Two New Species of Coccidia (Apicomplexa: Eimeriidae) from the Bearded False Chameleon chamaeleolis barbatus (Sauria: Polychridae) from Cinco Pesos, Pinar Del Rio, Cuba

J. Parasitol., 85(4), 1999 p. 719-722 ? American Society of Parasitologists 1999

FROMTHE BEARDED TWO NEW SPECIES OF COCCIDIA(APICOMPLEXA:EIMERIIDAE) BARBATUS(SAURIA:POLYCHRIDAE) FROM FALSECHAMELEONCHAMAELEOLIS CINCOPESOS, PINARDEL RIO, CUBA David Modry*,MilanVeselyt, and Bretislav Koudela* Departmentof Parasitology,Universityof Veterinaryand PharmaceuticalSciences, Brno,Czech Republic ABSTRACT: Parasitological examination of bearded false chameleons Chamaeleolis barbatus freshly imported from Cuba revealed the presence of 2 species of coccidia that are described as new. Oocysts of Isospora chamaeleolidis n. sp. are spherical to slightly subspherical, 16.1 (13-21) X 15.6 (13-19) ixm, with a brownish and bilayered wall -1.0-1.5 pLmthick; outer layer markedly pitted, 0.75-1.0 pxmthick. One, rarely 2, globular polar granules, 1.5 in diameter are present in the sporulated oocysts. Sporocysts are ellipsoidal, 10.8 (10-13) X 7.8 (7-9) u-m,with a smooth, colorless, and unilayered sporocyst wall. Stieda body and substieda bodies are present. A sporocyst residuum is present, consisting of small granules of irregular size scattered among the sporozoites. Oocysts of Eimeria chamaeleolidisbarbati n. sp. are broadly oval, 19.0 (17-21) X 15.7 (15-17) pLm,with a bilayered, colorless oocyst wall -0.75 thick; outer layer of oocyst wall is smooth, 0.5 ,um thick. One or 2, rarely 4, globular, irregular polar granules, -1.5 ,um in diameter, are present in sporulated oocysts. Sporocysts are broadly oval, 7.4 (7-8.5) X 6.1 (5.5-7) pum,with a smooth, colorless, and unilayered sporocyst wall, composed of 2 valves joined by suture; Stieda body and substieda bodies are absent.

The genus Chamaeleolis

Dumeril and Bibron, 1873, includes

4 species of unique arboreal lizards. All members are endemic to the island of Cuba. Due to local distribution and a highly secretive life, knowledge about the distribution and biology of Chamaeleolis spp. is limited to a few reports (Garrido and Schwartz, 1967; Garrido, 1982; Shwartz and Henderson, 1985; Hottmar, 1991). During a survey of fecal samples of freshly imported Chamaeleolis barbatus, oocysts of 2 species of coccidia were found and are described in this paper as new. MATERIALSAND METHODS A group of C. barbatus was imported from Cuba by a pet animal dealer in March 1998. All specimens were freshly caught by local people and originated from the type locality Cinco Pesos, Pinar del Rio Province. Animals were housed separately in glass terraria and fecal samples were collected for routine parasitological examination. Fresh fecal samples of 7 animals were placed in 2.5% aqueous (w/v) potassium dichromate (K2Cr207) solution to enhance oocyst sporulation and checked daily. To describe the stage of sporulation of expelled oocysts, fresh fecal samples were preserved in 5% formalin. Individual samples were concentrated by centrifugation flotation in modified Sheather's sugar solution (specific gravity 1.3) and examined and photographed using Nomarski interference-contrast microscopy. All measurements throughout are reported in micrometers (p.m), with means followed by ranges in parentheses.

DESCRIPTION Parasitological examination of fecal samples revealed the presence of coccidian oocysts and numerous nematode larvae in 6 of 7 (86%) animals. Based on morphological features of the sporulated oocysts, 2 species of coccidia were recognized and are described below as new.

Isospora chamaeleolidis n. sp. (Figs. 1, 2, 5) Oocysts spherical to slightly subspherical, 16.1 (13-21) x 15.6 (1319); oocyst shape index (length/width) 1.0 (1.0-1.1). Oocyst wall brownish and bilayered, -1.0-1.5 thick; outer layer markedly pitted, 0.75-1.0 thick. Micropyle and oocyst residuum absent. One, rarely 2, globular polar granule, 1.5 in diameter, present in sporulated oocysts. Received 27 October 1998; revised 9 February 1999; accepted 9 February 1999. * Institute of Parasitology, Academy of Sciences of the Czech Republic, Ceske Budejovice, Czech Republic. t Department of Zoology, Faculty of Natural Sciences, Palacky University, Olomouc, Czech Republic.

Sporocysts ellipsoidal, 10.8 (10-13) x 7.8 (7-9), with smooth, colorless, and unilayered sporocyst wall; shape index 1.4 (1.25-1.4). Stieda body present, knoblike, 0.75 high and 1.5 wide. Homogenous subglobular substieda body present, 1.0-1.25 high and 2.0-2.5 wide. Sporocyst residuum present as small granules of irregular size scattered among sporozoites. Sporozoites elongate, lying parallel within sporocyst. Each sporozoite with prominent spherical anterior and posterior refractile body and single, spherical nucleus located centrally.

Taxonomic summary Type host: Bearded false chameleon, C. barbatus Garrido, 1982. Site of infection: Unknown. Oocysts recovered from feces. Type locality: Limestone uplands at Ojo de Agua, Cinco Pesos, about 9 km NW San Crist6bal, Pinar del Rio Province, Cuba. Type specimens: Phototypes are deposited in the Parasitological Institute of the Czech Academy of Sciences in Cesk6 Budejovice (no. R 86-a/98). Symbiotype host (alcohol preserved) is deposited in the herpetological collection of Forschungsinstitut und Naturmusem Senckenberg, coll. no. SMF 77959. Sporulation: Exogenous. Oocyst became fully sporulated within 3 days at 22-25 C. Prevalence: Two of 7 (29%). Etymology: Specific epithet chamaeleolidis is derived from the generic name of the type host.

Eimeria chamaeleolidisbarbati n. sp. (Figs. 3, 4, 6) Oocysts broadly oval, 19.0 (17-21) X 15.7 (15-17), with bilayered, colorless oocyst wall -0.75 thick. Oocyst shape index (length/width) 1.2 (1.1-1.4). Outer layer of wall smooth, 0.5 thick. Micropyle and oocyst residuum absent. One or 2, rarely 4, globular irregular polar granules, -1.5 in diameter, present in sporulated oocysts. Sporocysts broadly oval, 7.4 (7-8.5) X 6.1 (5.5-7), with smooth, colorless, and unilayered sporocyst wall, composed of 2 valves joined by suture; sporocyst shape index 1.2 (1.0-1.4). Stieda body and substieda bodies absent. Sporozoites elongate, arranged head to tail within sporocyst. Each sporozoite possesses spherical anterior and posterior refractile body and nucleus situated between.

Taxonomic summary Type host: Bearded false chameleon, C. barbatus Garrido, 1982. Site of infection: Unknown, oocysts recovered from feces.

Typelocality: Limestoneuplandsat Ojo de Agua, Cinco Pesos, about 9 km NW San Crist6bal,Pinardel Rio Province,Cuba. Type specimens: Phototypes are deposited in the Parasitological Institute of the Czech Academy of Sciences in Cesk6 Budejovice (no. R 86-b/98). Symbiotype host (alcohol preserved) is deposited in the herpetological collection of Forschungsinstitut und Naturmusem Senckenberg, coll. no. SMF 77959. 719

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I

FIGURES 1-4. Nomnarski interference-contrast n. sp. and Eijmeria clhanaeleoli(NIC) photographs of the oocysts of Isospora chainaeleolidis note distinct polar granule (arrow), scale bar = 10 WLm.2. Oocyst wall of l. disbarbtlati n. sp., all in the same scale. 1. Isospora clalmaeleolidis, 4. Oocyst of E. chamaeleolidisbarboti 3. Eimer/eia ( Ilcl(i7iele(olidisvblrati. chliaiaieleoliis. with distinct polar granule (airTow).

-

FIGURRES5, 6.

Composite

line drawings

of sporulated oocyst.

Scale bar = 5 Am. 5. Iso.powr

clumaweleolidis.

6. Eiert-ia

(-IiaieleoliIisli,(rl)ibti.

MODRY ET AL.-NEW COCCIDIA FROM C. BARBATUS

FIGURES5, 6.

Sporulation: Exogenous, oocyst became fully sporulated within 2 days at 22-25 C. Prevalence: Six of 7(85%). Etymology: The specific epithet chamaeleolidisbarbati is derived as genitive from the full name of the host.

REMARKS AND DISCUSSION Regardless of the high diversity of iguanid lizards (sensu lato), data on coccidian parasites of this widespread lacertilian family are relatively limited. To date, 20 species of Eimeria and 6 species of Isospora were described and named from iguanid hosts sensu lato (Carini, 1940; Ruiz, 1959; Bovee and Telford, 1965; Bovee, 1966, 1969; Lainson, 1968; Duszynski, 1969; Clark, 1970; Pellerdy, 1974; Upton et al., 1989; Aquino-Shuster et al., 1990; Daszak and Ball, 1991, 1998; Bui et al., 1992; Mitschler et al., 1993; Cisper et al., 1995; Telford, 1998). The increasing numbers of coccidian species, described during last 2 decades, logically leads to partial or total overlap of morphological characters of some of them, making it difficult for taxonomists to separate them. Previous authors have repeatedly suggested the conservative character of morphological features, when compared to the evolution of host specificity (Barker et al., 1989) and therefore considered host systematics to be an important criterion for the recognition of new species (Duszynski, 1986; Duszynski and Wilber, 1997). We are aware of at least 8 families of iguanid lizards, and for the following differential diagnosis we include only coccidian species described and named from members of the Polychridae.

721

Continued.

Oocysts of I. chamaeleolidis n. sp. are most similar in size to those of Isospora reui Cisper, Huntington, Smith, and Powell, 1995, described from Anolis bahorucoensis from the Dominican Republic. Both species could be distinguished easily, based on the heavily pitted and smooth oocyst wall of L chamaeleolidis and L reui, respectively. Also, Isospora achiotensis Telford, 1998 from Anolis limifrons from Panama possesses a smooth oocyst wall and differs also in the absence of Stieda and substieda bodies. Four other species, i.e., Isospora guarocuyai Bui, Powell, Smith, Pamerlee, and Lathrop, 1992 from Anolis distichus, Isospora hendersoni Cisper, Huntington, Smith, and Powell, 1995 from Anolis armouri, Isospora limifronsi Telford, 1998 from A. limifrons, and Isospora tahuayoensis Upton, Freed, and Freed, 1989 from Anolis fuscoauratus possess a heavily pitted oocyst wall, but all of them are significantly larger. Of the 6 species of Eimeria reported from members of the Polychridae, 3 species have oocysts similar enough to be compared with E. chamaeleolidisbarbati. Oocysts of Eimeria intermedia Ruiz, 1959 described from Anolis intermedius from Costa Rica are more elongate and slightly larger, and sporocysts of this species are smaller and more spherical. Eimeria limifronsi Telford, 1997 described from A. limifrons from Panama possesses larger oocysts and sporocysts and differs also in the absence of polar granules. Oocysts of Eimeria sanctaluciae Daszak and Ball, 1998 from Anolis luciae from the Lesser Antilles are smaller and more

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CLARK,G. W. 1970. Eimeria ahtanumensis n. sp. from the northwestern fence lizard, Sceloporus occidentalis in Central Washington. Journal of Protozoology 17: 526-530. DASZAK,P, ANDS. J. BALL. 1991. Five new species of Eimeria (Apicomplexa: Eimeriidae) from lizards. Systematic Parasitology 20: 141-147. . 1998. Description of the oocysts of three new , AND. species of Eimeria (Apicomplexa: Eimeriidae) from iguanid lizards (Sauria: Iguanidae) of Central and South America. Memorias do Instituto Oswaldo Cruz 93: 471-475. D. W. 1969. Two new species of coccidia (Protozoa: EiDUSZYNSKI, meriidae) from Costa Rican lizards with a review of the Eimeria from lizards. Journal of Protozoology 16: 581-585. . 1986. Host specificity in the coccidia of small mammals: Fact or fiction? In Advances in protozoological research, Vol. 33, M. Bereczky (ed.). Symposia Biologica Hungarica, Akademiai Kiado, Budapest, Hungary, p. 325-337. , ANDP. G. WILBER. 1997. A guideline for the preparation of species descriptions in the Eimeriidae. Journal of Parasitology 88: 333-336. O. H., ANDA. SCHWARTZ.1967. Cuban lizards of the genus GARRIDO, LITERATURE CITED Chamaeleolis. Quarterly Journal of Florida Academy of Science AND H. L. SNELL.1990. A. L., D. W. DUSZYNSKI, AQUINO-SHUSTER, 30: 1-25. Three new species of coccidia (Apicomplexa) from the hood island . 1982. Description de una nueva especie cubana de Chamaelizard, Tropidurus delanois, from the Galipagos archipelago. Jourleolis (Lacertilia: Iguanidae) con notas sobre su comportamiento. nal of Parasitology 76: 313-318. Poeyana 236: 1-25. ANDI. BEVERIDGE. 1989. HostBARKER,I. K., M. G. O'CALLAGHAN, HOTTMAR,V. 1991. Zur Erztnachzucht von Chamaeleolis barbatus Garparasite associations of Eimeria spp. (Apicomplexa: Eimeriidae) in rido, 1982, im Terrariummit einer Ubersicht fiber die Gattung Chamaeleolis auf Kuba. Herpetofauna (Weinst) 13: 29-33. kangaroos and wallabies of the genus Macropus (Marsupialia: MaR. 1968. Parasitological studies in British Honduras, IV. Some cropodidae). International Journal for Parasitology 19: 241-263. LAINSON, BOVEE,E. C. 1966. Eimeria urosauris n. sp., a coccidium from the coccidial parasites of reptiles. Annals of Tropical Medicine and lizard Urosaurus graciosus Hollowell, in California. Journal of Parasitology 62: 260-266. R. R., R. L. MORRISON, S. R. GOLDBERG, ANDS. J. UPTON. Protozoology 13: 607-610. MITSCHLER, .1969. Eimeria umis n. sp., a coccidian parasite from the lizard 1993. Redescription of Eimeria sceloporis (Apicomplexa: EimeriUma notata Baird. Transactions of the American Microscopical idae) from a new host Sceloporus jarrovii (Sauria: PhrynosomatiSociety 88: 257-265. dae). The Texas Journal of Science 45: 356-358. 1965. Eimeria sceloporis and Eimeria mol, ANDS. R.TELFORD. L. P 1974. Coccidia and Coccidiosis. P. Parey, Berlin, HamPELLERDY, ochis sp. n. from lizards. Journal of Parasitology 51: 85-94. burg, Germany, 959 p. ANDA. LATHROP. RuIz, A. 1959. Eimeria intermedia n. sp., parasita de la lagartija Anolis D. D. SMITH,J. S. PARMERLEE, BuI, H. T, R. POWELL, 1992. A new coccidian parasite (Apicomplexa: Eimeriorina) from intermedius Peters. Revista de Biologia Tropical 7: 109-112. Anolis distichus (Sauria: Polychrotidae) in the Dominican Republic. 1985. A guide to the identification SCHWARTZ, A., ANDR. W. HENDERSON. of the amphibians and reptiles of the West Indies exclusive of HisJournal of Parasitology 78: 784-785. CARINI,A. 1940. Sombre um Eimeria de um lagarto de Goias. Archivos paniola. Milwaukee Public Museum, Milwaukee, Wisconsin, 165 p. S. R. 1998. Coccidian parasites of four Panamanian lizard de Biologia, Sao Paulo 24: 282. TELFORD, 1995. Four CISPER,G. L., C. HUNTINGTON,D. D. SMITH,ANDR. POWELL. species. Systematic Parasitology 40: 195-202. new coccidia (Apicomplexa: Eimeriidae) from anoles (Lacertilia: UPTON,S. J., P S. FREED, AND M. G. FREED. 1989. Two new species of coccidia (Apicomplexa: Eimeriorina) from Peruvian reptiles. TransPolychrotidae) in the Dominican Republic. Journal of Parasitology actions of the American Microscopical Society 108: 96-100. 81: 252-255.

rounded than those of E. chamaeleolidisbarbati. Additionally, all 3 of the aforementioned species of Eimeria are described distant and systematically from hosts that are geographically from C. barbatus. Eimeria schwartzi Cisper, Huntington, Smith, and Powell, 1995 from Anolis armouri, Eimeria avilae Cisper, Huntington, Smith, and Powell, 1995 from Anolis olssoni, and Eimeria anolidis Daszak and Ball, 1991 from Anolis carolinensis possess larger oocysts that, in the case of E. anolidis and E. avilae, also differ significantly in shape. Based on the oocyst shape index, both E. anolidis and E. avilae parasitize the gall bladder (Bovee and Telford, 1965; Telford, 1998). The same criterion suggests that E. chamaeleolidisbarbati probably develops in the intesand tine. Based on these observations, both I. chamaeleolidis are considered to be new species. E. chamaeleolidisbarbati