Slran CnnNrvonnCoNSERVATToN The Newsletlerand Journalot the IUCN/SSC Mustelid,Viverrid & ProcyonidSpecialist Group tucN Number14 April1996 SPECTESSuRVrvl,r- CoMMrsstoN l'lrlagas) civet fossd fossono)in the RanomafanaNational Park, lladagascar- Photo: Mafio Perschke Theproduction and distribution ol thisissue has been sponsored by '*t "BlijdorpZoo", Rotterdam, Holland, ffi .r1T' "Columbuszoo", Powell, Ohio, USA r#"L' and the "RoyalZoological Society of Antwerp",Antwerp, Belgium. ffitn 6ffimE Su,q.n CnnNrvoRECoNSERVATToN TheNewsletter and Joumal of theIUCNiSSC Mustelid,,Mverrid & ProcyonidSpecialist Group Editor-in-chief: Hany VanRompaey, Edegem, Belgium Associateeditor: Huw Gri1fiths,Hull, UniredKingdom Editorial board: AngelaGlatston, Rotterdam, Netherlands MichaelRiffel, Heidelberg,Germany Arnd Schreiber,Heidelberg, Germany RolandWirth, Miinchen, Germany The !icws erpressedin rhis publicationare those ol the authorsand do not necessarilyreltect rhosc ol thc IUCN. nor the IUCN/SSC Mustelid. Vivcnid & procyonid Spe.iatist croup. We afe particulirly grrteful to Walter RrsnNslen fbr reiding rhe manuscriptsand jnrtroving the English slylc. The ain of this tuhlication is 10offcr the nrembersofrhc ITJCN/SSCMV&PSG. and thosewho are con.enredwith muslclids,viverrids..tnd pr)cyonids. briefpapcrs. news items.ibsrracrs. and titles of recentliteralure. All readersare invircd 1o send material ro: Small CarnivoreConservation c/o Dn H. Van Rompaey Jan Verbertlei.15 2650Edegem Belcium I)rirred on recyclcd papef tssN 1019-5041 Mongoosesin th e TsimbazazaZoo and the RanomafanaNational Park, Madagascar MarioPERSCHKE In Scptember 1995, two Malagas) brold sniped mon- gooses16d1tdi.1is i?r.iala) ca,ne into the boranicalzoological garden Tsimbaza4' in Antananarivo,the capilal ol Madagascar. Thc couplc wa! caught in the easternrain forest of Zahamena (Dorth-earlN{idagascar). Clearly. this is the firsr rimc rhis species ha\ bcen kcpl in a zoo (R. Wirth. pers. comm.). Ihe richly conttu\trnSlycolouredanimnlslived nrrdouble panedglasscase nr the !i\,riunr. which was providcd wirh treebranches. sleeping borc \. ltoncs, rnd a u oor covenrg of dned lervcs. l hey { ere ted wirh picccs oi beef and ffeshlr- killed onc day old ohicks (two chicks per mongoose).Thc mrle died ,rfter a shon iime. bnt rhc lcmrle $,ai obviouslyhealthy and lcouldoblerve and photograph her in Octobcr rnd h'ovembef 1995.She was quitc lii.ndly. and $a\ activc during severalpans ofthe da}. UnlbrtunrGly. I heard thrt thc itmale has also died. 'f he pictureof lhebroxd sr.ipcdmongoo!e ff onTsimbaza7a Broad-stripedmongoose. G al i rlk\ it Jasdat a. in f \imbazazaZoo. Zoo appearedin thc Gennan journal Ahtitsptut. Zn (No. 3. Phoro:Marrc Perschke. 1995).It is probably tbe first rime thar a photographof a living indi!idualol rhis{ttracti!e species has been published (R. Wirh. Free'living rirg-triled mongooscs.Cdlidia.ll]gd4Jr,l,.'td'rr. nnd Malagasycivers, Fossafussaaa. can hoth be observed\rell andeasily in theRanomxfanr Natbnal Park( south,east Madagas In October and November 1995, there w€re arso oucf car. nofth'easl of Fianarantsoa).which wrs tiNnded in 1991. \pecresoi mongooses1i!ing rr TsimbazazaZoo: Malagasyring During the day. severalring-r.rilcd mongooscsstay in the sur- Iailed mongoosc. Calidia elqans .[,ganr (l,l) canght near rcundings of a snall crmp sile. close to its entrance.With the Anjozorobe. nonh-east of Antananarivo, also Galidi! (legans bcginning ofdafkness, up to two Malagasycivcls appeaf.which ,..id.nrd1t' (l.l) caughtnear Morondava wcsLcmMadagascar. are aitricled by fricd tieces of neat. ollered by rhe M.rd.rgrscln and Malag!sy narrow,striped mongoose (M&xaorl./lr tour leiders. Thc aniDal! have lost rheir shynessoi humans.and le.etnlinedta !1e.?,ltrcal4 (2.1). also cnughrncrr Morondava. e\'en o1 hrge groups of tou.ists. and will cone \ery closc. Thesewere alsokcpt in il(Nble panedglasscases in thc vivarium. each in two outer enclosurcs(modclled concretestructures wirh Wilhr bitofluck.in theearly nrornjnghours you may even cales. Iree branches,and small island! of plarrs). lt sccms Lhrr encounLcrthefoisa. C/1?ropr../a l.&rr. along onc ol rhepath! in both spccieshave been bred atthezoo lir sclcral yeafs.Offspring the.11.600ha-l.trge rain lbrcs! rese|ve. ofthe rng-tiiled nr('ngooscwere born at the end of lisl ycar. bur were probably crtcn b\r their parents. Egon-Kisch-Strasse,55, 13059Berlin, Germany Narrow srriped mongoose.Mungotidis rl?knlineard. in TsimbaTrTr Zoo. Photo: Nlffio Perschke. Molecular techniques and small camivore conservation IsabelleSCHON and Huw l. GRIFFITHS ''unilcrsal Intmduction (properly. versatilc") primcrs". Both techniquesto gelhef enabledDNA anallsis f.om poor quality specimcn!.with Zoological nsearch hasbeen transibrmcd with the advent no pnor sequenceknowledge and. norc imponantly. no con!en "cloDing". ol ncw molecular basediechniques. pxrriculafly those incorpo rional PCR and subscquentDNA sequenceanalysis rrlin! the polymerasechain rexctn,n (PCR)(Mullis & Faloona. havealso madepossible Lhc DNA equivalenlof allozyme clc.rro -microsrlellitcs. 1987i Saiki €r a/.. 1985).Howevcr. the zoological conmuniry is phoresis.i.e. and havefacilitared the improved !er) nuch dilided hetweenthose who hrvc firsr harrdexperience use oi RIiLPs fo.large scale screeninsprogmnrmes ol rhc usc and applicationofthese techniquesand those$ho arc asafe of their signilicancc. but have a limited undcrsrandingof DNA techniques rh. ways iD which they can hc applied to conservationbnllogy. The iinr ofthc prcscnta(icle is toprovide a basic levelreview on I)NA extraction hbora(ory molecular techniques, lo exphin the renninolog)" Thc majoritv of DNA lechniqucs rcqui.e a very small uscd. rnd 1o provide examples i11uslrltingtheir application lo lissuc/DNA samplethat neednot ncccssarilybe of good qualiiy. cdrse.\alion troblenrs. The review is di!ided into three parts: Nonerhelcss.stcps shouldbe rakento prevenrsrmple deSradarion pro!iding r hislorical background,erphining lone of rhe key asnuchaspossible,e.r. storingin95E eftanol o.a 20'Cfi€ezer. D\A lechniquesrsed (chronok)gically.by order ol lirst wide A range of nonlelhal sanpling .egilne! are possibleand should spruadusc by /oologists). .1ndsome cxrnrple\ ofrheir applicarion. ai$ay\ be coDlidered. Ear punchcs or hlood lamples nlay be collected (e.g. trom thc lails of voles).but sioce lnamnrdran red The basic problen lrcnrg non molecular biologisrs is blood cells havc no nu.leus. the volume ofsanple rnust he larger undchtanding the simple principlcs behind DNA-based rcchni thar a corrcspondingsrmple taken fion other \c.rcbmte grcups !ucs. coutled sith diiticuhies in knowing the technical jargon (bird!.Anphibi). elc.).WithDNA rechniquesit maynol evcnbe comnr()rly cmtloyed. To nnny the brsic teflninotogy used in necessaryto capturefic rnimal. Hairs (from "sticky trpc" trNl't 'noleculafwork ls complelcllrincomprehensible, and prolides an or elen lrc.al remains(Tabe.let & Bonver. 1992) conlain suffi- in\uflnounLablebaffier to apprecirtun ol findings presentedin cieDtDNA for PCR b.rse(ltechniques. Finally, it is worth noting researchreports.joum!1 pa['ers. and evenresearch proposals. we rhar hrge numbers of specimensltrc alailNble in museumsand hefe a\sume rhe najoriry of reade.sare falniliar wirh the brlic othcr archilal resourcesand rhcscmav also be usedas sourcesol {trr.rL're ofdeoryribonucleic rcid (DNA). Thosewho rre not, we DNA. Howevef. nruseumsreccive frcqucnt requestslbr tissue r.lcr to dre various s(Lrdentlcxtbooks availdle \hich olfcr sp€cinensrnd will often .equire a researchproposal bcfore they d.rrrl.d dcscnftrcns(e.g. Aiberrs ?t d1..1989:Wa$on ?/ d1.. are willing to releascpafticularly importaol specimens.Spccifi. t9N-rr. technicalproblems are associated wrth so-crllcd NncientDNA". patrcularly when lhe samtle is fonnaldehydepresened, so thrr Historicalbackground specializedcxtrrction protocols are required. Prior ro thc 198(is,the most sophisricrlcdtechnique avail- Tissue! selectedas DNA sourcc malcrial areprocessed to rble Io zoologists lbr gcnctic analyseswas allozlmc clecrro yield relalivelv purc DNA. Extracrionmust be undeiakcn with a phoresis.This operateson ihe principlc thrt vllriantsofthe sane certainamount ofcare, primNrily ro prevenrcross,conmminition protcin (albzynes) nigrate al dilicrent ratesacross a srarchgcl when a currenl is ,rpplied. Although allozymc elecrrophoresis prcvides a certain anxlunl of infornation at the alpha taxonomic scale(especially a1 spccies levelJ, ilis of lessuse li'r intrapopulation studiesbecause ofthc often limiteddegreeol vrridion (polymor- I'hi.n'' in!ol!ed. A mur{,,.nerJllim,rJlron r. rhe n{r.*ir} r,,u.e iiesh o. frozen material. With DNA brsed moleculartechniqucs thc major advantagesare thal much more variatbn mry be available,the sampleneed notbe fresh and rharonly very limited quantitiesof DNA are rcquired. _,,-rA. Hnu<rer.the lir{ D\A hJ,edrechnrque' $er. n.,rqu c -' flcxible. Reitriction tragncnl lcngth polytnorphisms"(RFLP' "DNA .:: and fingcrprintinS were among thc first available (all t* - :> - "?<1 theseterms willbe fully describedlater). Bolh sufferedto a degree *<.-_-----------------: in rhattheyrequired fairlycomplcx protocolsand the inlbrmation ._/4F|y."* rhar rhey provided was ofien dilficult to inrerpret.Since the lale 1980slwo key developrnentshave enabled researchers other ihan sI'ecializedmoleculff bnllogiststo useDNA techniques.The firsr was the advent of previously mentioned 'polymerase chain Fig. I Singlestepsofthe"polymerasechainreaclion"(PCR)(after