The Development and Expansion of in vivo Germline Editing Technologies in Arthropods: Receptor-Mediated Ovary Transduction of Cargo (ReMOT Control) and Beyond.

In the past 20 years, sequencing technologies have led to easy access to genomic data from nonmodel organisms in all biological realms. Insect genetic manipulation, however, continues to be a challenge due to various factors, including technical and cost-related issues. Traditional techniques such as microinjection of gene-editing vectors into early stage embryos have been used for arthropod transgenesis and the discovery of Clustered regularly interspaced short palindromic repeats and CRISPR-associated protein (CRISPR-Cas) technologies allowed for targeted mutagenesis and the creation of knockouts or knock-ins in arthropods. Receptor-Mediated Ovary Transduction of Cargo (ReMOT Control) acts as an alternative to embryonic microinjections, which require expensive equipment and extensive hands-on training. ReMOT Control's main advantage is its ease of use coupled with the ability to hypothetically target any vitellogenic species, as injections are administered to the egg-laying adult rather than embryos. After its initial application in the mosquito Aedes aegypti, ReMOT Control has successfully produced mutants not only for mosquitoes but for multiple arthropod species from diverse orders, such as ticks, mites, wasps, beetles, and true bugs, and is being extended to crustaceans, demonstrating the versatility of the technique. In this review, we discuss the current state of ReMOT Control from its proof-of-concept to the advances and challenges in the application across species after 5 years since its development, including novel extensions of the technique such as direct parental (DIPA)-CRISPR.

Evaluating invasion risk and population dynamics of the brown marmorated stink bug across the contiguous United States.

BACKGROUND: Invasive species threaten the productivity and stability of natural and managed ecosystems. Predicting the spread of invaders, which can aid in early mitigation efforts, is a major challenge, especially in the face of climate change. While ecological niche models are effective tools to assess habitat suitability for invaders, such models have rarely been created for invasive pest species with rapidly expanding ranges. Here, we leveraged a national monitoring effort from 543 sites over 3 years to assess factors mediating the occurrence and abundance of brown marmorated stink bug (BMSB, Halyomorpha halys), an invasive insect pest that has readily established throughout much of the United States. RESULTS: We used maximum entropy models to estimate the suitable habitat of BMSB under several climate scenarios, and generalized boosted models to assess environmental factors that regulated BMSB abundance. Our models captured BMSB distribution and abundance with high accuracy, and predicted a 70% increase in suitable habitat under future climate scenarios. However, environmental factors that mediated the geographical distribution of BMSB were different from those driving abundance. While BMSB occurrence was most affected by winter precipitation and proximity to populated areas, BMSB abundance was influenced most strongly by evapotranspiration and solar photoperiod. CONCLUSION: Our results suggest that linking models of establishment (occurrence) and population dynamics (abundance) offers a more effective way to forecast the spread and impact of BMSB and other invasive species than simply occurrence-based models, allowing for targeted mitigation efforts. Implications of distribution shifts under climate change are discussed. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Comparison of Sex Pheromone and Kairomone-Enhanced Pheromone Lures for Monitoring Oriental Fruit Moth (Lepidoptera: Tortricidae) in Mating Disruption and Non-Disruption Tree Fruit Orchards.

Oriental fruit moth, Grapholita molesta (Busck), populations were monitored using standard sex pheromone lures (OFM L2) and kairomone-enhanced lures to aid the interpretation of trap captures with enhanced relative to conventional lures. Initially, comparison of 10 different lures showed that a10X load of OFM pheromone, codlemone, terpinyl acetate, and acetic acid were key components of the most attractive lures (TRE11034 and 1123). Subsequent trapping studies in mating disruption and non-disrupted orchards in the United States and Spain compared trap captures with TRE1123 and OFM L2 lures. Compared to the OFM L2 lure, the TRE1123 lure captured more moths in mating disruption and non-disrupted orchards, caught female moths, improved the precision of mean population estimates, and led to greater resolution of generational flights. Suppression of trap captures in mating disruption versus non-disrupted orchards was similar with both lures. There were significant linear correlations between weekly trap captures with the two lures in the majority of mating disruption and non-disrupted orchards across locations and years. Furthermore, regression of the slopes of trap capture regressions (i.e., attractiveness of enhanced lures relative to sex pheromone lures alone) versus moth density (as measured by mean cumulative moth capture with TRE1123 and OFM L2 lures) exhibited a significant positive relationship in non-disrupted orchards, indicating enhanced lures were relatively more attractive under high population densities. This relationship was not significant in mating disruption orchards, likely due to the density independent, non-competitive mechanism of mating disruption for oriental fruit moth when using high-dose reservoir dispensers.

Survey for Adventive Populations of the Samurai Wasp, Trissolcus japonicus (Hymenoptera: Scelionidae) in Pennsylvania at Commercial Fruit Orchards and the Surrounding Forest.

The samurai wasp, Trissolcus japonicus (Ashmead) (Hymenoptera: Scelionidae), is an egg parasitoid associated with the brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae). Trissolcus japonicus is a candidate for classical biological control of H. halys populations. Since 2014, adventive populations of T. japonicus have been detected in 14 US states, in the Canadian provinces of British Columbia and Ontario, and in two European countries, Switzerland and Italy. Establishing baseline information about populations of T. japonicus is important, as this species is not host specific to H. halys and the potential ecological effects of the accidental introductions are not fully known. In this study, yellow sticky cards were deployed at commercial fruit orchards in nine counties in Pennsylvania separated by more than 400 km. Trissolcus japonicus was detected on cards in eight counties, and in two habitats, in the orchard and at the forest border. Other native species of Scelionidae known to attack the eggs of H. halys were also identified, including Trissolcus euschisti (Ashmead), Trissolcus brochymenae (Ashmead), and Telenomus podisi Ashmead (Hymenoptera: Scelionidae). These results are important baseline ecological knowledge for both T. japonicus, which appears to be established in orchards throughout Pennsylvania, and other native Scelionidae.

Nosema maddoxi sp. nov. (Microsporidia, Nosematidae), a Widespread Pathogen of the Green Stink Bug Chinavia hilaris (Say) and the Brown Marmorated Stink Bug Halyomorpha halys (Stål).

We describe a unique microsporidian species that infects the green stink bug, Chinavia hilaris; the brown marmorated stink bug, Halyomorpha halys; the brown stink bug, Euschistus servus; and the dusky stink bug, Euschistus tristigmus. All life stages are unikaryotic, but analysis of the consensus small subunit region of the ribosomal gene places this microsporidium in the genus Nosema, which historically has been characterized by diplokaryotic life stages. It is also characterized by having the reversed arrangement of the ribosomal gene (LSU -ITS- SSU) found in species within the "true Nosema" clade. This microsporidium is apparently Holarctic in distribution. It is present in H. halys both where it is native in Asia and where it is invasive in North America, as well as in samples of North American native C. hilaris collected prior to the introduction of H. halys from Asia. Prevalence in H. halys from mid-Atlantic, North America in 2015-2016 ranged from 0.0% to 28.3%, while prevalence in C. hilaris collected in Illinois in 1970-1972 ranged from 14.3% to 58.8%. Oral infectivity and pathogenicity were confirmed in H. halys and C. hilaris. Morphological, ultrastructural, and ecological features of the microsporidium, together with a molecular phylogeny, establish a new species named Nosema maddoxi sp. nov.

Generation of composites for bone tissue-engineering applications consisting of gellan gum hydrogels mineralized with calcium and magnesium phosphate phases by enzymatic means.

Mineralization of hydrogels, desirable for bone regeneration applications, may be achieved enzymatically by incorporation of alkaline phosphatase (ALP). ALP-loaded gellan gum (GG) hydrogels were mineralized by incubation in mineralization media containing calcium and/or magnesium glycerophosphate (CaGP, MgGP). Mineralization media with CaGP:MgGP concentrations 0.1:0, 0.075:0.025, 0.05:0.05, 0.025:0.075 and 0:0.1 (all values mol/dm3 , denoted A, B, C, D and E, respectively) were compared. Mineral formation was confirmed by IR and Raman, SEM, ICP-OES, XRD, TEM, SAED, TGA and increases in the the mass fraction of the hydrogel not consisting of water. Ca was incorporated into mineral to a greater extent than Mg in samples mineralized in media A-D. Mg content and amorphicity of mineral formed increased in the order A < B < C < D. Mineral formed in media A and B was calcium-deficient hydroxyapatite (CDHA). Mineral formed in medium C was a combination of CDHA and an amorphous phase. Mineral formed in medium D was an amorphous phase. Mineral formed in medium E was a combination of crystalline and amorphous MgP. Young's moduli and storage moduli decreased in dependence of mineralization medium in the order A > B > C > D, but were significantly higher for samples mineralized in medium E. The attachment and vitality of osteoblastic MC3T3-E1 cells were higher on samples mineralized in media B-E (containing Mg) than in those mineralized in medium A (not containing Mg). All samples underwent degradation and supported the adhesion of RAW 264.7 monocytic cells, and samples mineralized in media A and B supported osteoclast-like cell formation. Copyright © 2014 John Wiley & Sons, Ltd.

Attraction of the Invasive Halyomorpha halys (Hemiptera: Pentatomidae) to Traps Baited with Semiochemical Stimuli Across the United States.

A recent identification of the two-component aggregation pheromone of the invasive stink bug species, Halyomorpha halys (Stål), in association with a synergist, has greatly improved the ability to accurately monitor the seasonal abundance and distribution of this destructive pest. We evaluated the attraction of H. halys to black pyramid traps baited with lures containing the pheromone alone, the synergist methyl (2E,4E,6Z)-decatrienoate (MDT) alone, and the two lures in combination. Traps were deployed around areas of agricultural production including fruit orchards, vegetables, ornamentals, or row crops in Delaware, Maryland, North Carolina, New Jersey, New York, Ohio, Oregon, Pennsylvania, Virginia, and West Virginia from mid-April to mid-October, 2012 and 2013. We confirmed that H. halys adults and nymphs are attracted to the aggregation pheromone season long, but that attraction is significantly increased with the addition of the synergist MDT. H. halys adults were detected in April with peak captures of overwintering adults in mid- to late May. The largest adult captures were late in the summer, typically in early September. Nymphal captures began in late May and continued season long. Total captures declined rapidly in autumn and ceased by mid-October. Captures were greatest at locations in the Eastern Inland region, followed by those in the Eastern Coastal Plain and Pacific Northwest. Importantly, regardless of location in the United States, all mobile life stages of H. halys consistently responded to the combination of H. halys aggregation pheromone and the synergist throughout the entire season, suggesting that these stimuli will be useful tools to monitor for H. halys in managed systems.

Effects of orchard host plants (apple and peach) on development of oriental fruit moth (Lepidoptera: Tortricidae).

Studies were designed to examine the effects of host plants (apple, Malus domestica Borkh., and peach, Prunus persica L.) on the development of oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae). Oriental fruit moth larvae developed faster on peach than on apple, both on fruit as well as on growing terminal shoots. On fruit, these differences were shown to cause significant changes in both the rate (approximately 20-60 degree-days earlier emergence on peach than on apple) and patterns of adult emergence among several cultivars of peaches and apples. Slopes of female emergence plots varied by host in 2003, with emergence occurring over a longer period on peach cultivars than on apple cultivars (with one exception). Slopes of male emergence curves did not differ by cultivar in 2003. These host-driven effects could impact the efficacy of traditional pest management approaches and probably complicate efforts to predictively model G. molesta populations in mixed cultivar orchards. Such developmental effects may help to explain previously observed differences in patterns of pheromone trap captures in peach versus apple orchards. Host-associated effects should be incorporated into future models to develop more realistic predictive tools and thus improve integrated pest management efforts.

Effects of orchard host plants on the oviposition preference of the oriental fruit moth (Lepidoptera: Tortricidae).

Recently, the oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), has emerged as a major problem on apples (Malus spp.) grown in the mid-Atlantic and midwestern United States, despite its historically important and frequent occurrence as a peach (Prunus spp.) pest. It is possible that host-driven biological phenomena may be contributing to changes in G. molesta population dynamics resulting in outbreaks in apple. Studies were designed to examine the effects of host plants on oviposition behavior, in an effort to clarify the host association status of eastern U.S. populations and also to gain insight into how pest modeling and management efforts may be altered to take into account various host-associated effects. G. molesta adults exhibited ovipositional preference for nonbearing peach trees over nonbearing apple trees in close-range choice tests conducted in the field, regardless of the larval host origin. A significant preference for peach shoots over apple shoots was observed on six of 12 sampling dates with a wild G. molesta population at the interface of adjacent peach and apple blocks. Numbers of eggs found on apple fruit were higher after peach fruit were harvested and apple fruit began to approach maturity (during the flight period for third and fourth brood adults). Possible implications for population modeling and integrated management of G. molesta are discussed.

Comparative survival rates of oriental fruit moth (Lepidoptera: Tortricidae) larvae on shoots and fruit of apple and peach.

Studies were designed to examine the effects of host plants on oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), survival. G. molesta larval survival in the orchard was comparable between apple (Malus spp.) and peach (Prunus spp.) shoots, indicating that both host species can harbor large larval populations during the early season. G. molesta larvae used multiple shoots for feeding and development on peaches but usually only damaged single shoots in apple. Survival differences were present between peach and apple fruit, but this survival was affected by fruit maturity level. Generally, larval survival higher was in ripening peach fruit than in green, immature apple fruit. Larval survival varied among several apple cultivars, indicating that cultivar-level variability needs to be considered. These host-associated effects may impact efforts to predictively model G. molesta populations in commercial orchards where multiple host plant species, or different cultivars of the same species are often grown in proximity. Thus, host-associated dynamics should be included into future population models that underlie management programs.

Seasonal and cultivar-associated variation in oviposition preference of Oriental fruit moth (Lepidoptera: Tortricidae) adults and feeding behavior of neonate larvae in apples.

The Oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae) has become a pest of tree fruits since its introduction to the United States in the early twentieth century. Oriental fruit moth has historically been a major pest problem in peach production, and outbreaks in commercial apple (Malus spp.) orchards in the eastern United States were rare until the late 1990s. Recent outbreaks in Mid-Atlantic apple orchards have lead researchers to investigate host-associated effects on oriental fruit moth biology, behavior, and population dynamics. Studies were designed to assess cultivar level effects in apples on oviposition and larval feeding behavior of oriental fruit moth. In a mixed cultivar apple orchard, total oriental fruit moth oviposition and oviposition site preferences varied between cultivars. These preferences also varied over time, when sampling was repeated at various times of the growing season. Although most adult female oriental fruit moth preferentially oviposited in the calyx and stem areas of apple fruit, noticeable numbers of eggs also were laid on the sides of fruit, contradicting some previous reports. Oriental fruit moth females exhibited a strong ovipositional preference for fruit that were previously damaged by oriental fruit moth or codling moth, Cydia ponmonella (L.). The majority of newly hatched oriental fruit moth larvae were observed to spend <24 h on the surface of apple fruit before entry, and this behavior was observed on several apple cultivars. Neonate larvae exhibited a preference for entering fruit at either the stem or calyx ends, regardless of their initial site of placement. Our findings underscore the importance of adequate spray coverage and accurate timing of insecticide applications targeting oriental fruit moth.