The planthopper Lycorma delicatula (White) (spotted lanternfly; SLF) was introduced to North America from Asia. It was first found in southeastern Pennsylvania in 2014 and now, a decade later, has increased in abundance and spread into 18 eastern US states. To study naturally occurring fungal pathogens infecting SLF, eastern Pennsylvania sites were sampled every 1 to 2 wk in 2020 and 2021 during the adult life stage of L. delicatula to detect prevalence of infections by the fungi Batkoa major (Thaxt.) Humber (Entomophthorales: Batkoaceae) and Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Cordycipitaceae). To sample, living adult SLF were collected and reared and cause of death was diagnosed. In 2020, at the site hosting a co-epizootic of these 2 generalist pathogens in 2018, an epizootic caused by B. major was documented from 30 September to 21 October. Low levels of infection by both pathogens were detected at an additional 2020 site and both 2021 sites. Overall, there was a negative association of B. major infection with SLF density and no association with density for B. bassiana. Co-infections in individual SLF by both fungi were never documented, and there was an inverse relationship between prevalence of B. major and B. bassiana infections in the sampled populations. At the time that SLF for rearing were sampled, adult cadavers were also sampled. For B. major, infection rates of sampled cadavers and reared individuals were positively correlated, but higher infection rates were observed in cadavers than among reared SLF. For B. bassiana, no such pattern occurred.

Asian longhorned beetles, Anoplophora glabripennis (Motschulsky) (ALB), are native to China and Korea but invasive populations now threaten hardwood forests in North America and Europe where eradication is the main tactic. In North America, invasive ALB were first found in the New York City area in 1996 and, since then, infestations were detected in 5 additional states. Beginning in 1998, studies of fungal entomopathogens for ALB control were begun, followed by studies with entomopathogenic nematodes, Bacillus thuringiensis Berliner, and microsporidia. Studies with entomopathogenic fungi initially focused on a commercialized method used for application of Beauveria asiatica against adult cerambycids in Japanese orchards: cerambycids walk across bands covered with infective spores of entomopathogenic fungi that are attached around trees. For use in the United States, Metarhizium brunneum Petch F52 was grown in fungal bands and tested against adult ALB in US quarantine labs as well as in the field in China. Fungal infection reduced female longevity, fitness, and flight. Long-lived M. brunneum microsclerotia formulated in hydromulch were investigated as an alternative application method. Several species of entomopathogenic nematodes applied to ALB emergence holes showed promise for infecting and killing ALB larvae. The pathogens that have been investigated and developed are considered part of the arsenal of methods for ALB control and not "stand-alone" tactics. While no pathogens have been used operationally in the United States, information gained about these pathogens is available and could be used as situations warrant more tools for managing ALB.

Adult spotted lanternflies (Lycorma delicatula) were differentially susceptible to Beauveria bassiana when inoculated with conidia on the distal ventral abdomen versus distal forewings. More adults inoculated on the abdomens died of B. bassiana infections than those inoculated on the wings. Abdominal inoculants also died more quickly than wing inoculants. Due to the large dorsal forewings of these planthoppers, typically covering abdomen and thorax, we suggest that the wings can at least partially protect from dorsal sprays of an infectious Hypocreales; we hypothesize that spraying surfaces on which SLF stand could be more efficacious than spraying these insects with their bodies shielded by their wings.

The subphylum Entomophthoromycotina (Phylum Zoopagomycota) includes many arthropod pathogens, some of which are renowned for their abilities to alter host behavior prior to death and cause epizootics that impact host populations. The last checklist of arthropod-pathogenic species in this group was published in 1963 and consisted of 39 species in a single genus. Since then, more species have been named, and their taxonomy has changed extensively. We have constructed an updated checklist for species of Entomophthoromycotina in North America; this checklist includes species in the continental United States, Canada, and Mexico. Data were compiled based on available published literature and metadata available from the ARSEF culture collection, adjusting names based on current taxonomy. In North America, the arthropod-pathogenic Entomophthoromycotina now include 80 species belonging to 14 genera, within two classes, plus one species in a form genus. This checklist provides a current framework for future studies of the biodiversity of this group of fungi.

Serropalpus substriatus Haldeman (Coleoptera: Melandryidae) develops within the same trees as Sirex noctilio F. and Sirex cyaneus F. (Hymenoptera: Siricidae). This species is now reported emerging from red pine (Pinus resinosa Sol. ex Aiton) and balsam fir (Abies balsamea (L.) Mill.) (Pinaceae) in eastern North America. Numbers of Se. substriatus emerging from pines were always much lower than numbers of Sirex in trees, and 18% of Se. substriatus required 2 years before emergence. During first years of emergence, Se. substriatus was found in only 12 of 41 sampled red pines infested by Si. noctilio. Trees from which Se. substriatus emerged hosted higher densities of Sirex than trees without Se. substriatus. Comparing dually infested trees, Deladenus siricidicola Bedding, the parasitic/mycophagous nematode associated with Si. noctilio, was found parasitizing one of 141 (0.7%) Se. substriatus. This is the first report of a non-target impact of D. siricidicola in North America, but parasitism was very low in this uncommonly encountered woodboring beetle. In Europe, a related species, Serropalpus barbatus Schaller, has also been reported to be parasitized by D. siricidicola.

The fungal order Entomophthorales occurs worldwide, with most species infecting arthropods as pathogens. Species in this order can cause epizootics and change the behavior of infected hosts. Molecular data are available only for 20% of the known species, and distributions of species are seldom summarized. Significant diversity of hosts, poor molecular data availability, and poor resolution of the phylogenetic relationships within this fungal order suggest that the diversity of these fungi is not sufficiently described. The subfamily Erynioideae includes 111 arthropod pathogens, divided among six genera, with the genus Pandora being one of the most diverse genera. Sequences of 18S, 28S, and ITS for two species are used to place these Pandora species in a phylogenic tree of the subfamily; this tree also supports our synonymy of the genus Furia with Pandora. Among the two species specifically covered in this paper, Pandora gloeospora was observed during epizootics occurring in mushroom flies (Diptera: Sciaridae) on Agaricus bisporus cultures in Pennsylvania, Delaware, and Maryland (US) mushroom farms and also in Florida on Pleurotus sp. Outside the US, P. gloeospora was found infecting several Nematocera (Diptera) in Europe (France) and Asia (China). Pandora sylvestris n. sp. was collected during epizootics occurring in larvae of hickory tussock moths, Lophocampa caryae (Lepidoptera: Erebidae), in hardwood forests in Michigan and Vermont.

The entomophthoralean fungus Zoophthora rhagonycharum (Bałazy) S. Keller, previously recorded in Europe from Poland and Switzerland, is now reported in North America from New York State, United States of America. On both continents, this obligate insect pathogen is known only from resting spores found within dead, adult native soldier beetles (Cantharidae) of the genus Rhagonycha Eschscholtz, 1830. Resting spores have undulating, light brown episporia. In New York, columnar rhizoids attach cadavers tightly to the undersides of leaves in the understory of hardwood forests in late June and early July.

From 1897 until 1957, beetles of the genus Calosoma L. (Coleoptera: Carabidae) were repeatedly introduced to the Hawaiian Islands as predators of nutgrass armyworm, Spodoptera exempta (Walker), armyworm, Mythimna unipuncta (Haworth), and lawn armyworm, Spodoptera mauritia (Boisduval). Through examination of institutional voucher specimens, archival field notes, and the literature, we document failed biological control introductions of eight flight-capable Calosoma species: C. calidum (F.), C. frigidum Kirby, C. marginale Casey, C. peregrinator Guérin-Méneville, C. prominens LeConte, C. protractum LeConte, C. semilaeve LeConte, and C. senegalense Dejean. In contrast, the vestigially-winged C. anthracinum Dejean, a rarely collected Mexican species never intentionally introduced, is the only Calosoma currently resident in Hawaiʻi. Calosoma anthracinum was misidentified initially as C. semilaeve, and thereafter misidentified as Calosoma blaptoides tehuanacum Lapouge. Both C. anthracinum and C. protractum were collected sympatrically at Cuernavaca, Morelos, Mexico during Hawaiʻi-based explorations, and we hypothesize that they were introduced together as a species mixture in a 1923 release made at Waikiʻi, Parker Ranch, Hawaiʻi Island. Calosoma anthracinum was little known to taxonomists at that time due to its limited native distribution, rarity in institutional collections, and absence from taxonomic keys, leading to repeated misidentifications. Calosoma peregrinator and C. prominens, two abundant, sympatric species from the Sonoran and Chihuahuan Deserts, were also taxonomically confused over this period, resulting in mixed-species Hawaiian introductions. Three species that failed to establish upon introduction—C. marginale, C. peregrinator and C. semilaeve—have been intercepted as singletons or pairs of individuals decades later, without apparent establishment. Calosoma beetles' ecological requirements, dispersal abilities, introduction histories, and native ranges and associated climate are compared among species, the only unique characteristic of the successful colonist being flightlessness. The history of Hawaiian Calosoma introductions illustrates the need for robust and accessible classifications based on accurately determined specimens as well as comprehensive understanding of the biological attributes and habitat requirements of candidate biological control agents.

Biological control is often a key component of management strategies for invasive species. Yet, the effectiveness of biological control can be limited by a poor understanding of natural enemy ecology. To overcome this, habitat suitability models can predict distributions of invasive species and identify areas of potential overlap between invaders and natural enemies to guide biological control. Here we used data from a coordinated national monitoring network and a novel modeling method that incorporates physiology into correlative niche models to predict potential distributions of the brown marmorated stink bug, Halyomorpha halys, and two natural enemies (a parasitoid, Trissolcus japonicus, and a microsporidian, Nosema maddoxi) in current and future climates (2070s). We show all three species have broad similarity in habitat suitability, with especially high overlap in the mid-Atlantic and southeastern US where H. halys populations were first established. Each species will likely expand their range across the northern US in the future, but the overlap between species may decrease. In much of the central and southeastern US, H. halys may decrease its range over time, although natural enemies may be less impacted, and overlap between species may increase. Our study shows that biological control provided by T. japonicus and N. maddoxi could be key for managing H. halys given their overlapping niches, and our models can aid in delineating areas where biocontrol may be most effective. Our method of linking field data with correlative niche models can also be used for other insects.

Despite over a century of observations, the obligate insect parasites within the order Entomophthorales remain poorly characterized at the genetic level. In this manuscript, we present a genome for a laboratory-tractable Entomophthora muscae isolate that infects fruit flies. Our E. muscae assembly is 1.03 Gb, consists of 7810 contigs and contains 81.3% complete fungal BUSCOs. Using a comparative approach with recent datasets from entomophthoralean fungi, we show that giant genomes are the norm within Entomophthoraceae owing to extensive, but not recent, Ty3 retrotransposon activity. In addition, we find that E. muscae and its closest allies possess genes that are likely homologs to the blue-light sensor white-collar 1, a Neurospora crassa gene that has a well-established role in maintaining circadian rhythms. We uncover evidence that E. muscae diverged from other entomophthoralean fungi by expansion of existing families, rather than loss of particular domains, and possesses a potentially unique suite of secreted catabolic enzymes, consistent with E. muscae’s species-specific, biotrophic lifestyle. Finally, we offer a head-to-head comparison of morphological and molecular data for species within the E. muscae species complex that support the need for taxonomic revision within this group. Altogether, we provide a genetic and molecular foundation that we hope will provide a platform for the continued study of the unique biology of entomophthoralean fungi.

The fungal order Entomophthorales in the Zoopagomycota includes many fungal pathogens of arthropods. This review explores six genera in the subfamily Erynioideae within the family Entomophthoraceae, namely, Erynia, Furia, Orthomyces, Pandora, Strongwellsea, and Zoophthora. This is the largest subfamily in the Entomophthorales, including 126 described species. The species diversity, global distribution, and host range of this subfamily are summarized. Relatively few taxa are geographically widespread, and few have broad host ranges, which contrasts with many species with single reports from one location and one host species. The insect orders infected by the greatest numbers of species are the Diptera and Hemiptera. Across the subfamily, relatively few species have been cultivated in vitro, and those that have require more specialized media than many other fungi. Given their potential to attack arthropods and their position in the fungal evolutionary tree, we discuss which species might be adopted for biological control purposes or biotechnological innovations. Current challenges in the implementation of these species in biotechnology include the limited ability or difficulty in culturing many in vitro, a correlated paucity of genomic resources, and considerations regarding the host ranges of different species.

Insect pest species are generally expected to become more destructive with climate change because of factors such as weakened host tree defences during droughts and increased voltinism under rising temperatures; however, responses will vary by species due to a variety of factors, including altered interactions with their natural enemies. Entomopathogens are a substantial source of mortality in insects, but the likelihood of epizootics can depend strongly on climatic conditions. Previous research indicates that rates of infection of the spongy moth (Lymantria dispar) by its host-specific fungal pathogen, Entomophaga maimaiga, increase with environmental moisture and decrease as temperatures rise. High temperatures may have direct and indirect (due to the associated drying) effects on the fungus, but the interactive effects between temperature and moisture level on larval infection are unclear. Here, we test the hypothesis that warmer, drier conditions will decrease rates of infection of spongy moth larvae by E. maimaiga. We evaluated the effects of precipitation and temperature on larval mortality caused by E. maimaiga with a manipulative field experiment, conducted in one of the northernmost and coldest parts of the spongy moth’s non-native range in North America. We caged laboratory-reared spongy moth larvae in experimentally warmed open-air forest plots, exposing the larvae to soil inoculated with E. maimaiga resting spores during two consecutive trials. Caged larvae were exposed to three temperature treatments — ambient, 1.7 °C above ambient and 3.4 °C above ambient — and either supplemental precipitation (+173 mm per trial) or ambient precipitation. Opposite to our hypothesis, there was no significant effect of supplemental precipitation, nor an interaction between precipitation and temperature. There was, however, a significant positive effect of increasing temperature on the number of larvae infected. On average, in each respective trial, larval infection increased by 44% and 50% under the elevated temperature treatments compared to ambient temperature. Experimental warming may have increased infections because ambient temperatures at the field site were suboptimal for fungal germination. The results from this experiment suggest that, in colder portions of the spongy moth’s invasive range, increasing temperatures due to climate change may enhance the ability of E. maimaiga to help control populations of the spongy moth.

The generalist entomophthoralean insect pathogen Batkoa major was recorded causing epizootics in populations of a new invasive fulgorid in North America, the spotted lanternfly (Lycorma delicatula). We conducted studies on the basic biology and ecology of B. major using Galleria mellonella larvae exposed to conidial showers. Death of G. mellonella followed a diurnal cycle with most larvae dying within 4 h before or after the end of photophase. Time for initiation of rhizoid emergence also followed a diurnal rhythm and, on average occurred 3.6 h after host death. While B. major sometimes began producing rhizoids to attach cadavers to substrates while G. mellonella were alive (but moribund), often hosts were dead before rhizoids began emerging. On average, conidial discharge began 18.6 h after host death and was greater 4-8 h before the end of photophase, compared with 4-8 h after scotophase began. At 20 degrees C under high humidity, initiation of conidial discharge was 95% complete within 24 h after host death. To evaluate B. major activity by temperature, we tested percent conidial germi-nation over 24 h from 5 to 35 degrees C. When showered onto water agar, all primary conidia produced secondary conidia. At 20 and 25 degrees C, at 3 h >= 89% of primaries had produced and discharged secondaries and from 10 to 30 degrees C, secondaries were produced by over 75% of primary conidia within 12 h. When cover slips were placed over primary conidia to force production of germ tubes, germination was much slower, with >85% germination from 20 to 30 degrees C only by 24 h. Batkoa major therefore times host death and initiation of conidial discharge for night-time hours and conidial germination occurs within 24 h over a broad temperature range (10-30 degrees C).

Dimorphic nematodes in the genus Deladenus have been used or are being considered for use in biological control of the invasive Eurasian woodwasp, Sirex noctilio, which threatens pine (Pinus spp.) trees. Deladenus species that are parasitic on Sirex can kill woodwasp eggs and occupy these same eggs for their own dispersal. These nem-atodes also have mycophagous phases that feed on the white rot fungal symbionts of Sirex, Amylostereum species. The mycophagous stage of the Hungarian strain of Deladenus siricidicola developed for control of S. noctilio in Australia feeds exclusively on A. areolatum. The mycophagous stage of a North American Deladenus species being evaluated for biological control, D. proximus, feeds on either A. chailletii, or A. areolatum. Amylostereum species and strains associated with Sirex have differential impacts on survival and growth of these nematodes. We investigated whether differences in species and strains of Amylostereum influence the numbers of Deladenus ju-veniles adjacent to cultures, as this would impact potential parasitism of Sirex. Fungal species or strain did not influence persistence of juveniles in the fungal vicinity although retention could be influenced by the fungal strain consumed by parents. Investigating D. proximus, we tested whether the most common invasive strain of A. areolatum associated with S. noctilio in North America (IGS D) impacted nematode growth, compared with the common native Amylostereum chailletii. Deladenus proximus increased very slowly when feeding on A. areolatum IGS D, compared with A. chailletii; when provided A. areolatum IGS D, 55 eggs were produced after 4 weeks compared with 8.1 x 104 eggs after 2 weeks when A. chailletii was provided. In summary, behavior of Deladenus juveniles resulted in no or low avoidance of Amylostereum species and strains, regardless of poor growth when mycophagous forms fed on fungi on which growth was poor..

An epizootic caused by fungal pathogens occurred among Halyomorpha halys, brown marmorated stink bugs, while they were overwintering, with infections also occurring after overwintering. We report that one of the two pathogens responsible was Colletotrichum fioriniae (Marcelino & Gouli) Pennycook; a species well known as a plant pathogen and endophyte and which has only previously been reported naturally infecting elongate hem-lock scales, Fiorinia externa. To prove pathogenicity, H. halys adults challenged with conidia died from infections and the fungus subsequently produced conidia externally on cadavers.

In the eastern United States, populations of the invasive spotted lanternfly, Lycorma delicatula, are abundant and spreading. Four species of naturally occurring entomopathogenic fungi have previously been reported as infecting these planthoppers, with two of these causing epizootics. Nymphal- and adult-stage lanternflies in Pennsylvania and New York were surveyed for entomopathogenic fungal infections from October 2021 to November 2023, and assays were conducted to confirm the pathogenicity of species that were potentially pathogenic. Beauveria bassiana was the most abundant pathogen, but we report an additional 15 previously unreported species of entomopathogenic fungi infecting spotted lanternflies, all in the order Hypocreales (Ascomycota). The next most common pathogens were Fusarium fujikuroi and Sarocladium strictum. While infection prevalence by species was often low, probably impacted to some extent by the summer drought in 2022, together these pathogens caused a total of 6.7% mortality. A significant trend was evident over time within a season, with low levels of infection among nymphs and higher infection levels in mid- and late-stage adults, the stages when mating and oviposition occur.

The entomopathogenic fungus Beauveria bassiana is cosmopolitan and known to infect a variety of sap-sucking pests like aphids, mealybugs, and scales in the order of Hemiptera. In Fall 2017, spotted lanternfly (SLF) adults killed by the fungal entomopathogen B. bassiana were found in Berks County, Pennsylvania. In 2018-2020 we collected SLF and nearby non-target insects killed by Beauveria spp. from 18 field sites in southeastern Pennsylvania. We identified 159 Beauveria isolates from SLF and six isolates from non-targets. Five isolates of B. bassiana and one isolate of B. brongniartii were identified from the non-targets. Based on sequence data from the nuclear B locus (Bloc) intergenic region, all the isolates from SLF were identified as B. bassiana , but there were 20 different strains within this species, grouped into two clades. Three B. bassiana strains (A, B, and L) were found in most field sites and were the most prevalent. Representative isolates for these three strains were used in laboratory bioassays and were compared to a commercial B. bassiana strain (GHA). Strain B was inferior to A, L, and GHA against nymphs; strains A and L had greater efficacy than B and GHA against adults. We also quantified conidial production on SLF cadavers. This paper discusses the diversity of these B. bassiana strains in SLF populations and implications for biological control of this abundant invasive.