TThhee ZZoooloolgiocagliSoccaielty Society oofJfap anJapan ZOOLOGICAL SCIENCE 17: 259-263 (2000} @ 2000 Zoologica lSociety of Japan ANewCalcarobiotus (TardigraMdaac:robiotidaef)rom the lmperial Palace ot Japan Wataru Abei'2 'andMasatsune Takeda2 `Deparment of Biologic aSlciences ,Graduat Sechool of Science ,the Universi tofy foicsto, 2Department ofZbotogy} Nationat Science Museum, 3-23-f i-lyakunincho, Shinjuku-kuT,bkyo, 169-O073 .Lapan ABSTRACT-A new semiterrestrial tardigrade species of the genus Caicarobio tius dsescribe dfrom the Imperia Plalace of Japan as the thir dspecies of the genus which has been previousl kynown only from South Afric aT.he new species, Caicarobiotu ismperiali iss, closely related to C. occultus Dastych ,but can be distinguish ferdom the latt emrainly by the absence of a spine on the tip of each egg process ,and by having large rpores on egg surface, a narrower bucca ltube, and large crlaws. restrial tardigrades. Each specimen was mounted on a glass slide in INTRODUCTION asufficientamountofHoyer'smedium;polyvinyl-lactophenolwasused Since 1996, the Nationa lScience Museum, Tokyo, has aMsu ra rsuapyplie mtaetnettearry the m omuonutnitnigng m emdieadiu md;ri ecodv.er slips were sealed with been conducting biologic ailnvestigati oonf sthe lmperia lPal- The specimens were closely examined using a phase contrast ace of Japan ,which is centrally locate din Metropolitan and Nomarski different iianlterferen cmeicroscope (Zeis Asxiophot) Tokyo, l toccupies a vast area with various environments, not and illustrate dwith the aid of camera lucida ;specimens were mea- osntlryea mwse,ll -akenpdt o racrhtairfdisc,ia laTnhdi s npartuorjaelc htfaosr ebsete snb upt laalsnon ep duonnddesr, scouef rntethdaeg eubsu icrncagatl iao nbtu ebeteyew p(eiPeec nitehl ema il1cte9rono8mg1e,tt)e h,r o,wfa asny Ta lhgseiov pecatn lsictunrluadctteeud rx,ew,h iacnhd itsh teh 1e epnegrt-h the auspices of His Majesty the Emperor of Japan to obtain ln order to get a clear image of the egg shell, a computer-en- up-to-date knowledge of the fauna and flora ,and aiso hancing technique was utilized. A series of egg shell images at differ- to monitor thei rchanges in the lmperial Palace, which is sur- ent foca lpoint swas fir scatptured on a UN1X Workstatio nwith a CCD rreosuunldtsed wi lbl yb ep apvuebdl irsoahdes di n atnhdis ycoenacrre ,t2e00 0b,uilding $T.he first cwAeadrmeoebr eat hP cehono ntpnoerscethpeoadpr,e adtn od ona pZare iiMnsast c Aeoixudntito uposihs noghcto m mpaiunctre orEscpospe o.unsi nign Tk hjtee hstpe ers ioifnmttawegareres. During the investigation w,e collected many samples of As comparative materials, the holotypes and paratype sot mosses, lichen sa,lgae, and lea flitt efrrom various habitats to Catcatobiotu sfilme rDiastych ,1993 and C. occultus Dastych ,1993 obtain the tardigrades, Among the ta rdigrades extracted from depesited in the Natal Museum, Pietermaritzbur Sgo,uth Afric awere m(ofssaemsi M laweycr erob isootmiedae )s,p ewchiimcehns was i rne ctehnet lgye enruecst eCda lbcya Dtaosbtiyocthus e Nxaa mti inTeodhn.ea lStcypiee snpce ecMiumseensum, o fT otkhye on e(wN SspMecTie)s, are deposited in the (199 3b)a$ed on two new species from South Afric aA.fter close comparison with the type material of the two known spe- cies, the specimens from the lmperia lPalace of Japan were DESCRIPTION determine dto be a new species and are describe dhere, Genus Calcarobio tDuasstych ,1 993 Caicarobiotu ismperiatis ,new species MATERIALS AND METHODS [Japane snaeme: Mikado-ch6meimushi, new] (Fig s1, ,2, Table 1) Moss samples collected from artificially-mounded soil were swoearkeed pi icnk wead teurp uinnd aer P eat drii dsissehc atnidn g mliecrf otosvceronpieght .at tLhie vhiin ggthaersd itmgargandi-es 7ype series .HOLOTYPE: adult animal <NSMT Tg-45), ficati o<nX50 a>nd then placed in a small amount of water on a glass sex undet.; side of Hasuikebori Pond near the lmperia Hlouse- slide and gradual lhyeate d(abo u5t sec.) with a lighte frrom under- hold ,the lmperia lPalace ,Tokyo ,central Japan (35040's2''N, side.AIthoughthisheat-killingtechniquehasbeencommonlyusedin 139o45i30''E>; ca. 10 m altitude; extracted from a moss, the fixati oonf tree-livi onrg plant-parasitiz nienmgatodes to avoid the Plagiomniumacutum{Lindb.)Kop,,onartificially-moundedsoil; subsequent shrinkage ot specimens, i tis also effective with semiter- W. Abe; 14 Dec. 1 998. PARATYPES: one adult animal (NSMT ' Correspondi naugthor:Tel,FAX . ++8811--33--33336644--77113034;. T(gN-S46M>T ,Tsegx- 4u7nd, e4t8.,) ,soanmee e mdbarytoan aatse ho leoggty p(e Nt;wSoM aT dTulgt- 4an9i)m ,aalnsd E-mail:[email protected] one egg (NSMT Tg-50) ,same locali at$y above, 12 Mar. 1 999, NNIII-IE-leEcltreoncitcronic Library Service TThhee ZZoooloolgiocagliSoccaielty Society ooff JJaapapnan 260 W. Abe and M.Takeda tpt-Ay ,,-,,Ky, v c D B NZ x / F A 50Fm20Fm B ' ce,s//itg/'tei,tc"D10"rn C, D 10pam G GE,F H 5pam3"m H Fig. 1. Caicarobio tiumsperiabs n,ew species.A-D, F, Holotype (NSMTTg-45 E), ;Paratype (NSMT Tg-46); G, H, Paratype (NSMTTg-50 ()A,} Habitus ,ventral vievv. (B )Buccopharyngeal apparatus, ventral view (phary anrxtificial lsyhifted posteriorl y().C }Buccal armature, ventral view. (D )Buccal armature, dorsa lview, (E )Ciaws oi leg 11 .(F )Claws of leg IV, (G )Egg proces sl,ater avliew. (H )Egg surface, dorsal view, NSMT Tg-48 is mounted in polyvinyl-lactophe notohler,s in median rounded granules ,Buccal tube rigid, 32.4 pm long, Hoyer's medium. 5.1 pm in external diamete rD,istanc teo style tsupport inser- Holotype .Body elongate, length 353 pm excluding leg tion from posterio ernd of buccal cavity 26.0 pm, 80,3% of the IV ,width 8t pm; body width:body length 1,:4.3 .Body whitish length of the bucca ltube; turca developed normally, similar to transluce nitn lif ewi,thout pigmentation. Dorsa lcuticle smooth, that of Atlacrobiotu sspecies in size and shape. without pores. Eyespots absent, Pharynx elongate spherical, 34,3 pm long, 22.6 pm wide; Buccopharyngeal apparatus moderate in size, Mouth pharynx length:widt h1,:O.66. Apophysis in pharyn xwell- opening 9.8 ym in diameter ,situated terminall syu,rrounded developed; macroplacoid 1 as long as macroplacoid 111; by 10 perjbucca llametlae ,Circum-or aselnsory fiel dweli- macroplacoid 1 1shortest; macroplacoid 11 1with con$picuou$ developed .Anterior band ot buccal armature consists of incisi opnosterior lmyac;roplacoids 1-11 13.9, 3.1, and 3.9 pm minuscule and densely-distribu tgerdanules ;posterior band long ,respectively; microplacoid well-developed, 3.8 um long. consists of closely spaced teeth; each tooth rounded on the Legs normally developed ;cuticl eof leg IV covered with dorsa lside and elongated on the ventral ;dorsa lcrest well- denseiy-distri gbruatneudle sthat are fin eand barel ydiscern- marked, consists of three transverse bars ;ventral cre$t con- ible ;other legs without perceptib fgeranules .Cuticula rring sists of transverse structures, two latera lbar$ and two (="cuti cbuarl" ain rDastych ,1 993) in legs 1-1 1weIll-developed; NNIII-IE-leEcltreoncitcronic Library Service TThhee ZZoooloolgiocaglical SSoociceityety ooff JJaapapnan New Tardigradafromthe 1mpenalPalace 261 Fig, 2. Caicarobtotu simpenalis n,ew species, A, Holotype {NSMT Tg-45); B, C, Paratype (NSMT Tg-50). (A) Anterior body. (B )Egg sculpture (computer-enha n(cCe )Dde)ta.i lof egg shell surface, Scalesi A, B=20 um; C=5 vm. cuticular ring not restricted to ventral side of leg, but contin- 3.2 pm high, ca. 3,5 "m in diameter at base, and ringed con- ued to its dorsal $ide, being more clearly observed on ventral centricallywith three well-marked cuticular ridges; ti pof each side compared with dersa lside; on dorsa lside of ring, thin process smooth, without any structure; ca. 36 processes on cuticular sheet, attached to primar ybranch of each claw, forms circumference of egg and ca. 1 70 processe pser hemisphere. di$tin cactcessory point l;eg IV without cuticula rring, but with Surfacecoveredwithanetofirregularlyshapedporesthrough- cuticular sheet and accessory point, out; pores O,3-O.8 pm in diameter, Each claw with distin cstu,btriangular base that is armed Remarks. 1n the class Eutardigrad aes,pecially in the fam- with a pair of laterally-protr uspduersd. Intern aalnd external ily Macrebiotidae, the morphology of the egg shell is consid- claws similar to each other in shape, but external claw clearly ered to be an importan ttaxonomic character (Berto l&ani iarge trhan intern acllaw on Ieg$ l-11 c1l;aw IV large trhan claws Rebecchi ,1993) .Caicatobiotu ismperial iiss readily distin- 1-11 1i;ntern acllaws 1-11 I8,.2-9.0 pm long (inclu daiccnesg- guished from C. fitrr ibeyn the shape ot egg processes ,which se ry poin tand lunula )ex,ternal claws 1-l1 11O,,5-1 1 ,7 pm long, are conical with strongly elongated and sometimes bi-trifur- anterior and posterio rc[aws IV ,11,7 and 12.1 pm long, cate tip sin the latte srpecies. Calcarobio tiumpseriad sis very respectively. Each claw provide dwith weH-developed lunula; similar morphologically to C. occuitus, both in the adult ani- width of Iunula 2.7-3.1 pm on claws 1-11 15,.9 pm on claw IV; mal and egg, but can be distinguish efdrom the latte bry the edge of lunula smooth on claws 1-Il ld,entate on claw IV. followi n1g) the spiniform structure at the tip ef each process Cloaca situated near base of leg IV ,similar to that of on the egg shell is always absent, 2) the cuticula rridges on Macrobiotu sspecies in shape and size. each proces osf the egg $hel1 are well-marked, 3) the pore s[n Paraope animals. Body 258-314 pm long; eyespot$ the network on the egg shell surface are larger in size and always absent. Emb ryo with posteri ionrcisi oant macroplacoid irregular in shape, 4) the bucca ltube is narrower (pt=14,1- llL 15,9), and 5) the claws are larger relative to the length of the Paratype eggs. Whitish ,sphencal, 69 and 71 pm in bucca ltube, diameterexcludingprocesses;surfacecoveredwithsmallpro- cesses which are bell-shape du,niform in size and shape, ca. 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Our report from Japan suggests tha tCalcarobiot uiss not an endemic genus to South Afric ab,ut a taxon with a broad dis- REFERENCES tributi oWne, collected C. imperiat ifsrom a moss growing di- rectly on soil. Dastych ( 1993) also collected the two species Bertolan iR, Bisero Vvl (199 6Le)g and claw adaptations in soi ltardi- mainly from soil or moss growing on soil, Furthermore, grades, with erection ot two new genera of Eutardigrada, Cafailelcda otfo tbhieo Ntau tssipo. nwaal sln csotlilteuct otefe dA rgerceon-tElynv firroomn am seonitl saSalcmipleenc eisn, Be r tM1oa1lc5arIn oR2ib,9 i9Ro-et3bi0ed4cace h:Pi sLe (uc1 9l9o3h) eAx arepvoisdiionbi ou fas ntdhe X Mearcorboiboitouts u,1s hnuvieer ltBainodli Tsukuba, central Japan (M .Ite ,pers .comm,), These col]ec- group (Tardigra dMaa,crobiotida weit)h, some observations on tions suggest that Calcarobiotu sspecies prefe r{altho unogth the taxonomic characters of eutardigrades. Zool Scr 22: 127- exclusively) to liv ein soil or mos$ growing on soil. As men- 152Dastych tione dby Bertolani and Biserov ( 1996), the soil environment tri aHwla t(e1r-9b9e3a r)As n ferwo mg Seonuuts ha Anfdr ifco au(r Tneawr dspiecgiersa M doifat ts}Hea.mmibter Zreoso-l has been considered a$ a habitat of tardigrades rather Mus lns t90: 175-186 recently, and thus the geographica dlistribut iofo nthe species PilatoG(1981)Analisidinuovicaratterinellostudiodeglieutardigradi. livin ign soil is poorl yunderstood. I tis highl yprobable that Animalia 8: 51 -57 many additional localitie wsill be found by furthe irnvestiga- (ReceivJeudly26.1999fAcceptedSeptember14,1999) tions of soil environments. NNIII-IE-leEcltreoncitcronic Library Service