Search

Root weevils in the genus Otiorhynchus are an important pest in the nursery and small fruit production worldwide. The night-activity of the adult weevils obstruct timely monitoring and oviposition often starts before effective control measures are taken. The primary objective of this research goal was to develop an effective trap for monitoring that can be used in conjunction with the kairomone (Z)-2-pentenol and an effective means to kill the insects that enter the trap. A novel ruffle refuge trap (WeevilGrip) caught on average 4 to 5 times more weevils than a grooved board refuge in a field trial. Addition of the kairomone to the WeevilGrip further increased catches 52%. Linseed oil increased mortality to 59% and addition of Botanigard (ai Beauveria bassiana, strain GHA, Certis, BotaniGard WP 10–25%) increased mortality to 79%. The lure-refuge device consists of a flexible ruffle that can be wrapped around trees or placed on the soil within ground covers. This flexible shape maximizes contact with weevils compared to other available weevil trap designs. The WeevilGrip is an improved monitoring tool to support growers in integrated control strategies.

Since the arrival of Drosophila suzukii M. (Diptera: Drosophilidae), field applications of broad-spectrum insecticides have increased in berry crop production to protect susceptible fruits from infestation. Field studies were conducted from 2011 to 2013 to determine whether alternate row middle or border spray programs could manage D. suzukii as well as complete sprays and have less of an impact on non-target arthropods. Four raspberry sites and one blueberry site were alternate row middle and border sprayed, respectively. Adult and larva counts of D. suzukii were compared to conventional complete spray programs. Non-target arthropods were evaluated 7 d post-harvest. In both reduced spray trials, no differences in mean adult numbers and larvae of D. suzukii were detected between treatments. Mean counts immediately after sprays (i.e. 3–12 d after treatment) were also similar in complete and alternate row sprays. Both reduced spray strategies had significantly more Stethorus spp.; additionally, alternate row sprays had significantly more Psyllobora spp. These reduced pesticide strategies are additional tools to consider in D. suzukii IPM programs that can reduce the area sprayed, application time, and input costs while conserving natural enemies.

Drosophila suzukii (Matsumura) is an economic pest of small fruits and cherries that attacks intact ripening fruits. Host susceptibility may be influenced by characteristics such as flesh firmness, penetration force of the skin, total soluble solids (TSS, also known as °Brix), and pH. Improved knowledge of factors affecting fruit susceptibility is needed for developing thresholds and risk prediction models for IPM. A combination of laboratory and field studies was conducted to develop prediction and potential management tools. First, a direct bioassay was used to calculate the probability of oviposition in a given fruit based on various characteristics as determined across laboratory and field trials in Oregon and North Carolina, US. When multiple characteristics were evaluated simultaneously, oviposition probability consistently increased as penetration force decreased and pH increased. Oviposition probability sometimes increased as TSS increased. Second, raspberries and blueberries in unsprayed fields had substantially lower infestation in ripening fruit compared to ripe fruit. There was no or minimal infestation in green fruit. Third, given that skin penetration force influences oviposition, practices used to improve fruit quality were examined in laboratory no-choice cages for potential reduction of oviposition. Blueberry fruit sprayed with calcium silicate in the field had greater penetration force and firmness and reduced number of eggs laid by D. suzukii compared to untreated fruit. Other calcium-based treatments increased Ca content and firmness of fruit relative to untreated fruit. Timing of insecticide spray for D. suzukii might be delayed until fruit become susceptible.

Spotted wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), may utilize wild ‘Himalaya’ blackberry (HB) Rubus armeniacus Focke or other non-crop plants as refugia and possibly exploit adjacent field margins before colonizing cultivated fruiting crops. Studies were conducted to determine the role of field margins containing HB and their effect on D. suzukii activity, density and distribution in an adjacent commercial red raspberry crop. One-ha plots adjacent to field margins containing HB or known non-host (NH) grass crops were established in 2011 and 2012 and replicated three times. Each plot contained two transects with monitoring traps for D. suzukii in the field margin (0 m) and spaced approximately 10 (crop boundary), 40, 70 and 100 m into the adjacent crop (n = 10 traps/plot). Field margin vegetation was treated with a 10% chicken egg white mark solution weekly from pre-harvest until the end of harvest using a cannon sprayer. Adult D. suzukii were collected from traps weekly and analysed for the presence of the egg white mark using an egg white-specific enzyme-linked immunosorbent assay (ELISA). During both years, marked flies and total flies were captured in higher numbers in HB field margins, whereas virtually no flies were captured in field margins containing no known alternative host. Similarly, more flies were captured in the crop near HB than near NH. Spatial Analysis by Distance IndicEs (SADIE) and mean D. suzukii trap captures additionally displayed significantly higher fly densities in the raspberry field near HB than near NH. These results suggest that HB may contribute to elevated D. suzukii populations and pest pressure in comparison with field margins containing no known alternate host vegetation for D. suzukii. Having closely adjacent non-crop alternate host landscapes may result in increased D. suzukii pest pressure.

Drosophila suzukii Matsumura (Diptera: Drosophilidae) has caused significant economic damage to berry and stone fruit production regions. Markers that are systemic in plants and easily transferred to target organisms are needed to track D. suzukii exploitation of host resources and trophic interactions. High and low concentrations of the trace element, rubidium (Rb), and the stable isotope, 15N, were tested to mark D. suzukii larvae feeding on fruits of enriched strawberry plants grown in containers under greenhouse conditions. Fly marker content and proportion of flies marked 1, 7, and 14 d after emergence from enriched fruits and fly dry mass were analyzed. Nearly 100% of the flies analyzed 14 d after emerging from 15N-enriched plants were marked, whereas only 30-75% and 0-3% were marked 14 d after emerging from high and low Rb concentration plants, respectively. Rapid Rb decay, strong 15N persistence, and the economics of using these markers in the field to elucidate D. suzukii pest ecology are discussed.

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is a pest of small fruits and cherries, and has also been noted to infest a variety of wild, ornamental, and uncultivated hosts. Identify- ing alternative hosts is critical for pest management. Research objectives were to: 1) survey fruits in the field for natural infestation of D. suzukii, 2) determine the susceptibility of fruits in laboratory no-choice studies, and 3) evaluate short-range preference between simultaneously ripe alternative hosts and culti- vated fruits in laboratory choice studies. Field surveys identified new hosts or confirmed previously re- ported hosts including: Berberis aquifolium Pursh, Oregon grape; Cornus spp., dogwood; Cotoneaster lacteus W.W. Smith, milkflower cotoneaster; Elaeagnus umbellata Thunberg, Autumn olive; Frangula purshiana (de Candolle) A. Gray, cascara buckthorn; Lindera benzoin (L.) Blume, spicebush; Lonicera caerulea L., blue honeysuckle; Morus sp., mulberry; Phytolacca americana L., pokeweed; Prunus avium (L.) L., wild cherry;Prunus laurocerasus L., cherry laurel;Prunus lusitanica L., Portuguese laurel;Rubus armeniacus Focke, Himalaya blackberry; Rubus spectabilis Pursh, salmonberry; Sambucus nigra L., black elderberry; Sarcococca confusa Sealy, sweet box; Solanum dulcamara L., bittersweet nightshade; andSymphoricarpos albus (L.) S.F. Blake, snowberry. High fruit infestations were observed in S. confusa during April-May and Lonicera spp. in June before most commercial fruits ripen. From both field and laboratory studies, there was no evidence of susceptibility during the estimated ripe period Crataegus L. 'Autumn Glory,' hawthorn; Ilex crenata Thunberg, Japanese holly; Nandina domestica Thunberg, sacred bamboo; Rhaphiolepis umbellata (Thunberg) Makino, yeddo hawthorne; Rosa acicularis Lindley, prickly rose; Skimmia japonica Thunberg, Japanese skimmia; and Viburnum davidii Franchet, David's vibur- num. Lastly, laboratory choice tests identified that several fall-ripening alternative hosts were more sus- ceptible than 'Pinot noir' or 'Pinot gris' wine grapes. By understanding host use, growers can identify high-risk areas where coordinated action may reduce infestation ofD. suzukii in crops.

The recent arrival of Drosophila suzukii, an invasive pest of soft-skinned fruit with a wide host range, has resulted in increased production costs for growers and the need for additional insecticide applications each growing season. There are few effective organic insecticides for D. suzukii, and insecticide use in conventional farms may be disruptive to natural enemies, suggesting a need for effective biological control to combat D. suzukii. Commercially available natural enemies were evaluated for their potential use in augmentative releases, including: the predators Orius insidiosus and Dalotia coriaria; the entomopathogenic fungi Metarhizium anisopliae, Beauveria bassiana and Paecilomyces fumosoroseus; and the entomopathogenic nematodes Heterorhabditis bacteriophora, Steinernema feltiae and S. carpocapsae. This suite of natural enemies was chosen to target D. suzukii adults as well as larvae in hanging or dropped fruit. Of the cultured fungal strains tested, only M. anisopliae significantly decreased D. suzukii survival, but it had low residual activity and no effect on D. suzukii fecundity. O. insidiosus decreased D. suzukii survival in simple laboratory arenas but not on potted blueberries or bagged blueberry branches outdoors. D. coriaria did not decrease D. suzukii survival in infested blueberries in simple laboratory arenas. The nematodes tested showed low infection rates and were not able to affect D. suzukii survival. Although this suite of natural enemies showed limited ability to suppress D. suzukii under the tested conditions, these and related natural enemies are present as part of the endemic natural enemy community in agricultural fields, where they may contribute to D. suzukii suppression.

The yeast Candida parapsilosis is an increasingly common cause of systemic fungal infections among immunocompromised individuals, including premature infants. Adhesion to host surfaces is an important step in pathogenesis, but this process has not been extensively studied in this organism. A microfluidics assay was developed to test the ability of C. parapsilosis to adhere to immobilized host extracellular matrix proteins under physiological fluid shear conditions. Growth in mammalian tissue culture medium at 37°C for 3 to 6 h led to the induction of an adhesive phenotype at shear forces of 1 to 5 dynes/cm 2 in some isolates of C. parapsilosis . Glutamic acid, proline, and calcium appeared to be the minimally necessary requirements for increased adhesion in these assays. To determine whether genes homologous to the ALS gene family of C. albicans were important for the adhesive phenotype, the expression levels of 5 homologous C. parapsilosis genes were quantified by using quantitative PCR (qPCR) under conditions leading to increased adhesion. CPAR2_404800 ( CpALS7 ) and CPAR2_404780 showed increased expression levels compared to those in control yeast. The extent of adhesion was variable among different isolates, and linear regression identified the expression of CpALS7 but not CPAR2_404780 as having a strong positive correlation with adhesion. A homozygous CpALS7 deletion strain was deficient in adhesion, whereas the expression of CpALS7 in Saccharomyces cerevisiae resulted in increased adhesion. Together, these data provide strong evidence that CpAls7 aids in the adherence of C. parapsilosis to the extracellular matrix under shear forces and support its previously reported role in virulence.

Drosophila suzukii Matsumura (Diptera: Drosophilidae) utilizes ‘Himalaya’ blackberry, Rubus armeniacus Focke (Rosaceae), as a host and may invade berry and stone fruit crops from field margins containing this invasive weed. Laboratory and semi-field studies were conducted to determine (1) the persistence of protein marks including 10% chicken egg whites (egg albumin protein), 20% bovine milk (milk casein protein), and 20% soy milk (soy trypsin inhibitor protein) on topically sprayed D. suzukii, (2) protein retention on blackberry leaves, and (3) D. suzukii acquisition of protein afterexposureto markedblackberryleaves for upto 14 days afterapplication. Allflies and leaves wereassayedforthepresenceoftheproteinmarksusingprotein-specificenzyme-linkedimmunosorbent assays. Egg albumin, milk casein, and soy trypsin proteins persisted on 94, 49, and 25% of the topically marked D. suzukii, respectively, throughout the 14-day study period. Egg albumin was retained on 100% of treated leaves for 14 days, regardless of environmental conditions. At least 50% of flies exposed residually to egg albumin-treated leaves were marked for 3 days, regardless of exposure time and environmental conditions. However, increasing fly exposure time to treated leaves in April and June appeared to improve protein mark acquisition. Acquisition of protein by flies from treated leaves for milk casein was inconsistent, and poor for soy trypsin, despite detectable levels on treated leaves. Egg albumin had the longest and most consistent persistence on flies, leaves, and flies exposed to leaves in laboratory and semi-field studies, under a variety of environmental conditions andexposuretimes.

Environ. Entomol. 42(6): 1348Ð1355 (2013); DOI: http://dx.doi.org/10.1603/EN13148 ABSTRACT Drosophila suzukii (Matsumura), an invasive pest of small and stone fruits, has been recently detected in 39 states of the United States, Canada, Mexico, and Europe. This pest attacks ripening fruit, causing economic losses including increased management costs and crop rejection. Ongoing research aims to improve the efÞcacy of monitoring traps. Studies were conducted to evaluate how physical trap features affect captures ofD.suzukii. We evaluated Þve colors, two bait surface areas, and a top and side position for the sy entry point. Studies were conducted at 16 sites spanning seven states and provinces of North America and nine crop types. Apple cider vinegar was the standard bait in all trap types. In the overall analysis, yellow-colored traps caught signiÞcantly more sies than clear, white, and black traps; and red traps caught more than clear traps. Results by color may be insuenced by crop type. Overall, the trap with a greater bait surface area caught slightly more D. suzukii than the trap with smaller area (90 vs. 40 cm 2 ). Overall, the two traps with a side-mesh entry, with or without a protective rain tent, caught more D. suzukii than the trap with a top-mesh entry and tent.

Ornamental trees, shrubs, annuals, grasses, and other plants are cultivated and planted for aesthetic and utility purposes in urban landscapes, including yards, parks, gardens, sports fields, cemeteries, medians, and roadsides. Insecticides are widely used to control pests impacting these plants. Public concern about the adverse effects of chemical insecticides has created an environment in which biological, botanical, and less acutely toxic insecticides are regarded as desirable alternatives, especially to older insecticide active ingredients with unfavorable toxicology profiles. Several entomopathogens have been developed for managing pests in high-value ornamental crops. In this chapter we discuss prospects and progress that has been made with entomopathogens for managing pests of ornamental plants in the following groups: phytophageous mites, wood-boring beetles/caterpillars, root-feeding weevils, defoliating beetles/caterpillars, and sap-feeding insects, including aphids, whiteflies, and thrips.

Drosophila suzukii Matsumura (Diptera: Drosophilidae), a recent invasive pest of small and stone fruits, has been detected in more than half of the U.S. states, and in Canada, Mexico, and Europe. Upon discovery, several different trap designs were recommended for monitoring. This study compared the trap designs across seven states/provinces in North America and nine crop types. Between May and November 2011, we compared a clear cup with 10 side holes (clear); a commercial trap with two side holes (commercial); a Rubbermaid container with mesh lid and rain tent (Haviland), and with 10 side holes and no tent (modified Haviland); a red cup with 10 side holes (red); and a white container with mesh lid and rain tent (Van Steenwyk). Although fly catches among traps varied per site, overall, the Haviland trap caught the most D. suzukii, followed by the red, Van Steenwyk, and clear trap. The modified Haviland and commercial trap had low captures. Among five crop types in Oregon, a clear cup with mesh sid...

Black vine weevil (BVW), Otiorhynchus sulcatus (Coleoptera: Curculionidae, Fabricius), is a serious pest of cranberry, Vaccinium macrocarpon Ait. Larvae feed undetected within the soil and cause damage to roots and underground vines. We correlated damage caused by feeding larvae from known BVW egg densities. Two cultivars of potted cranberry vines, ‘Stevens’ and ‘McFarlin’, were inoculated with 0, 5, 10, 20, 40, and 80 eggs per pot. Root damage and canopy health were assessed. ‘Stevens’ exceeded ‘McFarlin’ in dry shoot weight, total shoot length, total leaf area, and dry root weight before egg treatments. Damage to underground vines increased with increasing egg density and more damage was found in ‘Stevens’ than ‘McFarlin’ at the highest egg densities. In August, plant water use and total shoot length in ‘McFarlin’ were significantly greater in plants treated with 0–5 eggs per pot compared with plants treated with 40–80 eggs per pot. The effect on total shoot length was more pronounced in October. ‘Stevens’ showed no response to increasing BVW density for up to 24 weeks. Destructive measurements showed decreased root weight in ‘McFarlin’ but not ‘Stevens’. Both cultivars showed a similar decrease in dry shoot weight, total shoot length, and percent green leaf area with increasing BVW egg density. Root damage increased as BVW egg density increased and this damage resulted in reduced plant water use for ‘McFarlin’. Reduced shoot growth and leaf area was recorded for both cultivars, although these effects were more apparent in ‘McFarlin’ and at an earlier stage than in ‘Stevens’.

Root weevils in the genus Otiorhynchus are cited as one of the most important pests in the major nursery and small fruit production areas throughout the United States, western Canada, and northern Europe. A major problem in combating weevil attack is monitoring and timing of control measures. Because of the night-activity of the adult weevils growers do not observe the emerging weevils in a timely manner and oviposition often starts before effective control measures are taken. Several vine weevil electroantennogram-active plant volatiles were identified from a preferred host plant, Euonymus fortunei. Main compounds evoking antennal responses on the weevils' antennae were (Z)-2-pentenol, (E)-2-hexenol, (Z)-3-hexenol, methyl benzoate, linalool, (E)-4,8-dimethyl-1,3,7-nonatriene, methyl eugenol, and (E, E)-alpha-farnesene. Several of these compounds were tested alone and in mixtures on attractiveness for the vine weevil Otiorhynchus sulcatus (F.) in field-grown strawberry in Oregon. O. sulcatus were attracted to (Z)-2-pentenol (approximately 3 x more than control) and a 1:1 ratio mixture of (Z)-2-pentenol and methyl eugenol (4.5 x more than control). This is the first report of field-active attractants for O. sulcatus which holds promise for the development of new monitoring strategies for growers in the near future.

BACKGROUND: The spotted wing Drosophila, Drosophila suzukii (Matsumura, 1931) (Diptera: Drosophilidae), is an invasive pest of small-fruit crops. Unlike most other Drosophila, this insect is able to oviposit into and damage ripe and ripening fruit, making it unmarketable. Because this is a new pest in the United States, it is necessary to identify registered insecticides to manage this insect effectively in conventional and organic production systems. RESULTS: The present laboratory bioassays and field trials identified a number of insecticides representing various modes of action that are effective in controlling D. suzukii. Products that performed well in the laboratory bioassay also performed well in the field, indicating that screening of new chemistries in the laboratory is a worthy exercise. Field application of pyrethoids, organophosphates or spinosyns provided 5–14 days of residual control of D. suzukii. The efficacy of the neonicotinoids as adulticides was not satisfactory compared with the other contact-mode-of-action chemistries. Based on the zero tolerance by the small-fruit industry and the individual effects mentioned above, neonicotinoids are not currently recommended for D. suzukii management. CONCLUSIONS: There are effective insecticides registered for controlling D. suzukii infestations in susceptible small-fruit crops. Copyright © 2011 Society of Chemical Industry

Spotted wing drosophila, Drosophila suzukii Matsumura, a native of eastern and southeastern Asia, is a pest of small and stone fruits. First detected in California in 2008, the insect is now found across the Pacific Coast states. Its penchant for attacking healthy, ripening fruit (as opposed to overripe and rotting fruit favored by other so-called “vinegar flies”) makes it a potential economic threat to a host of soft- and thin-skinned fruit crops including cherry, raspberry, blackberry, blueberry, strawberry, peach, plums, pluots, nectarines, juice grape, table grape, and wine grape. Coordinated research projects to determine host preference, seasonal phenology, biology, and management options are taking place among entomologists in Washington, Oregon, and California. A description of the pest and initial findings on its biology, life history, known and expected geographic range, management and monitoring techniques, and economic considerations are presented and discussed.

Filbertworm, Cydia latiferreana (Walsingham) (Lepidoptera: Tortricidae), is a key insect pest associated with hazelnuts, Corylus avellana L. (Fagales: Betulaceae), in North America. This study investigated the feasibility of entomopathogenic nematodes as an alternative strategy for filbertworm control. Laboratory and field experiments were conducted between October 2007 and May 2008 in Benton County, OR, to evaluate the ability of the nematode Steinernema carpocapsae (Weiser) (Rhabditida: Steinernematidae) to infect filbertworm larvae and pupae. The susceptibility of larvae with and without hibernacula as well as pupae to S. carpocapsae was tested in laboratory bioassays using nematode concentrations between 40 and 200 infective juveniles (IJs) per cm2. Percentage of nematode infection was not significantly affected by the presence of hibernacula (infection range, 90–92%) compared with larvae without hibernacula (80–95%), or by pupal stage (50–75%) compared with larvae (65–75%). In additional field trials, the effect of nematode rate, water application rate, and orchard floor cover on nematode efficacy was determined in October 2007 and May/June 2008. Sentinel filbertworm larvae in plots with either bare soil or debris (leaves, twigs, husks, and blank nuts) were treated with S. carpocapsae at rates ranging from 40 to 150 IJs per cm2 applied in 75 or 190 ml/m2 water. Average filbertworm mortality ranged from 2 to 11% and from 50 to 78% in October and May, respectively. Larval mortality increased with increasing nematode concentration, but was not significantly affected by water application rate or orchard floor cover. Our experiments suggest that S. carpocapsae can play a role in the control of filbertworm larvae overwintering on the orchard floor.

The ecology of fungal entomopathogens in the rhizosphere is an understudied area of insect pathology. The rhizosphere is the region of soil in which the release of root exudates influences the soil microbiota, and may provide a favorable environment for fungal entomopathogens. The objective of this review is to bring together the relatively scant data available to date on the subject of fungal entomopathogens colonizing the rhizosphere and to highlight the importance of these findings. Gaining a better understanding of the ecology of fungal entomopathogens in the rhizosphere will help in the development of successful microbial control strategies against root-feeding insect pests.

Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae) is one of the most damaging insect pests of corn. Studies were conducted to determine whether live larval counts obtained from corn whorls were predictive of the amount of larval tunneling that would result in the stalk of the plant 40 d later at the end of larval development. Whorls from plants treated with Dipel 10G (6,400 IU per whorl) and untreated controls, both infested with O. nubilalis neonates, were evaluated for the number of live larvae in 50 whorls 5, 7, 9, and 12 d after Bacillus thuringiensis (Berliner) (Bacillales: Bacillaceae) application. Forty days after larval infestation, 25 plants from each plot were split from tassel to base, and the length of larval tunneling was recorded. There was a strong relationship between numbers of live larvae in the plant whorl and the length of larval tunneling that resulted. While linear at each location, there was significant variation in the relationship among locations, indicating that comparisons could not be made between years or locations within a year. Blocks within a location on a given year did not vary significantly and reproducible results were obtained each year within a given location as well as on any of the whorl pulling dates evaluated. Because of its ease of use, predictability, and rapid return of results, we propose this technique as an additional method to shot-hole feeding and stalk splitting to evaluate the effectiveness of O. nubilalis management strategies.

The entomopathogenic fungus Metarhizium anisopliae (Metchnikoff) Sorokin (Hypocreales: Clavicipitaceae) is registered in the United States and The Netherlands for black vine weevil, Otiorhynchus sulcatus (Coleoptera: Curculionidae) control in container-grown ornamentals. These studies were conducted to determine the compatibility of M. anisopliae (F52) with a wide range of fungicides commonly applied to container-grown ornamentals for the management of soil-borne plant pathogens. The impact of fungicides on spore germination and mycelial growth were determined in vitro. In addition, M. anisopliae persistence in bulk and rhizosphere soil was determined 30 days following dual application of each fungicide at 7–28 days intervals as prescribed. A number of fungicides (thiophanate-methyl, dimethomorph, captan, triflumizole, triflozystrobin, pyraclostrobin, azoxystrobin) inhibited spore germination in vitro. A larger number of fungicides (fosetyl-AI, thiophanate-methyl, dimethomorph, captan, quintozene, triflumizole, fludioxanil, triflozystrobin, pyraclostrobin, fludiox-mefanox, iprodione, azoxystrobin, phosphorus acid/K-salts) inhibited mycelial growth in vitro. Only three fungicides (etridiazole, propamocard and mafanoxam) had no significant impact in vitro on spore germination or mycelial growth. While a number of fungicides had a detrimental impact in vitro, there was no impact on M. anisopliae populations in bulk soil following dual application of any fungicide. However, the fungicides captan and triflumizolet, which have a short reapplication interval, had a detrimental impact on M. anisopliae populations in the rhizosphere. As researchers develop rhizosphere competence as an alternative management strategy for black vine weevil, the fungicides captan and triflumizole should be avoided.

Nosema pyrausta, an entomopathogenic microsporidium, is an important population regulator of the European corn borer, Ostrinia nubilalis. This manuscript is a review of research on the relationships between N. pyrausta and O. nubilalis. N. pyrausta was described from O. nubilalis in Hungary in 1927 and from O. nubilalis in IA in 1950. It affects the basic biology of O. nubilalis by slowing larval development, reducing percentage pupation, and decreasing adult longevity, oviposition and fecundity. Infections are maintained in a population by vertical and horizontal transmission. Success of vertical transmission depends on intensity of infection. Horizontal transmission is dependent on stage of larval development at time of infection, quantity of inoculum, and host density. Abiotic and biotic factors coupled with N. pyrausta usually have an additive effect in decreasing the fitness of O. nubilalis, i.e., cold temperatures reduce fecundity and increase larval mortality, host plant resistance reduces the number of larvae per plant. Also, microbial and chemical insecticides are more effective in reducing plant feeding if the insect is infected with N. pyrausta. Predators in general feed on N. pyrausta-infected O. nubilalis with no decrease in fitness. Parasitoids do coexist with N. pyrausta, however, parasitoid fecundity is usually reduced when developing in a N. pyrausta-infected host. Previously unreported data are presented on the prevalence of N. pyrausta in O. nubilalis populations from many parts of the US. These data demonstrate that N. pyrausta continues to be present and fluctuate in populations of O. nubilalis as it has since its discovery in the US. Also, the dynamics of its presence remain similar through changes in corn production including crop rotations, reduced tillage and transgenic insect-resistant varieties.

Summary Pseudomonas fluorescens CHA0 and the related strain Pf-5 are well-characterized representatives of rhizosphere bacteria that have the capacity to protect crop plants from fungal root diseases, mainly by releasing a variety of exoproducts that are toxic to plant pathogenic fungi. Here, we report that the two plant-beneficial pseudomonads also exhibit potent insecticidal activity. Anti-insect activity is linked to a novel genomic locus encoding a large protein toxin termed Fit (for P. fluorescens insecticidal toxin) that is related to the insect toxin Mcf (Makes caterpillars floppy) of the entomopathogen Photorhabdus lumine- scens, a mutualist of insect-invading nematodes. When injected into the haemocoel, even low doses of P. fluorescens CHA0 or Pf-5 killed larvae of the tobacco hornworm Manduca sexta and the greater wax moth Galleria mellonella. In contrast, mutants of CHA0 or Pf-5 with deletions in the Fit toxin gene were significantly less virulent to the larvae. When expressed from an inducible promoter in a non-toxic Escherichia coli host, the Fit toxin gene was sufficient to render the bacterium toxic to both insect hosts. Our findings establish the Fit gene products of P. fluorescens CHA0 and Pf-5 as potent insect toxins that define previously unappreciated anti-insect properties of these plant-colonizing bacteria.

The objective of this study was to determine the susceptibility of the two primary direct insect pests of hazelnuts in Oregon to three species of entomopathogenic nematodes. The entomopathogenic nematodes (Heterorhabditis marelatus Pt. Reyes, Steinernema carpocapsae All and Steinernema kraussei L137) were used in laboratory soil bioassays to determine their virulence against filbertworm, Cydia latiferreana (Walsingham) (Lepidoptera: Tortricidae) and filbert weevil, Curculio occidentalis (Casey) (Coleoptera: Curculionidae). All three nematode species were infective in laboratory bioassays. Infectivity ranged from 73-100% and 23-85% for filbertworm and filbert weevil, respectively. Field results were similar to those found in the laboratory with filbertworm larvae being more susceptible to nematode infection.

The black vine weevil (BVW), Otiorhynchus sulcatus (F.) is a serious pest of nursery crops. The fungus, Metarhizium anisopliae (F52), is registered by the U.S. Environmental Protection Agency for BVW control. The objectives of this study were to determine the efficacy of a curative drench application of M. anisopliae for controlling BVW larval infestations in container-grown nursery plants and the effect of temperature on the rate of fungal growth and speed of kill. Trials evaluating the efficacy of M. anisopliae as a curative application were performed in the spring of 2004 and 2005 as well as the fall of 2006. Laboratory studies were performed to quantify the impact of temperature (10, 15, 20, 24 and 28C) on fungal growth and speed of kill. Metarhizium anisopliae applied in the greenhouse and outdoors in 2004 were 92 and 30% effective, respectively. Fungal applications to container-grown plant material maintained outdoors in the spring of 2005 were nearly 100% effective 28 days after application. Fall applications in 2006 provided statistically significant reductions in the number of live BVW larve per pot, but were not as effective as spring applications in 2005. The mean media temperature of containers maintained outdoors in the fall of 2006 dropped considerably (10–12C) over the course of the experiment and were likely the cause for the reduced efficacy. Laboratory experiments demonstrated that temperatures below 20C (68F) significantly slowed fungal growth and the speed at which M. anisopliae infected BVW larvae. In the field, drench applications of M. anisopliae were very effective at eliminating BVW larvae in container-grown nursery plants when media temperatures were adequate (> 15C (59F)). The use of M. anisopliae as a curative drench application has similar temperature-dependent limitations as the use of entomopathogenic nematodes for BVW control. Therefore, applications should occur as early in the fall as possible once egg laying has ended or in late spring just prior to pupation when media temperatures would be most conducive to fungal infection.

The purpose of this study was to characterize the behavior of black vine weevil larvae, Otiorhynchus sulcatus (F.), in the presence of two possible control options: the synthetic pyrethroid bifenthrin and the entomopathogenic fungus Metarhizium anisopliae (Metch.) Sorokin. Five third-instar black vine weevil were placed in two-choice soil olfactometers that allowed larvae to infest one of two pots. Larvae were allowed to choose between M. anisopliae (1 × 106 spores/g dry media) and untreated media, bifenthrin (25 ppm) and untreated media, as well as M. anisopliae– and bifenthrin-treated media. For all comparisons, experiments were conducted without plants in the system to test for innate responses, as well as with plants to test host–plant influence. Larvae were significantly deterred by bifenthrin without plants present in the system. No significant effect on larval preference was observed when M. anisopliae was present in media for trials without plants. M. anisopliae–treated media was preferre...

The black vine weevil (BVW), Otiorhynchus sulcatus (F.) is a serious pest of nursery crops, particularly in the Pacific Northwest. The fungus, Metarhizium anisopliae (F52), has recently been registered by the U.S. Environmental Protection Agency for BVW control. The objective of this study was to determine the persistence of M. anisopliae in five common soilless potting media components (coir, fir bark, hemlock bark, peat moss and perlite), which was measured as pathogenicity against BVW larvae. Fungal granules (½ lb/yd3) were incorporated with each media component at potting and fungal persistence determined for 133 days. Experiments were performed with and without plants to determine if the presence of a plant impacted fungal persistence. Overall, the fungus persisted well in all of the potting media components tested up to 133 days post application. Persistence was somewhat lower in the first experiment, likely due to fluctuating media moisture. In the second experiment, with more stable media moisture levels, the percentage of larval infection did not drop below 88% in any media at 133 days post application. It is likely that M. anisopliae will persist well and provide high levels of BVW larval control in most of the commercial potting media used in containerized nursery production, particularly those comprised primarily of the media components tested in these studies.

Laboratory soil bioassays were performed at economic field rates for in-furrow (3.85 × 106 spores/g dry soil) and broadcast (3.85 × 105 spores/g dry soil) applications with three isolates of Metarhizium anisopliae (F52, ATCC62176, and ARSEF5520) and one isolate of Beauveria bassiana (GHA). All isolates tested were infective to second instar Delia radicum (L.). The conditionally registered M. anisopliae isolate (F52) performed best killing an average of 85 and 72% of D. radicum larvae at the high and low concentration, respectively. The mean LC50 and LC95 of F52 against second instar D. radicum was 2.7 × 106 and 1.8 × 108 spores/g dry soil, respectively. The use of F52 in an integrated management program is discussed.

Wholesale container-grown ornamentals are often maintained at the nursery for at least two growing seasons and are subject to infestation by black vine weevil (BVW), Otiorhynchus sulcatus F., for several months each year. Therefore, a potting media amendment aimed at controlling BVW needs to persist for an extended period of time. These studies were conducted to determine the persistence and ecology of Metarhizium anisopliae (MetchnikoV) Sorokin incorporated into peat and bark-based potting media with and without a crab meal amendment in container-grown Picea abies ‘Nidiformis.’ Rooted cuttings of P. abies were planted into potting media amended with M. anisopliae (1 g of formulated product/L; »6 log10 CFU/g dry potting media). The fungal population in bulk potting media was quantitatively determined using selective media at 14, 21, 28, 35, 49, 63, 77, 91, 105, 119, 143, 175, 203, 231, 258, 287 and 342 days. The fungal population in the rhizosphere was quantitatively determined at 203, 231, 258, 287, and 342 days. M. anisopliae colonized the rhizosphere of P. abies and the fungal population in the rhizosphere was signiWcantly greater than in the surrounding bulk media. M. anisopliae persisted in the peat and bark-based potting media at 6.22 and 5.74 log10 CFU/g dry potting media for 342 days, respectively. Bioassays using bark and peat-based potting media inoculated with M. anisopliae at 6 log 10 CFU/g dry potting media resulted in 93.5% and 97.5% infection of last instar BVW, respectively. P. abies roots inoculated with M. anisopliae infected 76% of 2nd–3rd instar BVW. Inoculation of roots with M. anisopliae represents a novel method for delivering entomopathogenic fungi and would greatly reduce application costs. Factors associated with fungal biology outside the host may be more important than virulence in a laboratory bioassay.

The black vine weevil Otiorhynchus sulcatus (F.) is the primary insect pest of field and container-grown woody ornamentals in the Pacific Northwest (PNW). These studies were conducted to determine the natural occurrence of soil-borne entomopathogens in PNW nursery soils and determine their virulence to black vine weevil. Soil samples were collected July–September of 2002 from field-grown woody ornamental nursery stock in Oregon, Washington, and Idaho. Sample collection in each state took place in the major nursery production areas. A total of 280 samples was collected (Oregon, 170; Washington, 50; Idaho, 60). Entomopathogens were isolated using insect baiting (nematodes and fungi) as well as semiselective media (fungi). Bacillus thuringiensis Berliner was isolated through sodium acetate selection. Soil-borne entomopathogenic fungi occur widely throughout the major nursery production areas in the PNW. The entomopathogenic fungi Metarhizium anisopliae (Metchnikoff) Sorokin, Beauveria bassiana (Bals...

The black vine weevil, Otiorhynchus sulcatus (F.) (Coleoptera: Curculionidae) is a univoltine, polyphagous insect that is a nocuous pest of field and container grown ornamentals as well as small fruit crops world-wide (Moorhouse et al., 1992). In the USA, the environmental horticulture industry (floriculture and nursery crops) is the third largest value crop behind corn and soybeans (USDA fact sheets 2001, http://www.nass.usda.gov). Otiorhynchus sulcatus is thought to have a northern European origin and was first recorded in North America in 1835 (Smith, 1932). Oviposition occurs at night with eggs being either dropped on the soil surface or inserted into crevices on plants (Smith, 1932). Early instars begin by feeding on small roots while the later instars feed on larger roots, especially on the phloem and cambium tissues near the soil surface (La Lone & Clarke, 1981). Adults are nocturnal and mainly cause aesthetic damage to plants by notching the leaves. Adults are parthenogenic, so a single individual left unchecked can result in the infestation of an entire nursery. Because of the biology of O. sulcatus , the collection of larvae and adults from the field is very tedious and often requires countless hours of digging through field soil, container-grown plants or searching for adults after dark. Indeed, many chemical and biological control companies developing products for O. sulcatus larval control still rely on searching through infested plant material in order to gather the needed larvae for efficacy studies. Although an artificial diet initially developed by Shorey & Hale (1965) as modified by Shanks & Finnigan (1973) and Shanks (1980) (hereafter referred to as the ‘standard diet’) sustains larvae and is commercially available, there has been no documented technique for continuous mass rearing of O. sulcatus . Studies of weevil biology as well as the development of control strategies require a large, uniform, and predictable supply of insects of all life stages throughout the year. When considering field or potted plants for use in control efficacy or host plant resistant studies, other researchers have often found it useful to store insect eggs for several weeks for subsequent use; many thousands to millions of eggs were needed. The stockpiling of eggs for future use has been necessary with other reared species of Coleoptera (Branson, 1978; Fisher & Edwards, 2002). The egg hatch and viability of the strawberry root weevil, Otiorhynchus ovatus L. were not significantly reduced when it was stored for up to 4 weeks at 4 ° C (Fisher & Edwards, 2002). We have developed a technique for continuous mass rearing of O. sulcatus in the laboratory using a meridic diet. Since small changes in diet composition can often lead to dramatic changes in larval survival and growth, experiments were also performed to determine if O. sulcatus larval survival and development were increased on the improved diet described here, compared to the standard diet. Additionally, we determined if O. sulcatus eggs could be stored for up to 4 weeks at 4 ° C without significant reductions in larval survival and development.

The entomopathogenic fungus Beauveria bassianainfects a wide range of insects and survives as a soil saprophyte and a plant endophyte. The objective of our study was to determine the role of rainfall in dispersing B. bassiana(Balsamo) Vuillemin to the surface of corn (Zea mays) from soil with different levels of crop residue. Laboratory studies which simulated field tillage systems, had levels of crop residue which covered 0.53 (control), 34, 59 and 84% of the soil surface. Simulated rainfall in a raindrop tower at an intensity of 73 mm/h caused a significant increase in the mean number of B. bassianacolony forming units (CFU) isolated from the surface of corn plants. Plants receiving rain had a mean (±S.E.M.) of 8.8 ± 2.8 CFU per plant; controls had a mean of 0.03 ± 0.01 CFU per plant. The mg of soil collected from the surface of plants also was significantly influenced by rainfall. Plants receiving rain had a mean of 15 .7 ± 1.7 mg of soil per plant while controls had a mean of 3.4 ± 0.4 mg of soil per plant. Linear regression revealed highly significant negative relationships between the mean mg of soil and the mean number of CFU per plant, over the four levels of crop residue. The amount of soil and number of CFU per plant decreased significantly with increasing levels of crop residue. In field studies with conservation- and no-till systems, results were similar to those recorded in the raindrop tower. The mean number of CFU and mg of soil per plant were both higher in conservation-till plots than in no-till plots, where surface residue averaged 45 and 95%, respectively. Rainfall plays an active role in the dispersal of B. bassianafrom the soil environment to the surface of whorl-stage corn. Increased levels of crop residue reduce the amount of soil and fungal transfer to the surface of young corn. © 2002 Elsevier Science B.V. All rights reserved.

The corn rootworm complex is the most damaging insect pest of corn (Zea mays L.). This study was conducted to determine the efficacy of whorl and pollen-shed stage applications of a granular formulation of Beauveria bassiana (Balsamo) Vuillemin for control of adult western corn rootworm (Diabrotica virgifera virgifera Le Conte). The effect of application time (whorl-stage, pollen-shed) and plant surface exposed (leaves and leaf collars; silks; leaves, leaf collars, and silks) on level of beetle fungal infection were investigated. In addition, the number of colony forming units of B. bassiana in the corn leaf collar area was quantified. In the three years (1998-2000) of the study, application of B. bassiana at whorl-stage did not significantly increase beetle fungal infection. Beauveria bassiana applied to plants at pollen-shed in 1998 resulted in a significant increase in beetle infection with 51% of beetles from treated plants infected and 6.0% from control plants. Similar applications at pollen-shed in 1999 and 2000 resulted in very low infection levels. Beauveria bassiana application at pollen-shed stage significantly increased the number of colony forming units per leaf collar during all years of the study. Beetle infection with B. bassiana did not differ consistently among plant surface to which beetles were exposed for either application. Increased fungal load in leaf collars was not correlated with increased levels of adult infection. Increased rates of B. bassiana and application when beetles are present on the plants are likely needed to significantly increase infection rates.

In August 2008, the first detection of the spotted wing drosophila, Drosophila suzukii, to the North America mainland in California caused great concern, as the fly was found infesting a variety of commercial fruits. Subsequent detections followed in Oregon, Washington, Florida and British Columbia in 2009; in Utah, North Carolina, South Carolina, Michigan, and Louisiana in 2010; and in Virginia, Montana, Wisconsin, Pennsylvania, New Jersey, Maryland and Mexico in 2011. In Europe, it has been detected in Italy and Spain in 2009 and in France in 2010. Economic costs to the grower from D. suzukii include the increased cost of production (increased labor and materials for chemical inputs, monitoring and other management tools) and crop loss. An effective response to the invasion of D. suzukii requires proper taxonomic identification at the initial phase, understanding basic biology and phenology, developing management tools, transferring information and technology quickly to user groups, andevaluatingtheimpactoftheresearchandextensionprogramonaneconomic,social,andenvironmentallevel.AsD.suzukii continues to expand its range, steps mustbe initiated in each new region to educate and inform the public as well as formulate managementtactics suitable for the crops and growing conditions in each. c � 2011 Society of Chemical Industry

Nosema pyrausta is an obligate pathogen causing reduced fecundity and longevity of Ostrinia nubilalis. This study was conducted to determine the combined effects of N. pyrausta infection and temperature on O. nubilalis egg production and hatch. N. pyrausta-infected and noninfected O. nubilalis were maintained in two different temperature regimes. The first regime allowed females to oviposit under optimum conditions (27 degrees C, 65% RH, 16:8 (L:D)), while females in the second regime were held initially under the same humidity and light conditions, but a constant temperature of 16 degrees C for 1 week after which they were transferred to optimum ovipositional conditions. Studies were performed initially with O. nubilalis populations and later with individual mating pairs. In studies with O. nubilalis populations, the mean number of eggs laid per female under optimum conditions was 660, while N. pyrausta-infected females held initially at 16 degrees C laid 116 eggs per female. In studies with individual mating pairs, N. pyrausta infection reduced egg production per female 53 and 11% in the 16 and 27 degrees C temperature regimes, respectively, compared to noninfected females under optimum conditions. Exposure to 16 degrees C temperatures early in the ovipositional period had a more profound impact on reducing egg production in N. pyrausta-infected than noninfected O. nubilalis.

Only recently have investigations focused on re- lationships between B. bassiana and the corn plant. Field and greenhouse studies were conducted to determine the pro- Beauveria bassiana forms an endopytic relationship with clivity of Bacillus thuringiensis (Bt) (Berliner)-transgenic corn (Zea mays L.) to form an endophytic relationship with Beauveria bassiana, the corn plant and provides season-long suppression of and to evaluate the corn for possible plant pathological effects associ- O. nubilalis (Lewis and Cossentine, 1986; Lewis and ated with this relationship. Beauveria bassiana (Balsamo) Vuillemin Bing, 1991; Wagner and Lewis, 2000). Two areas of was applied as a granular formulation to two separate lines of corn, research must be investigated before acceptance by pro- expressing Bt events MON802 and MON810, and their corresponding ducers of B. bassiana as a management tool: (i) will isolines. There were no significant differences in levels of endophytism B. bassiana colonize Bacillus thuringiensis (Bt) trans- between transgenic events or their near-isolines. In greenhouse stud- formed corn plants as well as non-transformed plants, ies, B. bassiana was applied as a liquid seed treatment to Bt transgenic and (ii) is the endophytic relationship void of plant corn hybrids Pioneer 34R06 (event MON810) and Ciba Max 454 pathological symptoms in transformed plants and/or (event 176) and their near isolines at a rate of 2 10 10 conidia per their isolines? ml. There were no significant differences in seed germination or pres- ence of root pathogens in transgenic or isoline seeds soaked in a This paper reports the results of research to determine B. bassiana suspension. The same lines of corn were used in field the proclivity of B. bassiana to form an endophytic rela- experiments with treatments of seeds soaked in a suspension of B. tionship with Bt transgenic corn and whether endo- bassiana, a foliar application of a granular formulation of B. bassiana, phytism causes a plant pathology. Field studies were and corresponding untreated checks. Plants were sampled throughout conducted in 1994 and 1995 to determine the ability of the growing season and evaluated for growth of individual plant com- Bt-transgenic corn to form an endophytic relationship ponents, including sheaths, leaves, stem, husk, ear, plant leaf-to-stem with B. bassiana. Greenhouse studies were performed ratio and overall plant growth. There were no significant differences in 1997 to determine the effect of B. bassiana on seed in overall plant growth between the B. bassiana treatments or in the germination and plant growth on two lines of Bt corn growth of each plant component. The results of this study indicate and their genetic isolines. Field studies in 1997 were that B. bassiana readily forms an endophytic relationship with trans- genic and non-transgenic corn and causes no plant pathology. conducted to determine any possible pathogenic effects of B. bassiana on overall plant growth and dry matter ac- cumulation.

The influence of adjacent habitats, trap location, and trap height on capture numbers of predators and a parasitoid were monitored in Iowa cornfields using insect traps (Gempler's . AM No-Bait). Traps re fastened to a single wood lathe (122 by 3.8 by 1.3 cm) with wire ties at and 1 m above ground level. Traps at the 2 heights were placed in corn and their adjacent habitat. The adjacent habitat consisted of 1 of 3 types- herbaceous, wooded, and intermediate. Adjacent habitat did not significantly influence the number of Macrocentrus grandii Goidanich (Hymenoptera: Braconidae) captured but slid influence the capture of Coleomegilla maculata (De Geer) (Co-leoptera: Coccinellidae), Coccinella spetempunctata L. (Coleoptera: Coccinellidae), and Chrysoperla carnea Stephens( Neuroptera: Chrysopidae ) with increased captures from fields with the herbaceous and intermediate habitats. Distance that traps were placed in corn and trap height significantly influenced numbers of M. grandii and C. maculata captured. Significantly more M. grandii and C. maculata were captured from traps 11 m into corn than from traps 2.5 m into corn or in the adjacent habitat. Capture rates of M grandii also were significantly higher on traps 1 m above ground. Data from this research indicate that M. grandii is synchronized with its host Ostrinia nubilalis (Hubner), in central Iowa.

Background The serum activities of ALT and AST are key indicators of liver toxicity in the drug safety evaluation of laboratory animals and patients. To ensure that the full aminotransferase activity is measured, exogenous Pyridoxyl-5-Phosphate (P5P) cofactor is included in the assay reagent. Clinical pathology laboratories make a choice to use aminotransferase assays with or without the added P5P cofactor, and the impact of assay selection on safety assessment is not well understood. Objectives The objective of this report was to investigate the effect of aminotransferase assay selection on the detection of liver toxicity based on a literature review. Methods Literature in public databases was searched using combinations of the search terms alanine aminotransferase, aspartate aminotransferase, pyridoxyl-5-phosphate, holoenzyme, apoenzyme, enzyme inhibition, artifact, clinical pathology, toxicology, and safety assessment. Regulations or guidance documents published by health authorities specifying clinical pathology evaluation in nonclinical and clinical safety studies of biopharmaceuticals, chemicals, and devices were also reviewed. Results Aminotransferase testing is not standardized in safety assessment studies and consequently, laboratories use aminotransferase assays with or without P5P cofactor. Individual studies have demonstrated mean differences of approximately 10–20% in serum ALT activity in animal and human populations. The impact of aminotransferase testing without P5P on detection of toxicity and decision-making in drug development has not been systematically evaluated. Conclusions The use of different assays for measuring aminotransferase activity contributes to the variability in data between laboratories and studies. Standardizing aminotransferase assays is an avenue for improving the diagnostic performance in drug safety evaluation.

Ten strains representing four lineages of the Pseudomonas fluorescens group (P. chlororaphis, P. corrugata, P. koreensis, and P. fluorescens subgroups) were evaluated for toxicity to the tobacco hornworm Manduca sexta and the common fruit fly Drosophila melanogaster. The three strains within the P. chlororaphis subgroup exhibited both oral and injectable toxicity to the lepidopteran M. sexta. All three strains have the gene cluster encoding the FitD insect toxin and a ΔfitD mutant of P. protegens strain Pf-5 exhibited diminished oral toxicity compared to the wildtype strain. Only one of the three strains, P. protegens Pf-5, exhibited substantial levels of oral toxicity against the dipteran D. melanogaster. Three strains in the P. fluorescens subgroup, which lack fitD, consistently showed significant levels of injectable toxicity against M. sexta. In contrast, the oral toxicity of these strains against D. melanogaster was variable between experiments, with only one strain, Pseudomonas sp. BG33R, causing significant levels of mortality in repeated experiments. Toxin complex (Tc) gene clusters, which encode insecticidal properties in Photorhabdus luminescens, were identified in the genomes of seven of the ten strains evaluated in this study. Within those seven genomes, six types of Tc gene clusters were identified, distinguished by gene content, organization and genomic location, but no correlation was observed between the presence of Tc genes and insect toxicity of the evaluated strains. Our results demonstrate that members of the P. fluorescens group have the capacity to kill insects by both FitD-dependent and independent mechanisms.

An epizootic is defined as an outbreak of disease with an unusually large number of cases. A central question in insect pathology is: what are the factors that cause an epizootic? The question is addressed through epizootiology, the study of animal disease dynamics on a population level. The major factors influencing an epizootic can be divided into four basic components: the pathogen population, the host population, transmission, and the environment. The question pertaining to the causes of an epizootic is of great interest to all aspects of insect pathology, and particularly to microbial control efforts. Microbial control can be defined as the use of entomopathogens (viruses, fungi, bacteria, protists, or nematodes) for pest suppression. This chapter presents a summary and analysis of epizootiological principles and concepts of microbial control. The goal is to promote expanded studies in epizootiology, and foster research and implementation toward improved microbial control programs.

BACKGROUND: The spotted-wing drosophila, Drosophila suzukii Matsumura, is native to Asia and was first detected in the North American mainland and Europe in 2008–2010. Drosophila suzukii is a serious economic pest to stone and small fruits because the female lays eggs within ripening fruit on a plant before harvest, which can lead to crop loss. The aim of this study was to evaluate the susceptibility of blackberries, blueberries, cherries, grapes, raspberries and strawberries to D. suzukii among various ripeness stages and cultivars. RESULTS: In 26 no-choice and choice replicated laboratory cage tests on ripeness stages, fruits were generally susceptible to D. suzukii once fruits started to color. Few D. suzukii developed on green fruit, wine grapes or overripe blueberries. In seven cultivar tests, D. suzukii preferences ranged from no differences to fourfold differences for specific cultivars of blackberries, blueberries, raspberries and wine grapes. As brix levels increased, more eggs were laid or more D. suzukii developed on blackberries, blueberries, cherries, raspberries and strawberries. In a choice test of various fruit types, strawberries, raspberries, blackberries, cherries and blueberries were more susceptible to D. suzukii than green table grapes ('Thompson'). CONCLUSION: The results suggest that fruits may become susceptible to D. suzukii as they start to turn color, and that specific varieties of grapes and overripe blueberries have low susceptibility to D. suzukii. Copyright © 2011 Society of Chemical Industry

Understanding habitat selection of fungal entomopathogens is critical to improve the efficacy, persistence and cost of these fungi as microbial insecticides. This study sought to determine the prevalence of Metarhizium and Beauveria spp. isolated from the rhizosphere of strawberry, blueberry, grape and Christmas tree crops in the Willamette Valley of Oregon. Entomopathogenic fungi were assigned to thirteen species based on molecular phylogenetic criteria. Four species of Metarhizium were isolated including Metarhizium brunneum, Metarhizium guizhouense, Metarhizium robertsii, and Metarhizium flavoviride var. pemphigi. Nine Beauveria species were isolated including, Beauveria brongniartii, an undescribed species referred to as Clade C and seven phylogenetic species of Beauveria bassiana. Strawberries and blueberries were significantly associated with M. brunneum and Christmas trees with M. guizhouense and M. robertsii. Grapes were significantly associated with B. bassiana phylogenetic species Bbas-16. All of the Metarhizium isolates screened were pathogenic to Otiorhynchus sulcatus larvae in laboratory bioassays but only M. brunneum and M. robertsii caused significant levels of infection. The study results suggest that certain species of Metarhizium and Beauveria are significantly associated with the strawberry, blueberry and Christmas tree rhizosphere and could potentially provide better control of O. sulcatus.

A newly discovered microsporidium infecting the black vine weevil, Otiorhynchus sulcatus (F.) (Coleoptera: Curculionidae), provisionally placed in the genus Canningia, was studied to determine its impact on O. sulcatus. O. sulcatus populations from several locations were sampled and evaluated for microsporidiosis. A very low prevalence of the disease was observed in all locations surveyed (

Over 2,500 species of insects and mites are destructive to nursery crops in the United States (Johnson and Lyon, 1991). Likewise, there is a large and diverse number of nursery plant species used in different landscape settings and geographic regions. Nursery and landscape plants represent a complex challenge for insect control products because of the large number of plant species, insect pests, growing conditions, potting media, and production techniques (Martin, 1997).

The objective of this study was to determine the persistence of Metarhizium anisopliae (F52), measured as infectivity against black vine weevil larvae, in a soilless potting medium at six wholesale nursery locations across the Willamette Valley, Oregon. A granule formulation (0.30 and 0.60 kg/m3) was incorporated into media at planting and fungal persistence determined over two growing seasons. The fungus persisted in the potting media over the duration of the experiment with 50–60% of the larvae exposed to treated media becoming infected at the end of the experiment. The percentage of infected larvae gradually declined from ⩾90% on week 3 to 40–60% by week 19. Larval infection rebounded over the fall and winter months of 2004 to 75–80% followed again by a slow decline over the course of the second growing season.

OSI-7836 (4'-thio-beta-D-arabinofuranosylcytosine) is a novel nucleoside analog in phase I clinical development for the treatment of cancer. As with other nucleoside analogs, the proposed mechanism of action involves phosphorylation to the triphosphate form followed by incorporation into cellular DNA, leading to cell death. This hypothesis has been examined by measuring and comparing the incorporation of ara-C, OSI-7836, and gemcitabine (dFdC) into DNA of cultured cells and by investigating the role of deoxycytidine kinase in OSI-7836 toxicity. We report here additional studies in which the role of cell cycling on OSI-7836 toxicity was investigated and incorporation of OSI-7836 into DNA of xenograft tumors measured. The role of the cell cycle was examined by comparing the toxicity of OSI-7836 in A549 NSCLC cells that were either in log phase growth or had reached confluence. A novel validated LC-MS/MS assay was developed to quantify the concentrations of OSI-7836 in DNA from Calu-6 and H460 human tumor xenografts in mice. Results showed that apoptosis induced by OSI-7836 was markedly greater in cycling cells than in confluent non-cycling cells despite only a modest increase in intracellular OSI-7836 triphosphate concentration. The LC-MS/MS assay developed to measure OSI-7836 incorporation into DNA had an on-column detection limit of 0.25 fmol, a quantification limit of 0.5 fmol, and a sensitivity of approximately 0.1 pmol OSI-7836/micromol dThy. Concentrations of OSI-7836 in splenic DNA (0.4 pmol OSI-7836/micromol dThy) averaged fivefold less than the average concentration in Calu-6 and H460 xenograft DNA (3.0 pmol OSI-7836/micromol dThy) following a 400 mg/kg dose of OSI-7836. Concentrations of OSI-7836 in Calu-6 tumor DNA isolated 24 h following a dose of 400, 1000, or 1600 mg OSI-7836/kg were approximately 1.3, 1 and 1.3 pmol OSI-7836/micromol dThy, respectively. Concentrations of OSI-7836 in DNA from H460 and Calu-6 xenografts did not appear to increase during repeated administration of 400 mg OSI-7836/kg on days 1, 4, 7, and 10. The majority of OSI-7836 in DNA from Calu-6 and H460 tumors of mice dosed with 1600 mg/kg was located at internal nucleotide linkages, similar to dFdC and ara-C. In conclusion, cell cycling studies supported the hypothesis that OSI-7836 cytotoxicity is dependent upon DNA synthesis. A validated LC-MS/MS assay was developed that could quantify OSI-7836 in DNA from tissues. The assay was used to show that OSI-7836 was incorporated in internal linkages in tumor DNA in a manner that was dose-independent at the doses tested and did not appear to accumulate during repeated dosing. The results suggest that if DNA incorporation is a toxic event, the relationships between administered dose, DNA incorporation, and toxicity are complex.

OSI-211 (liposomal lurtotecan), was evaluated using several different dose schedules (1mg/kg, d1-5, 1.75 mg/kg d1, 3, 5 and 6 mg/kg d1, 8) in severe combined immunodeficient (SCID) mouse models of acute myelogenous leukemia (AML) and acute lymphocytic leukemia (ALL) with early treatment (ET, days 6-8) or late treatment (LT, days 15-19), examining early and advanced disease, respectively. Due to the aggressive nature of the Molt-4 model, the ET and LT were accelerated to day 3 or 4 and day 8 post-implant, respectively. For each model, 2 x 10(7) (KBM-3B) or 1 x 10(7) (Molt-4, HL-60 and CEM) leukemia cells were injected intravenously into the tail vein. Each control and test group consisted of eight animals. All three schedules (1mg/kg qd1-5, 1.75 mg/kg d1, 3, 5 and 6 mg/kg d1, 8) increased the life span of OSI-211 treated animals in each model, with a tendency toward improved efficacy with the 6 mg/kg d1, 8 schedule. As a result, the activity of the 6 mg/kg d1, 8 schedule is detailed for each model. ET significantly (P0.005) increased survival in the KBM-3B model with 86% long-term survivors (LTS). Using PRC analysis, human beta-globin gene sequences in one or several tissues were amplified in all but 3 LTS, suggesting minimal residual disease in 26 of the 29 LTS. LT also significantly (P0.005) improved average life span in the KBM-3B model, with an average ILS=196+/-11% and one LTS. Treatment of HL-60 leukemia animals significantly (P0.005) increased life span, with an ILS=213+/-9% and two LTS for ET, and with an ILS=219+/-4% and no LTS for LT. Treatment of Molt-4 animals, the most aggressive leukemia model tested, significantly (P0.005) increased life span, with an average ILS=181+/-3% and no LTS for ET and an average ILS=172+/-1% with no LTS for LT. In the CEM model, ET resulted in a significantly (P0.005) improved ILS=244+/-24% with one LTS. In comparison to OSI-211, treatment with DaunoXome, the liposomal formulation of daunorubicin, a drug with clinical efficacy in AML and ALL, had no effect on survival in the KBM-3B, nor Molt-4 A4 leukemia models when administered at its maximum or near maximum tolerated doses of 3mg/kg d1, 8. These data demonstrate that OSI-211 has potent antileukemia activity in preclinical SCID mouse AML and ALL leukemia models, supporting the clinical investigation of OSI-211 for hematological malignancies.