Sam R. Telford

Haemoparasites of Reptiles

The haemoparasites of reptiles include representatives of all taxonomic groups which parasitize the other vertebrate classes: trypanosomatid flagellates, haemogregarines, haemosporidia and filariid nematodes, as well as a heterogeneous assemblage of organisms or inclusions which possibly contains viruses, bacteria, rickettsiae, piroplasmids and probably artifacts of staining as well.

Gastrointestinal Helminths of 14 Species of Lizards from Panama with Descriptions of Five New Species

Abstract One hundred ninety-one specimens representing 14 species of lizards, Ameiva ameiva, Basiliscus basiliscus, Corytophanes cristatus, Diploglossus monotropis, Echinosaura horrida, Gonatodes albogularis, Gymnophthalmus speciosus, Iguana iguana, Lepidoblepharis sanctaemartae, Lepidophyma flavimaculatum, Leposoma rugiceps, Mabuya mabouya, Polychrus gutturosus, and Thecadactylus rapicauda, from Panama were examined for helminths. Two species of Digenea, Mesocoelium monas and Parallopharynx arctus, 1 species of Cestoda, Oochoristica gymnophthalmicola n. sp., 21 species of Nematoda, 19 species capable of completing their life cycle in lizards, Africana telfordi, Aplectana herediaensis, Cosmocercoides variabilis, Cruzia mexicana, Cyrtosomum longicaudatum, Macdonaldius grassi, Oswaldocruzia panamaensis n. sp., Oswaldofilaria brevicaudata, Ozolaimus cirratus, Parapharyngodon colonensis n. sp., Physaloptera retusa, Piratuba digiticauda, Skrjabinelazia galliardi, Skrjabinodon caudolumarius n. sp., Skrjabinodon crassicauda n. sp., Skrjabinodon scelopori, Spauligodon oxkutzcabiensis, Spinicauda spinicauda, and Strongyluris panamaensis, and 2 species utilizing lizards as paratenic hosts, Ophidascaris sp. (larvae) and Acuariidae gen. sp. (larvae in cysts), and 1 species of Acanthocephala (cystacanths), were found. Thirty-seven new host records and 13 new locality records are reported.

Characterization of an erythrocytic virus in the family Iridoviridae from a peninsula ribbon snake (Thamnophis sauritus sackenii)

A wild peninsula ribbon snake (Thamnophis sauritus sackenii) in Florida was found to have hypochromic erythrocytes containing two different types of inclusions: purple granular inclusions, and pale orange or pink crystalloid inclusions that were round, oval, rectangular, or hexagonal in shape. Transmission electron microscopy revealed hexagonal or pleomorphic, homogenous inclusions and enveloped particles morphologically consistent with a member of the Iridoviridae. Histopathology of the animal revealed necrotizing hepatitis consistent with sepsis. Consensus PCR was used to amplify a 628-bp region of iridoviral DNA-dependent DNA polymerase. Bayesian and maximum likelihood phylogenetic analysis found that this virus was distinct from other known iridoviral genera and species, and may represent a novel genus and species.

Hepatozoon Species of the Timber Rattlesnake in Northern Florida: Description of a New Species, Evidence of Salivary Gland Oocysts, and a Natural Cross-Familial Transmission of an Hepatozoon Species

Two species of Hepatozoon, i.e., H. sauritus and H. horridus n. sp., were present in 1 of 8 timber rattlesnakes, Crotalus horridus. The narrow gamonts of H. sauritus are 15.0–19.0 × 3.5–5.0 μm, with LW 58–86 μm2 and L/W 3.2–4.7, with a narrow, rounded anterior end. The spherical to slightly ovoid oocysts produce ovoid to elongate sporocysts, 21–43 × 12–24 μm, L/W 1.20–2.7, containing on average 22.1 (10–34) sporozoites. This is the first report of a natural cross-familial transfer of a Hepatozoon species. Gamonts of H. horridus n. sp. are 13.0–17.0 × 4.0–6.0 μm, with LW 63–102 μm2 and L/W 2.6–4.0, and have broadly rounded ends. The gamont cytoplasm is vacuolated. The spherical to ovoid oocysts form spherical to elongate sporocysts 14–45 × 11–25 μm, L/W 1.0–2.3, producing an average of 13.0 (8–21) sporozoites. The salivary gland in 1 of 5 mosquitoes dissected contained 1 mature oocyst.

A new Haemogregarina species of the alligator snapping turtle, Macrochelys temminckii (Testudines: Chelydridae), in Georgia and Florida that produces macromeronts in circulating erythrocytes.

Abstract Haemogregarina macrochelysi n. sp. (Apicomplexa: Haemogregarinidae) of the alligator snapping turtle, Macrochelys temminckii, is characterized by slender, recurved gamonts 29–35 × 3–4.5 μm, in which the anterior limb comprises 48–54% of the total length. The gamont nucleus, 5–7.5 × 2–5 μm, is situated at approximately midbody of the gamont. Meronts typical of Haemogregarina occupying erythrocytes have 3-8 small, compact nuclei and are 13–17 × 4.5–9 μm. Erythrocytic meronts that contain larger, nearly square or rectangular nuclei become rounded, and then undergo 7 or more nuclear divisions, which produce very large, usually ovoid to rounded meronts that may contain up to 150 nuclei or more within the thinly stretched host erythrocyte membrane. In tissues of the Placobdella spp. leech vectors, merogony occurs directly from sporozoites, forming merozoites that presumably are infective for the turtle host.

TWO ADDITIONAL HEPATOZOON SPECIES (APICOMPLEXA: HEPATOZOIDAE) FROM THE SOUTHERN BLACK RACER, COLUBER CONSTRICTOR PRIAPUS (SERPENTES: COLUBRIDAE), IN NORTHERN FLORIDA

Hepatozoon priapus n. sp. from Coluber constrictor priapus has robust gamonts with broadly rounded ends, 18.0 × 4.2 μm (17.0–20.0 × 3.5–6.0), with LW 76.4 μm2 (59–105) and L/W 4.31 (2.9–5.4). The nucleus is always present in second quarter of gamont, seldom extend into first quarter but often into third quarter, 6.0 × 3.0 (5.0–7.0 × 2.5–4.0), with LW 17.9 (13.7–21.0). Erythrocyte cytoplasm is always thin, appearing dehemoglobinized, with infected cells always distorted. Infected erythrocytes are much longer and wider than uninfected cells, with longer nuclei. Oocysts are spherical to ovoid, 92.5 × 86.0 (55–123 × 47–115) and L/W 1.08 (1.0–1.3), contain 14.0 (6–31) sporocysts. Sporocysts, which are also spherical to ovoid, 26.3 × 23.3 (19–50 × 16–38), LW 641.2 (320–1,500) and L/W 1.13 (1.0–2.2), contain 12.6 (5–18) sporozoites. Hepatozoon confusus n. sp., also from C. constrictor priapus, has slender gamonts with rounded ends, 15.6 × 4.1 (14.0–17.0 × 3.5–5.0), with LW 64.3 (52–80) and L/W 3.82 (2.8–4.4). The nucleus is always present in second quarter of gamont, commonly extending into first and third quarters, 5.0 × 2.7 (2.5–4.4 × 4.0–6.0), with LW 13.5 (11.0–16.5). Erythrocyte cytoplasm is sometimes thin, appearing partially dehemoglobinized, with infected cells usually distorted. Infected erythrocytes are longer than uninfected cells but similar in width, with erythrocyte nuclei longer. Oocysts are spherical to ovoid, 115.5 × 108.9 (52–278 × 50–278), with L/W 1.06 (1.0–1.2), and contain 25.0 (7–111) sporocysts. Sporocysts are spherical to ovoid, 27.6 × 25.2 (21–38 × 20–33), LW 701.3 (420–1,125) and L/W 1.09 (1.0–1.4), containing 20.2 (12–32) sporozoites.

A HAEMOCYSTIDIUM SPECIES FROM THE EAST AFRICAN GECKO LYGODACTYLUS CAPENSIS GROTEI

Haemocystidium lygodactyli n. sp. parasitizes Lygodactylus capensis grotei (Gekkonidae) in Tanzania. Mature gametocytes in acute phase of infection average 16.3 × 5.7 μm (11–20 × 4–9.5 μm), with LW 93.0 (62–140 μm2) and L/W ratio 2.94 (1.2–3.9). Gametocytes usually lateral, lateropolar, or halteridial in position. There was no significant sexual dimorphism in gametocyte dimensions. Nuclei discrete in both sexes at maturity, with a rounded nucleolus usually present in microgametocytes. In chronic infection, gametocytes were 18.1 × 8.7 μm (8–25 × 5–11 μm), with LW 156.8 μm2 (80–250) and L/W 2.16 (1.1– 3.6). When gametocytes from the chronic infection were compared with the same sex in acute infection, length did not differ, but differences were present between the same sex in each comparison of width, LW, and L/W. Macrogametocytes and microgametocytes in chronic phase were broader, larger, and less elongate and most commonly halteridial. Meronts were found only in endothelium and connective tissue of lung. Elongate to oval in shape, the larger meronts filled with nuclei were 12.2 × 6.9 μm (10.0 × 5.0–16.0 × 9.0), with LW 50–144 μm2 (85.1). In 1 initial infection followed for 49 days, apparently mature gametocytes appeared by day 28 postcapture. Binucleate parasites were present from day 14 throughout the course of infection, with their frequency increasing from 5% of immature parasites to 34% of mature gametocytes. Binucleate mature gametocytes were found in 1 other infection, where 14% had 2 nuclei. Sex ratio varied from 51 to 63% in favor of macrogametocytes.

REDESCRIPTION OF HAEMOPROTEUS MESNILI (APICOMPLEXA: PLASMODIIDAE) AND ITS MERONTS, WITH DESCRIPTION OF A SECOND HAEMOSPORIDIAN PARASITE OF AFRICAN COBRAS

Haemoproteus mesnili (Bouet 1909) Wenyon 1926 is redescribed from the spitting cobra, Naja nigricollis nigricollis, of Tanzania. Mature gametocytes in the acute phase of infection averaged 17.7 X 7.3 jim, with LW 128.1 jim-, and L:W ratio 2.52. Nuclei were visible in both sexes. Both sexes were heavily pigmented, with 31-62 black granules dispersed in macrogametocytes; 20-46 granules were often clumped or concentrated near ends of microgametocytes. The halteridial form was present in 28% of active-phase gametocytes, but in only 8% of those in chronic phase. A few large, possibly first generation, meronts were present in cardiac muscle; uninucleate parasites within parasitophorous vacuoles in splenic cells produced small rounded or ovoid meronts, 12.2 x 9.6 microm, with 12-16 deeply basophilic, square-to-rectangular cytomeres. Meronts with 17-32 cytomeres were 16.9 x 11.9 microm. Meronts, 20 x 16 to 26 x 22 microm, contained 51-57 cytomeres. Mature meronts were ovoid, 13.7 x 11.5 microm, with many rounded merozoites. Haemoproteus balli n. sp, found in an Egyptian cobra, Naja haje haje of Kenya, differs from H. mesnili in average gametocyte dimensions, 10.8 x 7.7 microm; LW, 83.2 microm2; L/W ratio, 1.42; absence of halteridial forms; sparse pigmentation (3-10 granules); and presence of a broad peripheral band, apparently chromatin, along one side of microgametocytes.Haemoproteus mesnili (Bouet 1909) Wenyon 1926 is redescribed from the spitting cobra, Naja nigricollis nigricollis, of Tanzania. Mature gametocytes in the acute phase of infection averaged 17.7 7.3 m, with LW 128.1 m2, and L:W ratio 2.52. Nuclei were visible in both sexes. Both sexes were heavily pigmented, with 31–62 black granules dispersed in macrogametocytes; 20–46 granules were often clumped or concentrated near ends of microgametocytes. The halteridial form was present in 28% of active-phase gametocytes, but in only 8% of those in chronic phase. A few large, possibly first generation, meronts were present in cardiac muscle; uninucleate parasites within parasitophorous vacuoles in splenic cells produced small rounded or ovoid meronts, 12.2 9.6 m, with 12–16 deeply basophilic, square-to-rectangular cytomeres. Meronts with 17–32 cytomeres were 16.9 11.9 m. Meronts, 20 16 to 26 22 m, contained 51–57 cytomeres. Mature meronts were ovoid, 13.7 11.5 m, with many rounded merozoites. Haemoproteus balli n. sp, found in an Egyptian cobra, Naja haje haje of Kenya, differs from H. mesnili in average gametocyte dimensions, 10.8 7.7 m; LW, 83.2 m2; L/W ratio, 1.42; absence of halteridial forms; sparse pigmentation (3–10 granules); and presence of a broad peripheral band, apparently chromatin, along one side of microgametocytes. Plasmodium mesnili was described by Bouet (1909) from a cobra, either a species of Naja or Sepedon haemachatus, in West Africa, from 2 localities: Odienne, Ivory Coast, and Gaoua, Upper Volta. Wenyon (1909) described Haemocystidium najae from Naja haje and Naja nigricollis from 2 sites along the River Sobat in Sudan. Leger and Leger (1914) reported P. mesnili from Sepedon haemochotes ( haemachatus, not haemocholis as stated by Wenyon, 1926), probably from the vicinity of their laboratory in Bamako, now Mali, but then part of Haut Senegal-Niger. Macfie (1919) reported the parasite from N. nigricollis at Accra, Gold Coast, now Ghana. Wenyon (1926) recognized the priority of Bouet’s 1909 description to his own of that year, and referred to this parasite as Haemoproteus mesnili, as did Garnham (1966) and Ball (1967). Battelli (1949) found H. mesnili in 2 species of cobras in Eritrea, and it was mentioned by Huygelen and Mortelmans (1958) as present in N. haje of Katanga Province, Congo. There appear to be no other reports of H. mesnili in the literature. Haemoproteus mesnili is reported only from erythrocytic stages, and nothing is known of its life cycle. It is a common parasite of N. nigricollis, the spitting cobra, in Tanzania, and examination of tissues from infected snakes has provided information on the vertebrate phase of the life cycle that justifies its inclusion within Haemoproteus. The species is redescribed below in detail. Reexamination of the parasite identified by Ball (1967) has demonstrated that it is not H. mesnili, and it is described as a second haemosporidian parasite of African cobras. MATERIALS AND METHODS Seven of 8 N. nigricollis specimens collected on, or around, the Sokoine University campus on the north slope of the Uluguru Mountains, Morogoro, Tanzania, from 1981 to 1985 were found to be infected by H. mesnili; in 3 of these snakes, H. mesnili was present in mixed infection with a Hepatozoon species. Blood samples were collected by cardiac puncture from snakes captured alive and from the heart in the case of road-killed cobras. One cobra was sampled twice over a period of 12 days. Slides were fixed in absolute methanol, and stained using Giemsa stain at dilution of 1:10 parts stain to distilled water, pH 7.0, for 1 hr. Some slides were stained by the May–Grunwald Giemsa proReceived 16 August 2006; revised 12 December 2006; accepted 12 December 2006. cedure. Impression smears of tissues were stained by Giemsa, but at a dilution of 1:20. Tissue samples were fixed in buffered formalin, sectioned by standard procedures at 5 m, and stained by H&E, or by Giemsa using the Kimsey (1992) technique. Slides were screened at 400, and parasites studied and photographed at 1,000, except where noted in figures. All measurements were obtained by a calibrated ocular micrometer and are reported in micrometers with means and SD followed by ranges in parentheses, or in the case of LW (length width) values, square micrometers. Comparisons were done by 1-way ANOVA on transformed data (log10 x 1), with significance P 0.05. Differences noted under ‘‘Remarks’’ below (smaller, larger, greater, less, etc.) are statistically significant; the term ‘‘similar to’’ indicates that significant difference was not present. Taxonomic characters employed were those used by Telford (1996). Sex ratios were determined from samples of 100 or more gametocytes. A host voucher series was deposited in the herpetology collection of the Florida Museum of Natural History (UF 59666, 59668–59669, 59671–59672). Neohapantotype slides were deposited in the U.S. National Parasite Collection, Beltsville, Maryland (USNPC).

NEW SPECIES OF SCHULZIA (NEMATODA: MOLINEIDAE) IN PTYCHOGLOSSUS FESTAE (SQUAMATA: GYMNOPHTHALMIDAE) FROM PANAMA

Schulzia ptychoglossi n. sp. (Strongylida: Molineidae) from the intestines of Ptychoglossus festae (Squamata: Gymnophthalmidae) is described and illustrated. Schulzia ptychoglossi n. sp. represents the fourth species assigned to the genus and is most similar to the Venezuelan species S. usu by possessing a cervical inflation that begins a short distance from the anterior end of the body. Schulzia ptychoglossi differs from S. usu in that ray 8 separates midway between the root and tip of the dorsal ray in S. ptychoglossi,, but separates close to the root of the dorsal ray in S. usu.

TWO PLASMODIUM SPECIES OF THE CROCODILE SKINK TRIBOLONOTUS GRACILIS FROM IRIAN JAYA, INDONESIA

Two undescribed Plasmodium species were present in 1 of 8 New Guinea skinks, Tribolonotus gracilis. Plasmodium tribolonoti n. sp. is characterized by rounded or oblong schizonts 6.1 × 5.3 μm (5–7 × 4–7) that produce on average 14.3 merozoites (10–21), in no particular arrangement of nuclei. All parasitized proerythrocytes and had no effect upon dimensions of host cells or their nuclei. Gametocytes, mostly erythrocytic, averaged 7.2 × 6.3 (6.5–9.0 × 5.5–7.5), with length × width (LW) 45.5 μm2 (38–63) and L/W 1.15 (1.0–1.5). Gametocytes are not sexually dimorphic in size or shape and had no effect upon dimensions of host erythrocytes or their nuclei. Schizonts of P. gracilis n. sp. averaged 4.3 × 3.5 (3–6 × 3–5), with 4.9 merozoites (3–8) usually arranged as a fan, and had no effect upon dimensions of host erythrocytes or their nuclei. Gametocytes averaged 5.9 × 5.5 (5.0–6.6 × 5.0–6.0), with LW 31.9 μm2 (25–40) and L/W 1.06 (1.0–1.2). Gametocytes are not sexually dimorphic in size or shape and had no effect upon dimensions of host erythrocytes or their nuclei.