Two new species and distribution records for the genus Bohayella Belokobylskij, 1987 from Costa Rica (Hymenoptera, Braconidae, Cardiochilinae)

Abstract Two new species of Bohayella Belokobylskij, 1987 from Costa Rica are described: Bohayella geraldinae Kang, sp. nov. and Bohayella hansoni Kang, sp. nov. These are new distribution records for the genus in the Neotropical region. In addition, a key to species of the genus Bohayella of Costa Rica is presented. The current work elevates the number of species included in Bohayella from nine to eleven.

Cardiochiles nigricans Mao, 1949(Mao 1949) was transferred into Bohayella by Dangerfield et al. (1999) and recorded as the first species of Bohayella in the New World. Mercado and Wharton (2003) transferred the species into Toxoneuron Say, 1836 because the first metasomal tergite (T1) of the species is different from T1 of other members of Bohayella. Subsequently, members of Bohayella have been restricted to the Old World and new species of the genus have not been reported from the New World.
The first author (IK) had the opportunity to examine Costa Rican cardiochiline specimens housed in University of Wyoming Insect Museum (UWIM). Using the key to world genera of the subfamily Cardiochilinae and other diagnostic characters of Bohayella (Dangerfield et al. 1999), nine Bohayella specimens were identified. The characters of New World Bohayella are discussed in detail in diagnosis and discussion sections of this paper. Other Costa Rican cardiochiline specimens borrowed from several institutions were examined, but no more specimens of Bohayella were discovered. As a result, the nine specimens of Bohayella were confirmed as two species based on morphological data. Herein, we describe two new species and present a key to species of the genus Bohayella of Costa Rica. Distribution maps for both species are included.

Materials and methods
Specimens for this project were provided by UWIM (University of Wyoming Insect Museum; 1000 East University Avenue, University of Wyoming, Laramie, Wyoming 82071-3354, USA). We conducted morphological analyses using a Leica MZ75 stereomicroscope. The morphological terms and terms of wings mostly follow Dangerfield (1995) and Dangerfield et al. (1999). Morphological terminology can be checked at the Hymenoptera Ontology website (http://portal.hymao.org/projects/32/public/ontology/) as well. Terms for sculpturing are based on Harris (1979). Color habitus images were taken using a Visionary Digital BK Plus imaging system (Dun, Inc.), equipped with a Canon EOS 5DS DSLR camera. Images were stacked via Zerene Stacker v.1.04 (Zerene Systems LLC.). All images were edited using Adobe Photoshop CS 6 (Adobe Systems, Inc). Body parts of each species were measured via Adobe Photoshop CS 6 (Adobe Systems, Inc). Each number in parentheses in species descriptions indicate 0.01 times the actual length, width, or height of each body part. For example, 42 and 124 in parentheses (42:124) indicate 0.42 mm and 1.24 mm, respectively. Distribution maps of two Bohayella species were produced using QGIS 3.10.0 (QGIS Development Team 2019). Google satellite maps were downloaded using the QuickMapServices plugin. The following abbreviations are used throughout the current paper: POL: distance between posterior ocelli, T1, T2 (second metasomal tergite), T3 (third metasomal tergite), T5 (fifth metasomal tergite), T6 (sixth metasomal tergite), T7 (seventh metasomal tergite), and T8 (eight metasomal tergite). Holotypes and paratypes are deposited in the UWIM.

Results
Bohayella Belokobylskij, 1987 Type species. Bohayella tobiasi Belokobylskij, 1987. Diagnosis (based on Dangerfield et al. (1999) with modifications and additions). Diagnostic characters of Bohayella based on Old World members were described in Belokobylskij (1987) (in Russian) and Dangerfield et al. (1999) (in English). The following are re-described or additional characters based on morphological characters of both Old World and New World members.

Key to species of the genus Bohayella of Costa Rica
Male. Body ~ 5.0 mm. Same as female except for the following characters: antenna 32-segmented, melanistic color does not reach the dorsal margin of foramen magnum.
Host. Unknown. Distribution. Bohayella geraldinae sp. nov. is known only from the La Selva Biological Station owned and managed by Organization for Tropical Studies (OTS) in Heredia, Costa Rica at an elevation of 100 m (Figs 1, 2). The station is located in the Caribbean lowlands, at a confluence of the Sarapiquí river and Puerto Viejo (McDade and Hartshorn 1994). According to Holdridge's life zone system (Holdridge 1967), the station is in the tropical wet forest region (Hartshorn and Peralta 1987), and the average annual precipitation in the area is ~ 4,000 mm (Sanford et al. 1994).
Male. Unknown. Host. Unknown. Distribution. Bohayella hansoni sp. nov. is known only from the Las Alturas Biological research station owned and operated by Stanford University in Las Alturas, San Vito, Costa Rica at the elevations of 1,500 m and 1,700 m (Figs 1, 5).
Etymology. This species is named in honor of Dr Paul Hanson, collaborator and professor at the Escuela de Biología, Universidad de Costa Rica. He worked tirelessly for many years collecting and sorting Costa Rican braconids from Malaise samples. SRS is very grateful for his dedication to Hymenoptera studies.

Discussion
Most genus-level diagnostic characters are shared by both Old World and New World members (B. geraldinae sp. nov. and B. hansoni sp. nov.). None of the New World members have a mostly black body, 5-segmented maxillary palpi, or apically infuscate forewings. The following characters are only shared by New World members: angled Rs vein of forewing (Figs 3F, 4A), pectinate hind tarsal claw with sharp apical tooth (Fig. 3E), and antero-posteriorly slightly expanded hind basitarsus (Fig. 3A).
Specimens of B. hansoni sp. nov. collected at altitudes above 1,500 m have more melanistic metasoma than specimens of B. geraldinae sp. nov. collected at a low altitude of 100 m (Figs 3F, 4B). The melanism associated with high elevation was confirmed not only in braconid wasps such as members of the genus Sendaphne Nixon, 1965(Nixon 1965) (Fernandez-Triana et al. 2014 and Meteorus pulchricornis (Wesmael, 1835) (Abe et al. 2013), but also in other hymenopteran insects such as members of a vespid species, Agelaia pallipes (Olivier, 1792) (de Souza et al. 2020) as well as an undescribed scelionid species of Lapitha Ashmead, 1893 (Mora and Hanson 2019). According to Abe et al. (2013), emerged adults of M. pulchricornis were more melanistic when cocoons were reared at lower temperatures, and the effects of the melanism resulted in increasing body temperatures and improved flight ability of adult M. pulchricornis. Melanism of B. hansoni sp. nov. at high elevations may induce similar outcomes as in M. pulchricornis. Further research is needed when enough live samples are available to confirm this.
The elevation of Costa Rica ranges from sea level to 3,819 m (Hanson and Gauld 1995). If additional sampling is conducted across the country and more species of Bohayella are discovered, altitudinal distribution patterns of members of Costa Rican Bohayella can be investigated in the future (e.g., Aguirre et al. 2018).