Comparative molecular analyses of Eimeria Schneider (Apicomplexa: Eimeriidae) species from rock ptarmigan in Iceland, Svalbard-Norway, and Japan.

The rock ptarmigan (Lagopus muta) has a Holarctic breeding distribution and is found in arctic and sub-arctic regions. Isolated populations and glacial relicts occur in alpine areas south of the main range, like the Pyrenees in Europe, the Pamir mountains in Central Asia, and the Japanese Alps. In recent decades considerable effort has been made to clarify parasite infections in the rock ptarmigan. Seven Eimeria spp. have been reported parasitizing rock ptarmigan. Two of those species, E. uekii and E. raichoi parasitizing rock ptarmigan (L. m. japonica) in Japan, have been identified genetically. Here we compare partial sequences of nuclear (18S rRNA) and mitochondrial (COI) genes and we detail the morphology of sporulated oocysts of E. uekii and E. raichoi from Japan, E. muta and E. rjupa, from the rock ptarmigan (L. m. islandorum) in Iceland, and two undescribed eimerian morphotypes, Eimeria sp. A, and Eimeria sp. B, from rock ptarmigan (L. m. hyperborea) in Norway (Svalbard in the Norwegian Archipelago). Two morphotypes, ellipsoidal and spheroidal, are recognized for each of the three host subspecies. Our phylogenetic analysis suggests that the ellipsoidal oocyst types, E. uekii, E. muta, and Eimeria sp. A (Svalbard-Norway) are identical and infects rock ptarmigan in Japan, Iceland, and Svalbard-Norway, respectively. Eimeria uekii was first described in Japan in 1981 so that E. muta, described in Iceland in 2007, and Eimeria sp. A in Svalbard-Norway are junior synonyms of E. uekii. Also, phylogenetic analysis shows that the spheroidal oocyst types, E. rjupa and Eimeria sp. B (Svalbard-Norway), are identical, indicating that rock ptarmigan in Iceland and Svalbard-Norway are infected by the same Eimeria species and differ from E. raichoi in Japan.

CORRECTING THE NAMES OF TWO HAEMOGREGARINA SPP. FROM LIZARDS IN EGYPT.

Two haemogregarine "species" names, Haemogregarina tarentannulari and Haemogregarina rawashi, were cited by Saoud et al. (1995) as having been described by Mohammed and Ramadan (1996, in press). However, the paper by Mohammed and Ramadan (1996) was never published and, therefore, these names and their authorities must be suppressed because they violate Chapter 3 (Criteria for Publication), Article 8 (What Constitutes Published Work) of the International Code of Zoological Nomenclature. The following new names are introduced to replace them based on the Principle of Priority (Chapter 6, Article 23, 23.1, and Chapter 11, Article 51, Recommendation 51E): Hepatozoon rawashi (Mohammed and Ramadan in Saoud, Ramadan, Mohammed and Fawzi, 1995) n. comb., with gamonts in the erythrocytes and meronts in the lungs and liver of the fan-footed gecko, Ptyodactylus hasselquisiti (Donndorff, 1798) from Egypt, and Haemogregarina tarentannulari (Mohammed and Ramadan in Saoud, Ramadan, Mohammed and Fawzi, 1995), with gamonts in the erythrocytes and meronts in the lungs and liver of the white-spotted wall gecko, Tarentola annularis (St. Hilaire, 1827) also from Egypt. This latter species was determined to be a junior synonym of Haemogregarina annularis El-Naffar, Mandour, and Mohammed 1991, which was later reassigned to the genus Hepatozoon based on their phylogenetic analysis of 18S rDNA gene sequences.

Coccidian parasites in the endangered Forest Musk Deer (Moschus berezovskii) in China, with the description of six new species of Eimeria (Apicomplexa: Eimeriidae).

We examined 674 fresh fecal samples from forest musk deer (Moschus berezovskii Flerov) in Sichuan and Shaanxi Provinces, China, for coccidian oocysts and 65% were infected with Eimeria spp. Previously, only four Eimeria species were known from Moschus spp. Here we describe six new Eimeria species. Eimeria aquae n. sp., in 38% deer, has ovoidal oocysts, 32.0 × 23.0 µm, micropyle (M) and scattered polar granules (PGs) of various sizes are present, sometimes oocyst residuum (OR) is present; ovoidal sporocysts, 14.1 × 7.5 µm, with Stieda body (SB) and sporocyst residuum (SR). Eimeria dolichocystis n. sp., in 11% deer; cylindroidal oocysts, 36.6 × 18.9, with a M, 1 PG and OR; ovoidal sporocysts, 13.9 × 7.7, with SB and SR. Eimeria fengxianensis n. sp., in 7% deer; ovoidal oocysts, 36.3 × 25.2, a M and PGs present but OR absent; ovoidal sporocysts, 13.9 × 7.3, with SB and SR. Eimeria helini n. sp. in 24% deer; subspheroidal oocysts, 27.0 × 24.1, OR and PGs often present, but M absent; ovoidal sporocysts, 13.5 × 7.7, with SB and SR. Eimeria kaii n. sp. in 26% deer; ovoidal oocysts, 33.2 × 20.7, M and PGs present, but OR absent; ovoidal sporocysts, 14.4 × 7.5, with SB and SR. Eimeria oocylindrica n. sp., in 17% deer; cylindroidal oocysts, 36.0 × 21.4, M and 1-2 PGs present but OR absent; ovoidal sporocysts, 13.8 × 7.7, with SB and SR. Eimeria dujiangyanensis n. nom. is proposed to replace E. moschus Sha, Zhang, Cai, Wang & Liu, 1994, a junior homonym of E. moschus Matschoulsky, 1947.

TITLE: Coccidies parasites du cerf porte-musc (Moschus berezovskii), une espèce en danger en Chine, avec la description de six nouvelles espèces d'Eimeria (Apicomplexa, Eimeriidae). ABSTRACT: Nous avons examiné 674 échantillons de matières fécales fraîches de cerf porte-musc (Moschus berezovskii Flerov) dans les provinces du Sichuan et du Shaanxi, en Chine, pour les oocystes de coccidies et 65 % étaient infectés par Eimeria spp. Auparavant, seules quatre espèces d'Eimeria étaient connues chez Moschus spp. Nous décrivons ici six nouvelles espèces d'Eimeria. Eimeria aquae n. sp., chez 38 % des cerfs, a des oocystes ovoïdes, 32,0 × 23,0 µm, micropyle (M) et granules polaires (PG) de différentes tailles présents, parfois des résidus d'oocystes (OR) présents; sporocystes ovoïdes, 14,1 × 7,5 µm, avec corps de Stieda (SB) et résidu de sporocyste (SR). Eimeria dolichocystis n. sp., chez 11 % des cerfs; oocystes cylindroïdes, 36,6 × 18,9, avec un M, 1 PG et OR; sporocystes ovoïdes, 13,9 × 7,7, avec SB et SR. Eimeria fengxianensis n. sp., chez 7 % des cerfs; oocystes ovoïdes, 36,3 × 25,2, M et PG présents mais OR absent; sporocystes ovoïdes, 13,9 × 7,3, avec SB et SR. Eimeria helini n. sp. chez 24 % des cerfs; oocystes subsphéroïdaux, 27,0 × 24,1, OR et PG souvent présents, mais M absent; sporocystes ovoïdes, 13,5 × 7,7, avec SB et SR. Eimeria kaii n. sp. chez 26 % des cerfs; oocystes ovoïdes, 33,2 × 20,7, M et PG présents, mais OR absent; sporocystes ovoïdes, 14,4 × 7,5, avec SB et SR. Eimeria oocylindrica n. sp., chez 17 % des cerfs; oocystes cylindroïdes, 36,0 × 21,4, M et 1-2 PG présents mais OR absent; sporocystes ovoïdes, 13,8 × 7,7, avec SB et SR. Eimeria dujiangyanensis n. nom. est proposé pour remplacer E. moschus Sha, Zhang, Cai, Wang & Liu, 1994, homonyme junior de E. moschus Matschoulsky, 1947.

Hyaloklossia Labbé, 1896 (Alveolata: Apicomplexa) in frogs: Description of a new species and proposing a new subfamily to accommodate these enigmatic parasites.

Hyaloklossia Labbé ,1896 (Alveolata: Apicomplexa) is a monotypic genus of renal coccidia found in anurans, particularly in the edible frog Pelophylax kl. esculentus (Amphibia: Anura: Ranidae), distributed in different parts of Europe. Here we propose a new Hyaloklossia species from the Tokyo daruma pond frog, Pelophylax porosus porosus. The coccidium detected in the renal tissue of P. p. porosus shared some morphological characteristics with the type species, Hyaloklossia lieberkuehni (Labbé, 1894), reported from P. kl. esculentus. However, in addition to size differences in several oocyst and sporocyst features between these parasites, phylogenetic analysis of gene fragments from two nuclear ribosomal loci and the mitochondrial cytochrome c oxidase subunit 1, exposed distinct genetic differences between H. lieberkuehni and our new species. Although our analysis validated the monophyly of Hyaloklossia with some members of the Toxoplasmatinae Biocca, 1957, Cystoisosporinae Frenkel et al., 1987, and Eumonosporinae Chou et al., 2021 (Sarcocystidae Poche, 1913), comparison of genetic differences between Hyaloklossia species from P. p. porosus and H. lieberkuehni revealed the presence of a greater number of polymorphisms than that observed when comparing inter-species (Heydornia spp., Besnoisita spp.) or inter-genus (Toxoplasma vs. Neospora, Neospora vs. Hammondia, and Neospora vs. Heydornia) variabilities among members of the Sarcocystidae. This indicates that Hyaloklossia, as re-erected and defined by Modrý et al. (2001, Int. J. Syst. Evol. Microbiol. 51, 767-772), with its homoxenous life cycle, requires placement in its own subfamily. Thus, we propose a new subfamily, Hyaloklossiinae n. subfam., to accommodate two species, H. lieberkuehni from Europe and Hyaloklossia kasumienesis n. sp. which we describe here from P. p. porosus in Japan.

Biodiversity of the Coccidia (Apicomplexa: Conoidasida) in vertebrates: what we know, what we do not know, and what needs to be done.

Over the last two decades my colleagues and I have assembled the literature on a good percentage of most of the coccidians (Conoidasida) known, to date, to parasitise: Amphibia, four major lineages of Reptilia (Amphisbaenia, Chelonia, Crocodylia, Serpentes), and seven major orders in the Mammalia (Carnivora, Chiroptera, Lagomorpha, Insectivora, Marsupialia, Primates, Scandentia). These vertebrates, combined, comprise about 15,225 species; only about 899 (5.8%) of them have been surveyed for coccidia and 1,946 apicomplexan valid species names or other forms are recorded in the literature. Based on these compilations and other factors, I extrapolated that there yet may be an additional 31,381 new apicomplexans still to be discovered in just these 12 vertebrate groups. Extending the concept to all of the other extant vertebrates on Earth; i.e. lizards (6,300 spp.), rodents plus 12 minor orders of mammals (3,180 spp.), birds (10,000 spp.), and fishes (33,000 spp.) and, conservatively assuming only two unique apicomplexan species per each vertebrate host species, I extrapolate and extend my prediction that we may eventually find 135,000 new apicomplexans that still need discovery and to be described in and from those vertebrates that have not yet been examined for them! Even doubling that number is a significant underestimation in my opinion.

Eimeria spp. (Apicomplexa: Eimeriidae) in Didelphis aurita Wied-Neuwied, 1826 (Didelphimorphia: Didelphidae) and description of a new species infecting this opossum.

Didelphis aurita Wied-Neuwied, 1826 is a marsupial well adapted to anthropogenic activity and commonly found in urban areas of Brazil. Among the gastrointestinal parasites found in this opossum, protozoa of the genus Eimeria are frequently detected. This study investigated the biodiversity of Eimeria species infecting D. aurita in Southeastern Brazil, and provides morphological data on a newly discovered species of Eimeria. From January to June 2019, 43 D. aurita were captured, and their fecal samples were collected and evaluated by salt flotation; positive samples were allowed to sporulate in 2.5% potassium dichromate (K2Cr2O7), and detailed morphological measurements were performed to determine the species present. Thirty-two of forty-three (74.4%) opossums were infected with from one to five Eimeria spp. Four of the eimerians were described and named previously by others: Eimeria auritanensis, Eimeria caluromydis, Eimeria gambai, and Eimeria philanderi. Additionally, sporulated oocysts of a species new to science were detected. Oocysts of this new Eimeria species are spheroidal to subspheroidal, 21.7 × 20.7 (20-23 × 19-23), length/width (L/W) ratio 1.05, with a highly refractile polar granule, but lacking a micropyle and oocyst residuum. Sporocysts are ovoidal, 10.6 × 8.0 (9-12 × 7-9), L/W ratio 1.3, with a small, Stieda body and a sporocyst residuum of diffuse granules. Of the infected opossums, 5/32 (16%) were infected with only one Eimeria sp., 6/32 (19%) with two, 15/32 (47%) with three, 5/32 (16%) with four and 1/32 (3%) with five Eimerians.

A novel coccidian (Apicomplexa: Eimeriidae) from Scotophilus leucogaster (Chiroptera: Vespertilionidae) in southern Saudi Arabia.

A novel species of coccidia, resembling a member of the genus Eimeria, was found in bats, Scotophilus leucogaster, collected in southern Saudi Arabia has been described on the basis of unsporulated oocysts and DNA sequencing of the Internal Transcribed Spacer 1 (ITS1) and partial 18S rDNA regions. Unsporulated oocysts of this form are ovoidal to spheroidal and had a 2-layered wall, 1.5-2.0 (1.9 ± 0.2); the outer layer was light blue with striations, and thicker than the inner, darker layer. No micropyle was present. Unsporulated oocysts (N = 150) measured 27.2 × 22.1 (25-30 × 20-25), length width ratio, 1.2 (1.1-1.4). There was no evidence of an oocyst residuum and/or polar granule. This parasite was detected in 2/7 (29%) S. leucogaster collected from southern Saudi Arabia. Oocysts incubated at 25 °C in 2.5% K2Cr2O7 did not sporulate after > 1 month. Unsporulated oocyst measurements were compared with other coccidian parasites of bats that discharge oocysts in their feces. Sequences of the ITS1 and the 18S rDNA regions obtained from the unsporulated oocysts grouped this coccidium from S. leucogaster with eimerian species from various rodent and squirrel species. It is critical that future investigators obtain fully sporulated oocysts of this coccidium for full description of the parasite recovered in our study so it can be correctly assigned to genus and given an accurate binomial.

Presence of eimerid oocysts in faeces of a quarantined dog in Iceland is explained by coprophagic behaviour prior to its importation. Case report.

BACKGROUND: All dogs imported into Iceland must undergo mandatory quarantine in a special station before introduction into the country. A faecal sample is collected from the first stool passed by the dog in this station and subsequently examined for the presence of intestinal parasite stages. CASE PRESENTATION: In May 2019 unsporulated oocysts were detected in faeces from a 7-year-old household dog that had been imported from Sweden. Most of the oocysts studied strongly resembled those of Eimeria canis Wenyon, 1923. As this species is not valid, the purpose of the present article was to identify the correct species and examine their possible origin. Studies confirmed the presence of two distinct unsporulated oocyst morphotypes in the faeces; measurements and photomicrographs confirmed their identification as Eimeria magna Pérard, 1925 and Eimeria stiedai (Lindemann, 1865) Kisskalt and Hartmann, 1907, both common parasites of European rabbits, Oryctolagus cuniculus (L., 1758). When the owner of the dog was questioned about the food administrated to the dog prior to its import to Iceland, it turned out that it had exclusively been fed dry dog food pellets. However, the owner also reported that on the morning prior to transportation to Iceland, the dog was allowed to move freely in a grassland area where rabbits are common and heaps of their faeces are present. Furthermore, the owner confirmed that the dog consumed rabbit faeces that morning. CONCLUSION: It is believed that this coprophagic behaviour can explain the detection of rabbit eimerids in the dog's faeces, and that such behaviour must be taken into consideration by veterinarians and other diagnostic personnel when they detect atypical cysts or eggs during coprological examinations.

The Coccidia (Apicomplexa) of the Archosauria (Crocodylia: Eusuchia) of the World.

The order Crocodylia (suborder Eusuchia) includes 27 species of alligators, caimans, crocodiles, and gharials that are cosmopolitan in distribution, inhabiting subtropical and tropical locations. Numerous surveys (many of them trivial, with small sample sizes) have reported a wide variety of blood and intestinal apicomplexans from 17/27 (63%) crocodilian hosts, but neither a summation for the group nor a revisionary systematic approach to species evaluation has ever been provided. Herein, we summarize information on the 16 species of apicomplexans that we consider to be valid, including 8 Eimeria, 1 Haemogregarina (that eventually may be transferred to Hepatozoon), 4 Hepatozoon, 2 Isospora, and 1 Progarnia species. In addition, there are 46 apicomplexan forms that we have relegated to species inquirendae and/or only partially identified forms that await further study. We hope this review provides a foundation for future research between parasitologists and herpetologists on parasitism of all reptiles and their relatives. Since many apicomplexans seem to be reasonably host-specific, knowledge of shared species and/or genera may assist herpetologists to better understand the phylogenetic relationships among the New World crocodilians and the direction of their dispersal in the New World.

PREVALENCE AND PARASITE COMPOSITION OF GASTROINTESTINAL PARASITES IN THE ENDANGERED TIBETAN ANTELOPE (PANTHOLOPS HODGSONII) FROM THE CALVING GROUND OF HOH XIL NATURE WORLD HERITAGE SITE, QINGHAI, CHINA.

Our objective was to provide baseline information on the gastrointestinal parasites of the female Tibetan antelope (Pantholops hodgsonii) on its calving ground at the Zhuonai Lake region, in the Hoh Xil Nature World Heritage site, Qinghai, China. On 3 July 2018, 238 freshly deposited fecal samples were collected from the calving grounds and analyzed by flotation technique to recover eggs, oocysts, and nematode larvae. All fecal samples demonstrated the presence of gastrointestinal parasites: 93% (221/238) had nematodes, 36% (86/238) had cestodes, and 99% (235/238) had coccidian oocysts. For each Tibetan antelope, mean (SD) parasite species richness was 3.4 (1.3). Coinfections with five parasite genera were found in 19% (45/238) of fecal samples. These results showed that prevalence of Trichostrongylus, Marshallagia, and Eimeria infections in these Tibetan antelopes were sufficiently high to suggest long-term monitoring be implemented because the climate there is becoming warmer and moisture is increasing, both presumably due to the influence of global warming which, in turn, may be contributing to increased infection risks with these parasites.

Isospora phainopepla n. sp. (Apicomplexa: Eimeriidae) from Phainopepla nitens (Swainson) (Passeriformes: Ptiliogonatidae) in the Joshua Tree National Park, USA.

Isospora phainopepla n. sp. is described from the faeces of Phainopepla nitens (Swainson) collected in Joshua Tree National Park, California, USA. To our knowledge, there are no metazoan or protist parasites described yet from any species in the Ptiliogonatidae of the Americas. Sporulated oöcysts of the isosporan are ellipsoidal, 30-35 × 22-29 (32.9 × 25.4) µm, with a length/width (L/W) ratio 1.20-1.35 (1.30); one rice-shaped polar granule is present, but both oöcyst residuum and micropyle are absent. Sporocyts are elongate-ellipsoidal, 22-27 × 11-14 (23.9 × 12.0) µm, with L/W ratio 1.97-2.02 (2.00); prominent Stieda and sub-Stieda bodies are both present, but para-Stieda body is absent. Sporocyst residuum of many scattered granules is present and the sporozoites have prominent anterior and posterior refractile bodies. This parasite is clearly new to science and represents the first species of Isospora hosted by a New World bird species from this small family of silky flycatchers (Ptiliogonatidae). This article was registered in the Official Register of Zoological Nomenclature (ZooBank) as 7F664B0D-DF0E-4CAD-8CEA-45D79C8BBF92. This article was published as an Online First article on the online publication date shown on this page. The article should be cited by using the doi number. This is the Version of Record.

Five new species of Eimeria Schneider, 1875 from the endangered Tibetan antelope Pantholops hodgsonii (Abel) (Artiodactyla: Bovidae: Caprinae) in the Hoh Xil Nature Reserve Area of Qinghai Province, China.

We examined faeces of 76 endangered Tibetan antelopes Pantholops hodgsonii (Abel) in May 2017, from the Hoh Xil Nature Reserve, Qinghai Province, China, and found 62/76 (82%) discharging oöcysts representing five new species of Eimeria Schneider, 1875. Oöcysts of Eimeria pantholopensis n. sp., found in 54/76 (71%) chiru, are subspheroidal/ellipsoidal, 15-22 × 12-19 (18.6 × 16.1) µm, with a length/width (L/W) ratio of 1.0-1.3 (1.2); micropyle cap and 1-3 polar granules are present, but oöcyst residuum is absent. Sporocysts are ovoidal, 7-11 × 4-6 (9.2 × 5.3) µm, with a L/W ratio of 1.6-2.0 (1.7); Stieda body and sporocyst residuum of small, scattered granules are present; each sporozoite contains 2 refractile bodies. Oöcysts of Eimeria wudaoliangensis n. sp. found in 52/76 (68%) chiru, are pyriform, 21-29 × 17-21 (24.9 × 19.0) µm, with a L/W ratio of 1.1-1.5 (1.3); micropyle, micropyle cap and 1-4 polar granules are present, but oöcyst residuum is absent. Sporocysts are ovoidal, 9-13 × 5-8 (11.7 × 6.7) µm, with a L/W ratio of 1.4-2.7 (1.7); Stieda body and sporocyst residuum of disbursed granules are present; sporozoites have a single large refractile body. Oöcysts of Eimeria hodgsonii n. sp. found in 20/76 (26%) chiru, are elongate-ellipsoidal, 25-32 × 18-21 (28.9 × 19.8) µm, with a L/W ratio of 1.2-1.7 (1.5); micropyle, micropyle cap and 1-3 polar granules are present, but oöcyst residuum is absent. Sporocysts are ovoidal, 11-14 × 6-7 (12.3 × 6.8) µm, with a L/W ratio of 1.7-2.1 (1.8); Stieda body and sporocyst residuum as group of large granules lying along the interface between intertwined sporozoites are present; sporozoites have 2 refractile bodies. Oöcysts of Eimeria schalleri n. sp. found in 49/76 (64.5%) chiru, are ellipsoidal, 26-36 × 19-25 (30.4 × 23.2) µm, with a L/W ratio of 1.2-1.5 (1.3); micropyle with micropyle cap and polar granules appearing as many diffuse tiny bodies are present, but oöcyst residuum is absent. Sporocysts are ovoidal, 12-16 × 7-9 (14.2 × 7.8) µm, with a L/W ratio of 1.6-2.1 (1.8); Stieda body and sporocyst residuum are present, the latter as a group of small dispersed granules between intertwined sporozoites; sporozoites with 2 refractile bodies. Oöcysts of Eimeria sui n. sp. found in 4/76 (5%) chiru, are ovoidal, 32-38 × 26-30 (36.6 × 28.6) µm, with a L/W ratio of 1.0-1.4 (1.3); micropyle and micropyle cap and 1-3 polar granules are present, but oöcyst residuum is absent. Sporocysts are ovoidal, 15-18 × 8-10 (16.7 × 8.9) µm, with a L/W ratio of 1.7-2.1 (1.9); Stieda body and sporocyst residuum are present, the latter as a group of dispersed small granules; sporozoites with 2 refractile bodies. Five of 62 faecal samples in which oöcysts were detected (8%) had a single species infection, 13 of 62 (21%) had two species, 28 of 62 (45%) had three species and 16 of 62 (26%) had four species.

The Coccidia (Apicomplexa: Eimeriidae) of Legless Lizards (Squamata: Lacertoidea: Amphisbaenia) of the World.

Legless lizards (Amphisbaenia) belong to a group of mostly legless squamates that include about 196 species. One genus ( Bipes) retains a pair of forelimbs, but all other 19 genera in the clade are limbless. They are widely distributed, occurring in the Middle East and the Caribbean and nearly all of the major continents (except Australia). Only 2/6 (33%) families, 3/20 (15%) genera, and 4/195 (2%) species in the Amphisbaenia clade of the Sauria have been examined for coccidia and 8 coccidia species are now known. Here, we summarize information on the 8 species of coccidia (3 Choleoeimeria, 1 Eimeria, 4 Isospora) reported from legless lizards of the world. In addition, Eimeria amphisbaeniarum Huntington, Cisper, Smith, Powell, Parmerlee Jr., and Lathrop, 1996, is placed in the genus Choleoeimeria. We speculate that another 380 intestinal coccidia infecting this unique reptilian lineage wait to be discovered.

Transgenic Eimeria magna Pérard, 1925 Displays Similar Parasitological Properties to the Wild-type Strain and Induces an Exogenous Protein-Specific Immune Response in Rabbits (Oryctolagus cuniculus L.).

Rabbit coccidiosis causes great economic losses to world rabbitries. Little work has been done considering genetic manipulation on the etiological agents, rabbit Eimeria spp. In this study, we constructed a transgenic line of Eimeria magna (EmagER) expressing enhanced yellow fluorescent protein (EYFP) and red fluorescent protein (RFP) using regulatory sequences of Eimeria tenella and Toxoplasma gondii. We observed the life cycle of EmagER and confirmed that the transgenic parasites express exogenous proteins targeted to different cellular compartments throughout the entire life cycle. EYFP was expressed mainly in the nucleus and RFP both in the nucleus and cytoplasm. Then, coccidia-free, laboratory-reared 40-day-old rabbits were primarily infected with either EmagER or wild-type strain oocysts and challenged with the wild-type strain. EmagER showed similar reproductivity and immunogenicity to the wild-type strain. Finally, we examined the foreign protein-specific immune response elicited by EmagER. Rabbits were immunized with either transgenic or wild-type oocysts. Immune response against parasite-soluble antigen, EYFP and RFP in spleen, and mesenteric lymph nodes were detected by quantitative real-time PCR. The relative expression level of IFN-γ, IL-2, and TNF-α were higher in EmagER-immunized rabbits than wild-type parasites-immunized rabbits after stimulation with EYFP and RFP. Our study confirmed that a specific immune response was induced by the exogenous protein expressed by EmagER and favored future studies on application of transgenic rabbit coccidia as recombinant vaccine vectors.

Endogenous Life Cycle of Eimeria melanomytis (Apicomplexa: Eimeriidae) from the Dusky Rice Rat, Melanomys caliginosus (Rodentia: Cricetidae: Sigmodontinae) in Costa Rica.

Endogenous stages of the life cycle of Eimeria melanomytis, infecting the peripheral epithelial cells of villi of the small intestine of experimentally infected young dusky rice rats, Melanomys caliginosus , were studied. Giemsa-stained mucosal scrapings and histological sections were examined for all the stages. Eimeria melanomytis has 3 generations of meronts (M), different in size, shape, and number of merozoites (m); and in size, shape, and location of the nuclei within the cytoplasm of the meronts. The 3 meront types, M1-M3, respectively, had 11-14 (m1), 7-10 (m2), and 20-30 (m3) merozoites. Macrogametocytes and microgametocytes, as well as macrogametes and microgametes, complete the sexual cycle forming the unsporulated oocysts. This parasite's endogenous development produced severe intestinal lesions in experimentally infected dusky rice rats.

Four New Species of Eimeria (Apicomplexa: Eimeriidae) from Emoia spp. Skinks (Sauria: Scincidae), from Papua New Guinea and the Insular Pacific.

Between September and November 1991, 54 adult skinks from 15 species were collected by hand or blowpipe from several localities on Rarotonga, Cook Islands, Ovalau Island, Fiji, and Papua New Guinea (PNG), and their feces were examined for coccidians. Species included 5 seaside skinks (Emoia atrocostata), 1 Pacific blue-tailed skink (Emoia caeroleocauda), 2 Fiji slender treeskinks (Emoia concolor), 15 white-bellied copper-striped skinks (Emoia cyanura), 1 Bulolo River forest skink (Emoia guttata), 6 dark-bellied copper-striped skinks (Emoia impar), 5 Papua five-striped skinks (Emoia jakati), 2 Papua slender treeskinks (Emoia kordoana), 3 Papua robust treeskinks (Emoia longicauda), 1 brown-backed forest skink (Emoia loveridgei), 3 Papua black-sided skinks (Emoia pallidiceps), 2 Papua white-spotted skinks (Emoia physicae), 2 Papua yellow-head skinks (Emoia popei), 1 Papua brown forest skink (Emoia submetallica), and 5 Fiji barred treeskinks (Emoia trossula) Species of Eimeria (Ei.) were detected from these Emoia (Em.) spp. and are described here as new. Oocysts of Eimeria iovai n. sp. from Em. pallidiceps from PNG were ellipsoidal with a bilayered wall (L × W) 26.5 × 18.1 µm, with a length/width ratio (L/W) of 1.1. Both micropyle and oocyst residuum were absent, but a fragmented polar granule was present. This eimerian also was found in Em. atrocostata from PNG. Oocysts of Eimeria kirkpatricki n. sp. from Em. atrocostata from PNG were ellipsoidal with a bilayered wall, 18.6 × 13.5 µm, L/W 1.4. A micropyle and oocyst residuum were absent, but a fragmented polar granule was present. This eimerian was also shared by Em. cyanura from the Cook Islands and Fiji, Em. impar from the Cook Islands, Em. loveridgei from PNG, Em. pallidiceps from PNG, Em. popei from PNG, and Em. submetallica from PNG. Oocysts of Eimeria stevejayuptoni n. sp. from Em. longicauda were subspheroidal to ellipsoidal with a bilayered wall, 18.7 × 16.6 µm, L/W 1.1. A micropyle and oocyst residuum were absent, but a fragmented polar granule was present. Oocysts of Eimeria emoia n. sp. from Em. longicauda from PNG were cylindroidal with a bilayered wall, 29.2 × 15.7 µm, L/W 1.9. A micropyle and oocyst residuum were absent, but a polar granule was present. These are the first eimerians reported from Emoia spp. and they add to our growing knowledge of the coccidian fauna of scincid lizards of the South Pacific.

Intestinal Parasites in the Critically Endangered Przewalski's Gazelle ( Procapra przewalskii ) in China, with the Description of a New Species of Eimeria (Apicomplexa: Eimeriidae).

We used a flotation technique to survey fecal samples from 27 Przewalski's gazelles ( Procapra przewalskii Büchner, 1891) for intestinal parasites. Samples were collected from the Qinghai Lake region, Qinghai-Tibet plateau, China, in April 2015. We report parasites belonging to two nematode (Nematodirus, Marshallagia) genera and one apicomplexan (Eimeria) genus; one of the latter was a new species from the critically endangered Przewalski's gazelle. Sporulated oocysts of the new eimerian were spheroidal; they had a thick, outer wall without a micropyle, but with a distinct polar granule and an oocyst residuum. Oocysts measured a mean of 18.6(±0.8)×17.3(±0.9) µm in length and width, with a mean 1.1(±0.1) length/width ratio and a spheroidal oocyst residuum that had a mean width of 2.0(±0.7) µm. Sporocysts were ovoid, measuring a mean of 9.4(±0.6)×5.5(±0.5) µm in length and width, with a mean 1.7(±0.2) length/width ratio. A Stieda body and sporocyst residuum were present, but a substieda body was absent. Sporozoites each had a small anterior refractile body and a larger, posterior refractile body; a small nucleus was visible between them.

A new species of Isospora Schneider, 1881 (Apicomplexa: Eimeriidae) infecting the intestine of the Mediterranean house gecko Hemidactylus turcicus Linnaeus (Reptilia: Gekkonidae).

A new species of Isospora Schneider, 1881 was discovered in the Mediterranean house gecko, Hemidactylus turcicus (Linnaeus) in Saudi Arabia. Both exogenous (sporulated oöcysts) and endogenous developmental stages (meronts, gamonts) were studied and measured. Sporulated oöcysts are spheroidal to slightly subspheroidal, 17-22 (18) µm wide, with a smooth, bi-layered oöcyst wall; micropyle, polar granule and oöcyst residuum are all absent. Sporocysts are ovoidal, 9-11 × 6-8 (10 × 7) µm, with both Stieda and sub-Stieda bodies, a dispersed granular sporocyst residuum, and four sporozoites. Endogenous stages develop extranuclearly in the cytoplasm of epithelial cells of the small intestine. Early uninucleate meronts are spheroidal, c.3-5 (4) µm wide. Multinucleated meronts are subspheroidal, 11-15 × 9-12 (13 × 10) µm, but mature meront and merozoites were not seen. Mature microgamonts, with up to 60 microgametes, are spheroidal, 11-15 (13) µm; macrogamonts are subspheroidal, with a prominent central nucleus, and measured 11-14 × 7-10 (12 × 8) µm.

A new species of coccidia (Apicomplexa: Eimeriidae) from Cordillera striped shrew-rat, Chrotomys whiteheadi (Rodentia: Muridae), from the Philippines.

During July 2011, a single Cordillera striped shrew-rat (Chrotomys whiteheadi) was collected from the Philippines and its faeces examined for coccidian parasites. It harboured an eimerian that we describe here as new. Oocysts of Eimeria macarthuri sp. n. were spheroidal to subspheroidal with a bi-layered wall and measured (length × width, L × W) 18.2 × 17.0 µm, with an L/W ratio of 1.1. A micropyle, oocyst residuum and polar granule were absent. Sporocysts were ovoidal, 9.0 × 6.4 µm, with an L/W ratio of 1.3. A nipple-like Stieda body was present as well as a substieda body. A granular sporocyst residuum was present. To our knowledge, E. macarthuri represents the only coccidian ever described from a rodent of the Philippines.