Eimeria and Klossia spp. (Protozoa: Sporozoa) from Wild Mammals in Borneo*

SYNOPSIS. During a survey of parasites of Sabah mammals, coccidia oocysts were found in 9 of 22 host species examined. Sporulated oocysts of Klossia sp. from Rattus whiteheadi are described. New geographic records are reported for Eimeria callosciuri, Eimeria sabani, and Eimeria tupaiae. New host and geographic records are reported for Eimeria separata from Rattus cremoriventer, for E. tupaiae and Eimeria ferruginea from Tupaia tana, for Eimeria nieschulzi from Rattus muelleri, and for E. sabani and Klossia sp. from R. whiteheadi.     

A Model for Taxonomic Work on Homoxenous Coccidia: Redescription, Host Specificity, and Molecular Phylogeny of Eimeria ranae Dobell, 1909, with a Review of Anuran-Host Eimeria (Apicomplexa: Eimeriorina)

ABSTRACT. We attempt to extend knowledge of anuran Eimeria , and to provide a model for a complex approach to studies on coccidia. New host and geographic records of coccidia in European Anura are provided. In the second part, Eimeria ranae Dobell, 1909 is redescribed from European terrestrial frogs of the genus Rana based on light microscopic and ultrastructural data on both exogenous and endogenous developmental stages, host specificity, and molecular phylogenetic data. Results of experimental transmissions show for the first time that the host specificity of E. ranae is restricted to the genus Rana and that isolates from tadpoles and adults are conspecific. Disappearance of infection during metamorphosis was confirmed experimentally, suggesting that infections in adults result from reinfections. Poikilotherm-host Eimeria species possessing a Stieda body (SB) are for the first time included in a molecular phylogenetic analysis. Eimeria ranae and Eimeria arnyi from a colubrid snake form together a well-supported clade, basal to other SB-bearing coccidia. The other analysed reptile–host eimerians, Eimeria tropidura and Choleoeimeria sp., which possess bivalved sporocysts and lack a SB, represent a distinct basal lineage of the eimeriid clade. The third part of the article reviews anuran-host Eimeria . Three distinct oocyst morphotypes, apparently correlating with the character of endogenous development, are recognized and characterized among anuran eimeriids.     

Eimeria species in the Pearson Island Rock Wallaby, Petrogale lateralis pearsoni

Four species of previously described Eimeria were identified from oocysts present in the faeces of Pearson Island rock wallabies, Petrogale lateralis pearsoni from Pearson Island, South Australia. Eimeria petrogale and Eimeria sharmani have been reported in P. lateralis; however, the presence of Eimeria godmani and Eimeria inornata expands the suite of eimerian species known in rock wallabies of the lateralis complex. These observations indicate that the host distribution of Eimeria species in rock wallabies is more diverse than previously thought, and supports the hypothesis that these coccidia have an ancient association with rock wallabies which persisted as the hosts radiated.     

Attempted Cross-Transmission of Coccidia Between Sheep and Goats and Description of Eimeria ovinoidalis sp. n.

SYNOPSIS.
Attempted infection of 2 young lambs with oocysts of Eimeria christenseni from a goat was unsuccessful. Negative results were obtained also when young kids were fed oocysts of Eimeria ninakohlyakimovae from sheep. There was no difficulty in infecting lambs with the sheep coccidium resembling E. ninakohlyakimovae nor goats with the goat coccidium E. christenseni. Oocysts from the goat measured 38.4 × 26.7 m, but were easily distinguished from Eimeria ahsata from the sheep by sporocyst size and shape, and from Eimeria ovina by oocyst size. Eimeria ninakohlyakimovae -like oocysts from sheep averaged 23.0 ×18.2 m and were morphologically indistinguishable from previously reported goat coccidia.
Since no cross infections of sheep and goats could be accomplished with oocysts of Eimeria sp. characteristic of one or the other host, I concluded that sheep coccidia previously known as E. ninakohlyakimovae are distinct from morphologically similar goat coccidia and therefore constitute a separate species. Since the name E. ninakohlyakimovae was first used for coccidia from the goat, the sheep coccidium is renamed Eimeria ovinoidalis with oocyst structure and endogenous stages similar to those previously described from the sheep.     

Coccidia (Protozoa: Eimeriidae) of the Alpaca Lama pacos*

SYNOPSIS. Three new species of coccidia are described from the alpaca Lama pacos from Peru. These are the first species of coccidia to be named from this host. The oocysts of Eimeria lamae n. sp. are ellipsoidal, occasionally ovoid, 30–40 by 21–30 μ (mean 35.6 by 24.5 μ) with elongate ovoid sporocysts 13–16 by 8–10 μ (mean 15.3 by 8.5 μ). The oocysts of Eimeria alpacae n. sp. are ellipsoidal, rarely ovoid, 22–26 by 18–21 μ (mean 24.1 by 19.6 μ), with ovoid sporocysts 10–13 by 7–8 μ (mean 11.0 by 6.8 μ). The oocysts of Eimeria punoensis n. sp. are ellipsoidal, occasionally ovoid, 17–22 by 14–18 μ (mean 19.9 by 16.4 μ), with ovoid sporocysts 8–11 by 5–7 μ (mean 9.2 by 6.1 μ).     

Attempted cross-transmission of coccidia between sheep and goats and description of Eimeria ovinoidalis sp. n.

Attempted infection of 2 young lambs with oocysts of Eimeria christenseni from a goat was unsuccessful. Negative results were obtained also when young kids were fed oocysts of Eimeria ninakohlyakimovae from sheep. There was no difficulty in infecting lambs with the sheep coccidium resembling E. ninakohlyakimovae nor goats with the goat coccidium E. christenseni. Oocysts from the goat measured 38.4 X 26.7 microns, but were easily distinguished from Eimeria ahsata from the sheep by sporocyst size and shape, and from Eimeria ovina by oocyst size. Eimeria ninakohlyakimovae-like oocysts from sheep averaged 23.0 X 18.2 microns and were morphologically indistinguishable from previously reported goat coccidia. Since no cross infections of sheep and goats could be accomplished with oocysts of Eimeria sp. characteristic of one or the other host, I concluded that sheep coccidia previously known as E. ninakohlykimovae are distinct from morphologically similar goat coccidia and therefore constitute a separate species. Since the name E. ninakohlyakimovae was first used for coccidia from the goat, the sheep coccidium is renamed Eimeria ovinoidalis with oocyst structure and endogenous stages similar to those previously described from the sheep.     

Eimeria fraterculae sp. n. in the kidneys of Atlantic puffins (Fratercula arctica) from Newfoundland, Canada: species description and lesions

Renal coccidiosis is reported for the first time in an auk (Alcidae). Infection was detected in seven of 50 nestling Atlantic puffins (Fratercula arctica) and a new species of coccidia, Eimeria fraterculae sp. n., is described. The structure and sporulation of oocysts are characterized. Meronts, gamonts, and developing oocysts were present in collecting duct epithelium of medullary cones. The predominant host response was hypertrophy of infected cells, tubule dilation, and a mild localized peritubular infiltration with mononuclear inflammatory cells.     

Disseminated visceral coccidiosis in Indo-gangetic flap-shelled turtles, Lissemys punctata andersonii

Eimerioriniid coccidia commonly infect vertebrates and might contribute to morbidity and mortality under captive conditions. The common genus Eimeria typically shows tissue specificity, usually being limited to the epithelium of the gut; disseminated infections are rare in vertebrates. Disseminated visceral coccidiosis was found in two wild-caught adult female Indo-gangetic flap-shelled turtles (Lissemys punctata andersonii) that died while in captivity at a zoo. Sporulated oocysts of Eimeria spp. were found in lung and liver of one turtle and in auditory canal, nasal mucosa, pharynx, lung, liver, kidney, spleen, and intestine of the second. Two distinct species of Eimeria were indicated for the latter case by polymerase chain reaction amplification and sequencing of a portion of the 18S rRNA gene; one species was present in nasal mucosa and liver, with a separate species in lung, spleen, and intestine. Severity of inflammation was correlated with coccidial density. Coccidia were in melanomacrophages in liver and spleen; in the interstitium of auditory canal, nasal mucosa, pharynx, lung, and intestine; and within the interstitium and epithelial cells of the renal tubules in kidney. We suggest these disseminated infections might have been facilitated by a compromised immune system.     

Patterns of coccidial prevalence in lizards of Mauritius

This paper reports prevalence of coccidial oocysts in fecal samples from 6 endemic and 2 introduced lizard species on Mauritius, an island nation in the Indian Ocean. Total coccidian prevalence was 54% (n = 341) for the endemic 'ornate day gecko,' Phelsuma ornata; 48% (n = 159) for the endemic 'Durrell's night gecko,' Nactus durrelli; 53% (n = 15) for the endemic 'Serpent Island night gecko,' N. serpensinsula; and 78% (n = 248) for the introduced gecko, Hemidactylus frenatus. These high prevalences may reflect lack of long-lasting immune response to coccidial infection. There were few significant differences in prevalence among age, island, sex, or body condition within species, suggesting that these coccidia are relatively nonpathogenic and have little effect on host fitness. Prevalence was higher in the dry season than the wet season. These data suggest other factors, such as low host immune response to reinfection, affect overall prevalence more significantly than the effect of humidity on oocyst survival on Mauritius. No coccidia were found in samples from the endemic 'Gunner's Quoin night gecko,' N. coindemirensis (n = 155), probably reflecting parasite extinction due to a host population bottleneck following historical introduction of rats. There was no evidence of competitive or facilitative interactions between Eimeria sp. and Isospora sp., but evidence of competition between 2 Eimeria species in the 'ornate day gecko,' Phelsuma ornata. No evidence was found of cross-species infection, suggesting that reptile coccidia have high host specificity and are, therefore, poor subjects for studies of parasite-mediated competition and the evolution of sex.     

Observations on the Endogenous Stages of Eimeria confusa Joseph, 1969 from the Grey Squirrel Sciurus carolinensis*

SYNOPSIS. Stages in the endogenous cycle of Eimeria confusa from the grey squirrel, Sciurus carolinensis, are described from mixed infections with another species, Eimeria lancasterensis. All corresponding stages were markedly different in the 2 species. In E. confusa infections, the parasites were located below the host cell nuclei of the epithelial cells of the villi of the jejunum and ileum. Mature schizonts were ellipsoidal, averaged 20.9 × 18.6 μm and had 18–30 merozoites. The mature microgamonts measured 34.3 × 24.7 μm and had hundreds of microgametes. Mature macrogametes were ovoid, averaged 31.3 × 25.6 μm, and contained 2 kinds of plastic granules.     

Sustainable coccidiosis control in poultry production: the role of live vaccines

The development of new methods of administering coccidiosis vaccines has facilitated their use in the hatchery and thereby improved prospects for the economic vaccination of broilers. The acquisition of protective immunity to Eimeria species is boosted by further exposure to infection after vaccination. Factors that affect the reproductive efficiency of non-attenuated and attenuated vaccines are considered and the key role that oocyst production plays in establishing and maintaining uniform immunity in a flock of chickens is discussed. In addition to immunisation, a possible advantage to the application of certain vaccines is that their use could repopulate poultry houses with drug-sensitive organisms. Theoretical rotation programmes in which the use of drugs is alternated with that of vaccines are described. Variability of the cross-protective immune response between strains of the same species should be considered during vaccine development and subsequent use. The significance of less common species of Eimeria, not included in all vaccines, also needs to be assessed. An important consideration is the occurrence of pathogens other than Eimeria (such as the bacterium Clostridium) in flocks given coccidiosis vaccines and the methods by which they might be controlled. More research is required into the relationship between bacterial and viral infections of poultry and coccidiosis vaccination. Vaccines need to be developed that are simple to apply and cost effective for use in areas of the world where small-scale poultry production is commonplace. In the near future it is likely that more live vaccines based upon oocysts derived from attenuated strains of Eimeria will be developed but in the longer term vaccines will be based on the selective presentation to the host of specific molecules that can induce protective immunity. This achievement will require significant investment from the private and public sectors, and, if successful, will facilitate the sustainable control of coccidiosis in poultry production.     

Disseminated visceral coccidiosis in a wild white-naped crane (Grus vipio)

Disseminated visceral coccidiosis (DVC) was unexpectedly recognized in a wild white-naped crane (Grus vipio) killed by phosphamidon insecticide. On gross pathologic examination, widely disseminated white nodules were found on the serosa of the pro-ventriculus, gizzard, and intestine, as well as on the surface and in the parenchyma of liver, spleen, and cardiac muscle. Microscopically, asexual stages of a coccidia were observed in some nodules. However, the species of coccidia could not be determined because no oocysts were found on fecal examination. This is believed to be the first reported case of DVC in a wild white-naped crane infected with Eimeria spp.     

Major histocompatibility complex variation and age‐specific endoparasite load in subadult European rabbits

Genes of the major histocompatibility complex (MHC) play a fundamental role in the vertebrate immune response and are amongst the most polymorphic genes in vertebrate genomes. It is generally agreed that the highly polymorphic nature of the MHC is maintained through host–parasite co‐evolution. Two nonexclusive mechanisms of selection are supposed to act on MHC genes: superiority of MHC heterozygous individuals (overdominance) and an advantage for rare MHC alleles. However, the precise mechanisms and their relative importance are still unknown. Here, we examined MHC dependent parasite load in European rabbits (Oryctolagus cuniculus) from a distinct population with low MHC diversity (three alleles, six genotypes). Using a multivariate approach, we tested for associations of individual MHC class II DRB constitution and the rabbits’ intestinal burden with nematodes and coccidia. Rabbits having a particular allele showed lower infestations with hepatic coccidia (E. stiedai). However, a comparison of all six genotypes in the population revealed that carriers of this allele only benefit when they are heterozygous, and furthermore, MHC heterozygosity in general did not affect individual parasite load. In conclusion, this study suggests an immunogenetic basis of European rabbit resistance to hepatic coccidiosis, which can strongly limit survival to maturity in this species. Our study gives a complex picture of MHC–parasite correlations, unveiling the limits of the classical hypotheses of how MHC polymorphism is maintained in natural systems.     

Eimeria chihuahuaensis sp.n. and Other Coccidia from Dipodomys spp. in El Paso County, Texas

SYNOPSIS. Dipodomys merriami Mearns and Dipodomys ordi Woodhouse were surveyed for coccidia in El Paso County, Texas. Infections with Eimeria chobotari. Eimeria dipodomysis and Eimeria balphae were 24.8%. 4.4%, and 11%, respectively, for D. ordi. Dipodomys merriami had an infection level of 23.8% with E. chobotari . Four animals concurrently harbored E. chobotari and E. balphae or E. dipodomysis or a new species Eimeria chihuahuaensis. Male and female host infection levels were not significantly different. The new species is described and photographs of 3 previously described Eimeria from Dipodomys are presented.     

Description of the two strains of turkey coccidia Eimeria adenoeides with remarkable morphological variability

Although oocyst morphology was always considered as a reliable parameter for coccidian species discrimination we describe strain variation of turkey coccidia, Eimeria adenoeides, which remarkably exceeds the variation observed in any other Eimeria species. Two strains have been isolated - the first strain maintains the typical oocyst morphology attributed to this species - large and ellipsoidal - while the second strain has small and ovoid oocysts, never described before for this species. Other biological parameters including pathogenicity were found to be similar. Cross-protection between these 2 strains in 2 immunization and challenge experiments was confirmed. Sequencing and analysis of 18S and ITS1 ribosomal DNA revealed a close relationship according to 18S and a relatively distant relationship according to ITS1. Analysis of 18S and ITS1 sequences from commercial turkey coccidiosis vaccines Immucox?-T and Coccivac?-T revealed that each vaccine contains a different strain of E. adenoeides and that these strains have 18S and ITS1 sequences homologous to the sequences of the strains we have isolated and described. These findings show that diagnostics of turkey coccidia according to oocyst morphology have to be carried out with caution or abolished entirely. Novel PCR-based molecular tools will be necessary for fast and reliable species discrimination.     

Detection of four important Eimeria species by multiplex PCR in a single assay

The oocysts of some of the recognized species of chicken coccidiosis are difficult to distinguish morphologically. Diagnostic laboratories are increasingly utilizing DNA-based technologies for the specific identification of Eimeria species. This study reports a multiplex polymerase chain reaction (PCR) assay based on internal transcribed spacer-1 (ITS-1) for the simultaneous diagnosis of the Eimeria tenella, Eimeria acervulina, Eimeria maxima, and Eimeria necatrix species, which infect domestic fowl. Primer pairs specific to each species were designed in order to generate a ladder of amplification products ranging from 20 to 25 bp, and a common optimum annealing temperature for these species was determined to be 52.5 °C. Sensitivity tests were performed for each species, showing a detection threshold of 1–5 pg. All the species were amplified homogeneously, and a homogenous band ladder was observed, indicating that the assay permitted the simultaneous detection of all the species in a single-tube reaction. In the phylogenic study, there was a clear species clustering, which was irrespective of geographical location, for all the ITS-1 sequences used. This multiplex PCR assay represents a rapid and potential cost-effective diagnostic method for the detection of some key Eimeria species that infect domestic fowl.