ISOSPORA TAIZII (APICOMPLEXA: EIMERIIDAE), A NEW COCCIDIAN PARASITE FROM THE YEMEN CHAMELEON (CHAMELEON CALYPTRATUS) (SAURIA: CHAMAELEONIDAE)

IN TAIZ CITY, YEMEN REPUBLIC

Badria M. Abdel-Wasae

Biology Department, Faculty of Science, Taiz University, Yemen Republic

INTRODUCTION:

                Coccidial parasites of the genus Isospora (Apicomplexa: Eimeriidae) are the cause of intestinal disease, widely distributed among reptiles (Amoudi, 1993; Modry et al., 1998; Paperna & Lainson, 2000 and McQuisstion et al., 2001), Family: Chamaeleonidae (Modry & Koudela, 1995; Modry et al., 1997 & 1999 & 2000 and 2001), birds (Ball & Daszak, 1997 and Upton et al., 2001), mammalian host species (Sayd & Kawazoe, 1998 and Mundt et al., 2003) and on humans which were given recently by (Frenkel et al., 2003 and Resiere et al., 2003).

                Coccidia infection causes a watery diarrhea, which is sometimes bloody. These protozoan parasites have asexual and sexual stages within the intestinal cells of their hosts and produce an environmentally resistant cyst stage, the oocyst. Infections are acquired by the ingestion of infective stage (sporulated oocysts) in contaminated food or water. The sporulated oocyst breaks open and release eight sporozoites. This sporozoite each finds an intestinal cell and begins to reproduce inside it. Ultimately, the cell is so full of what are called “merozoites” that it bursts releasing the merozoites, which seek out their own intestinal cells, and the process begins again. It is important to note how thousands of intestinal cells can become infected and destroyed as a result of accidentally swallowing a single oocyst.

                Only one species of Isospora was described from saurian hosts in Yemen chameleon by Modry & Koudela (1995). This paper describes the oocysts of Isospora taizii n. sp. from the faeces of the Yemen chameleon Chameleon calyptratus (Sauria: Chamaeleonidae), and compared their structure to those reported from other members of this family.

MATERIALS AND METHODS:

                The Yemen chameleons Chameleon calyptratus were captured alive. This animal is a large, aggressive species, the veiled chameleon occupies the wadis and agricultural lands of this otherwise arid region. The nominate form, Chameleon calyptratus is found in the more southern reaches of the distribution in Yemen and southwestern Saudi Arabia (Le Berre, 1994 and Barnett et al., 1999).

                The alimentary canal of each animal was opened. Smears from the intestinal contents were prepared from successive parts and examined microscopically. For determination of the sporulation time, oocysts were collected from faeces and strained through sieve. After staining, the suspension including the water used in washing the sieve, was allowed to undergo sedimentation after which the water was siphoned off. The sediment was then suspended in 2.5% Potassium dichromate solution (K₂Cr₂O₇) to sporulate. The suspension was then spread in a thin layer in petri dishes and kept at room temperature 25-28°C for sporulation. Sporulated oocysts were examined with a Zeiss universal photomicroscope with 100X Planapochromatic oil immersion objective lenses. The oocysts were measured with ocular micrometer. All measurements are in mm and given as means, followed by the range in parentheses and the shape-index (ratio of length /width).

RESULTS:

                During a limited survey for coccidian infections in some of the reptiles in Taiz region between March and July 2003, faeces samples of 28 Yemen chameleons were examined by cover slip floatation with Sheather’s solution (Levine, 1973). From the 28 Yemen chameleon, 18 animals (64.3%) were infected with Isospora taizii n. sp. detailed examination of sporulated oocysts revealed apparently new species of Isospora, which does not resemble either any other species described from chameleons or published descriptions of any species of Isospora found in other chameleon.

                Examination of freshly shedded, non-sporulated oocysts in the faeces collected from several individuals show the cytoplasm of the zygote was containing large granules mostly at the periphery and smaller ones in the center. No nucleus could be seen in freshly passed oocysts (Fig. 1). These oocysts were subspherical to ovoid in shape, measured 28 (25-32) mm in length and 22 (18-26) mm in width. The oocyst wall is bilayered, smooth and colourless approximately 1.2 mm thick. The outer layer was light and thin, while the inner layer was dark and thick.

                No micropyle or polar granules were observed. At the beginning of the sporulation process, the cytoplasm condensed (Fig. 2), divided, and two sporoblasts were formed without leaving an oocyst residuum. Sporoblasts developed forming sporocysts. As sporulation proceeded, four sporozites developed within each sporocyst (Figs. 3&4). Later on, the sporoblasts begin to elongate with the formation of sporocysts. These sporocysts were ellipsoidal in shape. Then, differentiation of sporozoites took place (Fig. 4). The sporocysts measured 14-15 mm in length and 10-12 mm in width, with a mean length× width 12.8×9.2 mm. The sporocyst wall is single layered, thin, smooth and colourless. A sporocyst residual body and a steida and substeida bodies were present. Each sporozoite contains spheroid anterior refractiel body and spheroid or ellipsoid posterior refractiel body, nucleus located between the refractile bodies. Fully sporulated oocysts are observed within 48 hours at 26°C.

Explanation of Figures

Figs. (1-4): Photomicrograph for events of sporoulation process for the subspherical to ovoid oocysts Isospora taizii n. sp. All photos. × 1600.

Fig. (1): Fresh non-sporulated oocyst with zygote (ZY) nearly filling the space of oocyst. The oocyst is enclosed with bilayered wall, outer layer (OL) and the inner layer (IL) the oocyst wall (OW).

Fig. (2): Two sporoblasts (SB) are found each with four sporozoites.

Figs. (3,4): Full sporulated oocysts of Isospora with two sporocysts (SPC) each with four sporozoites (SP). Notice: residual body (RB) in sporocysts, Steida body (ST) and substeida body in the sporocyst. Residuum bodies absent in the oocysts but present in the sporocysts.

Taxonomic summary of Isospora taizii n. sp. :

Host type: Yemen chameleon (Chameleon calyptratus) Sauria: Chamaeleonidae.

Location type: Taiz Governorate, Yemen Republic.

Prevalence: found in 18 of 28 (64.3%).

Sporulation time: 48 hours at 26°C.

Site of infection: small and large intestine.

Sporulation: Exogenous oocysts were passed unsporulated and fully sporulated oocysts were observed within 48 hours at 26°C in 2.5% K₂Cr₂O₇.

Etymology: The specific name is derived from the collection locality, Taiz Governorate.

DISCUSSION:

                About 65 named Isospora species have been described to date from saurians. Only 6 Isospora spp. were described from the Family: Chamaeleonidae (Chameleons) (Duszynski and Couch, 2003). Isospora mesnili from Chamaeleon chamaeleon (Sergent, 1902); Isospora jaracimrmani from Chameleon calyptratus (Modry and Koudela, 1995), Isospora tigris from Calumma tigris (Tiger chameleon) (Modry et al., 1997); Isospora wildi from Chamaeleon dilepis (Flap-necked chameleon) (Modry et al., 2000); Isospora munriyu from Chamaeleon jacksonii (Jackson’s shameleon) (Modry et al., 2000) and Isospora necasi from Chamaeleon melleri (Meller’s chameleon) (Modry et al., 2000). However, Modry et al., (1999) described one, Isospora chamaeleolidis n. sp., from Chaamaeleolis barbatus also, Modry et al., (2001) described one Isospora brygooi n. sp., from five new coccidian species infecting Madagascar chameleons of the genera Furcifer and Brookesia.

                Although, the oocyst shape of Isospora taizii n. sp. is subspherical to ovoid only similar with Isospora wildi and Isospora necasi (Modry et al., 2000). The absence of oocyst residuum body and the ellipsoidal shape of the sporocyst in the present study was similar to Isospora wildi; Isospora munriyu and Isospora necasi (Modry et al., 2000) and Isospora brygooi (Modry et al., 2001). In addition, the presence of sporocyst residuum, Stieda and substieda bodies was similar to four Isospora species from the chameleon (Isospora wildi; Isospora muriyu and Isospora necasi (Modry et al., 2000) and Isospora brygooi (Modry et al., 2001). However, the primary differences center on the size of the oocyst, Isospora taizii 28 (25-32)× 22 (18-26) mm, but in Isospora Isospora chamaeleolidis 16.1(13-21)×15.6 (13-19) mm (Modry et al., 1999) ; Isospora wildi 25 (22-28)× 21.4 (18-24) mm. ; Isospora necasi. 26.6 (21-30)×24 (20-27) mm. and Isospora muriyu 23 (21-25)×21.9 (21-23) mm (Modry et al., 2000), and Isospora brygooi 20.7 (17-24.5)× 19.3 (16-23) mm (Modry et al., 2001). Although, the surface of oocyst wall of the Isospora taizii is smooth double layer 1.2 mm thick; that of the oocyst of Isospora chamaeleolidis bilayered 1.0-1.5 mm thick; Isospora necasi is a velvetlike wall 2 mm; Isospora munriyu is a finely granulated wall 1.5 mm thick; Isospora brygooi is a slightly pitted wall, and Isospora wildi is a smooth wall 1 mm thick. So, it is obvious that the Isospora species found in the present host genus Chameleon calyptratus belong to a new species and it is suggested to be named Isospora taizii according to the locality.

Comparative data of Isospora sp. from Family: Chamaeleonidae (Chameleons)

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