Cutaneous Mycoses in Chameleons Caused by the Chrysosporium Anamorph of (Apinis) Currah Author(s): Jean A. Paré, Lynne Sigler, D. Bruce Hunter, Richard C. Summerbell, Dale A. Smith and Karen L. Machin Source: Journal of Zoo and Wildlife Medicine, Vol. 28, No. 4 (Dec., 1997), pp. 443-453 Published by: American Association of Zoo Veterinarians Stable URL: http://www.jstor.org/stable/20095688 . Accessed: 02/02/2015 19:24

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CUTANEOUS MYCOSES IN CHAMELEONS CAUSED BY THE CHRYSOSPORIUM ANAMORPH OF NANNIZZIOPSIS VRIESII (APINIS) CURRAH

Jean A. Par?, D.M.V., D.V.Sc, Lynne Sigler, M.Sc, D. Bruce Hunter, D.V.M., M.Sc, Richard. C. Summerbell, Ph.D., Dale A. Smith, D.V.M., D.V.Sc, and Karen L. Machin, B.Sc, D.V.M.

Abstract: A dermatophyte-like was isolated from skin biopsies of three different species of captive chameleon in which fungal elements had been observed by histologie examination. An adult Parson's chameleon {Chamaeleo parsonii) presented with vesicles that became crusty brown lesions on the limbs and body. Skin biopsies revealed fungal hyphae in the affected epidermis and underlying dermis. The lesions regressed fully after oral administration of itraconazole. An adult jewel chameleon {Chamaeleo lateralis) from the same private collection presented with localized black skin lesions and died while being treated with itraconazole. A pulmonary granuloma was also present in this chameleon at autopsy. Cultures obtained from skin and lung lesions yielded the same fungus. A third isolate was obtained from a skin biopsy of a Jackson's chameleon {Chamaeleo jacksoni) with deep ulcerative cutaneous lesions located at the base of the tail. The fungus, in all three cases, has been identified as the Chrysosporium anamorph of Nannizziopsis vriesii, a poorly known ascomycetous species recorded previously from the skin of a lizard and from soil, on the basis of its keratinolytic activity, resistance to cycloheximide, strongly restricted growth at 37?C, formation of cl?vate or pyriform single-celled or two-celled aleurioconidia, and alternate and fission arthroconidia. Key words: Chameleon, Chamaeleo, dermatomycosis, Chrysosporium, Nannizziopsis vriesii, itraconazole.

INTRODUCTION lesions in the Chamaeleonidae are Mucor circinel

loides in a two-lined chameleon {Chamaeleo bi Reviews of diseases in reptiles belonging to the taeniatus) and a common chameleon (C. chamae order Squamata (Lacertilia and Serpentes) indicate leon)43 Candida guillermondii in a Fischer's cha that mycoses of the skin may be underreported, that meleon (C fischeri) and a Jackson's chameleon (C. causative agents are often inadequately identified, an unidentified yeast in a Meller's cha and that it is often difficult to evaluate whether the jacksoni)43 meleon (C. melleri)4 Fusarium oxysporum in two isolated fungus is present as a contaminant or is flap-necked chameleons (C dilepis)443 and Asper involved in a pathologic process.4520283? Cutaneous g/Z/ws sp. in a Jackson's chameleon.15 mycoses in humans and other mammals are usually Dermatophytoses are fungal infections of the ke caused by dermatophytes, but most reports of skin ratinized tissues caused by a group of closely re infection concerning lacertilians and ophidians lated keratinophilic fungi known as dermato have been attributed to a wide variety of soil fungi, phytes.39 Dermatophyte species belonging to the including members of the genera Aspergillus,%X53243 genera Trichophyton and Microsporum commonly Candida,*243 Ciadosporium,n Fusarium,41943 Geotri cause skin disease in both humans and animals, but chum,2>4>'9>25<27>4]Monilia,'4 Mucor,20>32>40A3 Paecilomy ' ' true dermatophytosis has rarely been recorded in ces, P?nicillium32 Rhizopus43 Trichoderma,19-32 Tri lacertilians41732 and, to our knowledge, never in chosporon3243 Ulocladium43 and unidentified chameleons. The paucity of reports linking derma taxa.420 Fungi that have been associated with skin tophytes with cutaneous disease prompted Aus twick and Key mer to state that "The susceptibility of skins to infection does not From the Department of Pathology, Ontario Veterinary reptile fungal clearly College, University of Guelph, Guelph, Ontario NIG lie in the direct invasion of the keratinized layers . . . "4 2W1, Canada (Par?, Hunter, Smith); the University of Al by dermatophytes Reports of dermatophy berta Collection and Devonian Microfungus Herbarium, toses in ophidians are also rare. Trichophyton Botanic Garden, Edmonton, Alberta T6G 2E1, Canada mentagrophytes was recovered from a ball python (Sigler); the Ontario Ministry of Health, Laboratory Ser {Python regius) with dermal lesions,15 but most oth vices Branch, 81 Resources Road, Etobicoke, Ontario er reports involve species of keratinophilic fungi of M9P 3T1, Canada (Summerbell); and the Department of soil origin that may be isolated as contaminants and Veterinary Internal Medicine, Western College of Veteri for which demonstration of a role is dif nary Medicine, University of Saskatchewan, Saskatoon, pathogenic ficult. a Saskatchewan S7N 5B4, Canada (Machin). Present ad Trichophyton terrestre, geophilic fungus dress (Par?): Calgary Zoo, Box 3036, Station B, Calgary, that is usually considered nonpathogenic, has been Alberta T2M 4R8, Canada. isolated from the scales of an apparently healthy

443

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boa constrictor {Boa constrictor)4 and was associ to the Veterinary Teaching Hospital of the Ontario ated with progressive digital necrosis in eastern Veterinary College for evaluation of a skin lesion blue-tongued skinks {Tiliqua scincoides), in which of the left stifle. The lizard had been purchased numerous hyphae were seen histologically,17 but a from a pet store 2 mo earlier, quarantined for 6 wk, causal relationship could not be ascertained. Al and treated twice, at 10-day intervals, with fenben though fungal elements could be seen histological dazole (Safe Guard, Hoechst Canada Inc., Regina, or ly, whether they were primary etiologic agents Saskatchewan, Canada), 50 mg/kg orally, as ascarid secondary invaders of devitalized tissue could not eggs had been seen on a fecal flotation. The lizard be determined. An unknown Trichophyton species was fed crickets dusted with a multivitamin powder. was isolated from the cutis and muscles of a day It ate well, although it refused any other kind of gecko {Phelsuma sp.) with multiple nodular skin food, and drank well. Housing and husbandry met lesions.32 This was one of a group of recently im the minimal standards recommended for mainte ported geckos showing similar cutaneous lesions. nance of chameleons in captivity,13 except for in A survey of keratinophilic fungi from Australia adequate ultraviolet radiation exposure. The lizard demonstrated the presence of Microsporum cookei was presented in late September, 45 days after and Chrysosporium species from epidermal scales weather made it impossible for the owner to take it of a clinically normal monitor lizard {Varanus sp.) outside on a daily basis. and from the skink Egernia bungana.3X Chrysospor The chameleon weighed 368 g and was alert, re ium sp. was isolated from nodular subcutaneous le sponsive, and in good body condition. Abnormali sions in a corn snake {Elaphe guttata guttata)4523 ties were restricted to a 1 X 0.5 cm ovoid, de and C keratinophilum, C. tropicum, and Chrysos pressed area of gray devitalized skin on the crani porium sp. have been cited as agents of dermato olateral aspect of the left stifle. A thin crevice vis mycoses in lizards {Varanus salvator, Lacerta vir ible along the rim of the lesion exposed the dermis, idis, and Crotaphytus sp.).943 One report identified suggesting that a large vesicle had ruptured and col C. keratinophilum as an agent of deep infection in lapsed. Within a week, two small gray vesicles, 3? volving the lung and stomach in two iguanas.42 4 mm in diameter, appeared on the lateral aspect of This report describes three cases of skin infection the right elbow. The chameleon was anesthetized in captive chameleons. The same fungus was iso with isoflurane (AErrane, Anaquest, Mississauga, lated in pure culture, and fungal elements were Ontario, Canada) delivered through an induction demonstrated by histopathology of biopsied lesions chamber and then through a face mask. Biopsies in two of the cases. The causative agent was iden were taken from the edge of the stifle lesion and tified as the Chrysosporium anamorph (mitotic from one of the elbow lesions using a 3-mm dis stage) of Nannizziopsis vriesii (Apinis) Curran, a posable skin biopsy punch (Acu-Punch 3mm, Acu poorly known ascomycetous fungus first described derm Inc., Fort Lauderdale, Florida 33307, USA). under the name Rollandina vriesii Apinis3 for an The resulting skin defects were closed with a single isolate obtained from the skin and lungs of a lizard simple suture using polydioxanone monofilament {Ameiva sp.). The fungus was disposed subsequent (PDS 3-0, Ethicon, Inc., Somerville, New Jersey ly as Arachnotheca vriesii (Apinis) Samson38 and 08876, USA). The other vesicle was incised, and later as Nannizziopsis vriesii.12 Nannizziopsis vriesii the clear ex?date it contained was collected and is a member of the order , family Ony submitted for bacteriology. The patient was dis genaceae,12 and records to date indicate that it is charged from the hospital with oral trimethoprim known only from the original lizard isolate and one sulfamethoxazole (Apo-Sulfatrim, Apotex Inc., To other from soil in California. Variability in micro ronto, Ontario, Canada), 30 mg/kg once daily, scopic features suggests that this species could be pending the results of the biopsy. The culture was misidentified or confused with dermatophyte spe negative for any bacterial agent. Poor sectioning of cies such as Trichophyton mentagrophytes or T ter biopsied tissues made the histopathologic interpre restre or with geophilic, keratinophilic Chrysospor tation difficult, but marked heterophilic infiltration ium or Malbranchea species. Its propensity to form of the epidermis and dermis was noted. One week fission arthroconidia could suggest Geotrichum. after initiation of antibiotic therapy, the lizard was again presented with numerous new lesions on all CASE REPORTS limbs, the flanks, and tail. These were roughly cir cular, focal, brown encrustations that Case 1 protruding measured several millimeters in diameter. The ini

An adult male Parson's chameleon {Chamaeleo tial lesion on the stifle had thickened, and the de a parsonii) from a private collection was presented vitalized skin had become large thick scab. The

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Figure 1. Skin from a Parson's chameleon. Focal hyperkeratosis and coagulation necrosis of the stratum corneum are visible (arrow). Note the discontinuity of the dermal melanophore layer (open arrow). H&E, X40.

lizard was anesthetized and three more skin biop An oral liquid suspension of itraconazole (Jans were was ven sies taken. Blood collected from the sen Pharmaceutica, Beerse, Belgium; 10 mg/mL) a tral tail vein and submitted for complete blood was added to the treatment regimen (10 mg/kg once cell count. daily for 21 days). No new lesions appeared, and X 109 A leukocytosis (34.5 cells/L, suggested the preexisting ones regressed partially and dried reference in lacertilians: 12-22.5 X 109 cells/ range up. Antibiotic therapy was discontinued 1 week pri and a or 20.3 X 109 L7) lymphocytosis (59%, or to cessation of antifungal administration, 21 days cells/L) were the prominent h?matologie findings. after the initial visit. The owner noted that the cha Occasional micronTariae also were noted. On he meleon was more reluctant to accept the itracona matoxylin-eosin-stained sections, there was focal zole than the antibiotic. The lizard's appetite de hyperkeratosis with necrosis of the stratum cor clined over the last few days of medication, and the neum and heterophilic infiltration of the subjacent owner was required to hand-feed and, weeks later, epidermis and dermis (Fig. 1). Aggregates of mac force-feed the lizard. The patient was reevaluated rophages and diffuse heterophil infiltration within 45 days from the initial visit. Many skin lesions had the deeper layers of the dermis were consistent with and no new lesion had how a deep heterophilic cellulitis. A periodic acid disappeared, appeared; ever, a general loss of condition was noticed. The Schiff stain of the same section revealed fungal hy lizard appeared to favor the left hind leg and would phae within the affected stratum corneum and ad not as as it did Ra jacent epithelium (Fig. 2) and deeper layers of the grasp perches easily previously. The cell dermis. Hyphae were partly refractile and septate diographs suggested hepatomegaly. packed volume had from initial value with minimal branching. A frozen biopsy submitted (18%) dropped but was still within the reference for culture yielded a slow-growing Trichophyton (23%) suggested for lacertilians Elevated serum like fungus. The isolated fungus was referred to the range (16-45%).7 University of Alberta Microfungus Collection for levels of creatinine kinase (10,898 U/L) and aspar further study. tate aminotransferase activity (>800 U/L) were the

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Figure 2. Coagulation necrosis of the stratum corneum of a Parson's chameleon cutaneous epithelium. Note the fungal hyphae in the epidermis and dermis (arrows). PAS, X100.

only biochemical abnormalities noted. Microfilariae mg/kg once daily for 21 days. Six days after initi were again visualized on the blood smear. ation of therapy, the lizard was found dead in the The chameleon's appetite increased and its gen vivarium. Postmortem examination revealed a gran eral body condition improved in the months that ulomatous mass located in the caudal lung and ex followed. After ecdysis, only the scab on the stifle tending to the wall of the coelomic cavity and to remained. The chameleon died about 1 year after the kidney. Histologie examination revealed the initial presentation. Autopsy indicated that death presence of a severe deep heterophilic cellulitis ex was due to cholecystitis and septicemia, but no ev tending from the skin surface into the muscles of idence of mycosis was seen. the foot. Periodic acid-Schiff stain showed many hyphae in the affected tissues. Numerous hyphae Case 2 were also seen within the granuloma involving the An adult male jewel chameleon {Chamaeleo la lung and kidney (Fig. 4). A fungus identical to the teralis) purchased from the same pet store by the isolate from case 1 was cultured from the pedal same collector was presented for evaluation. The squames, the lungs, and the kidney and was for tiny lizard was alert and active. It weighed 17 g warded to the University of Alberta for further and appeared in good flesh. There were two local evaluation. ized areas of black discoloration of the skin. One Case 3 measured approximately 2 mm and involved the right upper lip at the mucocutaneous junction. The An adult male Jackson's chameleon was pre second lesion, slightly larger, was located on the sented to the Veterinary Teaching Hospital of the dorsal aspect of the digits of the left hind foot (Fig. Western College of Veterinary Medicine with a 2 3). A thin layer of exfoliated squames partially cov cm round scab on the left side, at the base of the ered the pedal lesion, and a sample was submitted tail. The lizard weighed 100 g, was alert and active. for fungal culture. The chameleon was sent home Blood analysis revealed a leukocytosis (32 X 109/L) on itraconazole oral suspension, at a dosage of 10 with a lymphocytosis (83%, or 26.6 X 109/L). El

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Figure 3. Black discoloration of the skin on the dorsal aspect of the left hind foot and digits of a jewel chameleon (arrows). Note the exfoliative white squames.

evated packed cell volume (57%) and total solids Anyang, Korea). A second culture of the wound (15 g/dl) values suggested dehydration. The area yielded Enterococcus faecalis and Staphylococcus was debrided and cultured. Large numbers of Clos sp. The antibiotics were changed to an amoxicillin tridium sp. were isolated as well as a fungus iden clavulinic acid oral suspension (Clavamox drops, tified as a Trichophyton sp. This isolate was iden Smith Kline Beecham Animal Health Inc., Missis tical to that of cases 1 and 2, and a frozen biopsy sauga, Ontario, Canada; 22 mg/kg b.i.d.), and ke was later forwarded to the University of Alberta. toconazole was continued. The chameleon was pre Fluids were administered intracoelomically, the sented 2 wk later with a recurrence of the hemi wound was bandaged, and the lizard was sent home penis prolapse. The area of necrotic skin was al on a trimethoprim-sulfamethoxazole oral suspen most circumferent around the base of the tail. The was sion (Novotrimel, Novopharm Limited, Calgary, hemipenis removed surgically under ketamine Alberta, Canada; 15 mg/kg b.i.d.) for 14 days. Ke anesthesia, and tolnaftate (Tinactin, Schering Can toconazole (Nizoral, Janssen Pharmaceutica Inc., ada Inc., Pointe Claire, Quebec, Canada) was to be Mississauga, Ontario, Canada; 25 mg/kg, Q2D, p.o. applied topically on a daily basis by the owner. This for 4 wk) was added to the regimen when fungal chameleon was then lost to follow-up. culture results were obtained. The lizard was re examination examined 2 wk later as a hemipenis had prolapsed Mycological through a ventral extension of the wound. The liz Isolates from cases 1 and 2 were deposited in the ard was anesthetized with ketamine hydrochloride University of Alberta Microftingus Collection as (Ketaset, Ayerst Laboratories, Montreal, Quebec, UAMH 7582 and 7583. The chameleon isolates Canada; 20 mg/kg), and the prolapsed hemipenis were compared with known strains of Nannizziop was gently cleansed with saline and was reduced sis vriesii in all tests. Isolates of N. vriesii included through the hole in the ventral tail base. Closure of the ex-type (culture derived from the type speci = = the wound was performed using 4-0 NovofiT (Davis men) (UAMH 3713 ATCC 22444 CBS 407.71 = & Geek, Cyanamid Medical Device Company Inc., IMI 149994), which had been isolated from the

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Figure 4. Chrysosporium anamorph of Nannizziopsis vriesii hyphae in a necrotic focus in the lung of a jewel chameleon. PAS, XlOO.

a skin and lungs of a lizard {Ameiva sp.) in Nether terns, Cockeysville, Maryland 21030, USA), mod lands by G. A. de Vries, and an isolate from soil ification of Sabouraud dextrose medium that has been in southern California (UAMH 3526). The case 3 used for the study of Chrysosporium species,103537 and isolate (UAMH 7861) was obtained later and not potato dextrose agar (Difco Laboratories, Detroit, included in all tests. Michigan 48232-7058, USA). Media used for the pro For observation of colonial features and growth motion of sporulation and development of sexual rates, isolates were grown at 25?C on phytone yeast stages included 10% Pablum cereal agar22 and oat extract agar (Becton Dickinson Microbiology Sys meal agar22 with incubation at 22? or 25?C for 2-3

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Figure 5. Colony of Chrysosporium anamorph of Nan on nizziopsis vriesii potato dextrose agar after 21 days at 6. or cl?vate sessile conidia formed 25?C (UAMH 7583). X0.8. Figure Pyriform on branched hyphae (UAMH 7582). X580. mo. In a subsequent attempt to obtain ascomata (sex ual fruiting bodies), isolates were grown at 30?C ? lowish white to pale yellow (3A2/4A3); flat or on 2?C Takashio agar,22 amended with 5% yeast ex slightly raised in the center (umbonate); dense, ex and on tract, oatmeal agar. Terminology for colony cept thin at margin; sometimes with concentric colors follows and zones Kornerup Wanscher.24 Because of of denser growth; powdery; and often show some similarities between Chrysosporium species and ing fine fissures or cracks, with or without small were dermatophyte species, isolates also subjected to droplets of clear ex?date on the surface, reverse a battery of physiologic tests performed commonly in uncolored. Colonies of the two N vriesii isolates the Isolates were are dermatophyte diagnostic. evaluated similar, except that they grow slightly faster, for their in the tests: responses following 1) growth, reaching diameters of 4.5-5.5 cm after 21 days. Co pH change, and clearing of milk solids on Bromcresol nidia (aleurioconidia) (Fig. 6) are pyriform (tear to or purple-milk solids-glucose agar212236 compared drop-shaped) cl?vate (club-shaped), single-celled on growth Sabouraud dextrose agar (Difco); 2) urease or rarely two-celled, and are borne sessile (i.e., activity in Christensen's urea broth22; 3) tolerance to formed on the sides of the not on ' directly hyphae, at 400 or at cycloheximide |xg ml by recording growth stalks) the ends of branched fertile hyphae. rates on tolerance to salt at con are mycosel agar2233; 4) Lateral and terminal conidia 3-12.5 |xm long centrations of and 7% and as 0, 3, 5, measured the and 1.5-2.5 |xm wide but are commonly 3-6 u,m sodium chloride concentration >50% in causing growth length and single-celled. Portions of the hyphae inhibition at 14 days'22; 5) requirements for vitamins frequently fragment to form arthroconidia, which thiamine and inositol on are by comparing growth Tri either alternate (i.e., separated from each other and 4 with on a or chophyton agars 2, 3, growth vitamin by cell cells that undergo lytic disintegration) free casamino acids or agar (Trichophyton agar 1) (Dif in chains and separated by fission at the septum to at and to co); 6) ability grow 37?C; 7) ability digest (i.e., schizolytic dehiscence). Arthroconidia (Fig. 7) hairs after 2-3 wk incubation.35 of are are Descriptions mi cylindrical and 4-9 u,m long and 1.5-3.5 croscopic features are based mainly on slide culture |xm wide. A feature common to all isolates is the preparations.34 formation of undulate, solitary, sparsely septate lat Colonies of the three chameleon isolates (Fig. 5) eral branches (Fig. 8). Rarely, these branches frag are moderately fast growing, reaching diameters of ment to form arthroconidia. cm on 4-4.5 phytone yeast extract and potato dex The chameleon isolates failed to fruit (form sex trose after 21 at or rates agar days 25? 30?C; growth ual fruiting bodies) under any growth conditions, were same on the mycosel agar. Colonies are yel but isolates of N vriesii formed fertile ascomata

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Figure 8. Undulate lateral branches (UAMH 7861). X460.

DISCUSSION

Regarding the taxon as "reminiscent" of Ar throderma Berkeley, Apinis described Rollandina Figure 7. Fertile hyphae bearing sessile conidia (arrow vriesii in the Rollandina Patouillard based on and to form arthroconidia head) fragmenting (arrow) a single isolate from the skin and lung of an Ame iva (UAMH 7582). X460. species.3 Ascomata of N. vriesii lack appendages are and composed of anastomosed asperulate hy phae that are constricted at the septa and in which individual cells are uniform in Asco characteristic for the species312 when grown on Tak fairly shape. mata of members of the Arthroderma ashio and oatmeal agars at 30?C. The N vriesii iso genus (family Arthrodermataceae, Onygenales) differ in that they lates appeared initially to have lost sexual vigor, are composed of hyphae in which the cells are typ since they failed to form ascomata even after sev ically dumbbell-shaped or ossiform (bone-shaped), eral months at lower temperature of incubation, but and hyphae may terminate in tightly coiled append once sexual reproduction was reestablished, isolates ages.12 Apinis also described an associated ana fruited also at 25?C on cereal agar. or morph asexual stage as "pyriform or cylindrical In the special tests, the chameleon isolates and to cl?vate conidia that are formed on two singly vege the isolates of N vriesii shared the following tative hyphae of the Sporotrichum or Chrysospor features: 1) strong urease 5-7 activity, usually by or ium type, in chains {Oidium type) 2-3 X 3-8 days; 2) digestion of hairs with formation of per |xm in diameter."3 Later workers rejected Rollan forating bodies; 3) greater than 50% growth inhi dina as a nomen confusum, and Curran transferred bition at 37?C; 4) no requirements for vitamins thi R. vriesii to the new genus Nannizziopsis Curran amine or inositol; 5) inhibition by salt at a within the family (Onygenales), pri concentration of 3% >50% (concentration causing on marily the basis of ascospore shape and wall reduction of colony diameter). On Bromcresol pur ornamentation.12 Ascospores of N. vriesii are glo ple-milk solids-glucose agar after 11 days, the cha bose, measuring 2.5-3 \xm in diameter, and punc meleon isolates showed moderate to profuse tate-reticulate, whereas ascospores of members of no in or trace indicated growth, change pH acidity the Arthrodermataceae are oblate (round in face a color to faint and clear by change yellow, strong view, flattened in side view) and smooth. The ana of the milk solids the of the ing beyond margin morph has been accommodated in Chrysosporium. colony. On this medium, the N vriesii isolates Since none of the chameleon isolates has fruited showed some of the milk profuse growth, clearing (i.e., formed fertile ascomata), they cannot be iden solids behind the and no in or colony, change pH tified with certainty as N. vriesii; however, charac trace indicated a color to faint alkalinity by change teristics of their conidia and physiology and their within the zone. purple clear isolation from lizards suggest a close relationship.

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Similar features include 1) limited thermotolerance of isolates involved in reptilian skin infections (restricted growth at 37?C); 2) the pyriform or cl? would be useful in delineating etiologic agents. or vate shape of the conidia, which are occasionally It is difficult to determine where when the 1-septate and borne sessile rather than on stalks; 3) chameleons became infected. The authors were in the presence of lateral curved or undulate branches; formed that another chameleon, bought by a dif 4) the tendency to form arthroconidia that are either ferent party from the same source as chameleons 1 alternate or in chains; 5) strong keratinolytic activ and 2 and around the same time, had been diag ity; and 6) similar physiologic features, including nosed elsewhere with fungal dermatitis and died urease activity, cycloheximide tolerance, absence of shortly thereafter. It is likely that the case 1 and 2 requirements for vitamins, and tolerance for salt at lizards were from the same shipment and were har a concentration of 3%. The chameleon isolates dif boring the organism at the time of purchase. This fered from N vriesii in having strongly powdery fungus may be part of the normal skin flora of cha colonies that grew slightly more slowly, and in their meleons. Unsanitary captive conditions, malnutri growth patterns on Bromcresol purple-milk solids tion, high humidity, overcrowding, and poor water glucose agar medium, with a clearing of milk solids quality are thought to predispose reptiles to fungal beyond the margin of the colony. The chameleon disease.20 Parson's chameleons and jewel chame isolates form a subset of a larger group of similar leons originate from Madagascar, and Jackson's fungi obtained from skin lesions in members of the chameleons from Africa.26 Parson's chameleons are order Squamata and currently under study by two notoriously short-lived in captivity, with one source of us (LS, RCS). Additional investigations of rela stating a longevity record of 11 months.6 We spec tionships will include molecular analyses. ulate that the stresses of capture, quarantine, and The isolation of the same fungus from three cha shipping contributed to undermine the immunity of meleons with cutaneous lesions in which fungal hy these lizards to fungal infection. It is probable that phae were observed by histology firmly establishes pulmonary infection occurred by inhalation from a this fungus as an etiologic agent of infection. The contaminated, poorly ventilated environment; how involvement of the fungus in deep mycosis as seen ever, hematogenous spread cannot be precluded. in case 2 is unusual and further establishes a link Microfilaremia in the Parson's and intestinal trem with N vriesii, which was isolated also from both atodiasis in the jewel chameleon also may have skin and lung. It seems likely that this chameleon contributed to lower hosts' resistance. fungus has been reported previously, but correla Lesions reported in reptiles with skin mycoses tions are difficult because many reports fail to il are variable and often poorly described. These in lustrate the fungus, and the isolates from these clude loosening of the horny part of the scales, nod cases rarely are deposited into culture collections. ules, abrasions, plaques, crusts, granulomas, and fo Previous reports concerning Trichophyton spe cal necrotic ulcers, perhaps reflecting different 17.32 cjes4,is. or Chrysosporium species459234243 could stages of disease progression. Focal black or gray have been dealing with the same fungus. Two of discoloration of the skin was observed in cases 1, the chameleon isolates were identified initially by 2, and 3. Bullous or vesicular lesions ("blister dis one laboratory as T. verrucosum and by a second ease") in reptiles are common but are usually as laboratory as T. terrestre, but they differed slightly sociated with bacteria, rarely with fungi.16 Affected in in vitro growth characteristics. The sessile co animals are often kept under suboptimal condi nidia of TV. vriesii are reminiscent of microconidia tions.16 The vesicular aspect of the early skin le of species of Trichophyton such as T mentagro sions in case 1 suggests that fungal disease must be phytes or T. terrestre, but N. vriesii fails to develop considered by the clinician when confronted with a macroconidia typical of Trichophyton species. bullous or vesicular skin condition in a chameleon. Moreover, the development of characteristic fruit The use of itraconazole in reptiles has not been ing bodies at 30?C on certain media confirms their well documented. Itraconazole is a triazole antifun distinction. Several reports link reptilian skin infec gal with a broader spectrum of activity, greater ef to a tions Geotrichum candidum,24i92527A] yeast-like ficacy at lower dose, and less toxic potential than hyphomycete in which hyphae fragment to form the more frequently used ketoconazole.29 Itracona cylindrical arthroconidia. Although G. candidum zole, like ketoconazole, inhibits ergosterol biosyn seems unlikely to be confused with the Chrysos thesis by binding to fungal, but not mammalian, porium anamorph of N vriesii when the character cytochrome P-450 enzymes. Ergosterol is an essen a istic lateral conidia predominate, there may be tial component of the fungal cell membrane. When preponderance of fission arthroconidia under some its synthesis is disrupted, a defective membrane re cultural conditions. Further mycological evaluation sults; intracellular accumulation of ergosterol pre

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4. P. K. and I. F. 1981. cursors may also contribute to eventual cell rup Austwick, C, Keymer. Fungi and In: J. E., and O. F Jackson ture.29 Itraconazole does not suppress cortisol syn actinomycetes. Cooper, (eds.). Diseases of the Reptilia, vol 1. Academic Press, thesis in humans and dogs29 and thus should not Harcourt Brace Jovanovich, London, England. Pp. 193 interfere with the animal's response to stress. Itra 231. conazole has been shown to be effective against 5. Austwick, P. K. C. 1982. Some mycoses of reptiles. various agents of systemic mycoses, yeasts, der VII Congress International Society for Human and Animal some matophytes, and protozoans. The dose of 10 Mycology. Pp. 383-384. was based on the for mg/kg/day suggested dosage 6. Barten, S. L. 1993. The medical care of iguanas and the In case with dog (1-10 mg/kg/day).29 1, therapy other common pet lizards. In: Vet. Clin. North Am. Small itraconazole appeared to resolve the lesions, al Anim. Pract.: Exotic Pet Medicine I. 23(6): 1213-1249. though transient anorexia and loss of condition 7. Beynon, P. H., M. P. C. Lawton, and J. E. Cooper were noted. Elevations in the creatinine kinase and (eds.). 1992. Manual of Reptiles. British Small Animal unex Glou aspartate aminotransferase activity remain Veterinary Association, Sherdington, Cheltenham, cestershire, U.K., III, p. 220. plained. The possibility of ongoing muscular dam Appendix 8. Borst, G. H. A., C. Vroege, F G. Poelma, P. Zwart, age from fungal growth under the seemingly qui W J. and J. C. Peters. 1972. escent stifle lesion was considered. Itraconazole Strik, Pathological findings on animals in the Royal Zoological Gardens of the Rot therapy was not reinstituted because of potential terdam zoo the years 1963, 1964 and 1965. Acta a during hepatoxicity in a patient with palpably and radi Zool?gica et Pathologica Antverpiensia 56: 3-19. ographically enlarged liver. It is likely that the 9. Bryant, W. M. 1982. Mycotic dermatitis in a col stress of and was also daily handling medicating lection of desert lizards. Am. Assoc. Zoo Vet. Proc. Annu. detrimental to this animal's and appetite general Meet. New Orleans, Louisiana. P. 4. health. The role of itraconazole in the demise of the 10. Carmichael, J. W. 1962. Chrysosporium and some in case 2 is as chameleon undetermined, hepatotox other aleuriosporic Hyphomycetes. Can. J. Bot. 40: 1137? was icity, the main adverse effect, not identified. 1173. The animal died too soon to assess the efficacy of 11. Cork, S. C, and P. H. G. Stockdale. 1994. Mycotic disease in the common New Zealand the antifungal therapy. The use of ketoconazole in gecko {Hoplodac N. Z. Vet. J. 42: 144-147. case 3 did not seem to alter significantly the pro tylus maculatus). 12. Currah, R. S. 1985. of the Onygenales: gression of the lesions, although the role played by Arthrodermataceae, Gymnoascaceae, Myxotrichaceae and the fungus is unclear. Onygenaceae. Mycotaxon 24: 1-216. Raising and maintaining the lizard's environment 13. de Vosjoli, P. 1990. The General Care and Main temperature in the upper zone of the species' pre tenance of True Chameleons, part I, Husbandry. Advanced ferred optimal temperature range should be gener Vivarium Systems, Lakeside, California. ally beneficial and, given N vriesifs limited ther 14. Dillberger, J., and M. Abou-Gabal. 1979. Mycotic motolerance, may be therapeutic. dermatitis in a black ratsnake. Mykosen 22: 187-190. 15. Frye, F. L. 1981. Biom?dical and Surgical Aspects Acknowledgments: We thank Arlene Flis and of Captive Reptile Husbandry. Veterinary Medicine Pub Linda Abbott (University of Alberta Microfungus lishing Co., Edwardsville, Kansas. Collection and Herbarium) for assistance with cul 16. Frye, F. L. 1991. Biom?dical and Surgical Aspects ture and photography, and Debbie Bateman and Gi of Captive Reptile Husbandry. Krieger Publishing Co., sela Kittler Microbi (Ontario Veterinary College Malabar, Florida. and Jamie and for ology Laboratory) Lynda Horgan 17. Hazell, S. L., G. J. Eamens, and R. A. Perry. 1985. their and commitment. L. S. acknowl cooperation Progressive digital necrosis in the eastern blue-tongued the Natural Sci edges financial assistance from skink, Tiliqua scincoides (Shaw). J. Wildl. Dis. 21: 186 ences and Engineering Research Council of Cana 188. da. 18. Jacobson, E. 1978. Diseases of the respiratory sys tem in reptiles. Vet. Med/Small Anim. Clin. 73: 1169? LITERATURE CITED 1175.

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