PAPERS & ARTICLES

TIMONEY, J. F., SHEORAN, A. & ARTIUSHIN, S. (1998) Detection of strani-gles carriers. Veteritnary Record 142, 648

TODD, T. G. (1910) Strangles. JournalolJComparative Pathological Tlherapeuitic-23, 212-229

TODHUNTER, R. J., BROWN, C. M. & STICKLE, R. (1985) Retropharyngealinfections in five horses. Journal of the American Veterinary Medical

Associationi 187, 600-604WILSON, D. A. & (CONSTANTINESCU, G,. M. (1999) Lymph nodes and

lymphatics. In Equine Surgery. Eds J. A. Auer, J. A. Stick. Philadelphia, W. B.Sauniders. pp 399-403

WOOD, J. L. N., DUNN, K. & CHANTER, N. (1993) Persistent infection withStreptococctus equi and the epidemiology of strangles. Veteritiary Recorcd 133,365

Pathology of sporotrichosis in 10 cats in

Rio de Janeiro

T. M. PACHECO SCHUBACH, A. DE OLIVEIRA SCHUBACH, T. CUZZI-MAYA, T. OKAMOTO,R. SANTOS REIS, P. C. FIALHO MONTEIRO, M. C. GUTIERREZ-GALHARDO, B. WANKE

Ten cats with sporotrichosis were examined clinically and pathologically. They were in very poor generalcondition, and had widespread ulcerated cutaneous lesions and respiratory signs. Gross internalabnormalities were found only in the lungs and lymph nodes. Histologically, an inflammatory infiltrate andyeast-like structures were observed in the skin, lungs, liver and lymph nodes. The spleen was congested andcontained fungal elements. No microscopical changes were observed in the pancreas, kidneys and heart.Sporothrix schenckii was isolated from all the skin samples and nasal swabs obtained in vivo, and from allthe samples of lung, liver, spleen, lymph nodes, heart and kidney taken postmortem.

SPOROTRICHOSIS is caused by the dimorphic fungusSporothrix schenckii which infects human beings and manyanimal species. Classically, the infection occurs by cutaneousinoculation and is limited to the skin and subcutaneous tis-sue. Occasionally, the inhalation of conidia may result in pul-monary sporotrichosis, but the organism rarely spreads toother organs (Rippon 1988a, Kauffman 1999).

Although the infection was induced experimentally in catsby De Beurmann and others (1909) and naturally acquiredfeline sporotrichosis was first described in 1952 (Singer andMuncie 1952), the disease is considered to be rare and is littleknown (Davies and Troy 1996). During the 12 years from 1987to 1998, only 13 cases of human sporotrichosis were recordedat the Evandro Chagas Clinical Research Institute in Rio deJaneiro; two of these patients said that they had been scratchedby a sick cat. However, during the two years from July 1998 toJuly 2000, cases of sporotrichosis were diagnosed in 66 people,117 cats and seven dogs. Fifty-two of the people reported hav-ing been in contact with an infected cat, and 31 ofthem reportedhaving been scratched or bitten (Barros and others 2001).

The description of feline sporotrichosis has been based onreports of isolated cases, usually with multiple skin ulcers,sometimes accompanied by clinical signs of systemic involve-ment (Freitas and others 1965, Rosser and Dunstan 1998).Owing to the difficulty in evaluating the involvement of inter-nal organs in vivo, a diagnosis of disseminated sporotrichosisis almost always made postmortem, either by the observationof the fungus in tissues or by its isolation in culture(Schamroth and others 1988). This paper describes the clin-ical and laboratory findings observed in 10 cats with wide-spread cutaneous lesions of naturally acquired sporotrichosis.

MATERIALS AND METHODS

Seven male and three female cats of mixed breed, rangingin age from six months to three years (median one year)

and from eight different households in Rio de Janeiro,underwent a general clinical and dermatological examina-tion. Blood samples were collected by venepuncture forroutine biochemical examination, the determination ofpacked-cell volume, white blood cell and differential bloodcell counts, and assays for feline immunodeficiency virus(FIv)-specific antibodies and feline leukaemia virus (FeLV)antigen (IDEXX) were carried out, following the manufac-turer's instructions.

Punch biopsy specimens of skin were obtained from theborders of active lesions after local anaesthesia with 2 per centlignocaine. Each specimen was divided into two portions: onewas fixed in 10 per cent buffered formalin, embedded inparaffin, and stained with haematoxylin and eosin, Gomori'smethenamine silver and periodic acid-Schiff for histopatho-logical examination; the second was kept in sterile saline, trit-urated and then seeded on to Sabouraud's dextrose agar andmycobiotic agar (Difco), incubated at 25°C, and observed forfour weeks for fungal growth. Suspected isolates were sub-cultivated on potato dextrose agar medium (Difco) at 25°Cfor macroscopic and microscopical studies, and dimorphismwas demonstrated by conversion to the yeast-like form onbrain heart infusion agar medium (Difco) at 37°C (Rippon1988a, Werner and Werner 1994). Swabs of the surface secre-tion of cutaneous ulcers and swabs of the nasal cavities werecollected and subjected to mycological examination asdescribed above.

The animals were treated orally once a day with 5 to 10mg/kg itraconazole.

Four of the cats died and were brought by their owners tothe Zoonosis Service, following biosafety regulations(Cardoso 1998). The other cats were euthanased with anintravenous overdose of pentobarbitone after being tran-quilised with a combination of ketamine and acepromazine.

The organs were examined macroscopically and samplesof lungs, liver, spleen, lymph nodes, heart and other organswere removed. Each specimen was divided into two portions:

Veteritnary Record (2003)152, 172- 175

T. M. Pacheco Schubach,VMD,T. Okamoto,Serviio de Zoonoses,A. de Oliveira Schubach,MD, PhD,M. C. Gutierrez-Galhardo, MD, PhD,ServiSo de DermatologicaInfecciosa,T. Cuzzi-Maya, MD, PhD,ServiSo de AnatomiaPatologica,R. Santos Reis,P. C. Fialho Monteiro,MD,B. Wanke, MD, PhD,ServiSo de Micologia,Instituto de PesquisaClinica Evandro Chagas-Fiocruz, Av Brasil 4365,21045-900 Rio de Janeiro,Ri, Brasil

The Veterinary Record, February 8, 2003172

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FIG 1: Cat in poorgeneral condition withwidespread ulceratedcutaneous lesionscaused by Sporothrixschenckii

FIG 2: Photomicrograph of skin showing numerous yeast-likeforms in the dermis. Gomori's methenamine silver. x 10

one was fixed in 10 per cent buffered formalin and embeddedin paraffin for histopathological examiniationi; the other waskept in sterile saline and subjected to mycologicalexamination as described above.

RESULTS

The cats were all in very poor general condition, wvith wide-spread ulcerated cutaneous lesions (Fig 1). At least one of thefollowing respiratory signs was observed in all of them: mod-erate inspiratory dyspnoea with stertorous breathing in nine,sneezing in five accompanied in four by a moderate, inter-mittent, bilateral, mucous niasal discharge secretion, and anoni-productive cough in two. Generalised lymphadeno-megaly was detected in three animals. None of them had ahigh rectal temperature and none had hepatosplenomegaly.

Histopathology of eight skin biopsies revealed a widelydistributed dermal inflammatory infiltrate consisting ofmononuclear and polymorphonuclear cells, predominantlymacrophages and neutrophils. Numllerous intracellular cigar-shaped, round or oval budding cells, sometimes exhibitingelongated buds measuring Up to 5 or 7 pm in diameter, sug-gestive of S sclhctickii, were observed in all the biopsies (Fig 2).

S scluickii was isolated from skin samples from all of the anIi-mals and from the nasal swabs taken from the five cats whichwere sneezing.

Aniaemiia, lcucocytosis with nCutrophilia, hypoalbumin-aemi.ia and hyperglobulinaemiiia were frequent abnormiialities.No chainges in liver en7zyme activities or urea concenitrationin plassma were observed.

Of the 10 cats, four were euthanased when the conditioniwas diagnosed, and the other six were examinied after anti-fuLntgal treatment; of these, four died three to 35 davs afterstarting the treatment, and in the other two the disease con-tinued to progress and they were euthanased after sevein and12 months of treatment.

Antibodies to iiv were detected in one of six ainimilalstested. This cat initially had an ulcerated lesion on its face thatregressed after two months of treatment with itraconazole.However, over the subsequent 10 moniths, despite the treat-ment being maintainied, the disease slowly progressed, newlesions developed on its face and limbs, its respiratory sys-tem became involved and its general condition deteriorated.

The condition of another cat, whiclh initially had anenlarged bridge of the nose and suffered from dyspnoiea and

FIG 3: (a) Macroscopicappearance of the lungsshowing small whitishdots on the pleuralsurface and on theparenchyma.(b) Macroscopic view ofthe enlargement of theretropharyngeal andmandibular lymphnodes.(c) Photomicrograph oflung tissue showing amixed infiltrate ofmononuclear andpolymorphonuclearcells in the alveolarwalls and some yeast-like organisms (arrows).Periodic acid-Schiff.x 40.(d) Photomicrograph ofa lymph node showingyeast-like forms, mainlyin groups. Periodic acid-Schiff. x 40

The Veterinary Record, February 8, 2003 173

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I \u)1 I,( \, \t I,it-I SI1-

Number of positive samples/number of samplesisolation in culture Histopathological examinationTissue

Lung 10/10 6/9Liver 10/10 5/9Spleen 9/9 2/10Pancreas 3/4 0/2Lymph node 8/8 6/10Heart /05Kidney 6/6 0/5

Stiee/tflg, XV1ssosend despite the cotitiltinoLS LlsC of itiracofla/oIC tot- sexemiionths.

Postmortem examination findings\LaCt oscop1IcAlls thelongII(Ss etC- 11o1t11llx act atecd hut hiadsmall wshitish clots tileac,ItIII(~It pprosamllatelvx 11tIIIi (Indiaetet onI thelenaIlF1 SotI fa-Ce and In the patrenchyma, simiilar- tothose Oh)sCetCCs Iedin 1milVr tuthetCn0ioSs (jf 3 a ). Sotme lymphHodeis wsete CHlat-ed ( [1 t 3b) TAp-ltI f-11tom rh cLotanCotSlCesion acltedv describted, thetre ssetc no e,ioss chanlces to othet

Histoloc,icalls, a tiit\dc intilt -ate Of m11n10noLCleat and pols11OtjphonneIClea Cells WSsasOhSetVCeI In theC IlungS, (IOCalsed InIthe alseolatil ssalls FiB, )c) Somec alveoli ssere tilled w5ith atii11orphIOL I)Sinophi[)llC'licmtet ial CoMpaItihie ssitli OedemIa tin,idaix eolat ocedema ).HistoloBicall sectionis of lixvet showsed amIld tot tmodcrate nflatii mto)t 'i-nfilttrate locatedi mainixvi

the portal splaces, an1d Inl somei cases aming the hecpatocs teCot cl,s. heC tiSSneC Of theC SPleen sxas congested. 'I'het- e T~S asmlatrked i-nfiltrtaont ott the parenchs mna itt the ls mph nodes hxma1"crophages conitatmng'l( nnmerICIonS paraite; atithet pattetrnof inflammlatioin ohsersved conisisted ott liv pet plasia of the

lx mlphoid tollicles wxith plasmla cells; inl this case, theti- e vxevlesserl patrasites wshich xxetre located miaiinlx inl the snhcapsularat ea (FI(ig ). Iti Othet- Oittgans, SL Cli as the pancrecas, kidtiexsandic heart , thercssciev- rio stgtiificat-it mictroscopical chlatigs.

lAlICe SliOWS theC tinmhibC sOf CaItS t1mm01sWiiclI S sicIcnkiixxVas isolated bix cnltnt eC ft otii ditteret-cit tiSSLIeS adicld the nntii11her s Iti xwhlicli flies xxere observed liistopatliologicallx.

DISCUSSION

FCeline sp(itotriclinsis has heeti cotisiicierd a tare disease, iitilx\occastotia(lls ieCiro' ttr(itsmiittedl to 1LIniii lieitigS (Dtavies atid'I'orNt 1996I lowchisx et, atittic eIaSitlig noILIetutI0CaiSeS itt telllti,caiIiiie anid 1lin1iiaii sporott icliosis hasve heeti occut ingII iiihoLISeholds Ini the c11 ityf lizii dce Jateirio aiic lCnatlixV tIIiCinncpalities. Thils epidclitic max hasve stat ted itisidlincislx hefotre1998 and its ot oiii atidc Canses aire uiiexplaiiiedl (Batini-s anid

Iti theC CLaSSHiCatiOti Ott hntIiialspoSOt-t lICIOSIS, theC disserniiiiatecl CLtaie1COIS t(0irtii is chatracterisecd hx tte cdissemiiinatiOtiitt theC tougHILS thi-on gli the lilood stiteaiii; 'It CauISeS lesionis allitsCer theC 1itegmeLilt, anid is atssic"tcitdi xxitli lesicins oif otliet-i)t ganl 5 O sxvstetiis, espiecially the skeletoni (Samnpaio aticl cotli-et-s 1 9 4 ). It has heeti litoposecd that there (iretwxxi-ititius ofclissetiiitiiatecd sporotr icliosis: the dlisseciniiated CnItan-eOitsfoi)tiii, etitirelx conintteci to the skini, anid a disseciiitiiated svstetiiilc tot iii ( Lynticland otliers 1 971). It hias ailsot heeti estah-lIshied that disseminailted spot citi iclliisis lesi(itis catiiciot liedetionistratecd cionclusivNelwsitliiout tiiici oscopic CxatIiiinationls ot tissties t(akeii eitlic ix Iibiopsx ori piistmiiortemi. A.seSuLtt, tie SubdixVisiOtII iiito dlissemiiiiatecd cLitatiCoLs atidc

dissemuiiatied systemiic sproritrichiisis mu1Lst he Sei-SinnslqjnCStIiitid Schamrtihtl anid ittletrs 1 988). Fotlloxwinig the rec-

FIG 4: Chest radiographof one of the cats withsporotrichosis showingan interstitial infiltrateat the base of the rightlung

itti11i-IiitilatiOlis tot- theC 110111C11clatni C itt (UltigI'l diseaSeS lixthe Intrite atiitnal Si),tcfClxiiIltiaiail\iii(lNxclga1 dissctiiitated iiixcisis is cicffiicci (a5 (1 uycosis sprcacd x ahlIioc lto itIxVitVxC tVOit 1ii 0otC io0u COtiltg dLitLts deep soilidiir atis,ta is, (1 pat ticli snit pC itt a sx stetiiic uS cit)Sst(Odds and others 1992)

I,ittIe is ktnxsnai(lit theC p)atlOigeneSIS itt Spii_ttiticlSitsiIiIICats DUnnStan (mnd ittliet 986), (itdl It Is pitssibile that it IsdilffeeitCltt om-11 tliila ttleIIaOt1i ii(l fS CCIS, 'at1ic teCj_ni tCs hOt ,cdetailecd Stndxy. Pc(istutti ( 1993 StndIciec a C(it xsTitli loc-lI'sedCLitaliCOIStuts lsciis oiii its face tici (1 driattitig nis(il IcsiOitt.BttLike atic otliet s (1982i aitid (Giti/alc/ ( (bli atic it)thies1 989 Cacli (clscribihd a cat ssitli mnIltpleI Cctlt(tieitCLS ICSIOitis

bimt iii ClinCal sigtls Ot' SxVStCtIi1C itixOitlx e Cilnt. III (ill itt theSetlii-cc cases, die animals rcspittdecd lto thietr(px.

InI the pt CSCtit Studx, the cdisscm invatcd tot-iwnxx(s icilminstr(tIed lixv the detecti(in itt S Sclhinckii ini the skin in xvixoiintdInI muiltiple OitrgaiiS piStt01tiin tti. Spot (idiC (iid ,1SnStisxsteaticrecpor-ts itt postmit rtciil Cxaminia(tionis pet tot tiicd oti tl(itLt alkiittCctcd ((its liaxve meCItiiotied theC dCtCCtioti Of theC fLinguLS lixvIiistitlitical tiiectliocls aiid/oi- lix CLutlttr InI theC hu,tigS, IlcartI,splceti, kidhievs, lxymph tiodces, lirain, adicrn il glands (aid lix et-

Wertiet amid (OtliC S 197 1, DLutnStall antd iithct s 1986). I13x CuL1lIxVatItig ti SSLIC Samlples t Ii 18 C\liCtit11ii-it(IN'1 itifectedi cats,Bat hiec and iitlier-s (1 clemottstrtc(Id that theC futt,(igutS hadspt c(id lto the xviscctra iniiriiuc oh- the c(iscs, cex Ciin the ahseiiccot clinical sigtis anid ma(croscopic cli(ingcs in the oirg(Itis, 'SuigIgeCStitig tlilat theC flttigutI smtiy sprecid sxystctiii(ilxy tiiote ft e-(LCjietItlx tli(iti has iCetI1 aSSctIIelCC1

lIII theC pi-CSCtit StUtdy, tii(iC titSCdtfiC CIjatigeCS xx--er tlstxC-Tcini the skiii, lutig1S alid lxynplh tiutdes. 'I'lic initlimiitiirtivpi-itcc'ss detected miicrOisc(tpicahxy inl thcse tISSLtCS, (ilidi iti theixvCr-, 55 (s cOnrsidcredc lt(- he cit miidcrtate ii-itci-sity. Hoxewcic, niohecpatusphetiltiiegalx itt c hiatliges Ilix et eii ti/lells d)t Pd tlasmutre(i cildicitritmiit xxvere itsex_SCIcl. TheC lxtllphiadcetnot-iegahdletected pocsttliriCotcii hias (ihtcadx lieeii ithbsesecd cliiitcahlx Ill

threeC C(tS. 'Ihie CLttaI1CLeItS lcsiotis atnd the lxytiphi iidchs hI-icllhigh dciisitx, cit p(r(sites, xxith tile cxCviiitLilI rCphacetiievit itttheinodal paretchixA bV(th anl CXuhetFaitt i1tfiltratC itt liiglilx pat (isitiscd miiacr(tphi(igs. The rCSP'irat(itrx s(igius deCteCCted xxT-eeCorrelated xxithi thec CpCtcscc itt intitrstitial pilecitiloilzi avidaolaIxrthi occheicia. The onds cat cxamiieiicd ridhidograpliicallx had(lit]inletsllh initiltratc at the bise itt its i- gIll lttxx11Whicli 55(Scotitif iedii( postmortem, disc lisintg tlic futiugal (ictioiogy ( Fi

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4). In the lungs, the round fungal elements were observed inthe alveolar walls where there were infiltrates of inflamma-tory cells. S schenckii was isolated from its lungs and nasal cav-ities. Larsson and others (1989) also observed respiratorysigns, but Freitas and others (1965) reported that, despitethe presence of extensive cutaneous lesions, there were noclinical signs of systemic involvement.

The present clinical and pathological observations suggestthat in cats the route of entry of S schenckii may be by inhala-tion, causing respiratory signs similar to those observed ininfections caused by Cryptococcus neoformans and the otherdimorphic pathogenic fungi that are agents of systemicmycoses (Rippon 1988b, Davies and Troy 1996). Recently,Schubach and others (2002) studied 148 cats with a clinicaldiagnosis of sporotrichosis. S schenckii was isolated from clin-ical samples of cutaneous lesions taken from all 148 cats, fromthe nasal cavities of 47, from the oral cavities of 33, and fromthe nails of 15 of the cats. These results reinforce this view andthe zoonotic potential of feline sporotrichosis.

The high parasite burden detected in the tissues examined,demonstrated by the visualisation of countless fungal struc-tures in the skin lesions and internal organs, and by the easewith which the fungus was isolated in culture, may beassociated with a higher susceptibility to S schenckii in cats.Co-infection with FIV may have contributed to the dissemi-nation of the disease in one case, but it was not possible tofind any associated predisposing factor in the other nine cats.Little is known about the prevalence of FIV infection inBrazilian cat populations. Souza and others (2002) studied126 cats from Rio de Janeiro and showed that 99 of them hadclinical signs of immunodeficiency and that the remaining 27were apparently healthy cats in domestic contact with theaffected animals. They found that 22 of the cats were sero-logically positive for FeLV, 21 were positive for FIV, and twowere positive for both viruses.

In six of the 10 cats, antifungal treatment did not preventthe disease from progressing or prevent the detection of fungiin their tissues postmortem.

ACKNOWLEDGEMENTS

The authors thank the staff and students of the ZoonosisService, Evandro Chagas Clinical Research Institute-Fiocruz:Virgilio Ferreira da Silva, Dilma Ferreira, Isabella ViannaPellon, Rodrigo de Almeida Paes (Mycology Service, EvandroChagas Clinical Research Institute-Fiocruz), and EleonoraCarregal (Image Diagnosis Service, Evandro Chagas ClinicalResearch Institute-Fiocruz) for their technical support; andHeloisa Diniz (Multimedia Service, Oswaldo Cruz Institute-Fiocruz) for her design support.

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RIPPON, J. (1988a) Sporotrichosis. In Medical Mycology - The PathogenicFungi and the Pathogenic Actinomycetes. Ed J. Rippon. Philadelphia, W. B.Saunders. pp 325-352

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Gutierrez-Galhardo and B. WankeOkamoto, R. Santos Reis, P. C. Fialho Monteiro, M. C. T. M. Pacheco Schubach, A. de Oliveira Schubach, T. Cuzzi-Maya, T.

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