether refuge plants in a seed mixture strategy could provide a comparable refuge population of D. saccharalis to a 'structured refuge' planting. RESULTS: Larvae of D. saccharalis showed the ability to move from infested plants to at least four plants away, as well as to adjacent rows, but the majority remained within the infested row. However, the number of larvae found on the non-Bt plants in the mixture plantings was not significantly reduced compared with the pure stand of non-Bt corn. CONCLUSION: The results of this study show that refuge plants in a seed mixture may be able to provide a comparable refuge population of D. saccharalis to a structured refuge planting.

Extended monitoring of resistance to Bacillus thuringiensis Cry1Ab maize in Diatraea saccharalis (Lepidoptera: Crambidae).

The sugarcane borer, Diatraea saccharalis (F.), is a major target of transgenic maize expressing Bacillus thuringiensis (Bt) proteins in South America and the mid-Southern region of the United States. During 2007-2009, a total of 986 feral individuals of D. saccharalis were collected from maize fields in six locations of Louisiana and Mississippi and examined for resistance to Cry1Ab maize using F 1/F 2 screens. Major resistance alleles to Cry1Ab maize in the populations sampled from non-Bt maize plants during 2007 and 2008 in Louisiana and 2009 in Mississippi were rare. From a total of 487 individuals collected from three locations in Louisiana in 2007 and 2008, only one individual was identified with major resistance alleles. In addition, no major resistance alleles were detected in 242 individuals collected from three locations in Mississippi in 2009. The frequency of major resistance alleles was estimated to be 0.002 with a 95% CI of 0.00025-0.0057 for the Louisiana populations and < 0.0061, with 95% probability, for the Mississippi populations. The resistance frequency estimated for the Louisiana populations in 2007 and 2008 was not significantly different from those reported previously for populations sampled in 2004-2006. However, among 200 individuals sampled from non-Bt maize plants in 2009 in Louisiana, six individuals were identified to possess major resistance alleles. The estimated major resistance allele frequency for the populations sampled from non-Bt maize plants in 2009 in Louisiana was 0.0176 with a 95% CI of 0.0072 to 0.0328, which was significantly greater than those estimated for the populations collected in 2004-2008. Similarly, the frequency of minor resistance alleles to Cry1Ab maize for the Louisiana populations collected in 2009 was also significantly greater than those estimated for the populations sampled before. In addition, two out of 57 feral individuals collected from Bt maize plants in Louisiana in 2009 were identified to carry major resistance alleles to Cry1Ab maize. Since 2010, transgenic maize expressing pyramided Bt genes has been planted in the US mid-Southern region and by 2011, pyramided Bt maize has replaced Cry1Ab maize as the dominant Bt maize for managing lepidopteran pests including D. saccharalis. The timely switching from single-gene Cry1Ab maize to the pyramided Bt maize should prevent further increases in Cry1Ab resistance allele frequency and thus ensure the continued success of Bt maize for managing D. saccharalis in the region.

Efficacy of genetically modified Bt toxins against insects with different genetic mechanisms of resistance.

Transgenic crops that produce Bacillus thuringiensis (Bt) toxins are grown widely for pest control, but insect adaptation can reduce their efficacy. The genetically modified Bt toxins Cry1AbMod and Cry1AcMod were designed to counter insect resistance to native Bt toxins Cry1Ab and Cry1Ac. Previous results suggested that the modified toxins would be effective only if resistance was linked with mutations in genes encoding toxin-binding cadherin proteins. Here we report evidence from five major crop pests refuting this hypothesis. Relative to native toxins, the potency of modified toxins was >350-fold higher against resistant strains of Plutella xylostella and Ostrinia nubilalis in which resistance was not linked with cadherin mutations. Conversely, the modified toxins provided little or no advantage against some resistant strains of three other pests with altered cadherin. Independent of the presence of cadherin mutations, the relative potency of the modified toxins was generally higher against the most resistant strains.

F2 screening for resistance to pyramided Bacillus thuringiensis maize in Louisiana and Mississippi populations of Diatraea saccharalis (Lepidoptera: Crambidae).

BACKGROUND: In the mid-southern region of the United States, sugarcane borer, Diatraea saccharalis (F.), is a major target pest of transgenic maize expressing Bacillus thuringiensis (Bt) proteins. Novel transgenic maize technologies containing two or more pyramided Bt genes for controlling lepidopteran pests have recently become commercially available. Insect resistance management (IRM) is an important issue in the sustainable use of Bt crop technologies. The objective of this study was to determine the frequency of resistance alleles in field populations of D. saccharalis to the new pyramided Bt maize technologies. RESULTS: A total of 382 F(2) family lines derived from 735 feral larvae/pupae of D. saccharalis collected from six locations in Louisiana and Mississippi during 2008 and 2009 were screened for resistance to three new Bt maize technologies: MON 89034, Genuity™ VT Triple Pro™ and SmartStax™. None of the 382 F(2) isoline families survived on the Bt maize leaf tissue for ≥ 12 days in the F(2) screen. The joint frequency for two- or three-gene resistance models with 95% probability in these populations was estimated to be < 0.0063 to MON 89034 and < 0.003 to VT Triple Pro™ and SmartStax™. CONCLUSION: These results suggest that the resistance allele frequency in D. saccharalis to the three pyramided Bt maize technologies is low in the mid-southern region of the United States, which should meet the rare resistance assumption of the currently used IRM strategy for Bt maize.

Suitability of different lepidopteran host species for development of Bracon hebetor (Hymenoptera: Braconidae).

Bracon hebetor Say (Hymenoptera: Braconidae) is a gregarious larval ectoparasitoid of several species of Lepidoptera that are associated with stored products. The suitability of 12 potential lepidopteran host species representing four families was studied in this study for the development and reproduction of B. hebetor. The Lepidoptera species used were the Indianmeal moth, Plodia interpunctella (Hübner), Mediterranean flour moth, Ephestia kuehniella (Zeller), almond moth, E. cautella (Walker), rice moth, Corcyra cephalonica (Walker), navel orangeworm, Amyelois transitella (Stainton), greater wax moth (laboratory reared and commercial), Galleria mellonella (Linnaeus) (all Pyralidae); tobacco budworm, Heliothis virescens (Fabricus), corn earworm, Helicoverpa zea (Boddie), beet armyworm, Spodoptera exigua (Hübner) (all Noctuidae); webbing clothes moth, Tineola bisselliella (Hummel) (Tineidae); and Angoumois grain moth, Sitotroga cerealella (Olivier) (Gelichiidae). Experiments were conducted using petri dishes (100 by 15 mm) as experimental arenas at 29 +/- 1 degrees C, 65 +/- 5% RH, and a photoperiod of 14:10 (L: D) h. B. hebetor females were introduced singly into arenas and given a full-grown host larva every day for 5 consecutive d. Paralysis of the host larvae and oviposition by B. hebetor females were significantly affected by host species. The cumulative fecundity in the 5-d period was highest on A. transitella (106.42 +/- 5.19) and lowest on T. bisselliella (9.64 +/- 1.28). The egg-to-adult survivorship and progeny sex ratio were also significantly affected by the host species. The highest percentage of parasitoid survival to the adult stage was on A. transitella (84.07 +/- 2.26) and zero on T. bisselliella. Egg to adult development time was shortest on E. cautella (9.75 +/- 0.25 d) and longest on G. mellonella (12.63 +/- 0.28 d). Results from these studies suggest that B. hebetor females can use a wide range of lepidopteran hosts for paralysis and oviposition. However, B. hebetor cannot necessarily develop and reproduce on all host species that it can paralyze and oviposit on, and optimum reproduction is with the stored-product pyralid hosts.

Effects of prior experience on host selection and host utilization by two populations of Anisopteromalus calandrae (Hymenoptera: Pteromalidae).

The effect of adult learning through an oviposition and host feeding experience was studied in two populations of Anisopteromalus calandrae (Hymenoptera: Pteromalidae), in choice and no-choice experiments in the laboratory using last instars of rice weevil, Sitophilus oryzae L., and lesser grain borer, Rhyzopertha dominica (F.), as hosts. In choice experiments, 20 wheat kernels infested by rice weevil and 20 by lesser grain borer were placed randomly in petri dish (15 by 100 mm) arenas. In no-choice experiments, 10 host-infested wheat kernels, either with rice weevils or with lesser grain borers, were mixed thoroughly in 500 g of uninfested wheat placed in a 0.945-liter glass jar (75 by 170 mm) arenas. A. calandrae females from a laboratory and field populations that were experienced with host larvae or naïve were introduced singly into each experimental arena and allowed to sting and oviposit for 24 h. An oviposition experience with rice weevil or lesser grain borer had a stronger effect on host preference in choice experiments compared with experiments with naïve wasps. Host-finding and parasitism rates were increased by prior experience with rice weevils in no-choice experiments. A. calandrae females clearly preferred rice weevil larvae for parasitization over larvae of lesser grain borer regardless of parasitoid population or prior experience. Also, A. calandrae females chose rice weevil larvae for producing female progeny and lesser grain borer for male progeny in choice experiments. The implications of the results of this study are discussed in relation to host preference and the selection of parasitoid populations for biological control programs in stored grains.

Suitability of five species of stored-product insects as hosts for development and reproduction of the parasitoid Anisopteromalus calandrae (Hymenoptera: Pteromalidae).

We investigated the ability of two populations of Anisopteromalus calandrae (Hymenoptera: Pteromalidae), to parasitize and develop on late instars of five different stored-product insects that typically complete their development inside seeds of grain or legume species or other dry commodity. The host species were the cigarette beetle, Lasioderma serricorne (F.); cowpea weevil, Callosobruchus maculatus (F.); rice weevil, Sitophilus oryzae (L.); lesser grain borer, Rhyzopertha dominica (F.); and Angoumois grain moth, Sitotroga cerealella (Olivier). Experiments were conducted in the laboratory in a no-choice design by using petri dishes (15 by 100 mm) as experimental arenas with 20 host larvae. A. calandrae females from populations originating in Georgia (GA) and Oklahoma (OK) were introduced singly into experimental arenas and allowed to sting and oviposit for 24 h. Parasitism by the OK population was greater than that for the GA population across all hosts. However, no or very low parasitism was found on Angoumois grain moth for either population in this experiment. The highest number of parasitoid progeny was recorded on cowpea weevil (15.9) followed by rice weevil (11.5) and cigarette beetle (10.8) for the OK population. A similar trend was observed in the GA population. The highest proportion of female progeny was produced on cowpea weevil (73.0%) by the OK population. Conversely, a higher proportion of female progeny was produced on rice weevil (64.6%) by the GA population than produced by the OK population. Parasitoid adults were significantly larger and heavier when they developed on cowpea weevil irrespective of parasitoid population. The possible application of these results for biological control of stored-product insects is discussed.