Guidelines for the Procedure of Obtaining Mammal Specimens as
Approved by the Mammal Society of Japan (Revised in 2009)
The Committee of Reviewing Taxon Names and Specimen Collections, Mammal
Society of Japan([ ]: notes by The Committee at 2015)
Today’s challenges faced by wildlife research communities are complex,
involving many issues on nature conservation and animal welfare. They also face various
issues in society relating to their accountability for the credibility of scientific studies andtheir results, while adhering to professional criteria through original publication. A
number of academic societies and related organizations whose members conduct
scientific study on animals have established policies and guidelines pertaining to thehandling of live animals and preserved animal specimens (e.g., Japanese Association forLaboratory Animal Science 1991; Japanese Society of Zoo and Wildlife Medicine 2003;
Nature Conservation Committee and Editorial Committee of the Ichthyological Societyof Japan 2004; Japan Ethological Society 2005; Science Council of Japan 2006). In the
Mammal Society of Japan (MSJ), The Committee of Reviewing Taxon Names andSpecimen Collections (“the Committee”) instituted the first edition of the Guidelines for
the Procedure of Obtaining Mammal Specimens (“Guidelines”) under the leadership ofDr. Kazuhiro Koyasu in 2001 (Mammalogical Society of Japan Committee on
Taxonomical Names and Collections, 2001; in Japanese). In the first paragraph, theoriginal edition explains in detail the background information that led to theestablishment of the Guidelines. Thereafter, our society has seen increasing nationwide
interest in environmental issues, animal welfare, and zoonotic diseases. On top of thisgeneral trend, major revisions to existing wildlife regulations as well as enactment of newdomestic legislation that directly affect wild mammal studies in Japan, such as the
Wildlife Protection and Hunting Law and the Invasive Alien Species Act, have made the
MSJ recognize the need for a timely revision to the Guidelines. Under thesecircumstances, in 2006 the Guidelines Revision Task Force was formed under the1Committee. Members of the Task Force are as follows, in alphabetical order of their lastnames: Masahiro Iwasa, Hideki Endo, Mariko Kageyama, Shin-ichiro Kawada, Kazuhiro
Koyasu, Motoki Sasaki, Keisuke Nakata, Masaharu Motokawa, and Yasushi Yokohata(Task Force Chair). The revised version was approved by the MSJ Council in July 2009.
The purpose of the Guidelines is to provide professional standards by whichhandling procedures of specimens in mammalogical research, including field collection
methods, can be rightly evaluated as endorsed by the MSJ. Given the fact that thecapturing of wild mammals in the field normally precedes preparation of specimens, theGuidelines address not only field research techniques but also ethical considerations fromthe standpoints of animal welfare and conservation. As far as the handling of live animals
is concerned, the MSJ has not yet adopted procedural guidelines as of 2009. Therefore,
similar guidelines developed by some other organizations (e.g., Japanese Association forLaboratory Animal Science 1991; Fukui 1991; Japanese Society of Zoo and Wildlife
Medicine 2003; Japan Ethological Society 2005; Science Council of Japan 2006; outside
Japan, Ad hoc Committee on Acceptable Field Methods in Mammalogy 1987; Bookhout
1996; Animal Care and Use Committee 1998; Gannon et al. 2007) should be referred towhere appropriate. Ethical aspects of conducting research using live animals are outlinedin Murakami and Saeki (2003).
The Guidelines are not intended to discourage researchers from attempting toundertake a project using a revised and improved methodology or from designing an
original research project using a new technique. Ultimately, each researcher must be
responsible for their decision as to which project design and method they adopt. Intended
users of the Guidelines are basically MSJ members and other non-member mammalogists.
However, it is highly expected that the Guidelines are of interest to other stakeholders,such as natural history museums with no mammal expert staff, policymakers andgovernment agencies that deal with legal regulations on mammals, foundations andsupport organizations that provide research grants to mammalogists, as well as to editors
and reviewers of academic journals that accept papers in the field of mammal research.

21. Value of Specimens in Mammalogy
Today, mammalogy encompasses a broad range of research hypotheses as well as
various approaches to test them. When the subject of research includes wild mammals, it
is often the case that you have to live-capture or kill animals in order to obtain necessary
data from the study material. The information obtained enables accurate identification of
species and an understanding of systematic and evolutionary relationships, genetic
phenomena, population dynamics, community structure and dynamics, comparative
anatomy and physiology, behavior, parasites and diseases, economic importance,
geographic and microhabitat distributions, ecology of mammals in their natural ormanaged environments, and other scientifically important phenomena (Ad hoc
Committee on Acceptable Field Methods in Mammalogy 1987). In reference to othermammalogical societies outside Japan, for instance, the American Society of
Mammalogists strongly recommends that researchers donate various types of mammal
specimens obtained during field studies to “systematic collections that meet minimal
standards” (Committee on Systematic Collections 1975, 1978; Systematic Collections
Committee 2004; see Appendix to the Guidelines for a Japanese translation of the latter,
which is omitted in this English version) as vouchers, after the completion of the study. A
collection in a museum that meets such standards serves as a permanent repository ofvoucher specimens and their associated data examined by different researchers. Voucher
specimens preserved in this manner subsequently constitute a systematic collection thatenables validation of past research, and that also serves as a resource available for present
and future research. Abe (1992a) goes into detail about the significance of voucher
specimens collected and preserved in mammalian research from the angles of taxonomic
information, ecological and physiological information, and other ancillary information.
Maximizing the scientific value of specimens entails adequate preparation,
documentation, and preservation of original specimens and their associated data. The
Guidelines aim at providing professional benchmarks to standardize procedures ofcollecting and preparing mammal specimens, and for documenting associated data. The
Guidelines are also meant for mammalogists to serve as a practical guide in preparing
specimens. Furthermore, topics of professional ethics and techniques related to wild
mammal collections discussed herein would be relevant to ecologists in general.

32. General Issues Concerning Mammal Collection in the Field
Sampling protocols for mammal specimens include not only traditional lethalmeans of collecting museum specimens, but also other nonlethal approaches such as
obtaining biopsy samples from live-captured individuals and releasing themafterwards. The Guidelines mainly take into account ramifications of adopting lethal
methods. Hamazaki (1998), Kishimoto (2002), and Kaneko and Kishimoto (2004)
point out “safety of animals,” “safety of workers,” and “minimal impact on theenvironment” as “three basic rules of animal collecting.” In this context, emphasis is
made on the need for compliance, significance of the rules, and some other points ofconsideration. Despite originally being proposed primarily with nonlethal methods in
mind, these three principles are also applicable to lethal methods in general.
Consulting the Handbook of Japanese Wildlife Care and Medicine Editorial
Committee (1996) and Bookhout (1996) should be required to reference the technicalaspects of stabilization, rescue, and external measurement of data collected from livecaptured wild mammals.
2–1. Laws on Field Collecting
The following discussion is based on Ikeda and Hanai (1988), one of the firstreview articles that outlined regulations and legal procedures that are relevant to field
mammal collectors in Japan. The topic is divided into three main sections, namely,
Wildlife Protection and Hunting Law, more commonly referred to as the “Wildlife
Protection Law (WPL)” (original Japanese title, Choju Hogo oyobi Shuryo ni
Kansuru Houritsu, subsequently changed to Choju no Hogo oyobi Shuryo no
Tekiseika ni Kansuru Houritsu) [currently, Wildlife Protect Management Law(WPML), Choju no Hogo oyobi Kanri narabini Shuryo no Tekiseika ni Kansuru
Houritsu]; Law for the Protection of Cultural Properties (Bunkazai Hogo-ho), andother relevant laws. WPL experienced a major revision in 2003, in which the term
“wildlife” (= Choju) was distinctly defined as “wild animals that belong to Aves andMammalia.” This definition allows for further inclusive application of the existing
law to all the Japanese mammals encompassing the groups that were previously
placed outside the scope of the law: order Insectivora [currently, Soricomorpha andErinaceomorpha], family Muridae (excluding pests caught by agricultural and forestry
workers in addition to three residential species: Mus musculus, Rattus rattus, and R.
norvegicus), and certain taxa of sea mammals (i.e., seals, sea lions [not including

Steller Sea Lions], and dugongs; see below for the other marine mammals that remain
exempt from said regulation) (Ishinazaka 2003; Yokohata 2003). In order to collectany wild mammal species regulated by the WPL for research purposes, either one of
the following two types of collection permits is required: Wildlife Collection Permit
issued under WPL Clause 1, Article 9, or Class-A Hunting License issued under WPL
Article 39 [currently, also see WPML Articles 9 and 39]. Note that the latter is more
restrictive in the conditions than the former in that a permit holder is allowed to catchonly those species listed as game animals using regulated tools and equipment only,
within a restricted hunting zone, for a limited period of the hunting season. The
following reference is useful, besides the aforementioned, in acquiring information
before applying for a new collection permit for scientific research purposes: Komaru(2001), Wildlife Conservation Administration Study Group (1992, 2003), and Wild
Bird and Mammal Management Study Group (2001). For further details, first consult
the appropriate governmental agencies responsible for issuing collection permits.
Hatakeyama (2004) and Sakaguchi (2007) recently added to the literature on thissubject.
Alongside the general laws discussed above, collection of Japanese mammals
that are listed as threatened species are now regulated under the Endangered Speciesof Wild Fauna and Flora Conservation Law that was issued in 1992 and has been
enforced since April 1, 1993 (Environmental Agency Wildlife Conservation
Administration Study Group 1993, 1995; Management and Coordination Agency
Administrative Inspection Bureau 1993). International regulations pertaining to theexport and import of specimens of endangered species are implemented by theConvention on International Trade in Endangered Species of Wild Fauna and Flora(CITES) as discussed in the section 6–2.
To summarize, collecting, importing, and/or exporting wild mammals are
subject to domestic and international regulations based on the wildlife laws andtreaties, such as those stated earlier; therefore, if certain species, locations, or
activities related to your fieldwork are subject to these laws, obtaining valid permits
issued by authorized agencies is mandatory. These laws and acts may be amended
with addition of new regulations in the future; hence, each researcher is responsible
for being versed in all the regulations that affect their study area and for acquiring
necessary licenses ahead of time (Rohlf 1995; Wildlife Management Study Group

2001; Wildlife Conservation Administration Study Group 2003; Hatakeyama 2004;
Sakaguchi 2007).
Among Japanese marine mammals, the collection of whales and Steller Sea
Lions is regulated under the Fisheries Act, and certain species of whales are also
stipulated in the Fisheries Resource Protection Law. In order to legally collect theseanimals for scientific study, a researcher must get permission from the Minister ofAgriculture, Forestry, and Fisheries based on the Enforcement Regulations of theFisheries Act, Article 1, or the Enforcement Regulations of the Fisheries Resource
Protection Law, Article 1. Likewise, collection of domestic sea otters and fur seals isbasically prohibited by the Sea Otters and Fur Seals Hunting Control Act. For
scientific collection of these groups of Japanese sea mammals, persons must obtain alicense from the Minister of Agriculture, Forestry, and Fisheries following Article 1of the Enforcement Regulations of the above law.
In addition to field collection permits, persons may also need to consider
seeking approval related to public and private land use. For instance, persons need toapply for a forest use permit granted by a district forest office as a requirement forentering a national forest, and in the case of a public land, they are obligated to obtain
permission from a land owner and manager ahead of time, before they collect wildlife
on their land. Furthermore, the Invasive Alien Species Act (issued in 2004, enforcedsince October 1, 2005) prohibits the raising, possession, and transferring of Invasive
Alien Species, including 16 species [currently 19 species and 2 hybrids] and 4 generaof mammals (as of 2008) in principle. Komaru (2001) provides general guidance on
biological fieldwork and relevant laws, which include a listing of contact information
for inquiries about the field sampling of organisms.
2–2. Ethical Issues on Field Collecting
Ethical issues regarding wildlife sampling are clearly illustrated by Nagorsen
and Peterson (1980). The possession of a valid collecting permit does not give thecollector the right to collect animals irresponsibly. Persons are supposed to conduct
fieldwork using the most humane method possible and strive to minimize impact on
the local biota without disturbing sampling sites. Do not engage in indiscriminate
mass collecting. This is not permissible, especially in circumstances where a large
number of animals are clustered in one area. If possible, it is advisable to live-capture
mammals, and as soon as the total number reaches the needed sample size, releaseexcess live animals to the wild intact. Such flexibility is only possible when live traps
are used, and there is no such option if persons conduct fieldwork using kill traps. In
fact, the sample size required for a certain study is subject to the study design.
Therefore, every investigator who conducts fieldwork is held accountable for aparticular sample size adequate for their study at their own discretion, and they shouldcollect no more specimens than are needed (Ad hoc Committee on Acceptable Field
Methods in Mammalogy 1987; Animal Care and Use Committee 1998; Gannon et al.
2007). On the other hand, it is also inadequate if too few animals are sampled because
this might not allow the investigator to achieve the study purpose and, subsequently,
will result in waste of animals (Gannon et al. 2007).
Friend et al. (1994) justifies with clarity the need for adopting lethal methods
in wildlife study as described in the following statement: “[The] collection of animals
often is an essential component of field investigations. These studies may involve
systematic zoology, comparative anatomy, disease assessments, food preference
studies, environmental contaminant evaluations, and numerous other justifiable causes
and scientific needs.” To evaluate a proposed study requiring animal collection,
Friend et al. (1994) further enumerates the following criteria of assessment: 1)
whether the proposed collection will provide scientific data that is not duplicative of
information already in the scientific literature, or information that is presently
available in accessible scientific collections and repositories; 2) whether suitable
information can be obtained from alternative methods that do not require taking live
animals; 3) whether methods of collection are acceptable and effective to minimize
the potential of trapping non-target species; 4) whether target animals can be killed as
quickly and cleanly as possible; and 5) whether appropriate methods are employed to
be as age-class specific as possible according to the purpose of the study. Finally,
specimens intended to be preserved in a museum to build a systematic collection must
be prepared with meticulous care, and scientific data associated with specimens must
be documented and archived using a standard method (see 4–1 through 4–7).

3. Animal Collecting Equipment and Techniques
3–1. Collection Using Kill Traps and Guns
When collecting mammals using kill traps, special attention should be paid to not
let animals suffer unnecessarily nor damage their body parts that are to be saved for the
study. An all-plastic snap trap model, commercially known as a “Panchu trap” in Japan,
is one of the most convenient and easiest to use for catching small mammals, such as
small rodents and terrestrial insectivores (Abe 1991a, 1992b; Murakami 1992; Yoneda etal. 1996). Other brands of snap traps include a Victor® trap and various other types ofmetal traps, but these models, excluding a Museum Special (made in USA) and a custombuilt metal snap trap (see Murakami 1992), are prone to crushing skulls (Imaizumi 1970),
and therefore, may require a slight adjustment to the device such as repositioning a bait
trigger plate. Murakami (1992) provides source information about these products. As forcatching fossorial moles, various types of mole traps are introduced, including a scissor
trap, a tube trap that comes with a wire loop for strangling, and a harpoon trap (Abe1992b, Yokohata 1998). Specifically for water shrews, a large-size snap trap with a bait
trigger plate, a tube trap attached with a mesh screen, as well as a “mujiri” cage
traditionally woven from reed or bamboo, have been used (Abe 1992b). Using these killtraps is considered an adequate means of collecting small animals that meet a research
purpose as well as an ethical standard.
Decisions regarding a trapping method and trap positioning should be made basedon a factor that maximizes the odds of catching the intended species and minimizes the
chance of trapping non-target animals. To avoid overlooking traps, it is important to markthe site of each trap station with a marker like colored tape. In addition, a collector isrequired to put a label on each trap that includes information such as the permit holder’sname, a name of the office issuing the permit, and a permit ID number. In case a trap is
too small to label, a collector must post a placard or a sign at the site. It is essential tocheck all the traps at least once a day, ideally early in the morning, to remove trapped
animals as soon as possible. Mammal carcasses tend to decompose rapidly at hightemperatures, and if they are left in traps long enough before being picked up, they could
be prone to ant damage or maggot infestation. Pitfall traps are particularly suitable forcollecting shrews (Abe 1992b; Koyasu 1998), but if this type of trap is used as a kill trap,8

it should contain sufficient water to ensure that the captured animal(s) do not suffer long
before drowning (Ad hoc Committee on Acceptable Field Methods in Mammalogy 1987).
Smaller-size “body-gripping traps” have been in use as kill traps for chipmunks
and tree squirrels (Abe 1991a) as an exceptional case, but generally body-gripping traps
and comparable foothold traps are known as typical non-humane models that do not kill
animals quickly. Therefore, alternative and more humane types of traps should be chosenwhen available. If there are no substitutes for steel jaw traps, it is recommended thatmodified traps with padded jaws be used (Kuehn et al. 1986; Nakazono and Doi 1989;
Ikeda 1989; Kaneko and Kishmoto 2004), and that traps be checked frequently, at least
twice a day (Ad hoc Committee on Acceptable Field Methods in Mammalogy 1987).
For mammals such as rabbits, hares, squirrels, and small- to medium-sizedcarnivores, shooting may be more humane than trapping (Nagorsen and Peterson 1980).
In order to use guns for scientific collecting by the researcher, they are responsible forobtaining a hunting license beforehand, specifically, the first-class gun hunting license foruse of firearms and air guns [currently, firearms only], or the second-class gun hunting
license for air guns only, as set forth in Article 39 of the WPL. On top of that, they mustabide by regulations on the possession and use of guns known as the Act for Controlling
the Possession of Firearms or Swords and Other Such Weapons, or more commonlyreferred to as the Swords and Firearms Control Law (Japanese title: Juhou Touken-rui
Shoji-tou Torishimari-hou or “Jutou-hou”). In addition, there is a law that specificallyregulates the possession and storage of ammunition called the Explosives Control Act(Kayaku-rui Torishimari-hou). Researchers must comply with these laws and gain
sufficient experience to use guns properly and safely.
For the scientific collecting of large cetaceans, a whaling gun loaded with an
explosive harpoon is used. A harpoon gun charged with an explosive called penthrite(pentaerythritol tetranitrate or PETN) is recognized by the International Whaling
Commission as one of the most humane killing techniques for whales (International
Whaling Commission 1981). In the case that an explosive harpoon did not kill a whale
instantly, a 9 mm (0.35 in) caliber or a larger caliber rifle may need to be used as asecondary means of finishing. Cetacean species are not subject to the WPL, and therefore,
regardless of the method of killing whales, a license is required for the possession of

9

firearms in accordance with Article 4, Paragraph 1, Item 2 of the Swords and Firearms
Control Law.
3–2. Collecting Using Live Traps and Nets
Live trapping is often a preferred method of sampling mammals in certain areas ofresearch such as karyotyping, biochemical and genetic studies, ectoparasites, and a markrelease-recapture approach in ecological studies. One of the merits of using live traps is toenable a researcher to capture only the target species and number of specimens needed
for study, and to release all the other unwanted animals to the wild unharmed. However,
it is important to choose the right type of live trap that works most efficiently for its
intended catch. For example, it is known that the trappability of Crocidura shrews is
significantly higher than that of Sorex shrews when Sherman-type live traps are used.
Live traps are usually made of box- or tube-shaped containers using various materials
such as aluminum, zinc, wood plates, wire mesh, and plastic. They are designed to workin a manner in which an animal that is lured into a trap container would come in contact
with a trigger inside to close its door in a set position, resulting in the live animal
contained in the trap. Commercially available brands include Sherman, Havahart,
Longworth, Penlon, and Tomahawk, and live traps can be ordered either directly fromthese overseas manufactures, through domestic retailers (see Murakami 1992), or byinternational mail order (e.g., Carolina Biological Supply Company; www.carolina.com).
Products similar in shape and function to those foreign brands are manufactured and soldin Japan as well (Murakami 1992; Yoneda et al. 1996). Penlon-model live traps made ofplastic are capable of trapping small to medium-sized mammals such as shrews and large
field mice (Shibanai and Izeki 1997), whereas wire-mesh traps are usually used to capture
live, larger-sized mammals such as nutrias (Miura 1992) and wild cats (Izawa 1990).
Simple techniques of collecting live moles include live-capture by ambush or with pitfall
traps, but tube-shaped live traps, such as the Nishi-method and the Konishi-method, aremore reliable and are often used by mole researchers (Abe 1992b; Yokohata 1998;
Kawaguchi 2004). An overview of live-trapping techniques for soricid species is found in
Abe (1992b), Koyasu (1998), and Motokawa (1998), whereas trapping techniques for
carnivores are summarized in Kaneko and Kishimoto (2004). Wooden box traps are10

preferably used for mustelids in order to keep trapped animals from damaging their teeth
in their attempt to break a metal trap open (Sasaki 1990). In the case of live-catchinglarge-sized mammals using foothold traps, the jaws must be cushioned or padded withrubber for softer contact as discussed in the preceding section (Ikeda 1989). Giant flying
squirrels can be caught using a modified sweep net (Baba 1988). Box traps or snare traps
are often used to catch live hares (Yamada et al. 1988).
It is important to choose a live trap of an appropriate size large enough to allow
for movement of the target species without its suffering inside the trap. During the coldseason, in order for the animal to avoid exhaustion due to decreased body temperature, it
is imperative to place a sufficient supply of food and nesting material inside the trap, andto cover the whole trap with heat insulation material. Each live trap should be regularly
checked at an adequate frequency to minimize accidental death or exhaustion of theanimal trapped inside. When live-capturing large-sized mammals using box-shaped traps,
persons may not need to visit traps so often if a radio transmitter is attached to the trap’s
door. It usually works best if a transmitter is kept turned on while the trap door is set open,
and the signal is automatically turned off as soon as the door is shut. The opposite
mechanism of keeping a radio device off when a trap is set open (and turning it on upon
the capture of an animal) is not recommended because of higher risk of mechanicalfailure. Live traps for nocturnal species should be set before dusk and checked as soon as
possible after dawn. They should be closed during the day after the morning check toprevent accidental capture of diurnal species (Animal Care and Use Committee 1998).
Live traps for diurnal species should be shaded, so as to avoid full exposure to the sunand should be checked frequently (Ad hoc Committee on Acceptable Field Methods in
Mammalogy 1987; Koganezawa 1989).
During routine walks, attention should focus on checking traps, so as to not
overlook any live animals caught in traps. Further, at the end of fieldwork, make sure all
traps are retrieved and temporary field markers, such as colored tape, are completely
removed from the study area. It is recommended to label all traps with sequential
numbers and keep track of them in numerical order during the initial setup, regularchecking, and final retrieval. In this way, one can reduce the chances of overlooking any

11captured animals during a regular trap inspection or leaving behind a trap at the end of atrapping session.
Highly frequent inspections are required when live-capturing small mammals
using pitfall traps (e.g., about once every hour for shrews). Bait should be placed in apitfall trap in order to keep trapped animals from starving to death. In addition, captured
animals experience a higher mortality rate when they are wet from rain, so it is necessary
to cover the top of a pitfall trap to provide shelter for the trapped animals (Pucek 1981).
To live-capture large-sized mammals, various types of live traps areconventionally used. For bears, box traps (box-shaped cages: Watanabe and Nozaki 1989;
Kaneko and Kishimoto 2004) and barrel traps (drum traps: Mano et al. 1990; Yoneda etal. 1996; Kaneko and Kishimoto 2004) are commonly used. For artiodactyls such as sika
deer, Japanese serow, and wild boar, box traps and net traps are known to be effective, as
well as a corral trap system, in conjunction with a metal fence structure, to drive animals
into an enclosure (Ito et al. 1989; Nakatani 1989; Kaji et al. 1991). Specifically for deer, amodified corral trap system is adopted, by which the deer would be enclosed by cloth
curtains that rise when the animals enter the enclosure (Uno et al. 1996). As for wild boar,
snare traps are also conventionally used (Nakatani 1989). When the two models of beartraps are compared, one of the disadvantages of using metal cages is that a captured bear
likely ends up getting its teeth damaged from biting the metal bars of the cage (Yoneda etal. 1996). In contrast, a bear is unlikely to be injured when captured in a barrel trap
(Mano et al. 1990). Likewise, when a metal fence corral trap is used for catching deer,
there is a higher chance that the animal would crash into the metal fence, leading to major
injury or death. On the other hand, an improved corral trap system outlined with soft
cloth screens is considered much safer for capturing deer (Uno et al. 1996). Today, snare
traps are deemed inappropriate to capture live wild boars because of higher injury andmortality rates (Nakatani 1989).
When live-catching large wild animals, it is essential to perform various necessary
procedures in as short a time period as possible after the capture or the animals may
become violent and be injured. It is also important to pay careful attention to each animal
that is captured and immobilized in order to avoid causing capture myopathy (CM). One
has to be prepared to be ready to administer a fast-acting treatment against CM symptoms

12

(Suzuki 1999). From a legal standpoint, a hunting license or a scientific collecting permit
is required for live-capturing wildlife, specifically a net hunting license and a trapping
license are required when using nets and live traps, respectively. Additionally, in the caseof using firearms and air guns, you must comply with laws and ordinances that regulate
the possession and use of these devices (cf. Ikeda and Hanai 1988). If an anesthetic is
applied in order to live-capture animals, it is mandatory to obtain permission from theEnvironment Minister to use an anesthetic designated as a deleterious medicine (cf. WPL
Article 15 [currently, Article 37]).
As far as bats are concerned, a collection permit is required to capture bats forscientific research purposes. In addition, permission must be obtained from theEnvironment Minister in order to use a mist net, because the law generally prohibits thesale, possession, or use of mist nets (cf. WPL Article 9, Paragraph 1, Item 3). Further, it
is required to present the above collection permit in order to buy a net (WPL Article 16,
Paragraph 2) [currently, WPML Article 16, Paragraph 2, Item 1]. The Bat Study andConservation Group of Japan (1998) provides useful information on how to purchase a
mist net, as well as techniques for using it to catch bats. It is important to adopt the mostappropriate capturing method for different species of bats depending on specific habitats
they live in (Mohri 1988; Kunz and Kurta 1988). Researchers generally should avoid
exploring a cave where a bat maternity colony is roosted during the period of parturitionand nurturing, so as not to disturb the breeding colony. Repeated disturbance ofhibernating bats may also lead to higher mortality (Ad hoc Committee on Acceptable
Field Methods in Mammalogy 1987) and must be avoided. There is a special device
known as a “One-Way Door” that can be installed to force a roost out of a residential
structure (Gelfand 1997; Tuttle and Hensley 2000). However, One-Way Doors should not
be installed during the parturition and nursing period as this may lead to higher mortalityof flightless pups, especially if this method is applied in the summer breeding months ofJune, July, and August. A collection permit is not required to build a bat house (a shed ora box designed to shelter a bat roost) as far as it is only meant for conservation andobservation of bats, and the project does not involve direct handling of bats. However,needless to say, one should get permission from a landowner and property manager prior
to building a bat house on their property. In addition, if fieldwork entails temporary or

13permanent capturing of bats from a bat house for data gathering, a collection permit isrequired to conduct such a scientific study. Choosing the right kind of bat house withrespect to its materials and size, and selecting a suitable location to install a bat house isimportant for its effective use. It is advisable to provide additional devices in the area that
protects bats from attacks by potential predators (Gelfand, 1997). Overall, bat researchrequires familiarity with the research methodology peculiar to this group of mammals,
including trapping techniques, and therefore investigators should be well versed in
ecological and behavioral study techniques (cf. Kunz 1988) for the specific group of batsof interest.
Encircling nets or hoop nets are usually used to live-capture smaller-sizedcetaceans, especially dolphins and porpoises. The former method has the advantage of
capturing a school of marine mammals effectively at one time, but necessitates operationof multiple vessels in coordination. The latter method takes advantage of the dolphin’s
habit of approaching a sailing ship to ride the bow wave (Ridgway 1972), but thetechnique is hardly adopted in Japan. In either way, full attention is needed during thelifting and transport of animals to an adequate aquarium facility to minimize undue stress.
When euthanizing a live mammal captured in the wild, the most ethical andhumane way must be followed (as a legal justification behind this statement, nonbinding
rules regarding animal euthanasia are stipulated in the Articles 23 and 24 [currently,
Articles 40 and 41] of the revised Act on Welfare and Management of Animals; see
Animal Welfare and Management Laws Study Group 2001). In general, euthanasia, or
mercy killing, means the act of putting an animal to death without inflicting prolonged
suffering by rendering it unconscious in the shortest time possible (Japanese Association
for Laboratory Animal Science 1991). Administratively, animal euthanasia “shall follow
chemical or physical procedures that are the most effective methods possible to trigger
the loss of consciousness without causing pain or distress to a sacrificed animal, resulting
in irreversible cessation of its cardiac or lung function, or follow other socially acceptable
methods in a normal manner,” according to the Guidelines on Methods of Sacrificing
Animals (Notice No. 40 of the Japanese Prime Minister’s Office, 1995 [currently, No.
105, 2007]; see also Asano et al. 2006). Various agents and methods of euthanasia are
known such as administering an overdose of anesthetics, ether, or carbon dioxide (Tajima

14

et al. 1979; Nakamura et al. 1984; Japanese Association for Laboratory Animal Science
1991; Japanese Prime Minister’s Secretariat Management Office 1996). Selecting atechnique should be based on an evaluation of the most humane method that, if at all
possible, would not subject the animal to pain or suffering while meeting the study
purpose. Administration of ether used to be one of the most frequently adopted
procedures, but today ether is not recommended because of its toxicity to humans andanimals, as well as its irritative, explosive, and flammable attributes. On the other hand,the use of carbon dioxide as a method of euthanasia can cause irritation of mucosal
linings in some species (Gannon et al. 2007). Euthanasia must be performed outside theperceptive range of other animals so as not to trigger fear or undue stress in other captive
individuals (Gannon et al. 2007). If you have any questions or uncertainty regarding
euthanasia techniques for particular study animals, it is recommended to consult
veterinarians or laboratory animal experts who are knowledgeable about euthanasia
protocols for domesticated or laboratory animals. The latest review on the ethicalperspective of animal experimentation is found in Kagiyama (2008).
4. Data Associated with Specimens
It is recommended to store field data associated with mammal specimens thatwere collected, examined, and preserved for systematic study, as well as for otherresearch purposes. For instance, field data in conjunction with specimens that wereoriginally captured for field surveillance can further provide a wide variety of biologicalinformation, such as geographic variation in morphology, age variation, age
determination, and age structure in a population, estimation of the breeding period basedon age determination, reproductive condition, litter size, diet and feeding habits, nutrition
status, quality of a population, and parasites (Abe 1991b). Moreover, in closely related
study areas, such as morphology, genetics, biochemistry, and parasitology, preservationof study materials as voucher specimens not only ensures the taxonomic underpinning ofa study, but also makes it possible to correspond with future taxonomic revisions.
Therefore, whatever the main study purpose of collecting mammal specimens could be,
documentation of basic field data for each specimen (e.g., collector’s serial number,species identification, sex, measurements, locality, date collected, method of collection,15

other remarks) should be accomplished as a part of the daily tasks of fieldwork, and suchprimary records should be archived together with physical specimens. However,uncertain data should never be recorded based solely on speculation even if aninvestigator is caught up with taking multiple specimen data. For example, if the sex of a
specimen cannot be determined in the field, it is better to indicate so with a question mark(?) in a field notebook or a field catalogue (Nagorsen and Peterson 1980).
4–1. Recording Data
It is most common that data associated with field collecting activities is recorded
in a notebook or a certain data form. Martin et al. (2001) recommends that field notes beorganized into three separate sections: 1) a diary journal, 2) field catalogue, and 3)
species accounts. In the journal, all collecting activities and daily observations are
documented as a field diary on a day-to-day basis. The field catalogue section is useful torecord specimen data, such as measurements, for each animal collected, and such data isrecorded in the order of the collector’s serial number assigned to each specimen. The
species accounts section includes detailed observational records on particular species thatare captured (Martin et al. 2001). It is important to archive a diary journal generated for
all types of fieldwork. It is also advisable to document a field catalogue on printed
catalogue sheets rather than a blank notebook (British Museum 1968; Nagorsen andPeterson 1980). Unless custom printed catalogue sheets are used for a field catalogue,commercial A4-sized horizontal-ruled data-recording sheets (e.g., KOKUYO Accounting
Pad SHIYO-26N) are recommended for use as catalogue data forms (catalogue sheets)given their ease of use and reasonable price. Today it is acceptable to record field data
electronically using spreadsheet software such as Microsoft Excel, but data on electronic
media should be backed up, ideally printed out on paper to be filed physically, to ensure
long-term preservation of the original data.
Data categories that appear as column headings on a catalogue sheet are as
follows:
Field Number–this is a unique number assigned to each specimen that is recorded on acatalogue sheet in the field. Unless one already follows their established specimen
numbering system, it is advisable to stick to the collector’s serial numbers, that is, a series

16

of the collector’s sequential numbers independent of field trips, localities, species, orcollection dates. The field number should be permanently written on a field tag attachedto each specimen with a pencil, India ink, or permanent ink, but never with a ballpoint
pen, felt-tip marker, or a fountain pen loaded with a water-based ink cartridge.
Catalogue Number–a number that is assigned and recorded once the specimen has beenaccessioned to an institutional collection; thus, catalogue numbers are not recorded in thefield.
Species Name–Record of species identification (Japanese name or scientific name) in thefield. Useful references for identification of Japanese mammals include coloredillustrations (Imaizumi 1960), keys (Maeda 1983; Yoshiyuki 1989; Abe et al. 1994, 2005),
and skull illustrations (Abe 2000).
Standard formats for recording Sex, Weight and Measurements, and Locality are
discussed below under the sections 4–3, 4–4, and 4–6, respectively. Information on
Reproductive Condition, Habitat, and Field Observations are documented either as three
separate data categories on a catalogue sheet or can be lumped together under “Remarks”(see sections 4–5 and 4–7).
When specimens are to be deposited in a museum collection, the documentation
associated with them should also be permanently archived by museums, including all
catalogues, field notes, photographs, and maps of collecting sites. As catalogue sheets
and field notes are often the only source of information for specimens, the catalogue andfield notes should be well organized, legible, and as accurate as possible (Nagorsen andPeterson 1980).
4–2. Specimen Labels
Choose the right kind of quality paper that is resistant to alcohol and formalinsolutions for specimen labels. Labels are generally attached to all different types ofpreparations: study skins, skulls, skeletons, and specimens in fluid. Writing thecollector’s initials preceding the collector’s field number on a specimen tag helps toprevent any possible confusion with specimens of another collector (Nagorsen andPeterson 1980). Refer to Abe (1991b) for data items to be recorded on a specimen label.
In essence, this means transcribing each data item as completely as possible from the17catalogue onto a label. Some museums, like the Royal Ontario Museum in Canada,
adopted a computerized cataloguing system, in which a specimen is originally labeledwith only a field number tag (Nagorsen and Peterson 1980). This approach is deemed
effective at institutions where a collection data management system is well establishedand flawless so that there is no chance of issuing duplicate labels. Determine data items
to be included on a specimen label as well as its format and style following examples
provided in references such as Okada (1940), British Museum (1968), Imaizumi (1970),
Nagorsen and Peterson (1980), Martin et al. (2001), Pucek (1981), Handley (1988), andYoneda et al. (1996). A set of skin, skull, and other associated specimens, such as tissue
samples obtained from an individual, should be assigned the same specimen number even
if they are stored at different locations. When molecular and cytological specimens or
host specimens for parasitological inspections are provided to outside institutions andresearchers, associated specimen numbers should always accompany the samples being
given. In published literature, numbers for voucher specimens, and tissue or parasite
samples should be cited for cross-reference.
4–3. Sex Determination
It is often possible to determine the sex of an animal based on an inspection of theexternal reproductive organs, such as the vaginal opening, scrotum, and penis, for
medium to large-sized mammals, as well as for small mammals if they are in the breeding
season (Imaizumi 1970; Abe 1991b). In the case of cetaceans, external reproductive
organs are housed within a genital slit, but it is possible to tell male from female based ona difference in the distance between the anus and the genital slit, which is relatively
longer in males than in females (Nagorsen and Peterson 1980). Similarly, rodents can besexed externally on the basis of the distance between the anus and the reproductive organ,
specifically, the clitoris or penis (Nakata 1986; Abe 1991b). On the other hand, juveniles
and small mammals in the non-reproductive season have poorly developed external
reproductive organs. Therefore, it is advisable to inspect the internal reproductive organsof a specimen to reduce the chance of false sex determination (Imaizumi 1970; Nakata
1986; Abe 1991b). In order to sex a specimen by its internal organs for reproduction, theventral side of the body has to be dissected, normally after it is skinned to prepare a study

18skin. Sex can be determined by verifying the presence of testes for males and the uterus
for females. Insectivores, such as shrews in a non-reproductive condition, demonstrate
fairly reduced internal reproductive organs. However, you can confidently differentiate
male from female soricomorphs by verifying the presence of testes located at each distal
end of a pair of vas deferens extending toward the right and left from the base of theurinary bladder for males and the presence of a T-shaped uterus positioned at the upperdorsal side of the urinary bladder for females (Abe 1991b). In order to sex fetuses orcarcasses of wild mammals, analytical techniques at an intracellular level (e.g.,chromosomes, DNA) are necessary (Dimmick and Pelton 1996). See Koyasu et al. (1995)
for examples of the application of such techniques in Japan.

4–4. Body Weight and External Measurements
Specimens should be weighed as promptly as possible before beginning
preparation. Weights are measured in grams for small mammals and in kilograms forlarger mammals. A scale is used for weighing small-sized animals, and a portable digitalscale (e.g., Tanita Mini Scale 1476 model) is convenient to use for animals that weigh up
to 100 grams. It is important to carry backup batteries with you whenever using a batterypowered scale in the field. The body weight of a large mammal is difficult to obtain in thefield. Nevertheless, the weight should be recorded whenever possible because of thepaucity of such field data for large animals (Nagorsen and Peterson 1980). Normally, a
portable mechanical spring scale is used for weighing large mammals in the field. When apower source is available, an electronic livestock scale is useful (e.g., Tru-Test
Distributers Limited [currently, Tru-Test Limited], New Zealand, AG and EZ models[currently, these models may be outdated and no longer available on the market.]).
Standard external measurements for small mammals are, in the North American
method (as established in Canada and the United States), Total Length (TL), Length of
Tail vertebrae (LTv), Hind Foot length (HF), and Ear length (E) (Corbet and Ovenden
1980; Nagorsen and Peterson 1980). In this system, hind foot length includes the extent
of the claw or nail without exceptions (Hfcu: Hind Foot cum unguis). In contrast, in
Europe including the United Kingdom, Head and Body length (HB) is more commonlymeasured instead of total length, and hind foot length is not supposed to include the19extent of the claw or nail (Corbet and Ovenden 1980; Nagorsen and Peterson 1980;
Pucek 1981; Imaizumi 1986). When measuring tail length, there are differences between
countries in defining its point of origin. In the United Kingdom and North America, thedistance from the root to the tip of the tail is measured (Imaizumi 1970, Corbet andOvenden 1980; Handley 1988; Burton 1991), whereas in the rest of Europe outside
United Kingdom (e.g., Poland), tail length is defined as the distance between the center of
the anus and the tip of the tail (Length of Tail anus: Lta) (Pucek 1981). In either system, atuft of hair extending beyond the end of the tail is excluded from the measurement.
Formerly in Japan, both the European method and the “amended” North American
method (tail length measured in the North American way versus hind foot length
measured in the European way) have been in use, but in more recent years, it is more
often the case where the amended North American system is introduced as a general
method (Imaizumi 1970; Abe 1991b; Abe et al. 1994; Yoneda et al. 1996). Therefore,
collectors who are planning to gather new measurement data, as well as any researchers
who intend to donate voucher specimens to a museum collection, should bear in mind
that field guides and textbooks published in Japan in recent years most likely follow theamended North American method in indicating the tail length and hind foot length
without the claw (Hind Foot sine unguis: Hfsu) as explained above. However, there arevery few countries other than Japan that have adopted the amended North American
method as a national standard for measuring mammals; thus, one should be aware that theamended North American method is not regarded as an international standard. On the one
hand, the British method is similar to the amended North American method in terms ofhind foot length as it excludes the claw and tail length with its origin taken at the root of
the tail vertebrae. On the other hand, the British method differs from the amended North
American method in the way the tail is measured without bending it toward the back(Corbet and Ovenden 1980; Burton 1991). In China, the European method is adopted as astandard for measuring mammals (Wang and Ganyun 1983). In the case of ungulates, it istechnically difficult to measure foot length exclusive of the hoof (homologous to claw),
making the North American method the only way to measure hind foot length ofungulates. That also means the North American method is the only system that enables
body measurements of terrestrial mammals, including ungulates, to be documented in a20

consistent manner. Technical instructions for measuring small mammals following theamended North American method are provided below (cf. Abe 1991b).
Total Length (TL): Completely extend the body on its back and measure the linear
distance from the tip of the snout to the end of the tail (excluding hairs at the tip of thetail). Loosen the muscles enough to stretch the carcass if it has undergone rigor mortis.
Length of Tail vertebrae (LTv): Bend the tail toward its back and hold it in an uprightposition against the body when laid out flat. Slide the edge of a ruler’s short sidebackward along the lower back of the animal until it naturally stops at the root of the tail.
Extend the tail straight along the long side of the ruler and read the scale at the terminalend of the tail (excluding hairs).
Head and Body length (HB): Take the difference of the TL value minus the LTv value
above.
Hind foot length sine unguis (Hfsu): Stretch the hind foot toes well and measure thedistance from the tip of the heel to the extremity of the longest toe (excluding the claw).
Ear length (E): Measure the length from the lowest edge of the intertragic notch, locatedbetween the tragus and the antitragus, to the furthest edge of the exterior front side of theauricle (excluding hairs).
Besides the basic measurements above, Fore Foot Length (FFL) and Fore Foot
Width (FFW) are commonly measured for soricomorphs. FFL is measured from the rearend of the forefoot sole to the tip of the longest finger excluding the claw. FFW is thelongest width across the forefoot sole. For chiropterans, Forearm length (FA), Tibia
length (Tib), Tragus length (TR), and Wing Span (WS) are usually also measured. FA is
the distance between the wrist joint and the elbow joint. Tib is measured from the knee
joint to the distal end of the tibia. TR normally refers to the tragus length from the base to
the tip along its inner edge. Please note that Handley (1988) defines two different types of21

measurements for tragus: Total Length of the Tragus and the Length of the Tragus Blade.
The former measures the tragus length along its outer edge, as opposed to the lattermeasuring the length along its inner edge. WS is the greatest width across the wings
when the bat is in a position with both wings spread out. For large-sized animals, Heightat the Shoulder (HS), chest girth, and other parts are measured. HS is measured straightfrom the upper edge of the shoulder blade to the bottom of the feet when the animal
stands on its four feet. In Japan, Yoneda et al. (1996) introduced standard externalmeasurements for cetaceans. The instructions in Yoneda et al. (1996) are almost the sameas the standard measuring system for cetaceans endorsed by the American Society of
Mammalogists. The only exception is that the body girth is measured at the umbilicus in
Yoneda et al. (1996), whereas the American standard method records body girth at its
largest. Follow Hamada (1986) for standard external measurements of the Japanesemacaque. Most of the published manuals provide illustrations and figures in conjunction
with instructions for recording external measurements of various groups of mammals(Nagorsen and Peterson 1980; Hamada 1986; Kitahara 1986; Abe 1991b; Geraci andLounsbury 1993; Abe et al. 1994; Yoneda et al. 1996; Martin et al. 2001; Asano et al.
2006), so one should refer to those diagrams and instructions when taking animal bodymeasurements until one becomes used to a set of metric data to be collected throughspecific methods of measuring each part of the body.
It should be noted that the morphometric values recorded by different
investigators could often lead to significant biases in parameters calculated from externaland skeletal measurements. Interobserver measurement error can be especially noticeableat body parts bearing significant individual variation or at those where the origin and/or
the end point of a particular metric are indistinct and difficult to define (Palmeirim 1998;
Blackwell et al. 2006). Therefore, as far as external measurements are concerned, it isrecommended that a researcher take measurements of the same part at least twice, and use
a mean value of variables for analysis (Blackwell et al. 2006).

4–5. Reproductive Condition Data
Observation of internal reproductive organs provides important biological data on
the reproductive condition of an animal. Based on such data, one can understand22

reproductive properties of a particular species in a specific region such as the yearly
number of breeding seasons, length of a breeding period, litter size, number ofparturitions per year, and age of reproductive maturity (Nagorsen and Peterson 1980).
Checking the size of the testes and visually inspecting the epididymal duct of
caudal epididymis are commonly used as a simple method of determining the
reproductive condition of males. For a fresh specimen that has not been dead long after
its collection, measure the longest and shortest diameters of a testis after opening theabdominal cavity. When it is difficult to measure the accurate dimensions of a testis,including one preserved in fluid, at least make a note on the condition of the testes as towhether they have descended to fill the scrotum, in other words, scrotal (+), or nonscrotal / abdominal (-). For small-sized mammals, the reproductive condition of males
can be determined by an inspection of the epididymal duct at the caudal epididymis: if theduct is visible, a positive indicator of sperm production and storage, the individual isreproductively active (+). Otherwise, mature sperms are not accumulating sufficiently inthe testes, and, thus, the animal was reproductively inactive (-). In order to determine thereproductive condition of a male with a high degree of certainty, it is effective to prepare
slides of a sperm smear or testis tissue for observation under the microscope. Whichever
method is adopted, it is recommended to observe and document the reproductive
condition of an animal immediately after its capture on a field catalogue or a field note.
As for the reproductive condition of females, it is obvious to tell that a pregnant or
lactating animal is in a reproductively active (+) condition for certain, but it is also a
reliable criterion to determine a female in active condition, or not by looking at thecondition of the vaginal orifice, whether it is open (+) or closed (-). Additionally, inspect
the condition of the uteri and document other reproductive criteria such as whether
fetuses are present (+) or absent (-), and if placental scars present (+) or absent (-), as wellas an external observation of the nipples, either developed (+) or not (-).
Methods of recording the reproductive condition of mammals as demonstrated
above are discussed further by Imaizumi (1970), Nagorsen and Peterson (1980), Nakata(1986), and Abe (1991b), among others.
4–6. Collection Locality Data

23

A detailed and accurate description of the collection locality of specimens is aprerequisite for all studies pertaining to an emerging field of biodiversity research, not tomention conventional study with a primary focus on geographic variation and variation
within a population of a species. For localities in Japan, it is required to record at least theprefecture, city, town or village, and a geographic feature such as a lake or a mountain.
Furthermore, exact geographic coordinates of the point of collection should be recordedin latitude and longitude at least down to the minute. Latitude and longitude can beobtained with reference to the Geospatial Information Authority of Japan (GSI; formerly
Geographical Survey Institute) standard base maps at a scale of 1:50,000 or 1:25,000.
Alternatively, use of a car navigation system, or another device equipped with aGeographic Positioning System (GPS), if available in the field, enables a researcher toread latitude and longitude of a collection site on the spot by taking advantage of the GPS
capability of such digital devices. Additionally, online topographic maps from the Map
Browsing Service (http://watchizu.gsi.go.jp) published by GSI are also of use. It is further
recommended to record a collection locality in reference to a 3rd Mesh code number
assigned to each 1km× 1 km land grid on a map of Japan developed for the National
Survey on the Natural Environment (Japanese Environment Agency 1997). For countries
outside of Japan, it is desirable to document a place name as an administrative unit aswell as its latitude and longitude according to the standard recording system in Japan as
described above. Ideally, it is best to map points of collection on a relatively large-scale
contour map (1:50,000 to 1:200,000) attached to a field catalogue or notebook.
4–7. Recording Additional Data
It is good to record habitat features at collection localities, as it provides anindication of the ecological distribution of a particular species. Useful habitat data thatshould be recorded in the field include the landscape, dominant vegetation, elevation,
condition of soil and other related information. The method of collection (e.g., trapping,
netting, salvage of carcasses) also should be recorded, but not necessarily for eachindividual specimen when all the specimens were obtained using the same method in one
field session. If that is the case, it is sufficient to summarize the specimen collection
method at the top of the first entry of a session in a field catalogue or notebook, making it

24

unnecessary to repeat the same information for the remaining data entries. Other
behavioral and ecological observations can also be recorded as supplementary data in afield catalogue or notebook. Specific examples include the time of specimen capture,unusual color phases (e.g., partial albinism, whitening on the tip of the tail), climaticconditions at the time of collection, vocal communication, and any field signs found left
behind. It may seem cumbersome to make notes of miscellaneous information, but suchdata often turns out to be an invaluable reference for natural history studies in the years tocome. Photographs of a collection site and the animals collected are particularly valuable
for extremely rare species in the wild or for an individual showing peculiar pelage
variation. Close-up photography is an effective means of documenting the facial region ofa live-captured mammal, including its facial parts that may shrink and no longer berecovered to their original forms once the animal is preserved as a specimen (Nagorsen
and Peterson 1980).
5. Preparation of Mammal Specimens
Generally, mammal specimens that are intended to be preserved in a collection
room are classified into three different categories determined by different types ofpreparation: 1) study skin and skull (which may include a partial post-cranial skeleton);2) complete skeleton including skull; 3) fluid-preserved whole body specimen. Each
preservation type presents strengths and limitations, and the choice of preparation should
be made based on a particular research purpose.
5–1. Preparation of Fluid-Preserved Specimens
This is a method to pickle and preserve a whole animal carcass in 70% ethyl
alcohol or 10% formalin solution (i.e., 4% formaldehyde aquatic solution) sealed in an
airtight container. It is necessary to take body measurements of a specimen before it isplaced in fixative because the chemical process of fixation causes hardening andshrinkage of the body. It is recommended to make a partial ventral incision in eachspecimen to facilitate infiltration of the fixative in to the body cavity (Abe 1991b).
Formalin solution is stronger than alcohol for effective specimen fixation, but is known to
break down into formic acid over time (in a matter of 1-3 years) leading to the25decalcification of teeth and bone tissue of specimens in the solution. In order to prevent
this undesirable degradation, specimens need to be transferred from the original 10%
formalin or other fixative solution after fixation, to 65–70% ethyl alcohol or 45–60%
isopropyl alcohol for permanent preservation (Nagorsen and Peterson 1980). If it isdifficult to switch the solutions, there is a way to neutralize the solution by adding
ammonia solution, marble fragments, or hexamethylenetetramine (Hayashi 1981). On theother hand, Bouin’s fixative solution requires special caution when used, because its mainchemical component, picric acid, is explosive. Specimens that are fixed and preserved
directly in ethyl alcohol solution, instead of being fixed initially with formalin, makes it
possible to subsequently obtain a tissue sample from such specimens for DNA analysis.
Whereas the method of directly preserving specimens in alcohol of exactly or almost
100% concentration is deemed optimal for preparing DNA samples, the compromise isthat it may cause shrinkage of the musculature, which results in deformation of theskeleton. For this reason, it is better to take and preserve DNA samples, such as minced
liver tissue, separately at the time of preparing fluid-preserved specimens. As for
specimen storage jars, a large, colorless, transparent glass container (a wide-mouth
container with airtight sealing like the one typically designed to brew plum wine) or asmaller glass container with a cap lined with an inner closure (usually referred to as a“mayonnaise jar,” available at scientific laboratory equipment and supply vendors) is
generally recommended. On the other hand, recycled food containers, such as emptied
powder coffee bottles, are inadequate and should never be used for storing specimens
because of their non-airtight sealing, which causes evaporation of fixative solution as
well as their metal lids, which are prone to rust. Labels associated with fluid-preserved
specimens should always be soaked in solution together with the specimens. Motomura(2009) discusses in detail the techniques of preparation, management, and photography of
fluid-preserved fish specimens. For the most part, the information therein is also
applicable to mammal specimens preserved in fluid.
5–2. Preparation of Skins
Standard techniques for preparing museum skins are discussed in the literature(e.g., Abe 1991b). Medium to large-sized mammal specimens are often preserved dry, as26

laid out hides or tanned skins for scientific use. On the other hand, for small-sized
mammals, there are three different types of preservation methods:
1) Flat skins: The skin is cut open along the ventral median line. Once removed from thebody, the inner surface of the skin is coated with a preservative that prevents hairs fromfalling off (e.g., borax powder or dried alum, or a mixture of equal parts of camphor andeither borax or alum), then the skin is stretched flat on a board, pinned, and left to dry.
2) Study skins: The skin is incised longitudinally on the posterior abdominal midline, but
not all the way anteriorly. Next, the skin is peeled off the body and cleaned by removing
fat and muscle from its interior. After a dry preservative such as those described above isapplied to the inner side, the skin is stuffed with cotton, to achieve just the right level ofhardness. The stuffed skin is then sewn up with a cotton thread to close its openings. The
skin is pulled into its natural shape, secured on a flat board for drying, and pinned at thefour feet. It is effective to replace the tail vertebrae and all four of its skeletal legs with
wire, bamboo rods, or gramineous grass rachises so as to keep the skin in shape andprevent breakage.
3) Flat skins of small mammals mounted on cardboard: The ventral side of the skin is cut
open transversely across the hind legs, and the hind leg bones are cut off without
damaging the skin. Finally, the whole skin is removed from the body. The interior of theskin is treated with a dry preservative as mentioned above. The skin is stretched over apaper body (cardboard; acid-free paper is recommended to avoid deterioration of the skinthrough contact), and the hind legs and tail are stapled onto the same cardboard withthread or staples made of stainless steel. The size of a paper body is based on the actual
body size of the animal. The cardboard also serves as a specimen label for writing fielddata and measurements.

Various manuals have been published that illustrate, in detail, step-by-step
techniques of preparing mammal skins along with diagrams (British Museum 1968;
Imaizumi 1970; Hashimoto 1979; Nagorsen and Peterson 1980; Pucek 1981; Honda

27

1985; Abe 1991b; Martin et al. 2001). One should refer to these references often until
becoming accustomed to preparing skins.

5–3. Preparation of Skulls and Skeletons
Once the skin is prepared following one of the methods illustrated in 5–2, one
should promptly prepare the skull. Cut the skull off from the skinned body at the joint of
the first vertebra (atlas), but care should be taken not to break the occipital region of theskull during the procedure. On the one hand, for relatively small mammals (soricomorphs,
murids, and bats) with fragile skulls that can easily be damaged, it is recommended to dry
the skulls without removing flesh. On the other hand, for animals larger than a squirrel, it
is better to remove the brain, eyeballs, tongue, and thicker layers of muscle from theskulls before drying. Skulls that are separated and dried in this manner during fieldwork
should be labeled immediately with the same number as that assigned to and recorded on
the specimen label attached to the matching skin before they are brought back to the lab,
along with the rest of the specimens upon completing fieldwork. If these dry skulls are
not immediately ready to be cleaned afterwards, they can be kept dry in insect-, dust-, andlight-proof containers for the time being. One of the easiest and most effective means ofcleaning a skull is to boil it in hot water to sufficiently loosen up muscle and soft tissue
attached to the skull. The softened tissue can then be removed by hand with a pair offorceps and brain tissue can be scooped out of the braincase through the foramen
magnum using various tools such as forceps, pins, and nails. If the facility is equippedwith a dermestid beetle colony, the larvae and adults can also be used to effectively toclean the skulls. When cleaning skulls of medium- and large-sized mammals, thefollowing procedure is recommended to prepare quality specimens: pretreat the skull byboiling it in hot water and removing most of the flesh, then soak the skull in an aquatic
solution of proteinase overnight to break down the soft tissue further; at last, wash theskull to clean it of soft tissue (Hachiya and Ohtaishi 1994). The enzyme brand known as
Tasinase N-11-100, which Hachiya and Ohtaishi (1994) originally recommended, is nolonger available. As a substitute, bioprase AL-15-FG (manufactured by Nagase ChemteXCo.) can be used at an optimum temperature of about 50°C, which is achieved by the useof a water heater with a thermostat. For large-sized mammals, aerobic bacteria can be28

also used instead of enzyme. Create a maceration bath heated to 37–38°C with constantaeration of small air bubbles pumped into the water. After soaking under these conditions
for a few days, the skull would be completely cleaned of soft tissue by the bacteria
proliferating in the system. The use of enzyme or aerobic bacteria requires good
ventilation as it generates a strong odor during processing. The use of caustic chemicals,
including bleach, pipe cleaning products, and strong alkaline agents (e.g., sodiumhydroxide, potassium hydroxide), should not be readily used as a way of removing flesh
from a skull. These substances are known to cause destruction and deformation of bone
as well as delamination of tooth enamel.
Whichever cleaning method is used, the defleshed skull may be bleached foraesthetic purposes. Soak small mammal skulls overnight, or for medium- to large-sized
mammals, soak for 3–5 days in approximately 10% hydrogen peroxide solution. Then,
wash the skulls thoroughly with water and allow them to air-dry. However, it isunnecessary to bleach specimens prepared specifically for research or museum
preservation. Bleaching is also usually unnecessary when the skull is cleaned withaerobic bacteria.
Preparation of the skeleton is conducted as follows: skin the specimen using oneof the methods described in 5–2, and cut open the abdomen to obtain data about its
reproductive condition (see 4–5). Next, remove all internal organs from the abdominal
and thoracic cavities (the remnant viscera can be a cause of decomposition). Dependingon the purpose of study, various samples are taken from the tissue and organs at this stage.
Subsequently, remove larger chunks of muscle from the remaining body and let theskeletal material dry. The method of preparing a skeletal specimen from the dry body isbasically the same as that used for the skull (although the larger the animal’s body mass,
the more difficult it becomes to process). Refer to Hachiya and Ohtaishi (1994) if
degreasing is needed to prepare skulls and skeletons. Ammonia water, which is safer to
use than hydrogen peroxide solution, is effective in degreasing and lightly bleachingskeletal material. However, if articulation, sutures, minute bones, and cartilage need to bepreserved intact, keep chemical treatment, like bleaching and degreasing, to the necessary
minimum.

29

6. Transport and Management of Mammal Specimens

6–1. Transportation Methods for Specimens and Data
The transportation methods demonstrated below are mainly those recommended
by Nagorsen and Peterson (1980) on the assumption that specimens and their data aregoing to be transported from a field collection site to a museum facility. Situations differ
significantly from one case to another, for example, when specimens are carried bycollectors themselves, as opposed to when they are shipped by a third party such as themailing service. There are also considerable differences between domestic and overseas
transport. Nonetheless, the following basic points apply to most of these various modes oftransportation. The methods recommended herein should also be applicable in cases
where completed specimens are moved between museums, among individual researchers,
as well as between museums and researchers.
Fluid-preserved specimens: A wet specimen properly fixed in formalin can be
kept in a moist condition during its transport by adding a very small amount of 10%
formalin solution in a tightly sealed packing container. The same approach can be taken
for specimens kept in a fixative or preservative other than formalin, but the key point is touse basically the same kind and strength of fixative or preservative as those solutions the
specimens are already fixed and preserved in. It is necessary to pack wet specimens alongwith cotton, cheesecloth, or newspaper in each container to fill the gap and prevent the
specimens from shaking, and at the same time, to keep moisture in the enclosure.
Cheesecloth is superior as filling, as it can cover the entire surface of a specimen andkeep it moistened effectively. When selecting a packing container for specimen transport,a wide-mouth plastic jar is ideal. If that is not available, a tightly sealed plastic bag is anacceptable substitute. On the other hand, a glass container is inappropriate for specimentransport. If specimens need to be transported in a glass jar, the whole container must be
packaged in a sturdy box together with enough cushioning material to fill the gap andprevent the glass from breaking. If plastic bags are used for packing, put the specimens
with cotton and fixative solution in an inner bag, and seal them tightly after removing air
from the bag. Next, seal the bag in another plastic bag, and place the double-bagged
content in a metal box (such as an emptied commercial cake box), with a matching lid

30

firmly secured in place with adhesive tape. If there is a large gap between the metal
container and the bagged content, make sure to fill it with cushioning and absorbent
materials such as cotton, cheesecloth, and used newspaper. The whole package wrappedin this manner is usually compact, and as far as it is handled with care to prevent leakageof the fixative solution, it can be shipped by regular parcel post or other delivery services
domestically. On the other hand, overseas transport requires different procedures
according to the policies and regulations of each specific country and region as well as
the present situation at each destination. For example, use of natural cotton in a package
may become a problem upon quarantine in some area, so attention should be paid tousing only synthetic cotton instead. Additionally, with regard to air parcels, the chemicalclass and quantity of solution contained in each package could be subject to strict
regulations.
Skins and skulls: It is ideal to transport skins in a container made of sturdy
plywood, but it is also acceptable to use a corrugated cardboard box for specimen
shipping purposes. First, line the bottom of a container with a sheet of cotton or similar
cushioning material as thick as about 5 centimeters deep. Evenly spread several dry study
skins above this bottom cotton layer. Line another flat layer of cotton on top of thespecimens for more skins to rest on. Repeat this layering until the container becomes full
to its top edge with the uppermost layer of cotton added to about 5 cm deep.
If skulls are completely dry and pest-free with no chance of infestation with
insects or their eggs, they can be shipped in the same container with dry skins. Otherwise,skulls must be shipped separately in a different sturdy container. If it is expected to takeseveral days or longer to transport skins, it may be necessary to treat the specimens byadding a paradichlorobenzene-based insecticide, commercially known as parazol, in thesame shipping container to repel moths and skin beetles, and prevent their eggs frombeing laid on skins. Alternatively, if ever possible, pretreat specimens to eradicate pests
by other non-chemical means such as by subjecting them to cold temperatures before
packaging them for transport. If the use of pesticides is necessary for a shipment of skins,
skulls should not be sent together with such a package because many museums use
dermestid beetles for skull and skeletal cleaning, and the presence of insecticide on theskulls may inhibit the beetles’ defleshing activity.

31

Skeletons: Skeletons must dry out prior to being enclosed in a shipment.
Skeletons of small-sized mammals can be transported together with matching skins in thesame container as long as insect repellents are not enclosed in the shipment. If that is thecase, skeletons should be packed in the bottom of a container first. Next, a layer of cottonis placed on top of the skeletons and, finally, skins should be placed evenly on the top
layer. In contrast, heavy skeletons from large-sized mammals need to be transported
separately in a sturdy wood-framed crate. If such large-sized skeletons are dehydrated, it
is recommended to wrap them with plastic bags just before transport to reduce thestrength of their odor. If skeletons of extra-large mammals are to be shipped, thereceiving museum should be notified ahead of time.
Data: Original copies of field catalogues and notebooks, as well as topographic
maps associated with specimens collected in the field should be mailed separately fromthe specimens themselves by express mail or airmail. In case they are mailed overseas
from a country that has an unreliable mailing system, the items should be shipped byregistered mail.

6–2. Regulations on Importation and Exportation
The Washington Convention, or officially the Convention on International Tradein Endangered Species of Wild Fauna and Flora (CITES), originally came into effect in1975. It has been applied to trade in Japan since November 1980. Under this treaty,
without legal permits, both imports and exports are banned for endangered and threatened
species that need special protection. This regulation applies to all internationaltransactions, and as long as a species is listed as threatened, not only a whole individual
body (whether live or dead), but also the parts thereof (including skulls, skeletons, tissue,
cells, and biochemical samples) and their artifacts (including taxidermy) are subject torestrictions. As long as either one of the countries of international trade is a CITES Party,
all wildlife trade is subject to strict monitoring as to whether the necessary permits havebeen acquired. Species under the CITES regulations in international trade are listed under
three different categories in the Appendices. Species listed in Appendix I are the mostendangered species. Species listed in Appendix II are species that are not necessarily
threatened with extinction today, but whose trade requires close monitoring in order to32

prevent them from becoming endangered. Species listed under Appendix III are species
for which either one of the CITES Parties recognizes a need for regulating trade within
their jurisdiction. Permits are required from both the importing and exporting countries
for international trade in the species listed in Appendix I. All species in Appendix I, as
well as domestic species designated as national endangered species of wild fauna andflora, including 4 species of mammals (i.e., Tsushima leopard cat Prionailurus
bengalensis euptilurus; Iriomote leopard cat Prionailurus bengalensis iriomotensis; Daito
flying fox Pteropus dasymallus daitoensis; and Amami rabbit Pentalagus furnessi[recently, Bonin flying fox Pteropus pselaphon was added]), are restricted for domestictransport (with mandatory applications and permits). Legal procedures are handled
differently on a case-by-case basis, for example, a transaction between museums versus
between a private party and a university. Therefore, it is necessary to consult with the
Wildlife Division, Nature Conservation Bureau of the Ministry of the Environment
before transporting these species. See the Environmental Agency Wildlife Conservation
Administration Study Group (1995) and other references for details. As for the species
listed in Appendix II, an export permit from the exporting country is required forshipping them overseas. As for the species listed in Appendix III, international traderequires an export permit from the exporting country only if that country lists thisparticular species in Appendix III, in addition to a certificate of origin. Any party
involved in wildlife trade must be well versed in the details of actions regulated in Japan
pertaining to CITES and the list of species in Appendices through the Wildlife
Conservation Issues Study Group (1988) and so on. Note that the CITES Conference of
the Parties holds a meeting approximately every two and a half years, in whichamendments to Appendix I and II are discussed and adopted (Kaneko 2001). Thus, it isimportant to obtain the most up-to-date information in order to ensure that the particularspecies of specimens that you intend to transport does not infringe on these regulations.
Refer to Kaneko (2001) regarding wildlife import clearance procedures following theWashington Convention, a list of CITES Management Authorities and Scientific
Authorities in Japan, as well as a list of Management Authorities of the ASEAN (=Association of Southeast Asian Nations) Parties. If there is anything unclear about theformalities concerning CITES, you should consult with the appropriate Scientific33Authorities and Management Authorities (for example, for terrestrial animals, contact the
Wildlife Division, Nature Conservation Bureau, Ministry of the Environment, and theTrade Licensing Division, Trade and Economic Cooperation Bureau, the Ministry of
Economy, Trade and Industry) (Kaneko 2001). On top of CITES, be aware of thedomestic regulations by the Wildlife Protection Law, Section 20, Paragraph 2[currently, WPML, Sections 25, 26 and 27], which control the export and import ofindividual mammals and their products (see Wildlife Protection and Management Study
Group 2001 for further discussion). Upon conducting fieldwork overseas, the local
country’s laws and regulations, as well as international treaties, must be followed inaddition to CITES (Ad hoc Committee for Animal Care Guidelines 1985).
6–3. Collection Management
The management of a systematic collection at a museum encompasses a wide
range of activities from the accessioning of specimens to the preservation treatment
necessary to prevent the deterioration of specimens in the collection (Wiley 1981).
Examples of published works in the form of field collection records, specimen catalogues,
or taxonomic reviews available in Japan are Maeda (1984, 1986), the Second Departmentof Anatomy, School of Dentistry, Aichi Gakuin University (1985, 1986), Miyazaki(1986), Shigehara (1986), Tomida and Sakura (1988, 1991), Yoshiyuki (1989), The First
Department of Anatomy, School of Medicine, Dokkyo Medical University (1992), Endo
(1996, 1997, 1998, 2000), Endo et al. (2001, 2002), Zholnerovskaya and Koyasu (1997),
and Yoshiyuki and Endo (2003). However, remarks on these publications are beyond themain scope of these guidelines. The Systematic Collections Committee of the American
Society of Mammalogists (2004) established “basic curatorial standards for systematiccollections of mammals” which are included in an appendix to these guidelines [Theappendix was deleted in this English version]. These standards, as well as Wiley (1981),
are useful references for those interested in learning more about museum collection
management.
7. Public Health34

Every individual mammal in the wild should be viewed as a potential carrier ofsome sort of zoonotic disease. Similar to those typically known as endemic, many of thediseases occur locally, so it is not realistic to make exhaustive lists of possible zoonoses
that could be transmitted during the process of collecting and preparing mammal
specimens (In Japan, a general list of zoonoses is found in Kamiyama (2004) and Kimura
and Kida (2004), whereas Yokohata (2009) covers diseases specifically transmitted via
wild mammals). Therefore, when researchers are ready to conduct field collecting, theyshould be concerned with any potential diseases that can be contracted from wildlife in
the study area, and educate themselves beforehand to learn more about them. If there is arisk of becoming infected with a disease, such as typhus or rabies from captured animals
during field activities in areas outside of Japan, it is necessary to consult a medical doctor
about options for vaccination against such diseases. Caution is advised, even in Japan, as
multiple zoonotic diseases are known to potentially infect researchers directly or
indirectly, through native animals such as echinococcosis, tsutsugamushi disease (scrub
typhus), Lyme disease, hemorrhagic fever with renal syndrome, and Japanese spotted
fever (e.g., Suzuki and Ikeda 1985; Arikawa and Hashimoto 1986; Arikawa 1996;
Takahashi 1998; Asano et al. 2006). Furthermore, in recent years these contagious
diseases, emerging infectious diseases in particular, are viewed as a threat not only tohumans and livestock but also to biodiversity (Daszak et al. 2000). It is also important for
researchers to act carefully so as to not serve as a source of infection to other people,
livestock, and wildlife as a transmitter of a pathogen.
When handling live mammals, your hands need to be protected with thick cottonor leather gloves, and you must pay attention not to be bitten. Fortunately, the chance ofrabies infection from wild animals in Japan is considered to be minimal. In contrast,
outside the country, mammals that are well known potential vectors of rabies include bats,foxes, mongooses, and skunks, just to name a few. Comparatively safer circumstances inJapan might contribute to the fact that most Japanese researchers tend to take fewer safety
precautions against the risk of rabies. Therefore, if it is necessary to catch animals forfieldwork overseas, as long as the occurrence of rabies has been reported from theparticular country or region, it is important to handle the animals carefully on thepresumption that every animal caught in the field is potentially infected with rabies. It is35most desirable to receive a rabies shot before conducting fieldwork in any country or
region with a high risk of rabies, even if vaccination is not a mandatory part of thecountry’s entry requirements. Including information on vaccination, each field researcher
must be equipped with sufficient knowledge of personal healthcare overseas prior to theirdeparture (see Miyazaki 1999). Refer to Constantine (1988) about health risks andnecessary handling measures, specifically pertaining to bat research.
When preparing mammal specimens, common sense precautions can help reduce
the risk of infection. It is recommended to wear latex gloves during dissections, and it iseven better to use a disposable paper mask to stay on the safe side. Animal excreta mustnot be touched by bare hands, as this can be a source of infection. Dissection utensils
should be disinfected using appropriate antiseptic after each use. The handling of roadkill
also requires precautions against possible infection. If you notice the following medical
symptoms subsequent to specimen collection or preparation, you must consult a medical
doctor and report a possible zoonotic infection to obtain an accurate diagnosis: a flu-like
symptom, a chronic respiratory problem, an enlarged lymph node, or signs of high fever,and vomiting or diarrhea. Incidentally, the Wild Animal Medical Center, Rakuno Gakuen
University, Hokkaido, and the Laboratory of Veterinary Pathology, Faculty of Applied
Biological Sciences, Gifu University, Gifu, serve as hubs of zoonoses research involving
wild animals in Japan. These institutions accept animal remains that are suspected ofbeing potential sources of infection.
Acknowledgements
Throughout the process of revising these procedural guidelines, valuablecomments and suggestions were received from various parties in addition to the membersof the Guidelines Revision Task Force listed in the introduction. The contributors include
the members of the Mammalogical Society of Japan Committee on Taxonomical Names
and Collections, including Dr. Hideki Endo, Committee Chair, as well as Drs. Hajime
Ishikawa and Junpei Kimura. The revised guidelines were approved by the MSJ Council
led by Dr. Sen-ichi Oda, President, and Dr. Kazuhiro Koyasu, then Secretary General.
We would like to express our profound gratitude to these dedicated parties.

36

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