Natural Enemies and Biological Control of Stink Bugs (Hemiptera: Heteroptera) in North America.

Stink bugs comprise a significant and costly pest complex for numerous crops in the US, including row crops, vegetables, and tree fruits and nuts. Most management relies on the use of broad-spectrum and disruptive insecticides with high human and environmental risks associated with them. Growing concerns about pesticide resistance in stink bugs are forcing pest managers to explore safer and more sustainable options. Here, we review the diverse suite of natural enemies of stink bugs in the US, noting that the egg and the late nymphal and adult stages of stink bugs are the most commonly attacked by parasitoids, whereas eggs and young nymphs are the stages most commonly attacked by predators. The effectiveness of stink bugs' natural enemies varies widely with stink bug species and habitats, influencing the biological control of stink bugs across crops. Historically, biological control of stink bugs has focused on introduction of exotic natural enemies against exotic stink bugs. Conservation and augmentation methods of biological control have received less attention in the US, although there may be good opportunities to utilize these approaches. We identify some considerations for the current and future use of biological control for stink bugs, including the potential for area-wide management approaches.

Effect of Breathable Row Covers and Ground Cover on Pest Insect Levels and Cucurbit Yield.

Organic control measures in muskmelon and squash production are part of an integrated pest management approach that can include using floating row covers, generalist predators, and ground cover. These are used in Kentucky, allowing for a reduction in insecticide use and diminished virus incidence while increasing yield. Commonly used row covers are made from spunbonded fabric that retains heat and must be removed at anthesis and kept off until the end of the season. Thus, a new farming regime containing breathable mesh covers which can be replaced after anthesis was tested for longer season insect exclusion across two growing seasons. Additionally, ground cover treatments, consisting of mulch or bare ground were tested for their effect on pest insect abundance and fruit yield. Pest insect numbers were usually lower in plots with mesh row covers and in some cases, mulch ground cover also contributed to lower pest numbers. A stronger impact on pest numbers was observed in melon than squash. Melon yield was always significantly higher in plots with mesh row covers and mulch ground cover. This trend was not observed with squash in 2014 but was true in 2015. In 2015, most plants under the fabric row covers died because of high temperatures immediately after transplanting highlighting the need for breathable mesh row covers.

Hidden Host Mortality from an Introduced Parasitoid: Conventional and Molecular Evaluation of Non-Target Risk.

Hidden trophic interactions are important in understanding food web ecology and evaluating the ecological risks and benefits associated with the introduction of exotic natural enemies in classical biological control programs. Although non-target risk is typically evaluated based on evidence of successful parasitism, parasitoid-induced host mortality not resulting in visible evidence of parasitism (i.e., nonreproductive effects) is often overlooked. The adventive establishment of Trissolcus japonicus, an exotic parasitoid of the introduced stink bug Halyomorpha halys, provides an opportunity to investigate the total impact of this parasitoid on target and non-target hosts in the field. We developed a new methodology to measure nonreproductive effects in this system, involving a species-specific diagnostic PCR assay for T. japonicus. We applied this methodology to field-deployed eggs of four pentatomid species, coupled with traditional rearing techniques. Nonreproductive effects were responsible for the mortality of an additional 5.6% of H. halys eggs due to T. japonicus, and were even more substantial in some of the non-target species (5.4-43.2%). The observed hidden mortality of native non-target species from an introduced parasitoid could change predictions about direct and indirect ecological interactions and the efficacy of biological control of the target pest.

Endosymbiotic Bacteria Are Prevalent and Diverse in Agricultural Spiders.

Maternally inherited bacterial endosymbionts are common in arthropods, but their distribution and prevalence are poorly characterized in many host taxa. Initial surveys have suggested that vertically transmitted symbionts may be particularly common in spiders (Araneae). Here, we used diagnostic PCR and high-throughput sequencing to evaluate symbiont infection in 267 individual spiders representing 14 species (3 families) of agricultural spiders. We found 27 operational taxonomic units (OTUs) that are likely endosymbiotic, including multiple strains of Wolbachia, Rickettsia, and Cardinium, which are all vertically transmitted and frequently associated with reproductive manipulation of arthropod hosts. Additional strains included Rickettsiella, Spiroplasma, Rhabdochlamydia, and a novel Rickettsiales, all of which could range from pathogenic to mutualistic in their effects upon their hosts. Seventy percent of spider species had individuals that tested positive for one or more endosymbiotic OTUs, and specimens frequently contained multiple symbiotic strain types. The most symbiont-rich species, Idionella rugosa, had eight endosymbiotic OTUs, with as many as five present in the same specimen. Individual specimens within infected spider species had a variety of symbiotypes, differing from one another in the presence or absence of symbiotic strains. Our sample included both starved and unstarved specimens, and dominant bacterial OTUs were consistent per host species, regardless of feeding status. We conclude that spiders contain a remarkably diverse symbiotic microbiota. Spiders would be an informative group for investigating endosymbiont population dynamics in time and space, and unstarved specimens collected for other purposes (e.g., food web studies) could be used, with caution, for such investigations.

Predation on stink bugs (Hemiptera: Pentatomidae) in cotton and soybean agroecosystems.

Stink bugs (Hemiptera: Pentatomidae) are significant pests of cotton and soybeans in the southeastern United States with annual control costs exceeding $14 million in these crops. Three of the most prominent stink bug pests are the southern green (Nezara viridula), brown (Euschistus servus) and green (Chinavia hilaris) stink bugs. To determine trophic linkages between generalist arthropod predators and these pests, species-specific 16S molecular markers were designed and used to detect the presence of prey DNA in predator gut-contents. Over 2700 predators were collected over two growing seasons in cotton and soybean in southern Georgia in 2011 and 2012 and screened for stink bug DNA. Trophic linkages were analyzed relative to prey availability, crop type and field location. The frequency of stink bug DNA in predator guts was negligible on E. servus (0.23%) and C. hilaris (0.09%). Overall gut content detection of N. viridula was 3.3% and Geocoris sp. (Hemiptera: Geocoridae), Orius sp. (Hemiptera: Anthocoridae) and Notoxus monodon (Coleoptera: Anthicidae) were the primary predators. This contrasts with previous studies that reported a much more diverse suite of predators consuming stink bugs with much higher frequency of gut-content positives. The discrepancy between studies highlights the need for replicating studies in space and time, especially if the goal is to implement effective and durable conservation biological control in integrated pest management.

Molecular gut content analysis of different spider body parts.

Molecular gut-content analysis has revolutionized the study of food webs and feeding interactions, allowing the detection of prey DNA within the gut of many organisms. However, successful prey detection is a challenging procedure in which many factors affect every step, starting from the DNA extraction process. Spiders are liquid feeders with branched gut diverticula extending into their legs and throughout the prosoma, thus digestion takes places in different parts of the body and simple gut dissection is not possible. In this study, we investigated differences in prey detectability in DNA extracts from different parts of the spider´s body: legs, prosoma and opisthosoma, using prey-specific PCR and metabarcoding approaches. We performed feeding trials with the woodlouse hunter spider Dysdera verneaui Simon, 1883 (Dysderidae) to estimate the time at which prey DNA is detectable within the predator after feeding. Although we found that all parts of the spider body are suitable for gut-content analysis when using prey-specific PCR approach, results based on metabarcoding suggested the opisthosoma is optimal for detection of predation in spiders because it contained the highest concentration of prey DNA for longer post feeding periods. Other spiders may show different results compared to D. verneaui, but given similarities in the physiology and digestion in different families, it is reasonable to assume this to be common across species and this approach having broad utility across spiders.

Land cover diversity increases predator aggregation and consumption of prey.

A lower diversity of land cover types is purported to decrease arthropod diversity in agroecosystems and is dependent on patterns of land use and fragmentation. Ants, important providers of ecosystem services such as biological control, are susceptible to landscape-level changes. We determined the relationships between land cover diversity and fragmentation on the within-field spatial associations of ants to pests and resulting predation events by combining mapping and molecular tools. Increased land cover diversity and decreased fragmentation increased ant abundance, spatial association to pests and predation. Land cover diversity and fragmentation were more explanatory than land cover types. Even so, specific land cover types, such as deciduous forest, influenced ant and pest diversity more so than abundance. These results indicate that geospatial techniques and molecular gut content analysis can be combined to determine the role of land use in influencing predator-prey interactions and resulting predation events in agroecosystems.

Development and application of molecular gut-content analysis to detect aphid and coccinellid predation by Harmonia axyridis (Coleoptera: Coccinellidae) in Italy.

Despite their positive effect in reducing pest populations, exotic generalist predators sometimes become invasive and contribute to the displacement of indigenous species in the same trophic level. Although laboratory experiments have linked intraguild predation (IGP) to these interactions, field evidence and quantification of IGP are still lacking for most systems. The recent establishment of the exotic Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) in Italy raises concern about the detrimental effect that the ladybird could have on native coccinellids. Here we assessed, under laboratory conditions, the acceptability and suitability of eggs of 2 native ladybirds, Adalia bipunctata L. and Oenopia conglobata (L.), as prey items for H. axyridis larvae. Then we developed primers for molecular gut-content analysis to detect predation by H. axyridis on the 2 ladybirds and on the aphid Eucallipterus tiliae L. Species-specific 16S primers were developed for the 3 species and laboratory feeding trials were conducted to quantify the rate of prey DNA breakdown in the gut of H. axyridis. Moreover, to field evaluate primers, H. axyridis 4th instars (n = 132) were systematically collected from linden trees in northern Italy and screened for the presence of prey DNA. Seventy-three percent and 7% of field collected H. axyridis were positive for aphid and coccinellid DNA, respectively. Predation upon aphid and A. bipunctata was lower than predicted if density dependent consumption was expected, while predation upon O. conglobata was significantly higher. Here, we provided the first evidence of IGP among feral populations of H. axyridis and indigenous ladybird beetles, occurring in Italy.