Kenna Mackenzie

Comparative pollination effectiveness among bees (Hymenoptera: Apoidea) on lowbush blueberry (Ericaceae: Vaccinium angustifolium)

Abstract The pollination effectiveness (floral visitation rate, percentage of flowers pollinated, and pollen deposition) of indigenous and introduced bees visiting lowbush blueberry (Vaccinium angustifolium Aiton) was studied in Nova Scotia from 1992 to 1994. Floral visitation rate alone was not a good indicator of pollination effectiveness, as not all floral visits resulted in successful pollination events. As a group, pollen-harvesting taxa pollinated >85% of flowers visited as compared with under 25% for nectar foragers. Equivalencies derived from floral visitation rates and pollination percentages show that the most effective pollen-harvesters, Bombus spp. queens and Andrena spp., would pollinate 6.5 and 3.6 flowers, respectively, in the time it would take a nectar-foraging honey bee, Apis mellifera L., to pollinate a single flower. Average pollen deposition for nectar-foragers (A. mellifera and Megachile rotundata F.) did not exceed 13 tetrads per visit, which was significantly less than all pollen-harvesters. Among pollen-harvesters, Bombus spp. workers, M. rotundata and Halictus spp. deposited moderate stigmatic loads (34, 28, and 26 tetrads, respectively), whereas Bombus spp. queens and Andrena spp. deposited >45 tetrads per single visit. Pollination equivalencies show A. mellifera would have to visit a flower four times to deposit the same amount of pollen as single visits by Bombus spp. queens or Andrena spp.

Trapping Phyllophaga spp. (Coleoptera: Scarabaeidae: Melolonthinae) in the United States and Canada using sex attractants.

Abstract The sex pheromone of the scarab beetle, Phyllophaga anxia, is a blend of the methyl esters of two amino acids, L-valine and L-isoleucine. A field trapping study was conducted, deploying different blends of the two compounds at 59 locations in the United States and Canada. More than 57,000 males of 61 Phyllophaga species (Coleoptera: Scarabaeidae: Melolonthinae) were captured and identified. Three major findings included: (1) widespread use of the two compounds [of the 147Phyllophaga (sensu stricto) species found in the United States and Canada, males of nearly 40% were captured]; (2) in most species intraspecific male response to the pheromone blends was stable between years and over geography; and (3) an unusual pheromone polymorphism was described from P. anxia. Populations at some locations were captured with L-valine methyl ester alone, whereas populations at other locations were captured with L-isoleucine methyl ester alone. At additional locations, the L-valine methyl ester-responding populations and the L-isoleucine methyl ester-responding populations were both present, producing a bimodal capture curve. In southeastern Massachusetts and in Rhode Island, in the United States, P. anxia males were captured with blends of L-valine methyl ester and L-isoleucine methyl ester. Resumen La feromona sexual del escarabajo, Phyllophaga anxia, es una mezcla de los ésteres metílicos de dos aminoácidos, L-valina y L-isoleucina. Se condujo un estudio de campo usando diferentes mezclas de los dos componentes en 59 sitios de Estados Unidos y Canada. Más de 57,000 machos de 61 especies dePhyllophaga fueron capturados e identificados. Tres de los resultados más importantes incluyen: (1) el extenso uso de los dos componentes [de las 147 especies de Phyllophaga (sensu stricto), en Estados Unidos y Canada, fueron capturados machos de cerca del 40% de ellas.]; (2) para la mayoría de las especies, la respuesta intraespecífica de los machos a las combinaciones de los dos aminoácidos fue consistente entre años diferentes, y en todos los sitios geográficos; y (3) un inusual polymorfismo de la feromona fue descrito para P. anxia. Poblaciones de algunos sitios fueron atrapados sólo con valina, mientras que poblaciones de otros sitios fueron atrapados sólo con isoleucina. También se encontraron sitios donde las poblaciones responden a ambos componentes, valina e isoleucina, produciendo una curva de captura bimodal. En el sureste del estado de Massachusetts y en Rhode Island, en Estados Unidos, machos de P. anxia fueron atrapados en trampas con mezclas de valina e isoleucina.

Mulch type and moisture level affect pupation depth of Rhagoletis mendax Curran (Diptera: Tephritidae) in the laboratory

Mulching can be beneficial for organic highbush blueberry production, but its effects on insect pests have received little attention. For pests that pupate in soil, depth may affect pupation success due to differences in temperature and moisture, mortality factors such as predation, or efficacy of controls such as insecticidal soil drenches. We examined how mulch type and moisture affect pupation depth for Rhagoletis mendax Curran (blueberry maggot), an important pest of blueberries. In laboratory studies, pupation depth was measured in wood waste compost, uncomposted pine needles, soil and sand, each at dry, field moisture levels and wet, 99% water holding capacity conditions. Pupation occurred more deeply but with greater variability in pine needle mulch compared to compost mulch, soil, or sand. Approximately 50% of maggots pupated on the surface of wet soil, however, maggots burrowed more deeply in wet than in dry pine needles. Moisture level did not significantly affect pupation depth in compost or sand. Some larvae unexpectedly escaped the stacks of cups used to assess pupation depth or died before forming a puparium. Less than 70% of stacks with dry pine needles contained pupae, and fewer pupae were recovered from wet than dry compost and soil. These results suggest that mulch material and/or moisture levels can have significant impacts on R. mendax pupation depth with potential implications for its management.

Optimization of duplex real-time PCR with melting-curve analysis for detecting the microsporidian parasites Nosema apis and Nosema ceranae in Apis mellifera

Abstract Honey bees, Apis mellifera (L.) (Hymenoptera: Apidae), are parasitized by the microsporidians Nosema apis (Zander) and Nosema ceranae (Fries). Molecular techniques are commonly used to differentiate between these parasites because light microscopy is inadequate. Our objectives were to (i) adapt the previously published duplex polymerase chain reaction (PCR) targeting the 16S rRNA gene of N. apis (321APIS-FOR, 321APIS-REV) and N. ceranae (218MITOC-FOR, 218MITOC-REV) using qualitative real-time PCR assay with SYBR® Green I dye (R-T PCR) and DNA melting-curve analysis, and (ii) determine whether the two Nosema species can be detected simultaneously in honey bees. Total spore counts and purified total genomic DNA were obtained from 37 bee samples (19 individual workers and 18 pooled samples of 15 workers) collected in Nova Scotia, Prince Edward Island, and Newfoundland, Canada. Overall, the prevalence of Nosema species was 86.5% (32/37 samples of bee DNA), based on conventional PCR and the optimized R-T PCR assay. The melting-curve analysis showed three groups of curve profiles that could determine the prevalence of N. apis, N ceranae, and co-infection (21.9%, 56.2%, and 21.9%, respectively). The duplex R-T PCR assay was efficient, specific, and more sensitive than duplex conventional PCR because co-infection was identified in 5.4% (n = 2) more samples. Sequencing of R-T PCR products confirmed the results of the melting-curve analysis. Duplex R-T PCR with melting-curve analysis is a sensitive and rapid method of detecting N. apis, N. ceranae, and co-infection in honey bees.

Characterization of early floral damage by cranberry tipworm ( Dasineura oxycoccana Johnson) as a precursor to reduced fruit set in rabbiteye blueberry ( Vaccinium ashei Reade)

The cranberry tipworm, Dasineura oxycoccana, is an important insect pest of rabbiteye blueberries, Vaccinium ashei, in the southeastern United States, with annual losses from infestation and damage exceeding

Ground and Rove Beetles (Coleoptera: Carabidae and Staphylinidae) are Affected by Mulches and Weeds in Highbush Blueberries

20 million USD. Using rabbiteye flowers from a range of developmental stages, we examined floral tissues microscopically to determine specific areas targeted by D. oxycoccana, as well as the impact of oviposition and larval feeding. Our results indicate that oviposition by D. oxycoccana occurs early in the growing season, as larvae were found exclusively between bracts of expanding inflorescences and the base of developing flowers. When bracts were removed, tissue necrosis resulting from larval feeding was found encompassing the calyx tube region near the pedicel. Such damage potentially leads to premature floral bud abscission, or aesthetically compromised fruit when mature. In the field, evidence of D. oxycoccana infestation would be undetectable until overlying bracts had abscised, and damage had already progressed to an extreme state.

Kaolin Affects Blueberry Maggot Behavior on Fruit

ASTRACT Biological control of insects by predators may be indirectly influenced by management practices that change the invertebrate community in agroecosystems. In this study we examined effects that mulching and weeding have on predatory beetles (Carabidae and Staphylinidae) and their potential prey in a highbush blueberry field. We compared beetle communities in unweeded control plots to those that were weeded and / or received a single application of compost or pine needle mulch. Compost mulch and weeding significantly affected the carabid community while the staphylinid community responded to compost and pine needle mulches. Effects because of mulch tended to intensify in the year after mulch application for both families. Estimates of species richness and diversity for Carabidae and Staphylinidae were similar in all plot types, but rarefaction curves suggested higher Carabidae richness in unmulched plots despite fewer individuals captured. Carnivorous Carabidae, dominated by Pterostichus melanarius, were most frequently captured in compost plots both years, and omnivores were most frequently captured in unweeded compost. Density of millipedes, the most abundant potential prey, was generally greater in mulched plots, whereas seasonal abundance of small earthworms varied among mulch types. Our results have potential implications for biological control in mulched highbush blueberries depending on beetle consumption rates for key pests and how rates are affected by alternative prey.

Laboratory studies of insecticide efficacy and resistance in Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) populations from British Columbia, Canada.

Abstract This study assessed the effects of Surround (kaolin) on several behavioral parameters of female blueberry maggot, Rhagoletis mendax Curran (Diptera: Tephritidae). First fruit visited, walking, cleaning, and oviposition behavior were quantified in two-choice and no-choice assays where females encountered Surround-treated and untreated fruit of highbush blueberries (Vaccinium spp.). In two-choice assays, females had a propensity (68%) to first visit untreated blueberries. In two-choice and no-choice tests, number of walking bouts and duration of walking bouts were significantly shorter on the Surround-treated than on untreated fruit. Few oviposition attempts on fruit were observed, irrespective of treatments and assays. Chromameter measurements showed significant modification of the blueberry skin color parameters lightness, hue, and saturation between untreated fruit compared with fruit dipped once or twice in a suspension of Surround. Oviposition trials with field-treated fruit showed that blueberries treated with Surround had fewer oviposition scars than the control, and this was more pronounced with weekly applications of Surround. Uneven coating of the fruit by Surround in field applications may have resulted in higher acceptance rates by flies than in blueberries individually dipped and used in laboratory trials.

Emergence of Blueberry Maggot Flies (Diptera: Tephritidae) From Mulches and Soil at Various Depths

BACKGROUND Spotted wing drosophila (SWD), Drosophila suzukii (Matsumura), control can be improved with better knowledge of insecticide efficacy, timing of sprays and rotations with registered products during the crop year. An efficacy bioassay was applied to study the dose response of adult laboratory-reared SWD collected from a cherry orchard in British Columbia (BC), Canada, to 11 insecticides. RESULTS Age- and sex-specific toxicities with the most effective insecticide, malathion, determined that older male SWD were significantly more susceptible than females. The selection for polygenic resistance to malathion using repeated exposure to sublethal concentrations with both the original and an additional blueberry-collected population of laboratory-reared SWD determined no significant difference in susceptibility after 30 generations. CONCLUSION Possible reasons reported in D. melanogaster studies where resistance did not occur after insecticide selection included avoidance and associated microbial complexes. The low level of malathion resistance observed in the selected SWD populations and a field-collected population is useful information in the development of insecticide resistance management programmes.

Investigation of a Possible Sexual Function Specialization in the Lowbush Blueberry (Vaccinium angustifolium Aiton. Ericaceae)

ABSTRACT Control of blueberry maggot, Rhagoletis mendax Curran, typically is achieved with insecticides targeting adult flies before females oviposit in ripening fruit. Management strategies targeting other life stages have received less attention. We tested effects of compost or pine needle mulches on emergence of blueberry maggot flies under laboratory and field conditions. Few flies emerged from pupae that were buried under 20 cm of pine needles in all experiments, but burial in 20 cm of compost did not always result in low fly emergence. Burial of pupae in 5 cm of compost or pine needles did not reduce fly emergence compared with 1 cm in soil. Low emergence with increased mulch depth appeared to be primarily because of failure of flies to ascend to the surface after they exited puparia. Low emergence also was associated with high moisture levels causing rotten, discolored pupae, particularly in the laboratory in compost. No flies emerged from pupae buried in 1 cm of pine needles in the field. In this case no flies exited puparia, likely because high temperatures (>30°C) at the surface killed pupae. Thus, mulch application under highbush blueberries (Vaccinium corymbosum L.) after maggots drop from berries can reduce emergence success of flies from buried pupae, but the level of control will depend on mulch depth and may vary with rainfall and temperature.