Catalogue of herpetological specimens of the Ewha Womans University Natural History Museum (EWNHM), Republic of Korea

Abstract The herpetology collection of the Ewha Womans University Natural History Museum (EWNHM) represents one of the oldest and largest institutional collections in the Republic of Korea. The specimens deposited in the EWNHM represent a major historical collection of the native herpetofauna, both in species diversity and time span. However, the full inventory of the herpetology collection has never been conducted and thus the collection has received little attention from researchers. Here, the first full account of the herpetology specimens held at the EWNHM is provided, with voucher information for all documented specimens to make the collection accessible for future studies.


Introduction
Natural history collections are an invaluable repository for modern biological research. These collections have broad applications including the detection of faunal changes, species decline, biogeography, systematics, and species discovery (Shaffer et al. 1998;Graham et al. 2004;Wójcik et al. 2010;Lister et al. 2011). A series of specimens collected over time also serves to shed light on changes in genetic diversity and morphology that occurred during that time period (Babin-Fenske et al. 2008;Cornetti et al. 2018). In order to conduct such collection based research, a full inventory of specimens held at the museum is a valuable resource as it saves time locating specimens, designing studies and managing collections.
In the Republic of Korea, early efforts to collect herpetological specimens were conducted by foreign researchers. Thus, vouchered specimens collected during this time were deposited in natural history museums outside of the country (American Museum of Natural History 2020; e.g., a series of Bombina orientalis vouchers collected in early 1950s). Although such specimens are a valuable historical collection, they usually do not fully encompass the native herpetofauna in taxa, time span, and geographic locations (American Museum of Natural History 2020; e.g., herpetological specimens historically collected in Korea). Moreover, natural history collections within the country are poorly known to researchers outside the institution where the collection is located and thus received little attention in herpetological research.
Ewha Womans University Natural History Museum (EWNHM; Sabaj 2016) was established in 1969 as the first natural history museum for the country (EWNHM website 2014). Although small in size, the herpetology collection of EWNHM is of great historical value for herpetological research, with some specimens dating back to early 1950s being collected during the Korean War.
Despite this great value, a complete catalogue of the collection has not been available. Although a public database (Korean Natural History Research Information System; NARIS) provides information on EWNHM specimens, this database is only a partial representation of the collection. Also, this database uses a different cataloging system from EWNHM, leading to voucher inconsistencies and potential problems of locating specimens. Therefore, a full herpetological inventory of the collection at EWNHM following a consistent format is needed for future utilization of specimens. Here, we provide the first complete catalogue of the herpetological collection of EWN-HM, using consistent voucher system throughout specimens. In doing so, we also changed degrading labels to prevent the loss of important information and updated the nomenclature of species if the taxon underwent taxonomic revisions between the time of initial labelling and our cataloging effort. Also, we applied a new and consistent voucher system throughout specimens to resolve confusion of conflicting voucher systems. Although this means yet another change of voucher system applied to specimens, this catalogue can serve as a reference point towards reducing confusions originating from multiple conflicting voucher systems.

Description of the collection
The herpetological specimens of the museum EWNHM are located in the wet collection room. The herpetology collection was roughly divided into reptiles and amphibians. For some species of anurans, 50 to 100 individual specimens were contained per glass jar (e.g., Bombina orientalis, Glandirana emeljanovi, Pelophylax nigromaculatus), with specimens packaged individually in small plastic bags. These specimens were mostly collected in one location on the same date, comprising voucher series of specimens. In some cases, specimens of one anuran species collected from two different locations were held in the same glass jar. In this case, specimens from two locations were divided into two by separate plastic bags. Some amphibian species were held in smaller glass jars with fewer number of individual specimens per container (< 25 individual specimens). We also found jars that contained two or more amphibian species collected from the same location. In this case, we either separated the species into separate jars or separated them into small plastic bags containing paper labels with the appropriate species information.
Specimens of lizards were housed in small individual containers or packed into small number of specimens (< 20 individuals per container) representing voucher series. Specimens of snakes and turtles were individually contained in glass jars. All specimens were preserved in 1% formalin solution at the beginning of collection and preservation process (Jaewon Ryu, pers. comm.). The use of preservative solution has not been changed over the years.

General protocol for processing specimens
We used a table to mount a tripod and camera (Nikon D50; Nikon Corporation, Tokyo, Japan) in order to photograph the specimens. The table was covered with white synthetic fabric to provide a white background for photography. A 15-centimeter ruler was taped above the fabric to be used as a scale of reference. We used a whole-face respirator (3M Company, Minnesota, USA) to prevent potential toxic inhalation of preservative fluids. We also used industrial-grade wiper (Yuhan-Kimberly, Ltd, Seoul, Korea) to remove fluid spillage. For photography, we mounted the camera on a tripod with the lens facing vertically downwards. Each specimen was photographed in dorsal, lateral and ventral angles. Specimens of eggs and amphibian larva were not photographed to prevent potential damage caused by the handling process.

Labelling protocol
As most of the Korean herpetofauna have undergone significant nomenclatural changes over the past 50 years, and as some of the labels showed signs of degradation, it was necessary to update the labels. In doing so, we kept the original labels alongside the new labels with updated nomenclature.
For labeling, we first prepared a general label for each specimen or series of specimens. This label contained key information about the specimen(s), including scientific name, Korean common name, geographic location of collection, collection date, and collector(s). The label was printed on regular A4 papers and was 8.8 × 5.7 cm in size. All relevant information were written on the labels using a pencil. The new labels were fully immersed in preservative fluid following previous collection maintenance practice of EWNHM.
Alongside the general label, it was necessary to assign an unique voucher number to each specimen that did not currently have one, according to the nomenclature rules of the museum. The preexisting voucher system for the museum collection is such as EWNM-AR-XXXX, "XXXX" being the serial number. However, this previous system was not consistently applied throughout all of the specimens held in the collection and needed to be corrected. Moreover, the voucher system represented in NARIS is different from the preexisting voucher system of EWNHM. Thus, in order to apply a consistent voucher number system to each and all herpetological specimens, the decision was made by the museum to discard both previous EWNHM and NARIS voucher systems and to use a new protocol for the herpetology collection. The new numbering protocol is comprised of museum code EWNHM, followed by a taxon code ANIMAL, and a four-digit serial number starting from EWNHM-ANIMAL 5279. Despite the change of protocol, the previous EWNM-AR voucher labels were retained alongside the new EWNHM-ANIMAL voucher labels for the traceability of information.

Georeferencing protocol
One of the main issues concerning the use of natural history collections is determining the area of collection with reasonable accuracy (Newbold 2010). The collection labels, especially of historical collections, usually contain text-only geographic information of different extent and accuracy (Garcia-Milagros and Funk 2010). This was also the case with specimens deposited in EWNHM. Therefore, each specimen was georeferenced using the collection information available, with the locality of collection being recorded as latitude and longitude (in decimal degrees) describing the midpoint. For localities within a named town or city, the center of the settlement or named district was used as the midpoint. Google Maps was used along with the 'What's here?' tool to display the latitude and longitude once the midpoint had been found. For those specimens collected in more rural localities, the midpoint of the named location was also used as in the urban ones.

Catalogue
We organized the catalogue in Order -Family -Species order. For each species, English and Korean common names are also given alongside the current scientific name.
The nomenclature used in the original labels is also given. Location and collector information (written in either Korean or Chinese characters) are directly translated.
Collection locality and collection date (see Table 1 and Table 2 for georeferenced GPS coordinates, and Fig. 1 for locations on the map) Juv.
Egg ( (1843), considered as distinct genus by Fei et al. (1990) and supported by Frost et al. (2006). ( (Frost et al. 2006). Remarks: Rana coreana was demonstrated to be distinct from R. amurensis by Song et al. (2006), and the labels were updated accordingly.  Fei et al. (1990). The presence of this species in the Republic of Korea was confirmed by Yang et al. (2000).  (Yang et al. 2000); however, distinguishing between the two species based on this method was not always possible during cataloging procedure. Therefore, it is likely that some of the specimens identified as R. uenoi by us are actually R. huanrenensis, and vice versa. Moreover, there is a collection of Rana froglet specimens that could not be identified and could be a mix of froglets of R. uenoi, R. huanrenensis, and R. coreana. Also, froglets of Pelophylax nigromaculatus and Glandirana emeljanovi were mixed with these specimens but identified. The specimens of Rana coreana have been labelled as Rana amurensis coreana.
Remarks: this specimen was labelled as "a neonate of Lycodon rufozanatus". However, direct comparisons of head shape, tail length, and body shape with another specimen of neonate L. rufozonatus (EWNHM-ANIMAL 6544) suggested that the specimen is not L. rufozonatus but instead belongs to the genus Chrysopelea (Somaweera et al. 2015).

Sibynophis chinensis (Günther, 1889)
Chinese many-toothed snake; 비바리뱀; one specimen Günther ACLG (1889) Hoge (1981), which was supported by results demonstrated by subsequent authors (Knight et al. 1992;Parkinson, 1999). Meanwhile, species assignment in the updated labels is based on the original description by Stejneger (1907 as Agkistrodon blomhoffii brevicaudus) because the name Agkistrodon (= Gloydius) halys used in former labels is only applicable to populations of Gloydius in Central Asia (Wüster et al. 1997). ( Shin et al. (2020b). Remark: generic assignment in the updated labels as explained above for G. brevicaudus. Although some authors consider G. saxatilis as valid, here we treat that name as a synonym of G. intermedius following Orlov and Barabanov (1999). Voucher EWN-HM-ANIMAL 6454 is the first reported specimen of G. intermedius with dicephalism (Shin et al. 2020b). (Emelianov, 1929 Remark: generic assignment in the updated labels and invalidity of the name "Agkistrodon halys" applied to East Asian Gloydius species as explained above for G. brevicaudus. The species name used in the updated labels follows the original species description by Emelianov (1929 as Ancistrodon blomhoffi ussuriensis Remarks: we were unable to identify these two specimens because the number of femoral pores (a characteristic that is clearly different between T. amurensis and T. wolteri) were not clearly visible. Pholidosis characteristics alone were insufficient to make a clear diagnosis at species level.

Family Scincidae Gray, 1825
Scincella vandenburghi (Schmidt, 1927 Remark. generic assignment in the updated labels is based on the original description of Trachemys by Agassiz (1857) following the resurrection of Trachemys from synonymy with Pseudemys by Iverson (1985). This taxonomic treatment has been supported by subsequent studies (Siedel and Smith 1986;Rhodin et al. 2017).

Pelodiscus sp.
One specimen EWNHM-ANIMAL 6613; Loc: no data; Leg: no data. Remark: morphological characteristics used to identify Pelodiscus species (Farkas et al. 2019) were insufficient to correctly identify this specimen at the species level.

Missing specimens and specimens not documented in the catalogue
Some specimens that were known to have been deposited in the EWNHM are not documented here because they were not found in the museum. For example, we failed to locate the holotype of Karsenia koreana (EWNHM 80314;Min et al. 2005) and the holotype, paratype series, and other associated specimens of Onychodactylus koreanus (holotype EWNHM 80316; paratype series EWNHM 80315, EWNHM 80317-80318; associated specimens EWNHM 80319-80320, EWNHM 80321-80322, EWNHM 80323-80325, EWNHM 80326; EWNHM 80327-80328; Poyarkov et al. 2012). Our attempts to communicate with species authors did not yield any information regarding the whereabouts of these specimens. At this point, it is uncertain whether these specimens are truly lost or on loan without traceability. However, at-     tempts to designate new type materials should be reserved until the exact whereabouts and status of these specimens are known.
In addition, we were unable to document nine specimens of Dryophytes japonicus because the specimen jars could not be opened. This was due to crystalized preservative fluid around the inner wall of the lids. Also, we did not document specimens of Lithobates catesbeianus and Pelophylax nigromaculatus that had been used for dissection samples. These specimens had been used in the medical school before being deposited in the museum, and are of indeterminate origins with no collection data available.

Conclusions
This catalogue is the first complete inventory of herpetology specimens deposited in the EWNHM. In total, the collection is comprised of 1554 specimens representing all native Korean terrestrial reptiles except one chelonian (Mauremys reevesii), all native anuran species, three of six native salamander species, and some exotic and invasive species (Figs 2-6). Some Korean herpetofauna, such as Hynobius unisacculus (Min et al. 2016) were not included in the collection because they were recently described and no collection have been made by the institution to acquire vouchers. The specimens of non-native species are most likely from live exhibits of the museum or from laboratory experiments (e.g., specimens of L. catesbeianus used in laboratory dissections). In taxonomic diversity, the herpetological collection of EWNHM contain 17 amphibian species across 12 genera and eight families and 22 reptile species across 16 genera and seven families. The specimens were collected between 1951 and 2013. Although the sampling interval is not even, the time span covered by the collection is one of the broadest among collections of Korean herpetofauna. Therefore, the EWNHM collection represents one of the most significant research collections of Korean reptiles and amphibians.
Natural history collections are a valuable resource for a number of reasons and by cataloging the collection at EWNHM, the specimens held there are now accessible to researchers for perpetuity. In recent times, natural history collections have proved valuable resources in order to trace the origins and spread of disease (Ouellet et al. 2005) as well as to reconstruct the genetic diversity in long-extinct populations (Wandeler et al. 2007). With advancing molecular methods, formalin-fixed specimens may soon be able to provide the wealth of knowledge that we can currently extract for those fixed in ethanol. As we shift towards a more holistic approach to conservation, supported by global projects, it is hoped that the above catalogue can aid in herpetofauna conservation for decades to come. recommended by the journal, Dr. Philipp Wagner and Dr. Luis M.P. Ceríaco, and the subject editor Dr. Johannes Penner for comments that greatly improved the manuscript.