Standard Operating Procedure and Manifesto of the Arkansas State University Museum Of

Standard Operating Procedure and Manifesto of the Arkansas State University Museum of Zoology (ASUMZ) Herpetology Collection

Created May 2013

Abstract—Natural history museums are an important part of scientific research even though many people may never see what goes on there. Even some scientists disagree about the proper role of natural history collections in the future of science. However, these collections continue to provide new information as long as we maintain these collections. The Herpetology Collection at Arkansas State University is the largest collection of its kind in the state of Arkansas, and many important publications have resulted from the specimens therein. For this collection to continue to be valuable to science (and to ASU) it must be maintained and taken care of properly. This standard operating procedure will provide a guide for processing animals and maintaining the collection so that it will remain a viable part of the research community.

INTRODUCTION Purpose—Natural history museums serve many purposes in addition to being repositories of the world’s biodiversity. Some of these uses have been discussed by Paolo Viscardi, deputy keeper of natural history at the Horniman Museum in London, UK. Writing a guest blog on 12 April 2011 on The Guardian website (http://www.guardian.co.uk/science/punctuated-equilibrium/2011/apr/12/2) , Viscardi lists these reasons for museums:  Providing base-line data against which to compare modern data and produce predictive models  Safeguarding type specimens for taxonomists and other scientists  Housing voucher specimens from research, showing when and where a species occurred  Providing a “snapshot” of a species or community in a particular space and time  Providing specimens for DNA studies and the like This is not a conclusive list by any means. But within the field of herpetology there is currently some debate concerning the role of collecting specimens for museums and how much is enough. Many researchers today have ethical concerns about collecting, even for voucher specimens providing a geographic or size record. In those instances they prefer to submit photographic vouchers. However, there are drawbacks to using photographic vouchers. The Society for the Study of Amphibians and Reptiles publishes geographic distribution records and stipulates that photographs are only to be used when the organism could not be collected due to protection status of that species, it was found in an area where collection is prohibited, or it is not feasible to preserve and house the specimen such as with large turtles or crocodilians (http://ssarherps.org/pages/HRinfo.php). It is also the policy of the Herpetology Collection at Arkansas State University (ASU) to give preference to actual specimens, though photographs may occasionally be accepted. In addition to the reasons listed above, collections can be used for numerous other studies that may not be conceived of until many years after the collection of the organism. James Stuart (pers. comm.) of the New Mexico Department of Game and Fish offers additional reasons for the utility of museums:  Dietary and reproductive studies  Identification guides and reference materials  Studies of disease and contaminants  Education—many students first learn about the animals by seeing preserved specimens

The Herpetology Collection at ASU has been used extensively for research purposes using both specimens previously preserved as well as collecting fresh specimens for research that are later preserved and accessioned into the museum. Many theses, dissertations, and publications have been the result of the Herpetology Collection, and the topics of study have been quite diverse. For a partial sampling of the recent studies utilizing specimens from the Herpetology Collection, see Appendix A. The most important publication arising from the specimens in the collection is The Amphibians and Reptiles of Arkansas (Trauth et al., 2004). This book is the definitive publication on Arkansas’ reptile and amphibian fauna and is widely regarded as one of the best and most informative state guides published. This guide could not have been published without the existence of the Herpetology Collection at ASU (Trauth, pers. comm.).

Current Holdings and History—The Herpetology Collection of ASU currently (summer 2013) houses ca. 33,000 specimens of reptiles and amphibians. This makes it the largest collection of reptiles and amphibians in the state of Arkansas and similar in size to the herpetology collections at the Virginia Museum of Natural History (ca. 10,300 http://www.vmnh.net/collections), Mississippi Museum of Natural History (ca. 15,000 http://www.mdwfp.com/seek-study/bio-collections/amphibans-reptiles.aspx), Auburn University Museum of Natural History (ca. 40,000 http://aumnh.org/research-collections/amphibians- reptiles/), and the Georgia Museum of Natural History at the University of Georgia (ca. 46,000 http://naturalhistory.uga.edu/index.php? page=content/researchcollections/collections#herpetology). However, the collection at ASU is considerably smaller than the collections at the Louisiana State University Museum of Natural Science (ca. 81,000 http://appl003.lsu.edu/natsci/lmns.nsf/$Content/Herpetology? OpenDocument), North Carolina Museum of Natural Sciences (ca. 200,000 http://naturalsciences.org/research-collections/research-specialties/amphibians-reptiles), and the Florida Museum of Natural History (223,000-largest in the southeastern US- http://www.flmnh.ufl.edu/herpetology/collections.htm). The vast majority of specimens in the collection at ASU are from Arkansas. However, there are specimens from other states, particularly Texas, Alabama, Georgia, and Missouri. The first documented specimen was collected in 1963. Other specimens existed before that time going back to the 1950’s (Hanebrink, 1993), however, there are no existing records for those specimens and those may have solely been used for teaching purposes. There are many specimens in the teaching collection, yet those specimens are neither tagged nor catalogued. Teaching specimens are not used for research purposes, and they are not reflected in the number of total specimens in the Herpetology Collection. The first curator of the Herpetology Collection was Dr. Earl Hanebrink who began teaching at ASU in 1958 (Hanebrink, 1993). Many of the early specimens in the collection are the result of class field trips taken by students of Dr. Hanebrink and Mr. William Byrd (Hanebrink, 1993; Trauth, pers. comm.). The current curator of the collection is Dr. Stan Trauth. The collection grew rapidly during the tenure of Dr. Trauth as specimens were actively being collected to document geographic ranges and other aspects of their natural history in preparation of The Amphibians and Reptiles of Arkansas by Trauth et al. (2004).

Organization—The Herpetology Collection is generally organized taxonomically. Amphibians are housed beginning in the row of shelves immediately inside the collection room beginning with salamanders followed by frogs. Lizards are housed next, followed by snakes, and finally turtles. Within each taxa, the genera are separated unto themselves with the shelves bearing the name of the genus. In some instances, specimens of one genus may appear on a shelf with another genus due to space constraints. Likewise, there may rarely be situations with more than one species in the same jar. However, species in general are only stored in jars with conspecific specimens. In addition to the general collection, there are certain special collections. The Texas collection, Alabama collection, Georgia collection, and Lyon College collection are housed on the same row of shelves as the salamanders. Some specimens from National Park Lands are housed on a shelving unit of their own. Also, large specimens (i.e. large turtles) are stored in 50 gallon drums and stored in a separate room due to space constraints. A teaching collection is also maintained. Most teaching organisms are kept in the teaching lab, however, some teaching specimens are housed in the collection room. On rare occasions when a specimen is only represented by a photographic voucher, the photos are laminated with a museum tag and filed in the filing cabinet.

PROTOCOL Loan Policy—Specimens in the Herpetology Collection may be loaned to other researchers if those researchers are unable to visit the collection. All loans require a specific research question that is to be addressed using the animals in loan. A copy of the loan form must accompany any specimens in loan and another must be kept on file with the Curator. The loan form is to state the research question to be investigated in addition to the name and address of the loanee and the condition of the specimens at the beginning of the loan period. At the conclusion of the loan period, the loanee must state on the loan form the condition of the animals being returned. Unless otherwise agreed upon and stated in the loan form, all loans will be for a period of six months. Loanees may request an extension if needed. Extensions are granted at the judgment of the Curator of the Herpetology Collection. Copies of completed loan forms in addition to additional blank copies are kept in the file cabinet in the collection room. Instructions for shipping amphibians and reptiles are also kept on file in the Herpetology Collection.

Accessioning—Information regarding the processing of reptiles and amphibians for accession into museums can be found in a variety of places and can be general information or standard practices for only one or a few steps in the process. Good starting places include the works by Hall (1962) and Pisani (1973) as well as the newsletters by the American Society of Ichthyologists and Herpetologists located at http://asih.org/curationnews.

Processing animals: Any animals captured should have been collected according to the laws governing the city or state from which it came. Animals should also be handled with care and ethical concern for the animal prior to death. Prior to the preservation process, the animal should be killed according to methods widely accepted. The preferred method for killing reptiles at ASU is with an overdose of sodium pentobarbital injected into the heart or as near to the heart as possible. Other accepted methods are overdose of MS-222 injected intracoelomically, freezing (though this method, while effective, is controversial and is usually only used in conjunction with other methods; additionally, freezing damages tissues due to formation of ice crystals in the cells and may cause tissues to be unacceptable for histological examination), and decapitation with pithing of the brain (though this method is most effective only with trained personnel). The preferred method for killing amphibians at ASU is immersion of the animal into a solution of chloretone until the animal is beyond revival. Other methods include overdose of sodium pentobarbital (as described above), immersion into a solution of MS-222, application of benzocaine to the head and body of the animal, and decapitation with pithing by trained personnel. It should be noted that removal of the head from the body is often not desirable for preserved animals as it reduces the utility of that specimen for research. Animals should each receive a unique museum number. For larval amphibians or egg masses, museum grade label paper should be used with all information written in pencil or indelible ink. The label is then placed inside the jar with the specimen(s). All other specimens will have a museum tag tied to their body. Snakes should have the tag tied around the body in the upper 1/3 of the body. Lizards and salamanders should have the tag tied around the chest just below the front limbs. Frogs and turtles should have the tag tied around the leg just above the knee on the hind limbs. All tags may be tied using a square knot with the excess tag thread trimmed down to ca. ½ inch. Collection/preservation information from that specimen should be placed in the appropriate catalogue. Relevant information takes on the form of: Museum Number | species name | Gender (if known) | Location (written as state abbreviation: county; specific location such as distance to intersection, TRS, or GPS | collector’s name(s) | preparer’s name(s). Any additional notes should be placed at the bottom of the page. Post-mortem, tissue samples should be taken (especially for uncommon species) and stored in 100% ethanol (EtOH) in the tissue vial cabinet. Parafilm should be used over the lid and top of the vial to prevent evaporation of fluid from the vial. The museum number of the animal should be written on a small piece of museum grade paper using pencil or indelible ink and placed inside the vial with the tissue. For lizards, salamanders, and frogs, a toe clip is an acceptable tissue sample. Acceptable tissues from turtles include toe nail clips or skin removed from an inconspicuous area of the body such as the hind leg area. A clip of the belly scales may be clipped from snakes. All tissue samples should be taken prior to the preservation process to remain useable for DNA studies. After tissue samples have been collected and stored, reptiles should be injected with 10% formalin throughout their body to preserve internal tissues. Amphibians will absorb formalin across their skin making injection unnecessary. Specimens should then be placed in a container for formalin fixation. Immersion in formalin is possible, however for most species, being covered with paper towels soaked in excess formalin is sufficient. Specimens should be placed in the desired position, covered by the paper towel, and soaked in enough formalin to thoroughly cover the animal and paper towel. Turtles should have hemostats applied to their tails and feet to maintain them outside the shell during fixation. Turtles should then be hung upright into a bucket of formalin until fully immersed. Each container of formalin should be closed to prevent evaporation during fixation. All specimens in fixation should remain for at least 48 hours to allow for proper fixation of tissues. Following fixation, the specimen may be rinsed if desired before transferring to an appropriate jar of EtOH. Ethanol concentrations for long term storage of specimens should be no less than 70%. Concentrations higher than 70% may be desirable in some circumstances if there is the possibility of water from the specimen’s body diluting the EtOH to an undesirable level. The ideal ratio of specimen volume to EtOH volume within the jar is ca. 25% specimen volume to 75% EtOH volume. Due to space constraints, this is often not possible. In situations where the volume of EtOH is much below 75%, the concentration of EtOH may be increased to 90% EtOH to compensate. Yearly, the catalogue should be transferred to an electronic file (such as MS Excel) and sent to the Arkansas Game and Fish Commission as required for renewal of Scientific Collecting Permits. The electronic file should be saved in multiple locations under the purview of the Curator.

Deaccessioning—Specimens in the Herpetology Collection are only to be deaccessioned under the oversight of the Curator. Reasons for deaccession may include damaged specimens not salvageable or removal of one or more specimens to another museum. Any damaged specimens that can not be salvaged are to be disposed of by the director of ASU’s Environmental Health and Safety. All reasonable efforts should be made to salvage any specimens or parts thereof. Identifying photos may be taken if possible to verify the species, particularly in the case of county records. Any specimens disposed of or moved to another museum must be noted in the appropriate catalog book at the bottom of the page where that specimen is listed as well as in the ‘Notes’ section for that specimen in any electronic database. Additionally, a loan form stating the action taken and location of specimens should be completed and kept on file.

General Maintenance—Periodic maintenance is vital to the upkeep of a research museum. Jars should be checked every three to six months to verify sufficient EtOH. Jars with insufficient fluids should be topped off as soon as noticed. Additionally, weak shelves may need to be replaced. Any leaks should be cleaned up and the source of the leak identified. If a jar or bucket is leaking then the specimens therein should be transferred to a new container. Research areas within the collection room should be cleaned as soon as possible after use to maintain a tidy workspace. Storage is limited within the collection room, however objects should not be placed on top of shelving units as this prevents the free flow of water from the sprinkler system in the case of a fire. Living organisms are maintained in the collection room, but are not part of the research collection. These animals are used for outreach and education purposes and are also under the oversight of the Curator. Feeding or clean-up of these organisms should occur periodically depending on the needs of that species. Upon the death of these organisms, they are to be added to the teaching collection rather than the research collection.

Checklists—All checklists associated with the Herpetology Collection are under the oversight of the Curator. The catalogues of information for the specimens are kept within the collection room and are only to be removed under the authority of the Curator. Additionally, any work conducted with the collection should be logged on the appropriate sheet on the door. Feeding of any live animals should also be logged on the log sheets by that animal’s terrarium/aquarium. Loaned specimens or equipment should also be logged appropriately according to the Curator and kept on file.

CONCLUSION The utility and importance of natural history museums can not be overstated. They are vital repositories for invaluable specimens and scientific data. The fact that relatively few people see the specimens does not detract from their importance because the benefits of the museums are far-reaching into society, often unknowingly by the lay-person. Museums provide opportunities for a myriad of scientific studies that can advance our understanding of the world around us and help us in determining how to better protect our environment. Regarding the Herpetology Collection at ASU, it’s importance lies in the fact that it is the largest reptile and amphibian collection in the state of Arkansas, numerous studies have been published from the use of it’s specimens, numerous students have learned about reptiles and amphibians from the specimens housed therein, and many lay-people benefit from the museum through the publication of The Amphibians and Reptiles of Arkansas (Trauth et al., 2004) which would have been impossible to produce without the use of the museum specimens. The faculty, staff, and students of Arkansas State University, as well as the citizens of Jonesboro, AR should feel proud that such a collection exists at their school. The collection should be cherished and maintained in such a way that it will be a thriving part of the research atmosphere of ASU for generations to come. This operating procedure will help in accomplishing that goal. LITERATURE CITED

Hall, E. R. 1962. Collecting and preparing study specimens of vertebrates. University of Kansas Museum of Natural History Miscellaneous Publication 30:1-46. Hanebrink, E. 1993. A history of biological sciences at Arkansas State University. Arkansas State University Printing Services: Jonesboro, AR. Pisani, G. R. 1973. A guide to preservation techniques for amphibians and reptiles. Society for the Study of Amphibians and Reptiles Herpetological Circular 1:1-22. Trauth, S. E., H. W. Robison, and M. V. Plummer. 2004. The amphibians and reptiles of Arkansas. University of Arkansas Press: Fayetteville, AR. Appendix A. Partial list of publications utilizing specimens from the Herpetology Collection at Arkansas State University. Arrangement is by chronological order and only includes the years following the publication of The Amphibians and Reptiles of Arkansas by Trauth et al. (2004).

McCallum, M. L. and S. E. Trauth. 2006. An evaluation of the subspecies Acris crepitans blanchardi (Anura, Hylidae). Zootaxa 1104:1-21.

McAllister, C. T., C. R. Bursey, S. E. Trauth, and D. B. Fenolio. 2006. Helminth parasites of the grotto salamander, Eurycea spelaea (Caudata: Plethodontidae), from northern Arkansas and southern Missouri. Comparative Parasitology 73:291-297.

Trauth, S. E. and M. M. Mary. 2005. Seasonal incidence of sperm within the spermathecae of Ouachita dusky salamanders (Desmognathus brimleyorum) in Arkansas. Journal of the Arkansas Academy of Science 59:173-177.

Mary, M. M. and S. E. Trauth. 2006. Histology and histochemistry of caudal courtship glands in three Arkansas plethodontid salamanders. Herpetological Review 37(3):280-284.

McCallum, M. L., S. E. Trauth, and B. Neal. 2006. Tail-coiling in ringneck snakes: flash display or decoy? Herpetological Natural History 10(1):91-94.

Trauth, J. B., R. L. Johnson, and S. E. Trauth. 2007. Conservation implications of a morphometric comparison between the Illinois chorus frog (Pseudacris streckeri illinoensis) and Strecker’s chorus frog (P. s. streckeri) (Anura: Hylidae) from Arkansas, Illinois, Missouri, Oklahoma, and Texas. Zootaxa 1589:23-32.

Milanovich, J. R., T. McKay, and S. E. Trauth. 2008. Diet of the western slimy salamander (Plethodon albagula) from two mountain ranges in Arkansas. Southwestern Naturalist 7(2):327-330.

McCallum, M. L., J. L. McCallum, and S. E. Trauth. 2009. Predicted climate change may spark box turtle declines. Amphibia-Reptilia 30:259-264.

Trauth, S. E. and D. M. Sever. 2011. Male urogenital ducts and cloacal anatomy. Pp. 411-475. In. Reproductive Biology and Phylogeny of Snakes. (eds., R. D. Aldridge and D. M. Sever). CRC Press, Boca Raton, FL.

Stanley, J. W. and S. E. Trauth. 2011. Nerodia rhombifer (Diamondback Watersnake). Intersexuality. Herpetological Review 42:296.

McCallum, M. L., C. Brooks, R. Mason, and S. E. Trauth. 2011. Growth, reproduction, and life span in Blanchard's Cricket Frog (Acris blanchardi) with notes on the growth of the Northern Cricket Frog (Acris crepitans). Herpetology Notes 4:24-35.

Sawyer, J. A. and S. E. Trauth. 2011. Seasonal Activity, Population Characteristics, and Age Estimation in the Aquatic Salamander, Siren intermedia nettingi (Goin). Journal of the Arkansas Academy of Science 65:117-125.

Trauth, S. E. 2012. Morphology of Rathke’s glands in the snapping turtle, Chelydra serpentina, with comments on the presence of multilaminar lamellar bodies in turtles. Journal of the Arkansas Academy of Science 66:164-172.