JHR 87:503-521 (2021) ge, JOURNAL OF eeertennacnn nn doi: 10.3897/jhr.87.7619 | RESEARCH ARTICLE ) I Tymenopter a 4 https://jhr.pensoft.net Te Inarasional Society of ymenopreriss, RESEARCH Psix striaticeps (Dodd) (Hymenoptera, Scelionidae): an Old World parasitoid of stink bug eggs arrives in Florida, USA Sarah Birkmire!, Cory Penca’, Elijah J. Talamas’, Matthew R. Moore’, Amanda C. Hodges' | Department of Entomology and Nematology, University of Florida, Gainesville, FL, USA 2. USDA-APHIS, Miami, FL, USA 3 Florida Department of Agriculture and Consumer Services, Division of Plant Industry, Gainesville, FL, USA Corresponding author: Elijah J. Talamas (elijah.talamas@fdacs.gov) Academiceditor: Zachary Lahey | Received 11 October 2021 | Accepted 8 November 2021 | Published 23 December 2021 http://zoobank.org/CA77B398-3072-4E25-9D3F-E966A61 CD6FF Citation: Birkmire S, Penca C, Talamas EJ, Moore MR, Hodges AC (2021) Psix striaticeps (Dodd) (Hymenoptera, Scelionidae): an Old World parasitoid of stink bug eggs arrives in Florida, USA. In: Lahey Z, Talamas E (Eds) Advances in the Systematics of Platygastroidea HI. Journal of Hymenoptera Research 87: 503-521. https://doi.org/10.3897/ jhr.87.76191 Abstract Psix striaticeps (Dodd) is an egg-parasitoid wasp previously known only from the Old World. We report this species from twelve counties in Florida, which are the first records in the Western Hemisphere. It was collected in yellow cylinder traps and reared from the eggs of three stink bug species: Nezara viridula L.., Chinavia marginata (Palisot de Beauvois), and Piezodorus guildinii (Westwood). A COI barcode analysis found a 100% match between the Floridian population and a specimen from South Africa. The prospects of using Ps. striaticeps as a biological control agent against exotic stink bugs are discussed. Keywords biological control, adventive species, host records, DNA barcoding Copyright Sarah Birkmire et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 504 Sarah Birkmire et al. / Journal of Hymenoptera Research 87: 503-521 (2021) Introduction Parasitoid wasps are important regulators of host populations, and for this reason are often used for biological control in agricultural ecosystems (Tillman 2011; Ogburn et al. 2016; de Freitas Bueno et al. 2020). Augmentative control of exotic pests using native parasitoids is sufficient in some cases, and others may require classical biological control: the introduction of exotic, coevolved parasitoids. We initiated a study to char- acterize the parasitoid fauna associated with two exotic stink bug pests in Florida, the southern green stink bug Negara viridula L. (1758) and the brown marmorated stink bug Halyomorpha halys (Stal, 1855) (Hemiptera, Pentatomidae), in order to determine if augmentative control is an option for management for these pests. Nezara viridula is a polyphagous pest, first reported in the United States by Distant (1880) and first detected in Florida in 1920 (Drake 1920). It affects the production of many food and fiber crops but prefers legumes such as soybean (Panizzi and Meneguim 1988). It is considered to be a pest in a number of states, but has a distribution mostly limited to the southern United States (Turnipseed and Kogan 1976). The species has been detected in states outside of this region including Texas, California and Washing- ton (Hoffman et al. 1987; Equivel et al. 2018). Numerous egg-parasitoids have been reported from JN. viridula in the southeastern United States (Tillman 2011), but no detailed studies have been conducted to characterize its parasitoid fauna in Florida. Halyomorpha halys is a well-known, widespread invasive pest in North America (Haye and Weber 2017). The species is broadly polyphagous, recorded from over 200 species of host plants (Bernon et al. 2004; Lee et al. 2013; Haye et al. 2014; Bergmann et al. 2016). The first detection of the species in the United States occurred around 1996 in Allentown, Pennsylvania (Hoebeck and Carter 2003). The initial detection of Hal. halys within an agricultural system in Florida occurred in a peach orchard in Lake County during 2016, and increasing numbers of Hal. halys were collected through trapping efforts in 2017 and 2018 (Penca and Hodges 2018). During 2018, detec- tion of eggs and nymphs at one of the field trapping locations resulted in a limited distribution pest status declaration for Lake County by regulatory personnel at the Florida Department of Agriculture and Consumer Services, Division of Plant Industry (FDACS-DPI). Natural enemies of Hal. halys were reviewed by Abram et al. (2017), but Hal. halys was not established in Florida at the time, and consequently a direct as- sessment of its egg parasitoids in the state was not included. Tillman (2011) and Tillman et al. (2020) characterized the parasitoids attacking eges of N. viridula and Hal. halys in Georgia and Alabama, which are adjacent to Flor- ida. Tillman (2011) reported six species associated with NV. viridula eggs in agricultural ecosystems including Hadronotus obesus (Masner, 1983), Trissolcus basalis (Wollaston, 1858), 77. brochymenae (Ashmead, 1881), Tr. thyantae Ashmead, 1893, Telenomus po- disi Ashmead, 1893 (Hymenoptera, Scelionidae), and Ovencyrtus spp. (Hymenoptera, Encyrtidae). These species were all reared from naturally occurring stink bug egg mass- es found in corn, peanut and cotton farmscapes in Georgia. Sentinel egg masses were deployed by Tillman (2020) in areas where wild populations of Hal. halys are present, utilizing both fresh and frozen Hal. halys egg masses. Frozen egg masses do not have Psix striaticeps arrives in Florida 505 the risk of introducing nymphs into the field and can increase the likelihood of suc- cessful parasitism by inhibiting the immune response of the eggs against parasitoid eggs and larvae (Haye et al. 2015). Eleven parasitoid species, including two new as- sociations, emerged from Hal. halys egg masses deployed by Tillman et al. (2020) and included Ovencyrtus spp., Acroclisoides sinicus (Huang & Liao, 1988) (Hymenoptera, Pteromalidae), Anastatus reduvii (Howard, 1880), A. mirabilis (Walsh & Riley, 1869) (Hymenoptera, Eupelmidae), Te. podisi, Tr. brochymenae, Tr. eddesae (Fouts, 1920), Tr. euschisti (Ashmead, 1893), 77. solocis Johnson, 1985 and Had. obesus. Except for A. mirabilis, Tr. solocis, and Had. obesus, each of these parasitoid species have been re- covered from wild Hal. halys egg masses (Tillman, unpublished data). Trissolcus japonicus (Ashmead, 1904) is an egg parasitoid that coevolved with Hal. halys and is adventive in the United States (Talamas et al. 2015). This species is not yet approved for release by federal agencies, but it has been used as a biological control agent, via within-state releases, in states where it has established on its own. (Jentsch 2017; Lowenstein et al. 2019; Milnes and Beers 2019). Trissolcus japonicus was recently recovered in North Carolina (stopBMSB.org), indicating that it is slowly but steadily moving south from the mid-Atlantic region. Because 1. halys populations are still low in Florida, detection of Tr. japonicus in the state might enable management early enough to avoid the large-scale impacts that have afflicted other regions. Although we have yet to find 77 japonicus in Florida, our surveys nonetheless de- tected an adventive parasitoid wasp that may be of consequence. Materials and methods Sentinel egg masses Sentinel egg masses of Hal. halys and N. viridula were deployed at three agricultural sites in Lake County, Florida, between March and August of 2021. Deployments were conducted weekly from March to June with a shift to bi-weekly deployments starting in July due to a shortage of eggs. For all but one week where egg mass availability was low, 120 egg masses were deployed per week. Of the 120 egg masses, approximately half were Hal. halys eggs and half N. viridula eggs. The first site, located in Okahumpka, FL (28.744666N, — 81.895032E), includes 8 acres where uncertified organic practices are utilized. The farm is surrounded by woodlands, and over 120 varieties of crops are rotated within the farm seasonally. The second site, located in Clermont, FL (28.629214N, — 81.76725E), includes 69.4 acres of muscadine grapes, but the study was conducted in a 25.4-acre plot of the vineyard. The vineyard used integrated pest management practices that include conventional pesticides. The vineyard is bordered by a thin strip of forest and is adjacent to a lake. The third site, located in Lady Lake, FL (28.927383N, — 81.904568E), is a 2.5-acre orchard of peach trees within a residential community. It has a border of weedy plants and hardwood trees. The past management style was conventional with some organic practices. However, management practices did not occur during this study. 506 Sarah Birkmire et al. / Journal of Hymenoptera Research 87: 503-521 (2021) The WN. viridula egg masses were sourced from a colony at the University of Florida's Ento- mology and Nematology Department. The stink bugs were reared in mesh cages (39.9 cm x 39.9 cm x 61.0cm). They were fed a diet of tomatoes, peanuts, baby carrots, and sweet corn on a Monday, Wednesday, Friday schedule. ‘The photoperiod in the rearing rooms was 16L:8D, and the average temperature and relative humidity were 24.5 °C and 42%, respectively. The Hal. halys egg masses were sourced from a quarantine colony at FDACS-DPI and the New Jersey Department of Agriculture (NJDA) Phillip Alampi Beneficial In- sect Lab. The FDACS-DPI colony of H. halys was reared in round, mesh-ventilated, polystyrene containers of various sizes. The temperature and relative humidity were +1 °C and 5% of the conditions in Medal et al. (2012), respectively. The colony was fed a diet of whole sweet corn, snap peas, peanuts, and organic baby carrots. The NJDA colony of Hal. halys was reared in quarantine according to protocols by Dorsey and Lovero (2014) and was fed a diet designed by Cohen (2000). Eggs were shipped to FDACS-DPI and were either directly frozen or reared to adulthood accord- ing to the protocols described above. All egg masses were frozen at -80 °C prior to their deployment and were glued us- ing Elmer’s glue to 1.5 x 1.5 cm squares of waterproof paper (Rite in the Rain, Tacoma, WA). The sentinel egg mass cards were then pinned to the undersides of the leaves of plants within the crop field and at its periphery. If the egg masses could not be pinned directly to plants, they were clipped to orange and yellow flags which were staked ap- proximately 12 cm above the ground. These measures were taken to reduce damage to plant products that would eventually be sold. The egg masses were left in the field for 72 hours, then returned to the lab where they were recounted, and predation damage was documented. ‘The egg masses were then placed into small petri dishes sealed with parafilm and stored in a quarantined room. The temperature and relative humidity in this room were 26.5 °C and 48%, respectively. The photoperiod in this room was 16L:8D. The egg masses were checked daily for parasitoid emergence, which often occurred between 20—25 days of storage and were stored for a total of 30 days before being discarded. Reared parasitoids were stored in 95% ethanol prior to identification. Collection of wild stink bug egg masses A single, wild stink bug egg mass, unidentified at the time, was collected on a squash plant in Lake County, on May 25", 2021. The egg mass was brought back to the lab for assessment of parasitism and identification of the stink bug species via COI barcoding. Eggs, nymphs and adults of the redbanded stink bug Piezodorus guildinii (West- wood, 1837) were collected from an unmanaged planting of hairy indigo (/ndigofera hirsuta L.) located at the University of Florida Plant Science Research and Education Unit in Marion County, Florida (29.4079530 N, 82.1459287 W). Eggs were hand collected and placed into 10-dram plastic vials until eclosion, after which the emerged Pi. guildinii nymphs were transferred to a larger cage for rearing. This was done to prevent accidental introduction of egg parasitoids into the main Pi. guildinii colony as the dark color of Pi. guildinii eggs makes it difficult to identify parasitism. Collections were made throughout the spring and summer of 2018. Psix striaticeps arrives in Florida 507 Yellow cylinder trap Yellow, 3D-printed cylinder traps (Figure 1) were deployed in Florida by the Coop- erative Agricultural Pest Survey (CAPS Program) throughout the year to monitor for exotic pests, especially psyllids in a variety of agricultural ecosystems (Snyder et al. 2019). These traps also serve well for collecting other insects that are attracted to yel- low, and residues were screened for parasitoid wasps. The preservative in the collection tube (Figure 1B) was propylene glycol (Prestone LoTox) with salt. Figure |. Yellow cylinder trap employed by CAPS Program in Florida A exterior view of trap; the inside of the funnel is coated with Insect-a-Slip B interior of trap where insects are preserved in a vial containing propylene glycol and salt. Material examined The collection data and images associated with the specimens examined were depos- ited in the database of the Museum of Biological Diversity, The Ohio State University (mbd-db.osu.edu), and a DarwinCore archive file is provided in Suppl. material 1. Collecting unit identifiers associated with specimens are listed by county below. Psix striaticeps: 54 females, 3 males, deposited in Florida State Collection of Ar- thropods (FSCA). Florida: Alachua Co. (FSCA 00095801, 00095797); Brevard Co. (FSCA 0009579 1-00095792, 00095830); Hardee Co. (FSCA 00034061, 00034693, 00034696); Hillsborough Co. (FSCA 00095795—00095796, 00095798, 00095800); 508 Sarah Birkmire et al. / Journal of Hymenoptera Research 87: 503-521 (2021) Lake Co. (FSCA 00094219, 00094237); Lee Co. (FSCA 00095788—000-5789); Mar- ion Co. (FSCA 00095892-—00095894); Miami-Dade Co. (FSCA 00095831); Orange Co. (FSCA 00034688—00034689, 00095793—00095794, 00095813—00095823); Pi- nellas Co. (FSCA 00095826—00095827, 00095790); St. Johns Co. (FSCA 00095799, 00095803—00095809, 00095811—00095812, 00095828—00095829, 00095891, 00095896—00095899); St. Lucie Co. (FSCA 00095824—00095825). Morphological identification Specimens of Psix striaticeps were identified using the key to Psix species by Johnson and Masner (1985). Molecular analysis Genomic DNA was nondestructively isolated from the whole adult specimens using the Qiagen DNeasy kit (Hilden, Germany) as described by Giantsis et al. (2016). The host eggs were pulverized prior to analysis and genomic DNA was isolated using the Qiagen DNeasy kit (Hilden, Germany) as described by Giantsis et al. (2016). Poly- merase chain reaction (PCR) was carried out to amplify the DNA barcode region of the Cytochrome Oxidase Subunit I (COI) using the LCO/HCO primers of Folmer et al. (1994). For the host eggs, the PENT_F2/HCO primer was used as described by Gariepy et al. (2014). The PCR was performed in a 25 ul reaction volume using the KAPA HiFi Hotstart Ready Mix (Roche) per the manufacturer’s standard protocol. PCR conditions were as follows: 95°C for 2 min, followed by 32 cycles of 95 °C for 30 s, 50 °C for 40 seconds, 72 °C for 1 min with a final extension at 72 °C for 7 min. All PCR products were electrophoresed through an agarose gel (1.5%) and were puri- fied and prepared for sequencing using BigDYE Xterminator (Applied Biosystems) per the manufacturer’s protocol. Products were then sequenced in both directions at FDACS-DPI, Gainesville, Florida. Chromatograms were then trimmed in Sequencher 5.4.6. and assembled into contigs. Amino acids were translated using MEGA7 (Kumar et al. 2016). Data was deposited in GenBank under accession numbers OK157284— OK157305, OK327010—OK327011 (Table 1). Both GenBank (Altschul et al. 1990) and BOLD (Ratnasingham and Hebert 2007) were queried, and sequences for the Psix group of genera (Psix Kozlov & Lé, Paratelenomus Dodd, Nirupama Nixon) barcodes were downloaded from BOLD. Based on the phylogenetic topology of Chen et al. (2021), a species of Telenomus was selected as an outgroup for the neighbor-joining analysis. DNA barcodes were aligned using MUSCLE with default settings as implemented in MEGA7 (Tamura et al. 2013). Neighbor-joining of DNA barcodes was performed in MEGA7 using K2P distance (Kimura 1980) with 10,000 bootstrap replicates. The resulting Ne- wick tree file (Suppl. material 2.) was altered in the Interactive Tree of Life (Letunic and Bork 2021) portal to collapse terminal clusters and apply bootstrap support values to branches. Psix striaticeps arrives in Florida 509 Results We collected specimens of Ps. striaticeps from twelve counties in Florida (Table 1, Figure 14). In two counties, Ps. striaticeps was reared from stink bug egg masses, in nine it was collected in yellow cylinder traps, and in one it was collected by an unspecified meth- od. The specimens that emerged from eggs of Pi. guildinii (Figure 10), Marion County, 2018, are the earliest record of the species in Florida. In Lake County, 2021, Ps. striaticeps emerged from a sentinel NV. viridula egg mass and a wild egg mass that was determined by its COI sequence to be Chinavia marginata (Palisot de Beauvois, 1817) (Figure 9). Table |. Locality data, collection method, collecting unit identifiers, and GenBank accession numbers for P striaticeps and C. marginata collected in Florida. Species County Collecting method Collecting Unit Identifier Genbank accession number C. marginata Lake hand collected (egg mass) FSCA 00095810A OK157305 Ps. striaticeps Lake hand collected (in egg mass) FSCA 00095810B OK157288 Alachua yellow cylinder trap FSCA 00095797 OK157292 Brevard yellow cylinder trap FSCA 00095792 OK157299 Brevard yellow cylinder trap FSCA 00095791 OK157300 Hillsborough yellow cylinder trap FSCA 00095796 OK157293 Hillsborough yellow cylinder trap FSCA 00095795 OK157294 Lake ex. wild C. marginata egg mass FSCA 00094237 OK157284 Lake ex. wild C. marginata egg mass FSCA 00093806 OK157286 Lake ex. sentinel NV. viridula egg mass FSCA 00094219 OK157285 Lee yellow cylinder trap FSCA 00095789 OK157295 Lee yellow cylinder trap FSCA 00095788 OK157296 Marion ex. wild Piezodorus guildinii egg mass FSCA 00095892 OK157303 Marion ex. wild Piezodorus guildinii egg mass FSCA 00095893 OK157304 Miami-Dade not specified FSCA 00095831 OK157289 Orange yellow cylinder trap FSCA 00095793 OK157301 Orange yellow cylinder trap FSCA 00095794 OK157302 Pinellas yellow cylinder trap FSCA 00095790 OK157290 Pinellas yellow cylinder trap FSCA 00095827 OK157291 St. Johns yellow cylinder trap FSCA 00095799 OK157287 St. Lucie yellow cylinder trap FSCA 00095824 OK157297 St. Lucie yellow cylinder trap FSCA 00095825 OK157298 Figures 2-3. Psix striaticeps, habitus, lateral view. Images are presented at identical scale size to illustrate intraspecific size variation 2 female (FSCA 00095801) 3 female (FSCA 00095798). 510 Sarah Birkmire et al. / Journal of Hymenoptera Research 87: 503-521 (2021) Morphological identification We identified specimens as Ps. striaticeps based on the following characters in the key to species by Johnson and Masner (1985): radicle black; frons without sub- median carina; width of frons greater than height of compound eyes; metas- cutellum (dorsellum) protruding weakly; ventral lip of metascutellum punctu- late; posterior margin of T2 punctulate; lateral sulci on S2 widely separated; area between sulci on S2 covered with large seta-bearing punctures (Figures 4—8). P, otf 5 { \ Wy i, rs 0.2mm y - Figures 4-8. Psix striaticeps, female (FSCA 00095790) 4 head, anterolateral view 5 head and mesosoma, lateral view 6 mesosoma, posterior view 7 metasoma, posterodorsal view 8 metasoma, ventral view. msct: metascutellum; r: radicle; S2: second metasomal sternite; T2: second metasomal tergite. Psix striaticeps arrives in Florida Sil > 4 ™" } ae ; ‘ 7 my me ; He 3 % \ : cod / ‘ H way ae &. ’ mk fe , ge , —> a i eourtee' a} pa A A eee a 9 | oO Oe Rh! eee OF Re eer > CS Su iit ai ip Seer : : d Figures 9-10. Wild, parasitized egg masses 9 Chinavia marginata (FSCA 00095810) 10 Piezodorus guildinii (FSCA 00095895). DNA barcoding Our analysis of COI sequences retrieved two clades of Ps. striaticeps that differ from each other by approximately 2.5% K2P distance (Suppl. material 3.). One of the specimens in BIN:BOLD:ADV1998 was identified by Taekul et al. (2014), and the specimens that match the Floridian population were previously unidentified (BIN:BOLD:ADX9028) (Figure 11). The Floridian specimens are identical to each other and to one of the specimens from South Africa (KMPUD3236-19). The wild egg mass of C. marginata (Figure 9) was identified by a BOLD que- ry that matched its COI sequence to specimens from Costa Rica (99.8%; BIN BOLD:AEE5175). Chinavia marginata occurs in Florida, the West Indies, and the southwestern US south to northern South America (Rolston 1983). Lo Te) Te) °o = ee N N oO Oo oO =) =) Telenomus alecto ; Paratelenomus saccharalis Psix sp. QO. é * re Psix watshami OO Nirupama sp. 0.8 Paratelenomus anu Psix tunetanus Psix saccharicola Psix sp. Psix striaticeps Figure I 1. K2P Neighbor-joining tree of COI for Psix group of genera. Bootstrap support values over 80 are labelled on the branches. 512 Sarah Birkmire et al. / Journal of Hymenoptera Research 87: 503-521 (2021) Identification of Nearctic Psix The two species of Psix that are now found in the Nearctic can be separated by sculp- tural differences on the frons: lateral to the central keel, the frons in Ps. striaticeps is rugulose (Figure 12) and in Ps. tunetanus it is smooth (Figure 13). sla Figures 12-13. 12 Psix striaticeps, female (FSCA 00095790), head, anterior view 13 Psix tunetanus, female (FSCA 00095802), head, anterior view. ck: central keel. Discussion During the past decade, classical biological control has been actively pursued for three invasive pentatomoid bugs in the United States, Megacopta cribraria F., 1798, Hal. halys, and Bagrada hilaris (Burmeister, 1835). For each pest, scelionid egg parasitoids tested in quarantine have been found to be adventive prior to approved release. Paratelenomus saccharalis (Dodd, 1914), a parasitoid of M. cribraria, was detected in the southeastern United States by Gardener et al. (2013), and reared from WM. cribraria egg masses in Georgia, documenting the first occurrence of this species in the western hemisphere. This was followed by 77. japonicus (Talamas et al. 2015), a parasitoid of Hal. halys, and then by parasitoids of B. hilaris eggs, Tr. hyalinipennis Rajmohana & Narendran, 2007 and Gryon aetherium Talamas, 2021 (Ganjisaffar et al. 2018; Hogg et al. 2021, Talamas et al. 2021). Before this recent spate of introductions, Johnson and Masner (1985) reported Ps. tunetanus (Mineo & Szabo 1979) in the southwestern United States. Psix tunetanus is originally an Old World species, found in Saudi Arabia, Tunisia, the Ivory Coast, and Gambia. It was suggested by Johnson and Masner (1985) that it arrived in the New World via shipments of Mediterranean plants into the region during the 20" century. We suspect that the introduction of Ps. striaticeps followed a similar pathway. The uniformity of the COI barcodes from Florida specimens of Ps. striaticeps suggests a single introduction. Psix striaticeps arrives in Florida 513 Phenology Johnson and Masner (1985) reported that Ps. striaticeps was collected year-round in Zimba- bwe, suggesting that it is multivoltine and possibly polyphagous. Our data on Ps. striaticeps in Florida supports these ideas. Specimens were collected in yellow cylinder traps continu- ously from December 2020 to August 2021. The females of Ps. striaticeps in Figures 2-3 are presented at identical scale size, and the specimen in Figure 2 is 1.5 times as long as the specimen in Figure 3. This size variation is consistent with Ps. striaticeps parasitizing eggs of various stink bug species, which vary in the size of their eggs, directly affecting the size of the parasitoid (Medal and Smith 2015). In Florida, Ps. striaticeps has been reared from the eges of Pi. guildinii, N. viridula, and C. marginata, indicating that it is at least oligophagous. eae ps Ea Figure | 4. Distribution map indicating the counties (red) in Florida where Ps. striaticeps has been collected. Establishment The twelve counties where we identified Ps. striaticeps span peninsular Florida from north to south and include localities on the Atlantic and Gulf coasts (Figure 14). The earliest record is from 2018 in Marion County, followed by a single specimen col- lected in a hemp plantation in Miami-Dade County in 2019. The broader distribu- tion that we detected in 2021 demonstrates the importance of continuous monitoring programs, such as CAPS, and their value for detecting a range of insects. Given that Ps. striaticeps is widespread in Florida and has been in the state for at least three years, we consider this species to be well-established. 514 Sarah Birkmire et al. / Journal of Hymenoptera Research 87: 503-521 (2021) Pest management The establishment of Ps. striaticeps in Florida has implications for stink bug manage- ment and ecology. It offers the prospect of biological control of N. viridula, from which it has been recorded previously (Johnson and Masner 1985), but the nontarget effects of Ps. striaticeps are yet unknown. Johnson and Masner (1985) also reported Acrosternum acutum (Dallas, 1851) (Pentatomidae) and Clavigralla scutellaris (West- wood, 1842) (as Acanthomia brevirostris Stal, 1873) (Coreidae) as hosts, indicating that its host range includes at least two superfamilies of true bugs. It has yet to be determined if Ps. striaticeps can parasitize Hal. halys eggs, and if it does, there is the possibility of its use for biological control of this species, particularly because there may be a suitable match in the phenologies of these species. Halyomorpha halys typically has two generations per year in mild climates, but in warmer climates, such as Florida, the species can give rise to several more generations per year (Hoffman 1933; Kistner 2017). Part of this effect may be that warmer climates tend to be at lower latitudes and have less drastic changes in daylight hours, and Hal. halys nymphs have been found to develop faster with a 12L:12D photoperiod than 15L:9D (Musolin et al. 2019; McDougall et al. 2021). Another factor is the year-round availability of host plants. Sweet corn, grown throughout the winter, is one of the top grossing crops in Florida and is a known host of Hal. halys (Poplin 2013). Tomatoes also have two cropping sea- sons in Florida, providing a second, economically important host for H. alys (Sargent et al 2014; Zoebel et al. 2016). Because the data from this study and from Johnson and Masner (1985) suggest that Ps. striaticeps is multivoltine as well, it may be able to provide control throughout the year. The logical next step is to determine the host range of Ps. striaticeps. This includes determining if it can parasitize Hal. halys eggs, if it targets beneficial stink bugs such as Euthyrhynchus floridanus (L., 1767) and Podisus maculiventris (Say, 1831), and how it might impact other native species of true bugs. Acknowledgments Sarah Birkmire and Amanda Hodges were supported by the University of Florida, IFAS Entomology and Nematology Department, University of Florida, IFAS Entomology and Nematology Department DPM Program, and USDA-APHIS-PPQ Plant Protec- tion Act funding for projects AP19PPQFO000C508 and AP20PPQFO000C390: Characterization of Brown Marmorated Stink Bug Establishment in Florida to Support Biological Control and Mitigation. Personnel from the University of Florida, Entomol- ogy and Nematology Department, DPM Program and Biosecurity Research and Exten- sion Laboratory that supported this work through insect rearing, preparation of sentinel egg masses, checking sentinel egg masses for parasitoids, and field collection included: Jennifer Carr, Heather Kalaman, Adam Pitcher, Jordyn Diaz, and Cameron Zuck. Sarah Birkmire is a co-enrolled MS in Entomology and Nematology and DPM student. This research is a component of her MS thesis in Entomology and Nematol- Psix striaticeps arrives in Florida 515 ogy, and committee members include: Dr. Norman Leppla, Dr. Cindy McKenzie, Dr. Lance Osborne, and Dr. Glynn Tillman. Elijah Talamas and Matthew Moore were supported by the Florida Depart- ment of Agriculture and Consumer Services-Division of Plant Industry (FDACS- DPI). We have many employees of FDACS-DPI to thank: Cheryl Roberts and Lynn Combee assisted in generating sequence data; Sara Ferguson, Alejandro Montoya, Susan Halbert and Natalie McGathey sorted and point mounted parasitoids. Julieta Brambila (USDA-APHIS-PPQ) also sorted samples. We also express our thanks to members of the CAPS Program who collected samples: Krystal Ashman, Robert Le- ahy, Scott Weihman, Max Carfagno, and Douglas Gaskill. Additional FDACS-DPI employees partnered with the University of Florida who should be acknowledged include Dr. Eric Rohrig, Dr. Mrittunjai Srivastava, Daniel Freed, Dr. Nicole Benda, and Stephanie Dowling. References Altschul, SK, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. Journal of Molecular Biology 215: 403-410. https://doi.org/10.1016/S0022- 2836(05)80360-2 Ashmead W (1881) On the Aphididae of Florida, with Descriptions of New Species. The Ca- nadian Entomologist 13(11): 220-225. https://doi.org/10.4039/Ent13220-11 Ashmead WH (1893) A monograph of the North American Proctotrypidae. Bulletin of the United States National Museum 45:1—472. https://doi.org/10.5479/si.03629236.45. 1 Ashmead W (1904) Descriptions of new Hymenoptera from Japan-1. Journal of the New York Entomological Society 12: 65-84. Bergmann EJ, Venugopal P.D., Martinson HM, Raupp MJ, Shrewsbury PM (2016) Host Plant Use by the Invasive Halymorpha halys (Stal) on Woody Ornamental Trees and Shrubs. PLoS ONE 11(2): e0149975. https://doi.org/10.1371/journal.pone.0149975 Bernon G (2004) Biology of Halyomorpha halys, the brown marmorated stink bug (BMSB). United States Department of Agriculture, Animal and Plant Health Inspection Service, Center for Plant Health Science and Technology. Report No.: T3P01. Burmeister H (1835) Handbuch der Entomologie. Zweiter Band, Berlin, 400 pp. Chen, HC, Lahey Z, Talamas EJ, Valerio AA, Popovici OA, Musetti L, Klompen H, Polaszek A, Masner L, Austin AD, Johnson NF (2021) An integrated phylogenetic reassessment of the parasitoid superfamily Platygastroidea (Hymenoptera: Proctotrupomorpha) results in a revised familial classification. Systematic Entomology 46: 1088-1113. https://doi. org/10.1111/syen.12511 Cohen, AC (2000) New olidic production diet for Lygus hesperus Knight and L. lineola- ris (Palisot de Beauvois). Journal of Entomological Science 52: 301-310. https://doi. org/10.18474/0749-8004-35.3.301 de Freitas Bueno, A, Braz EC, Favetti BM, de Barros Franca-Neto, Silva GV (2020) Release of the egg parasitoid Telenomus podisi to manage the Neotropical Brown Stink Bug, Euschistus 516 Sarah Birkmire et al. / Journal of Hymenoptera Research 87: 503-521 (2021) heroes, in soybean production. Crop Protection 137: e105310. https://doi.org/10.1016/j. cropro.2020.105310 Distant WL (1880) Insects: Rhynchota. Hemiptera-Heteroptera (Volume I). In: Godman FD, Salvin O (Eds) Biologia Centrali-Americana. London, 1-88. Dodd AP (1914) Some proctotrypoid egg-parasites of sugar cane insects in Java. The Canadian Entomologist 46: 293-294. https://doi.org/10.4039/Ent46293-8 Dorsey TW, Lovero A (2014) NJDA Non-Diapause Laboratory Rearing and Colony Mainte- nance of the Brown Marmorated Stink Bug, Halyomorpha halys. hetps://www.northeast- ipm.org/neipm/assets/File/BMSB-Resources/ BMSB-IWG-Dec-2014/5-NJ DA-Non-Dia- pause-Laboratory-Rearing-Dorsey-Lovero-Mayer. pdf Drake CJ (1920) The southern green stink bug in Florida. State Plant Board of Florida Quar- terly Bulletin [V: 41-93. Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research 32: 1792-1797. Esquivel JF, Musolin DL, Jones WA, Rabitsch W, Greene JK, Toews MD, Schwertner CE, Grazia J, McPherson RM (2017) Nezara viridula (L.) In: McPherson JE (Eds) Invasive Stink Bugs and Related Species (Pentatomoidea). 364 pp. https://doi.org/10.1201/9781315371221-7 Fabricius JC (1798) Entomologia systematica emandata et aucta. Secundum classes, ordines, genera, species adjectis synonimis, locis, observationibus descriptionibus. Hafniae, impen- sis Christian Gottlieb Proft., 349 pp. Fouts RM (1920) Some new parasites, with remarks on the genus Platygaster (Hymenoptera). Proceedings of the Entomological Society of Washington 22: 61-72. Ganjisaftar F, Talamas EJ, Bon M-C, Gonzalez L, Brown BV, Perring TM (2018) Trissolcus hya- linipennis Rajmohana & Narendran (Hymenoptera, Scelionidae), a parasitoid of Bagrada hilaris (Burmeister) (Hemiptera, Pentatomidae), emerges in North America. Journal of Hymenoptera Research 65: 111-130. https://doi.org/10.3897/jhr.65.25620 Gardner W, Blount J, Golec J, Jones W, Hu X-P, Talamas E, Evans R, Dong X, Ray C, Buntin G, Gerardo N, Couret J (2013) Discovery of Paratelenomus saccharalis (Dodd) (Hymenop- tera: Platygastridae), an Egg Parasitoid of Megacopta cribraria F. (Hemiptera: Plataspidae) in its Expanded North American Range. Journal of Entomological Science 48: 355-359. https://doi.org/10.18474/0749-8004-48.4.355 Gariepy, TD, Haye T, Zhang J (2014) A molecular diagnostic tool for the preliminary assess- ment of host-parasitoid associations in biological control programmes for a new invasive pest. Molecular Ecology 23: 3912-3924. https://doi.org/10.1111/mec.12515 Gariepy TD, Talamas EJ (2019) Discovery of Trissolcus japonicus (Hymenoptera: Sce- lionidae) in Ontario, Canada. The Canadian Entomologist 151: 824-826. https://doi.org/10.4039/tce.2019.58 Giantsis [A, Chaskopoulou A, Bon MC (2016) Mild-Vectolysis: A nondestructive DNA extrac- tion method for vouchering sand flies and mosquitoes. Journal of Medical Entomology 53: 692-695. https://doi.org/10.1093/jme/tjv236 Haye T, Wyniger D, Gariepy TD (2014) Recent range expansion of brown marmorated stink bug in Europe. In: Miller G, Pospichil R, Robinson W (Eds) Proceedings of the Eighth International Conference on Urban Pests. Kft. OOK Press, Veszprém, , 309-314. Psix striaticeps arrives in Florida 517 Haye T, Gariepy T, Zhang J, Fischer S (2015). Can native egg parasitoids adopt the invasive brown marmorated stink bug, Halyomorpha halys (Heteroptera: Pentatomidae), in Europe? Journal of Pest Science 88: 693-705. https://doi.org/10.1007/s10340-015-067 1-1 Haye T, Weber DC (2017) Special issue on the brown marmorated stink bug, Halyomorpha halys: an emerging pest of global concern. Journal of Pest Science 90: 987-988. https://doi. org/10.1007/s10340-017-0897-1 Hoebeck ER, Carter ME (2003) Halyomorpha halys (Stal) (Heteroptera: Pentatomidae): a poly- phagous plant pest from Asia newly detected in North America. Proceedings of the Ento- mological Society of Washington 105: 225-237. Hoffman WE (1933) A pentatomid pest of growing beans in South China. Peking Natural History Bulletin 5: 25-26. Hoffman MP, Wilson LT, Zalom FG (1987) The Southern Green Stink Bug, Nezara viridula Lin- naeus (Heteroptera: Pentatomidae); New Location. Pan-Pacific Entomologist 63: 333-334. Hogg BN, Hougardy E, Talamas E (2021) Adventive Gryon aetherium Talamas (Hymenoptera, Scelionidae) associated with eggs of Bagrada hilaris (Burmeister) (Hemiptera, Pentatomidae) in the USA. In: Lahey Z, Talamas E (Eds) Advances in the Systematics of Platygastroidea III. Journal of Hymenoptera Research 87: 481-492. https://doi.org/10.3897/jhr.87.73778 Howard LO (1880) Two new species of Eupelmus, with remarks on E. (antigaster) mirabilis, Walsh. Canadian Entomologist 12: 207-210. https://doi.org/10.4039/Ent12207-10 Huang DW, Liao DX (1988) A new genus and a new species of Pteromalidae (Hymenoptera: Chalcidoidea). Acta Entomologica Sinica 31: 426-428. Jentsch P (2017) Expanding the Range of the Samurai Wasp, Trissolcus japonicus, in New York Orchards. NYSHS Fruit Quarterly 25: 31-36. Johnson NF (1985) Systematics of New World Trissolcus (Hymenoptera: Scelioni- dae): Species related to YZ basalis. The Canadian Entomologist 117: 431-445. https://doi.org/10.4039/Ent1 1743 1-4 Johnson NE, Masner L (1985) Revision of the genus Psix Kozlov & Lé (Hymenoptera: Sce- lionidae). Systematic Entomology 10: 33-58. https://doi.org/10.1111/j.1365-3113.1985. tb00562.x Jones WA (1988) World review of the parasitoids of the Southern Green Stink Bug, Nezara viridula (L.) (Heteroptera: Pentatomidae). Annals of the Entomological Society of America 81: 262-273. https://doi.org/10.1093/aesa/81.2.262 Khrimian A, Shearer PW, Zhang A, Hamilton GC, Aldrich JR (2008) Field trapping of the in- vasive brown marmorated stink bug, Halyomorpha halys, wish geometric isomers of methyl 2,4,5-decatrienoate. Journal of Agricultural and Food Chemistry 56: 197-203. https://doi. org/10.1021/jf072087e Kimura M (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution 16: 111-120. https://doi.org/10.1007/BF01731581 Kistner EJ (2017) Climate change impacts on the potential distribution and abundance of the Brown Marmorated Stink Bug (Hemiptera: Pentatomidae) with special reference to North America and Europe. Environmental Entomology 46. https://doi.org/10.1093/ ee/nvx157 518 Sarah Birkmire et al. / Journal of Hymenoptera Research 87: 503-521 (2021) Kumar S, Stecher G, Tamura K (2016) MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution 33: 1870-1874. https:// doi.org/10.1093/molbev/msw054 Lee DH, Short BD, Joseph SV, Bergh JC, Leskey TC (2013) Review of the biology, ecology, and management of Halyomorpha halys (Hemiptera: Pentatomidae) in China, Japan, and the Republic of Korea. Environmental Entomology 42: 627-641. https://doi.org/10.1603/ EN13006 Letunic I, Bork P (2021) Interactive Tree of Life ((TOL) v5. An online tool for phylogenet- ic tree display and annotation. Nucleic Acids Research 49: W293-W296. https://doi. org/10.1093/nar/gkab301 Linnaeus C (1758) Systema naturae per regna tria naturae, secundum classes, ordines, genera, species, cum characteribus, differentiis, synonymis, locis. Tomus I. Edito decima, refor- mata, 1-824. Holmiae. [Salvius] https://doi.org/10.5962/bhl.title.542 Linnaeus C (1767) Systema naturae per regna tria naturae: secundum classes, ordines, genera, species, cum characteribus, differentiis, synonymis, locis. vl., pt. 2 Vindobonae [Vienna]: Typis loannis Thomae: 533-1327. https://doi.org/10.5962/bhl.title. 156772 Lowenstein DM, Andrews H, Hilton RJ, Kaiser C, Wiman NG (2019) Establishment in an Introduced Range: Dispersal Capacity and Winter Survival of Trissolcus japonicus, an Ad- ventive Egg Parasitoid. Insects 10: e433. https://doi.org/10.3390/insects10120443 Masner L (1983) A revision of Gryon Haliday in North America (Hymenoptera: Proc- totrupoidea: Scelionidae). The Canadian Entomologist 155: 123-174. https://doi. org/10.4039/Ent115123-2 McDougall RN, Ogburn EC, Walgenbach JE, Nielsen AL (2021) Diapause Termination in In- vasive Populations of the Brown Marmorated Stink Bug (Hemiptera: Pentatomidae) in Re- sponse to Photoperiod. Environmental Entomology. https://doi.org/10.1093/ee/nvab089 Medal J, Smith T, Fox A, Santa Cruz A, Poplin A, Hodges A (2012) Rearing the Brown Mar- morated Stink Bug Halyomorpha halys (Heteroptera: Pentatomidae). Florida Entomologist 95: 800-802. https://doi.org/10.1653/024.095.0339 Medal J, Smith T (2015) Reversible polyphenism in Trissolcus japonicus (Hymenoptera: Plat- ygastridae) induced by stink bug egg size. Journal of Entomological Science 50: 363-366. https://doi.org/10.18474/JES15-16.1 Milnes JM, Beers EH (2019) Trissolcus japonicus (Hymenoptera: Scelionidae) causes low levels of parasitism in three North American pentatomids under field conditions. Journal of In- sect Science 19: e15. https://doi.org/10.1093/jisesa/iez074 Musolin DL, Dolgovskaya MY, Protsenko VY, Karpun NN, Reznik SY, Saulich AK (2019) Photoperiodic and temperature control of nymphal growth and adult diapause induction in the invasive Caucasian population of the brown marmorated stink bug, Halyomorpha halys. Journal of Pest Science 92: 621-631. https://doi.org/10.1007/s10340-019-01080-1 Ogburn EC, Bessin R, Dieckhoff C, Dobson R, Grieshop M, Hoelmer KA, Mathews C, Moore J, Nielsen AL, Poley K, Pote JM, Rogers M, Welty C, Walgenbach JF (2016) Natural en- emy impact on eggs of the invasive brown marmorated stink bug, Halyomorpha halys (Stal) (Hemiptera: Pentatomidae), in organic agroecosystems: A regional assessment. Biological Control 101: 39-51. https://doi.org/10.1016/j.biocontrol.2016.06.002 Psix striaticeps arrives in Florida 519 Panizzi AR, Meneguim AM (1988) Performance of nymphal and adult Nezara viridula on selected alternate host plants. Entomologia Experimentalis et Applicata 50: 215-223. htt- ps://doi.org/10.1111/j.1570-7458.1989.tb01195.x Penca C, Hodges A (2018) First report of brown marmorated stink bug (Hemiptera: Pentato- midae) reproduction and localized establishment in Florida. Florida Entomologist 101: e708. https://doi.org/10.1653/024.101.0413 Poplin AV (2013) A Florida perspective on host preference, early detection, and identification of the brown marmorated stink bug, Halyomorpha halys (Stal). PhD Thesis, University of Florida. Rajmohana K, Narendran TC (2007) A systematic note on the genus Trissolcus Ashmead (Hy- menoptera: Scelionidae) with a key to species from India. Records of the Zoological Survey of India 107: 101-103. Ratnasingham S, Hebert PDN (2007) BOLD: The Barcode of Life Data System. Molecular Ecology Notes 7: 355-364. https://doi.org/10.1111/j.1471-8286.2007.01678.x Rolston LH (1983) A revision of the genus Acrosternum Fieber, subgenus Chinavia Orian, in the western Hemisphere (Hemiptera: Pentatomidae). Journal of the New York Entomo- logical Society 91: 97-176. Sargent C, Martinson HM, Raupp MJ (2014) Traps and Trap Placement May Affect Loca- tion of Brown Marmorated Stink Bug (Hemiptera: Pentatomidae) and Increase Injury to Tomato Fruits in Home Gardens. Environmental Entomology 43: 432-438. https://doi. org/10.1603/EN13237 Say T (1831) Descriptions of new species of heteropterous Hemiptera of North America. New Harmony, 39 pp. Singh RN, Saratchandra B (2002) Biological control of the pentatomid stink bug, Eocanthecona furcellata (Wolff.), by using their parasitoid, Psix striaticeps Dodd, in sericulture. Interna- tional Journal of Industrial Entomology 5: 13-22. Singh RN (2002) Studies on the biological attributes of scelionid egg parasitoid Psix striaticeps (Dodd) for the control of stink bug Canthecona furcellata (Wolff) in sericulture. Interna- tional Journal of Industrial Entomology 4: 117-120. Singh RN, Sinha SS (1995) Influence of Eocanthecona furcellata (Wolff) (Heteroptera: Pentato- midae) egg age on the progeny production of its parasitoid Psix striaticeps (Dodd) (Hyme- noptera: Scelionidae). Journal of Biological Control 9: 99-101. Snyder J, Dowling S, Rohrig E, Halbert S, Ramadugu C, Simmons G, Mizell R, Henderson R (2019) Field assays of 3D-printed Asian Citrus Psyllid (Diaphorina citri) trapping systems. International Research Conference on HLB, Riverside, California. Stal Cacis5 5) Ofversigt af Kongl. Vetenskaps-akademiens forhandlingar tolfte argangen. Stock- holm, 402 pp. Stal C (1873) Enumeratio Hemipterorum. Bidrag till en forteckning 6fver alla hittills kanda Hemiptera, jemte systematiska meddelanden. 3. Konglika Svenska Vetenskaps-Akade- miens Handlingar 11: 1-163. Taekul C, Alejandro A, Valerio AD, Austin HK, Norman FJ (2014) Molecular phylogeny of telenomine egg parasitoids (Hymenoptera: Platygastridae s.l.: Telenominae): evolution of host shifts and implications for classification. Systematic Entomology 39: 24-35. https:// doi.org/10.1111/syen.12032 520 Sarah Birkmire et al. / Journal of Hymenoptera Research 87: 503-521 (2021) Talamas E, Herlihy M, Dieckhoff C, Hoelmer K, Buffington M, Bon M-C, Weber D (2015) Trissolcus japonicus (Ashmead) (Hymenoptera, Scelionidae) emerges in North America. Journal of Hymenoptera Research 43: 119-128. https://doi.org/10.3897/JHR.43.4661 Talamas EJ, Bremer JS, Moore MR, Bon M-C, Lahey Z, Roberts CG, Combee LA, McGathey N, van Noort S, Timokhov AV, Hougardy E, Hogg B (2021) A maximalist approach to the systematics of a biological control agent: Gryon aetherium Talamas, sp. nov. (Hymenoptera, Scelionidae). In: Lahey Z, Talamas E (Eds) Advances in the Systematics of Platygastroidea III. Journal of Hymenoptera Research 87: 323-480. https://doi.org/10.3897/jhr.87.72842 Tamura K, Nei M (1993) Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Molecular Biology and Evolu- tion 10: 512-526. https://doi.org/10.3897/JHR.43.8560 Tillman PG (2011) Natural Biological Control of Stink Bug (Heteroptera: Pentatomidae) Eggs in Corn, Peanut, and Cotton Farmscapes in Georgia. Environmental Entomology 40: 303-314. https://doi.org/10.1603/EN10154 Tillman G, Toews M, Blaauw B, Sial A, Cottrell T; Talamas E, Buntin D, Joseph S, Balusu R, Fadamiro H, Lahiri S, Patel D (2020) Parasitism and predation of sentinel eggs of the invasive brown marmorated stink bug, Halyomorpha halys (Stal)(Hemiptera: Pentatomi- dae), in the southeastern US. Biological Control 145: e104247. https://doi.org/10.1016/j. biocontrol.2020.104247 Turnipseed SG, Kogan M (1976) Soybean Entomology. Annual Review of Entomology 21: 247-282. https://doi.org/10.1146/annurev.en.21.010176.001335 Walsh BD, Riley CV (1869) The joint worm. (/sosoma hordei, Harris.) American Entomologist 1: 149-158. Westwood JO (1837) A Catalogue of Hemiptera in the Collection of the Rev. EF W. Hope, with short Latin Descriptions of New Species. London, 1-46. Wollaston TV (1858) II. — Brief diagnostic characters of undescribed Maderian insects. Annals and Magazine of Natural History 1: 18-28. https://doi.org/10.1080/00222935808696865 Zobel ES, Hooks CRR, Dively GP (2016) Seasonal Abundance, Host Suitability, and Feeding Injury of the Brown Marmorated Stink Bug, Halyomorpha halys (Heteroptera: Penatomi- dae), in Selected Vegetables. Journal of Economic Entomology 109: 1289-1302. https:// doi.org/10.1093/jee/tow055 Psix striaticeps arrives in Florida 52 Supplementary material | DarwinCore Authors: Elijah J. Talamas Data type: occurences Explanation note: DarwinCore archive of specimen collection data. Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODDbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/jhr.87.76191.suppl1 Supplementary material 2 Tree Authors: Matthew R. Moore Data type: Tree file. Explanation note: Unedited tree file of COI analysis. Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODDbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/jhr.87.76191.suppl2 Supplementary material 3 Matrix Authors: Matthew R. Moore Data type: Spreadsheet. Explanation note: Matrix of genetic distance calculations. Copyright notice: This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODDbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited. Link: https://doi.org/10.3897/jhr.87.76191.suppl3