Augmentative biological control for squash bug (Hemiptera: Coreidae) using the egg parasitoid, Hadronotus pennsylvanicus (Hymenoptera: Scelionidae).

The squash bug, Anasa tristis (De Geer) (Hemiptera: Coreidae), is a serious pest of cucurbit crops across the United States. Conventional growers commonly use broad-spectrum insecticides to manage squash bugs, however organic growers lack these effective chemical tools and must rely on alternative management strategies. Biological control of A. tristis is largely understudied, specifically the potential of natural enemy, Hadronotus pennsylvanicus (Ashmead) (Hymenoptera: Scelionidae), as an augmentative biological control agent. For this reason, we performed early-season field releases of H. pennsylvanicus on organic farms in southeastern Virginia to test if this would improve A. tristis egg parasitism. We chose organic vegetable farms growing summer squash (Cucurbita pepo L.) as release sites and nearby Virginia Tech Agricultural Research Extension Centers (AREC) as no-release sites. Parasitoids were reared in the lab and deployed as parasitized egg masses (~2-3 females wasps/plant) in June 2020 and 2021. Before parasitoid deployment, host eggs collected from release and no-release sites displayed low levels of H. pennsylvanicus parasitism in 2020 (<21%) and 2021 (<8%). In both years, the percentage of A. tristis eggs parasitized within 2 weeks post deployment was significantly greater at release sites (~60%) than at no-release sites (~14%). High rates of H. pennsylvanicus parasitism (>72%) were further observed at release sites 4, 6, 8, and 10 weeks following parasitoid deployment. Our study demonstrates that releases of lab-reared H. pennsylvanicus can increase A. tristis egg parasitism rates and subsequently decrease successful nymph hatch rates in early summer squash plantings.

Both the Squash Bug Anasa tristis and Horned Squash Bug Anasa armigera (Hemiptera: Coreidae) are Attracted to Vittatalactone, the Aggregation Pheromone of Striped Cucumber Beetle.

Vittatalactone, the aggregation pheromone of the striped cucumber beetle, Acalymma vittatum (F.) (Coleoptera: Chrysomelidae), is attractive to two species of squash bugs (Hemiptera: Coreidae), the squash bug Anasa tristis (DeGeer) and horned squash bug Anasa armigera (Say). In field trapping experiments in Maryland and Virginia, clear sticky traps baited with 1 mg of a synthetic 8-isomer mix of vittatalactone captured ~9× more of female A. tristis and of both sexes of A. armigera, whereas male A. tristis were not significantly attracted, compared to unbaited traps. A. armigera showed a distinct dose-response to vittatalactone lure loading in the late season, and this species was more attracted than A. tristis, based on comparison to captures from underneath wooden boards emplaced in adjacent fields. Results suggest that vittatalactone could be a 'keystone semiochemical' in colonization of cucurbit hosts by specialist herbivores, and may offer the opportunity for multi-species behavioral control as a component of integrated pest management in cucurbit crops.

Effect of Plastic Mulch Colors on Anasa tristis (Hemiptera: Coreidae) Population Dynamics in Summer Squash, Cucurbita pepo (Cucurbitales: Cucurbitaceae).

The squash bug, Anasa tristis (De Geer), is a serious pest of cucurbit crops across the United States, especially within summer squash (Cucurbita pepo L.) systems. Using their piercing sucking mouthparts, squash bugs feed on both leaf tissue and fruits, often leading to leaf necrosis, marketable fruit loss, and even plant death. To date, the relationship between squash bug presence and plasticulture has not been adequately investigated. This 2-yr study evaluated the effects of white, black, and reflective plastic mulch colors on the occurrence of all squash bug life stages and marketable zucchini yield in Virginia. In both years, A. tristis adults and egg masses were more numerous on zucchini plants grown in white and reflective plastic mulch compared to bare ground plants. Greater nymphal densities and marketable fruit yield were observed in certain plastic mulch treatments versus the bare ground treatment, yet these differences were not consistent in both years. Contrary to the repellency effects reflective mulches have on other cucurbit insect pests, our research suggests that reflective and other plastic mulch colors can negatively impact squash bug management, especially in regions with high A. tristis pressure. Our study offers new insights for cucurbit growers to use when considering whether they should implement plasticulture in their growing systems.

Prediction of Immunotherapy Response in Melanoma through Combined Modeling of Neoantigen Burden and Immune-Related Resistance Mechanisms.

PURPOSE: While immune checkpoint blockade (ICB) has become a pillar of cancer treatment, biomarkers that consistently predict patient response remain elusive due to the complex mechanisms driving immune response to tumors. We hypothesized that a multi-dimensional approach modeling both tumor and immune-related molecular mechanisms would better predict ICB response than simpler mutation-focused biomarkers, such as tumor mutational burden (TMB). EXPERIMENTAL DESIGN: Tumors from a cohort of patients with late-stage melanoma (n = 51) were profiled using an immune-enhanced exome and transcriptome platform. We demonstrate increasing predictive power with deeper modeling of neoantigens and immune-related resistance mechanisms to ICB. RESULTS: Our neoantigen burden score, which integrates both exome and transcriptome features, more significantly stratified responders and nonresponders (P = 0.016) than TMB alone (P = 0.049). Extension of this model to include immune-related resistance mechanisms affecting the antigen presentation machinery, such as HLA allele-specific LOH, resulted in a composite neoantigen presentation score (NEOPS) that demonstrated further increased association with therapy response (P = 0.002). CONCLUSIONS: NEOPS proved the statistically strongest biomarker compared with all single-gene biomarkers, expression signatures, and TMB biomarkers evaluated in this cohort. Subsequent confirmation of these findings in an independent cohort of patients (n = 110) suggests that NEOPS is a robust, novel biomarker of ICB response in melanoma.

Host Kairomones Influence Searching Behavior of Trissolcus japonicus (Hymenoptera: Scelionidae), a Parasitoid of Halyomorpha halys (Heteroptera: Pentatomidae).

The brown marmorated stink bug, Halyomorpha halys (Stål), is a highly polyphagous species native to Asia that has become a serious invasive agricultural and nuisance pest across North America and Europe. Classical biological control host range evaluations have revealed egg parasitoid Trissolcus japonicus (Ashmead) to be the primary candidate biocontrol agent for field release against H. halys. However, these evaluations only provide us with the physiological host range of T. japonicus. Other Trissolcus species have demonstrated that contact kairomones from different host species elicit varied responses in the parasitoids' host foraging behaviors. To assess T. japonicus response to host kairomones, mated naive females were exposed to leaf surfaces contaminated with adult kairomones from its preferred host, H. halys, or from a native nontarget host, Podisus maculiventris (Say) (Heteroptera: Pentatomidae). Red maple, apple, and soybean were used as plant substrate treatments. The wasp's residence time on the leaf surface, linear walking velocity, and angular walking velocity were observed and measured using Noldus EthoVision XT tracking software. Within each leaf treatment, T. japonicus displayed stronger behavioral responses on leaves contaminated with contact kairomones from H. halys. The parasitoid resided on H. halys contaminated leaves for approximately twice as a long as it did on P. maculiventris contaminated leaves. Further, both species' kairomones elicited significant decreases in parasitoid walking velocity on all tested substrate types. Overall, our study suggests that kairomone-based behavioral studies can be used to further evaluate the host specificity of T. japonicus and can be an invaluable supplement to classical biocontrol host range testing regimes.

PATH-SCAN: a reporting tool for identifying clinically actionable variants.

The American College of Medical Genetics and Genomics (ACMG) recently released guidelines regarding the reporting of incidental findings in sequencing data. Given the availability of Direct to Consumer (DTC) genetic testing and the falling cost of whole exome and genome sequencing, individuals will increasingly have the opportunity to analyze their own genomic data. We have developed a web-based tool, PATH-SCAN, which annotates individual genomes and exomes for ClinVar designated pathogenic variants found within the genes from the ACMG guidelines. Because mutations in these genes predispose individuals to conditions with actionable outcomes, our tool will allow individuals or researchers to identify potential risk variants in order to consult physicians or genetic counselors for further evaluation. Moreover, our tool allows individuals to anonymously submit their pathogenic burden, so that we can crowd source the collection of quantitative information regarding the frequency of these variants. We tested our tool on 1092 publicly available genomes from the 1000 Genomes project, 163 genomes from the Personal Genome Project, and 15 genomes from a clinical genome sequencing research project. Excluding the most commonly seen variant in 1000 Genomes, about 20% of all genomes analyzed had a ClinVar designated pathogenic variant that required further evaluation.

An integrated approach to inferring gene-disease associations in humans.

UNLABELLED: One of the most important tasks of modern bioinformatics is the development of computational tools that can be used to understand and treat human disease. To date, a variety of methods have been explored and algorithms for candidate gene prioritization are gaining in their usefulness. Here, we propose an algorithm for detecting gene-disease associations based on the human protein-protein interaction network, known gene-disease associations, protein sequence, and protein functional information at the molecular level. Our method, PhenoPred, is supervised: first, we mapped each gene/protein onto the spaces of disease and functional terms based on distance to all annotated proteins in the protein interaction network. We also encoded sequence, function, physicochemical, and predicted structural properties, such as secondary structure and flexibility. We then trained support vector machines to detect gene-disease associations for a number of terms in Disease Ontology and provided evidence that, despite the noise/incompleteness of experimental data and unfinished ontology of diseases, identification of candidate genes can be successful even when a large number of candidate disease terms are predicted on simultaneously. AVAILABILITY: www.phenopred.org.

Cloning and analysis of Trypanosoma cruzi lanosterol 14alpha-demethylase.

Trypanosoma cruzi infection, transmitted by insect vectors or blood transfusions, is an important cause of morbidity and mortality in many Latin American countries. Treatments are toxic and frequently ineffective in curing patients with chronic manifestations of the infection (Chagas disease). Potentially exploitable chemotherapeutic targets of T. cruzi are enzymes of the sterol biosynthesis pathway. In particular, the P450 enzyme, lanosterol 14alpha-demethylase, has been implicated as the target of azole antifungal drugs that have potent anti-T. cruzi activity. In the work reported here, the T. cruzi lanosterol 14alpha-demethylase (Tc14DM) gene was cloned by degenerate PCR. The gene was found to be expressed in both insect and mammalian life-cycle stages of the parasite. Tc14DM was able to complement the function of the homologous gene in yeast (erg11) as demonstrated by restored ergosterol production in an erg11-deficient yeast strain. When the yeast strain was co-transfected with the P450 reductase gene from Trypanosoma brucei, the amount of ergosterol production was increased, indicating that the endogenous yeast P450 reductase was an inefficient partner with Tc14DM. Heterologous expression of Tc14DM in the baculovirus/Sf9 system resulted in a 52kDa product. The protein was observed to have the characteristic absorbance spectra of a P450 enzyme. A typical Type II binding spectrum was produced when the imidazole compound, ketoconazole, was mixed with the Tc14DM, demonstrating that ketoconazole binds the enzyme.