Effect of Temperature, Leaf Wetness Period, and Cultivar Susceptibility on Boxwood Blight Disease Development and Sporulation.

Boxwood blight causes great losses to the boxwood nursery industry and landscapes in 30 states in the United States. Understanding the epidemiological factors governing disease development will be important for disease forecasting and design of best management practices. We evaluated the effect of leaf wetness period (lwp) and temperature on lesion development and sporulation on three boxwood cultivars under controlled conditions to develop predictive models for disease development. We conducted detached leaf assays at 18 to 27°C and various lwp with the cultivars Buxus sempervirens 'Suffruticosa' (highly susceptible), B. sempervirens × B. microphylla var. koreana 'Green Velvet' (moderately susceptible), and B. microphylla var. japonica 'Winter Gem' (less susceptible). Detached leaves were inoculated with 200 conidia in 50 µl of suspension and disease incidence was recorded at 3 to 13 days postinoculation (dpi). Cultivar, lwp, temperature, and most interactions significantly influenced disease development. A minimum of 5 h of leaf wetness was required for any disease. Lesion development increased most rapidly between 12 and 15 h and continued to increase to about 21 h of leaf wetness. Temperatures between 21 and 25°C were optimal for lesion development. There was about a 7-day lag between appearance of lesions and maximal incidence of sporulation. The two less-susceptible cultivars had fewer lesions than Suffruticosa under the same infection conditions; in addition, leaf lesions of Winter Gem exhibited delayed sporulation and sporulation from a smaller proportion of symptomatic leaves. Response surfaces were developed for each cultivar to predict the disease incidence using the lwp and dpi. Our findings will help refine disease forecast models to improve management of boxwood blight.

What's under the Christmas Tree? A Soil Sulfur Amendment Lowers Soil pH and Alters Fir Tree Rhizosphere Bacterial and Eukaryotic Communities, Their Interactions, and Functional Traits.

In this study, we describe the legacy effects of a soil sulfur amendment experiment performed 6 years prior and the resulting alterations to the rhizosphere communities of fir trees on a Christmas tree plantation. The pH of bulk soil was ∼1.4 pH units lower than that of untreated soils and was associated with reduced Ca, Mg, and organic matter contents. Similarly, root chemistry differed due to the treatment, with roots in sulfur-amended soils showing significantly higher Al, Mn, and Zn contents and reduced levels of B and Ca. 16S rRNA and 18S rRNA gene sequencing was pursued to characterize the bacterial/archaeal and eukaryotic communities in the rhizosphere soils. The treatment induced dramatic and significant changes in the microbial populations, with thousands of 16S rRNA gene sequence variants and hundreds of 18S rRNA gene variants being significantly different in relative abundances between the treatments. Additionally, co-occurrence networks showed that bacterial and eukaryotic interactions, network topology, and hub taxa were significantly different when constructed from the control and treated soil 16S and 18S rRNA gene amplicon libraries. Metagenome sequencing identified several genes related to transport proteins that differentiated the functional potentials of the communities between treatments, pointing to physiological adaptations in the microbial communities for living at altered pH. These data show that a legacy of soil acidification increased the heterogeneity of the soil communities as well as decreasing taxon connections, pointing to a state of ecosystem instability that has potentially persisted for 6 years. IMPORTANCE We used sulfur incorporation to investigate the legacy effects of lowered soil pH on the bacterial and eukaryotic populations in the rhizosphere of Christmas trees. Acidification of the soils drove alterations of fir tree root chemistry and large shifts in the taxonomic and functional compositions of the communities. These data demonstrate that soil pH influences are manifest across all organisms inhabiting the soil, from the host plant to the microorganisms inhabiting the rhizosphere soils. Thus, this study highlights the long-lasting influence of altering soil pH on soil and plant health as well as the status of the microbiome.

Phytophthora abietivora, A New Species Isolated from Diseased Christmas Trees in Connecticut, U.S.A.

A number of fir species (Abies) are produced as Christmas trees around the world. In particular, Fraser fir (Abies fraseri (Pursh) Poir.) is popular as it yields high-quality Christmas trees in temperate North America and Europe. A Phytophthora sp. causing root rot on Fraser fir was isolated from a Christmas tree farm in Connecticut, U.S.A., and found to be new to science according to morphological and molecular phylogenetic analysis using multilocus DNA sequences from ITS, Cox1, ß-Tub, Nadh1, and Hsp90 loci. Thus, it was described and illustrated as Phytophthora abietivora. An informative Koch's postulates test revealed that P. abietivora was the pathogen causing root rot of Fraser fir.

Tracking Pesticide Residues to a Plant Genus Using Palynology in Pollen Trapped from Honey Bees (Hymenoptera: Apidae) at Ornamental Plant Nurseries.

Worldwide studies have used the technique of pollen trapping, collecting pollen loads from returning honey bee (Apis mellifera L.) (Hymenoptera: Apidae) foragers, to evaluate the exposure of honey bees to pesticides through pollen and as a biomonitoring tool. Typically, these surveys have found frequent contamination of pollen with multiple pesticides, with most of the estimated risk of acute oral toxicity to honey bees coming from insecticides. In our survey of pesticides in trapped pollen from three commercial ornamental plant nurseries in Connecticut, we found most samples within the range of acute toxicity in a previous state pollen survey, but a few samples at one nursery with unusually high acute oral toxicity. Using visual sorting by color of the pollen pellets collected in two samples from this nursery, followed by pesticide analysis of the sorted pollen and palynology to identify the plant sources of the pollen with the greatest acute toxicity of pesticide residues, we were able to associate pollen from the plant genus Spiraea L. (Rosales: Rosaceae) with extraordinarily high concentrations of thiamethoxam and clothianidin, and also with high concentrations of acephate and its metabolite methamidophos. This study is the first to trace highly toxic pollen collected by honey bees to a single plant genus. This method of tracking high toxicity pollen samples back to potential source plants could identify additional high-risk combinations of pesticide application methods and timing, movement into pollen, and attractiveness to bees that would be difficult to identify through modeling each of the contributing factors.

Sucrose Improves Insecticide Activity Against Drosophila suzukii (Diptera: Drosophilidae).

The addition of sucrose to insecticides targeting spotted wing drosophila, Drosophila suzukii (Matsumura), enhanced lethality in laboratory, semifield, and field tests. In the laboratory, 0.1% sucrose added to a spray solution enhanced spotted wing drosophila feeding. Flies died 120 min earlier when exposed to spinosad residues at label rates enhanced with sucrose. Added sucrose reduced the LC50 for dried acetamiprid residues from 82 to 41 ppm in the spray solution. Laboratory bioassays of spotted wing drosophila mortality followed exposure to grape and blueberry foliage and/or fruit sprayed and aged in the field. On grape foliage, the addition of 2.4 g/liter of sugar with insecticide sprays resulted in an 11 and 6% increase of spotted wing drosophila mortality at 1 and 2 d exposures to residues, respectively, averaged over seven insecticides with three concentrations. In a separate experiment, spinetoram and cyantraniliprole reduced by 95-100% the larval infestation of blueberries, relative to the untreated control, 7 d after application at labeled rates when applied with 1.2 g/liter sucrose in a spray mixture, irrespective of rainfall; without sucrose infestation was reduced by 46-91%. Adding sugar to the organically acceptable spinosyn, Entrust, reduced larval infestation of strawberries by >50% relative to without sugar for five of the six sample dates during a season-long field trial. In a small-plot field test with blueberries, weekly applications in alternating sprays of sucrose plus reduced-risk insecticides, spinetoram or acetamiprid, reduced larval infestation relative to the untreated control by 76%; alternating bifenthrin and phosmet (without sucrose) reduced infestation by 65%.

Use of Early Ripening Cultivars to Avoid Infestation and Mass Trapping to Manage Drosophila suzukii (Diptera: Drosophilidae) in Vaccinium corymbosum (Ericales: Ericaceae).

Use of early ripening highbush blueberry cultivars to avoid infestation and mass trapping were evaluated for managing spotted wing drosophila, Drosophila suzukii (Matsumura). Fourteen highbush blueberry cultivars were sampled for spotted wing drosophila infestation. Most 'Earliblue', 'Bluetta', and 'Collins' fruit were harvested before spotted wing drosophila oviposition commenced, and so escaped injury. Most fruit from 'Bluejay', 'Blueray', and 'Bluehaven' were also harvested before the first week of August, after which spotted wing drosophila activity led to high levels of blueberry infestation. In a separate experiment, damage to cultivars was related to the week in which fruit were harvested, with greater damage to fruit observed as the season progressed. Attractant traps placed within blueberry bushes increased nearby berry infestation by 5%, irrespective of cultivar and harvest date. The significant linear reduction in infestation with increasing distance from the attractant trap suggests that traps are influencing fly behavior to at least 5.5 m. Insecticides applied to the exterior of traps, compared with untreated traps, revealed that only 10-30% of flies visiting traps enter the traps and drown. Low trap efficiency may jeopardize surrounding fruits by increasing local spotted wing drosophila activity. To protect crops, traps for mass trapping should be placed in a perimeter outside fruit fields and insecticides need to be applied to the surface of traps or on nearby fruit to function as an attract-and-kill strategy.

Clothianidin and Imidacloprid Residues in Poa annua (Poales: Poaceae) and Their Effects on Listronotus maculicollis (Coleoptera: Curculionidae).

Competitive enzyme-linked immunosorbent assay was used to quantify the amounts of the neonicotinoids clothianidin and imidacloprid in Poa annua L. clippings from treated golf course fairways. Average clothianidin residues 7 d after application ranged from 674 to 1,550 ng/g tissue in 2012 and 455-2,220 ng/g tissue in 2013. Average clothianidin residues the day of application ranged from 17,100-38,800 ng/g tissue in 2014. Average imidacloprid residues 7 d after treatment ranged from 1,950-3,030 ng/g tissue in 2012 and 7,780-9,230 ng/g tissue in 2013. Average imidacloprid residues the day of application ranged from 31,500-40,400 ng/g tissue in 2014. Neonicotinoid or bifenthrin-neonicotinoid combination products applied in field plots in 2012 did not significantly reduce the numbers of larvae relative to the untreated control. However, in 2013, statistically significant reductions in the numbers of larvae recovered from treated field plots were associated with the presence of bifenthrin alone or when used in combination with neonicotinoid active ingredients. Listronotus maculicollis (Kirby) adults caged on neonicotinoid-, bifenthrin-, and bifenthrin-neonicotinoid-treated P. annua turf plugs fed on P. annua leaves, but mortality was only highly significantly different between treated and untreated foliage when weevils were placed on treated foliage the day after treatment and allowed to feed for 7 d. The modest degree of population suppression with bifenthrin in these experiments may not be adequate to justify the continued use of these products due to the increased risk of insecticide resistance and disruption of biological control.

Comparison of a synthetic chemical lure and standard fermented baits for trapping Drosophila suzukii (Diptera: Drosophilidae).

We determined the attractiveness of a new chemical lure compared with fermented food baits in use for trapping Drosophila suzukii Matsumura, spotted wing drosophila (Diptera: Drosophilidae), in Connecticut, New York, and Washington in the United States and at Dossenheim in Germany. The chemical lure (SWD lure) and food baits were compared in two types of traps: the dome trap and a cup trap. Regardless of trap type, numbers of male and female D. suzukii trapped were greater with the SWD lure compared with apple cider vinegar (ACV) baits at the Washington and New York sites, and were comparable with numbers of D. suzukii captured with a wine plus vinegar bait (W + V) at Germany site and a combination bait meant to mimic W + V at the Connecticut site. Averaged over both types of attractants, the numbers of D. suzukii captured were greater in dome traps than in cup traps in New York and Connecticut for both male and female D. suzukii and in Washington for male D. suzukii. No such differences were found between trap types at the Washington site for female and Germany for male and female D. suzukii. Assessments were also made of the number of large (>0.5 cm) and small (<0.5 cm) nontarget flies trapped. The SWD lure captured fewer nontarget small flies and more large flies compared with ACV bait in New York and fewer nontarget small flies compared with W + V in Germany, although no such differences were found in Washington for the SWD lure versus ACV bait and in Connecticut for the SWD lure versus the combination bait, indicating that these effects are likely influenced by the local nontarget insect community active at the time of trapping. In New York, Connecticut, and Germany, dome traps caught more nontarget flies compared with cup traps. Our results suggest that the four-component SWD chemical lure is an effective attractant for D. suzukii and could be used in place of fermented food-type baits.

Susceptibility of Cimex lectularius (Hemiptera: Cimicidae) to pyrethroid insecticides and to insecticidal dusts with or without pyrethroid insecticides.

Relative increases of bed bug, Cimex lectularius L., populations are probably due in large measure to their resistance to pyrethroids, which have been used extensively against urban pests. A Connecticut population of bed bugs was assessed for sensitivity to pyrethroids and exposed to commonly-used commercial insecticides applied to various substrates on which the residues were allowed to age for 0-24 wk. Type I and type II pyrethroids differed in toxicity when applied at a high dosage (1 microg) per bed bug. Some type II pyrethroids (cyfluthrin, lambda-cyhalothrin, cis-cypermethrin, and deltamethrin) caused > 80% mortality, whereas exposure to type I pyrethroids caused < 5% mortality over 72 h (with one exception, pyrethrins caused 23% mortality). Dust products were not affected by residue aging; mortality response over time of exposure closely fit (R2 > 0.95) an exponential rise to a maximum model from which the survival half-life (S1/2) was calculated directly. Tempo Dust (Bayer Environmental Science, Montvale, NJ) killed bed bugs relatively quickly, as did Syloid 244 (Grace Davison, Columbia, MD) and Drione (Bayer Environmental Science, Montvale, NJ) on hardboard and mattress fabric substrates (S1/2 < 1 d); DeltaDust (Bayer Environmental Science, Montvale, NJ) provided a relatively slow kill (S1/2 approximately equal to 3.5 d). The sprayable pyrethroids, Cyonara 9.7 (Insecticide Control solutions, Pasadena, TX) and D-Force HPX Aerosol 0.06% (Waterbury Companies, Waterbury, CT), displayed reduced residual toxicity as they aged; the mortality was < 50% on some substrates after 4 d. Desiccant dusts, with their physical mode of action and long residual activity, appear to be superior to sprayable pyrethroid products for killing bed bugs.

Optimizing a basal bark spray of dinotefuran to manage armored scales (Hemiptera: Diaspididae) in Christmas tree plantations.

The armored scales Fiorinia externa Ferris and Aspidiotus cryptomeriae Kuwana (Hemiptera: Diaspididae) are increasingly damaging to Christmas tree plantings in southern New England. The systemic insecticide dinotefuran was investigated for selectively suppressing armored scale populations relative to their natural enemies in cooperating growers' fields in 2008 and 2009. Banded soil application of dinotefuran resulted in poor control. However, a dinotefuran spray applied to the basal 25 cm of trunk resulted in its absorption through the bark, translocation to the foliage, and good efficacy. The basal bark spray did not significantly impact the activity of predators Chilocorus stigma (Say) or Cybocephalus nipponicus Enrody-Younga and in 2009 showed a dosage-dependent improvement in the percentage of scales parasitized by Encarsia citrina Craw. A field dosage-response factorial experiment revealed that a 0.25% (vol:vol) addition of a surfactant with dinotefuran did not enhance insecticidal effect. Probit-transformed scale population reduction relative to the untreated check was subjected to linear regression analysis; reduction of scale populations was proportional to the log of insecticide dosage, whereas basal bark spray efficacy declined in proportion to the cube of tree height. The regression equation can be used to optimize dosage relative to tree height. Excellent efficacy resulted from basal bark spray application dates of 28 April (prebud break) to mid-June, but earlier spray timing within that treatment window had fewer crawlers discoloring new growth with their short-lived feeding. A basal bark spray of dinotefuran is well suited for integration with natural enemies to manage armored scales in Christmas tree plantations.

Pyrethroid resistance mediated by enzyme detoxification in Listronotus maculicollis (Coleoptera: Curculionidae) from Connecticut.

In 2009, pyrethroid resistance was confirmed for seven "annual bluegrass weevil" Listronotus maculicollis Kirby (Coleoptera: Curculionidae) adult populations from southern New England. The mechanisms responsible for conferring this resistance were unknown. In this study, topical application bioassays with bifenthrin and bifenthrin combined with synergists affecting three detoxification systems were conducted on four field-collected adult populations of L. maculicollis from Connecticut to determined whether cytochrome P450 monooxgenases (P450s), glutathione S-transferases (GSTs), and/or carboxyl-esterases (COEs) mediated metabolic detoxification. Because a susceptible L. maculicollis laboratory strain does not exist, the most susceptible field-collected population (New Haven) provided a baseline against which all other populations were compared. In the population with the lowest resistance (Norwich), only detoxification by P450s was significant. Detoxification in the population with the second highest level of resistance (Stamford) involved both P450s and GSTs. Detoxification in the population with the highest level of resistance (Hartford) involved P450s, GSTs, and COEs. This study suggests that enzyme-mediated metabolic detoxification plays an important role in annual bluegrass weevil pyrethroid resistance.

Pyrethroid resistance in populations of Listronotus maculicollis (Coleoptera: Curculionidae) from southern New England golf courses.

Topical application bioassays of bifenthrin and lambda-cyhalothrin were conducted on field-collected populations of Listronotus maculicollis Kirby (Coleoptera: Curculionidae) from eight southern New England golf courses, six in Connecticut, one in Massachusetts, and one in Rhode Island. Concentrations versus mortality regression lines were estimated to compare the LD50 values and resistance ratios for each insecticide and each population. The LD50 ranges for bifenthrin and lambda-cyhalothrin were 1.80-244.67 ng per insect and 0.52-159.53 ng per insect, respectively. The field-collected strains showed low to high levels of resistance to bifenthrin (6.1-135.9-fold) and lambda-cyhalothrin (28.7-306.8-fold). This is the first report of insecticide resistance in this species. The loss of pyrethroid efficacy is making control of Listronotus maculicollis more challenging.

Metarhiziopsis microspora gen. et sp. nov. associated with the elongate hemlock scale.

A sporodochial fungus collected from the elongate hemlock scale, Fiorinia externa (Ferris) in Coventry, Connecticut, is described. This fungus has characteristics of both Metarhizium and Myrothecium but develops setae surrounding white to buff sporodochia and dry conidia in chains, a combination of characters found in neither genus. Phylogenetic analyses of the complete small subunit ribosomal DNA (ssu), partial ef1-alpha, and complete 5.8S ribosomal DNA and internal transcribed spacers (ITS) 1 and 2 shows that the fungus is allied with a subclade within Cordyceps including the species C. agriota, which places this fungus in the Hypocreales, Clavicipitaceae sensu lato or the newly erected Ophioclavicipitaceae. Morphological observation and molecular analysis indicate that this fungus is sufficiently different from Metarhizium and Myrothecium to warrant the erection of a new anamorphic genus. Therefore Metarhiziopsis microspora gen. et sp. nov. is proposed.

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

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 147 Phyllophaga (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.