Life cycle and biology of Eimeria lettyae sp. n. (Protozoa: Eimeriidae) from the northern bobwhite, Colinus virginianus (L.)

A new species of coccidium, Eimeria lettyae sp. n. (Protozoa: Eimeriidae) was recovered from feces of the northern bobwhite, Colinus virginianus (L.), from Pennsylvania and Florida. Oocysts measured 21.1 microns (16.4 to 25.8) by 17.2 microns (14.1 to 21.2); index (L/W ratio) = 1.22. Oocysts lacked a micropyle, residuum, and polar granules. Sporozoites penetrated the upper 1/2 of the villi, then moved to the lamina propria at the base of the villi. There were five asexual generations, all of which developed above the nucleus of the host cell. Meronts measured 9.4 X 7.0 microns, 18.6 X 11.2 microns, 11.8 X 10.1 microns, 7.1 X 6.2 microns, and 20.2 X 12.8 microns, respectively. These matured at 32, 40, 48, 56, and 72 hr postinoculation (PI) and contained 12, 50+, 24 to 36, 12 to 24, and 50+ merozoites, respectively. Infection was most intense in the duodenum although some gamonts were found in the ileum and ceca. The prepatent period was 88 to 91 hr PI. Sporulation time was 18 hr at 25 C. The peak of oocyst production was broad and extended from 4 days PI through 14 days PI. Oocysts were passed for at least 67 to 76 days PI. Eimeria lettyae sp. n. did not infect chickens (Gallus domesticus), domestic turkeys (Meleagris gallopavo), ring-necked pheasants (Phasianus cochicus), chukar partridge (Alectoris graeca), or Japanese quail (Coturnix coturnix). Immunizing bobwhite with E. lettyae sp. n. did not protect against challenge with E. dispersa. Immunizing bobwhites 25 times with 10(2) or 10(3) sporulated oocysts of E. lettyae did not entirely eliminate oocyst production following challenge with the same species.     

Pathogenicity of Eimeria lettyae Ruff, 1985 in the northern bobwhite (Colinus virginianus L.)

Sporulated oocysts of Eimeria lettyae were administered orally to 5-day-old or 18-day-old northern bobwhites (Colinus virginianus, L.). Five-day-old bobwhites were more susceptible based on higher mortality and reduced weight gain. A dose of 5 X 10(5) oocysts produced 25-43% mortality in 5-day-old bobwhites, but none in 18-day-old bobwhites. A dose of 1 X 10(6) oocysts/bobwhite produced 83-100% mortality in 5-day-old bobwhites, and 17-83% mortality in 18-day-old bobwhites. Body weight gain was reduced significantly with a dose of 1 X 10(5) oocysts or greater in 5-day-old bobwhites and with a dose of 5 X 10(5) oocysts or greater in 18-day-old bobwhites. Infection in all age groups reduced concentrations of plasma pigment and plasma protein, but did not affect packed cell volumes. No grossly visible lesions were present in the intestine although there was a shortening of the villi in the duodenum. In mature bobwhites, infection with E. lettyae did not cause mortality, but did reduce egg production and fertility.     

Oocyst production of Eimeria lettyae in northern bobwhites following low-dose inoculations

Inoculation of northern bobwhite quail ( Colinus virginianus ) with low doses of Eimeria lettyae oocysts stimulates a protective immune response, suggesting immunization may be an option for controlling coccidiosis. However, the oocyst production of inoculated birds could be considerable, leading to subsequent outbreaks. To determine the oocyst production following inoculation with E. lettyae, we orally infected 12-wk-old bobwhites with 100, 1,000, or 10,000 sporulated oocysts. Fecal materials were collected on days 5-9 post-inoculation, and total oocyst production was counted in McMaster chambers. Oocyst production/bird was 49.75, 89.5, and 436 × 10(6) for 100, 1,000, or 10,000 oocysts administered, respectively. Estimated oocysts produced/oocyst administered was 49.75, 8.95, and 4.36 × 10(4) for 100, 1,000, or 10,000 oocysts administered, respectively. These findings not only illustrate the crowding effect of larger oocyst inocula but also illustrate the fecundity of E. lettyae at low doses. This suggests that successful immunization of bobwhites against coccidiosis with live vaccines might require attenuated strains with reduced reproductive potential.     

Eimeria dispersa and Eimeria gallopavonis: Infectivity, Survival, and Development in Primary Chicken and Turkey Kidney Cell Cultures

SYNOPSIS. Eimeria dispersa (turkey strain) and Eimeria gallopavonis sporozoites were inoculated into primary cultures of chicken kidney (CK) and turkey kidney (TK) cells. Eimeria dispersa sporozoites were more infective in either cell type than those of E. gallopavonis : at 4 hr, the percentage of infection was 67-98 for E. dispersa but only 23-56 for E. gallopavonis . E. dispersa also survived better in culture: at 2 days, losses of E. dispersa in both cell types were only 4-19%, whereas losses of E. gallopavonis were 35-47% in TK cells and 60–95% in CK cells. However, E. gallopavonis developed further than E. dispersa . Location and increase in numbers of intracellular stages at 4 days indicated that E. dispersa proceeded through 2 schizogonic generations before development stopped.     

Year-long presence of Eimeria echidnae and absence of Eimeria tachyglossi in captive short-beaked echidnas ( Tachyglossus aculeatus )

The short-beaked echidna ( Tachyglossus aculeatus ) is 1 of 5 extant species of monotreme, found only in Australia and Papua New Guinea. The aim of this study was to identify the species of coccidia present and establish a range of subclinical Eimeria spp. (Coccidia: Apicomplexa) oocyst shedding in echidnas from eastern Australia over 18 mo. The coccidia were detected in 89% (49/55) of fecal samples from 12 long-term monitored and healthy captive echidnas, 75% (3/4) of 4 healthy long-term captive echidnas, 83% (5/6) of 6 short-term captive echidnas, and 60% (6/10) of 10 wild echidnas. Echidnas captive for 4 to 23 yr shed 100-46,000 oocysts g(-1) of E. echidnae and remained clinically healthy during this study. Sub-adult and adult wild, and short-term captive, echidnas shed oocysts of both E. echidnae and E. tachyglossi . The lack of coccidia in juvenile short-beaked echidnas suggests these animals are probably non-immune and should not be placed in environments heavily contaminated with oocysts. In addition, no oocysts were found in captive long-beaked echidnas ( Zaglossus bartoni bartoni , n = 2) housed at Taronga Zoo. This study represents an important step in understanding the host-parasite interaction between coccidia and short-beaked echidnas.     

Prevalence and identity of coccidia in pen-raised wild turkeys

One hundred nineteen pen-raised wild turkeys (Meleagris gallopavo) from 12 locations in nine states in the United States were examined for coccidia by sugar flotation of intestinal contents and mucosa or by subinoculating the contents into uninfected domestic turkeys. Seventy-eight (66%) of the turkeys were positive for coccidia. There were no differences in the frequency of coccidia among adult, sub-adult or juvenile turkeys. More females (75%) were infected than males (48%). The species of coccidia from 30 of the turkeys were identified based on microscopic examination of oocysts, fresh scrapings, stained sections and inoculations of bobwhites (Colinus virginianus). The frequency of each species was Eimeria meleagrimitis (97%), E. gallopavonis (47%), E. meleagridis (27%), E. dispersa (17%), E. innocua-E. subrotunda (13%), E. adenoeides (7%) and an undescribed species (3%). Of the 30 turkeys in which the species of coccidia was determined, 30% had a single species infection, 40% had two species, 20% had three species and 10% had four species.     

Eimeria species (Apicomplexa: Eimeriidae) infecting Peromyscus rodents in the southwestern United States and northern Mexico with description of a new species

Of 198 deermice (Peromyscus spp) collected from various localities in the southwestern United States and northern Mexico, 106 (54%) had eimerian oocysts in their feces when examined. These included 50 of 106 (47%) Peromyscus truei, 34 of 54 (63%) Peromyscus maniculatus, 4 of 17 (24%) Peromyscus leucopus, and 18 of 21 (86%) Peromyscus eremicus. The following Eimeria were identified from infected mice: Eimeria arizonensis and Eimeria langebarteli from P. truei; E. arizonensis, Eimeria peromysci, and Eimeria delicata from P. maniculatus; E. arizonensis and Eimeria lachrymalis n. sp. from P. eremicus; and E. langebarteli from P. leucopus. Of the 106 Peromyscus found positive for Eimeria, 97 (91.5%) harbored only a single eimerian species at the time of examination. Sporulated oocysts of E. lachrymalis n. sp. were ellipsoid, 27-35 X 17-21 (30.8 +/- 1.7 X 19.1-0.9) micron, possessed a smooth wall and one polar granule, but lacked a micropyle and an oocyst residuum. Sporocysts were teardrop-shaped, 9-13 X 6-10 (10.9 +/- 0.9 X 7.9 +/- 0.5) micron, and had a Stieda body and sporocyst residuum, but no substieda body. Prepatent periods in experimental infections were 3-6 days after inoculation (DAI) for E. arizonensis (hosts: P. eremicus, P. maniculatus, P. truei); 4-5 DAI for E. peromysci (host: P. maniculatus); 6-9 DAI for E. langebarteli (hosts: P. truei, P. leucopus); and 8-10 DAI for E. lachrymalis (host: P. eremicus). Patency in these infections lasted 6-11 days for E. arizonensis, 5-10 days for E. peromysci, 14-40+ days for E. langebarteli, and 19-50+ days for E. lachrymalis. Eimeria lachrymalis appears to produce occult infections in P. eremicus that can be reactivated upon inoculation of the host with E. arizonensis.     

Variation in Eimeria oocyst count and species composition in weanling beef heifers

Rectal fecal samples were collected daily on 10 consecutive days in November 2004 from 11 weaned beef heifers to assess daily variation in fecal oocyst count and species composition. Subsequent samples were collected from the same animals on 15 April 2005 and 9 June 2005. Oocyst numbers were determined by the modified McMaster's test, and species were identified by examination of oocysts recovered with the Wisconsin sugar flotation technique. Soil samples were collected from the heifer pasture on 8 June 2005, and oocysts were quantified and identified to species. Mean fecal oocyst counts varied little at all sampling dates ranging from 134-377 oocysts/g. Ten Eimeria spp. were identified in fecal samples collected in November and April and 11 in June. Eimeria bovis was the most common species identified at all samplings. Mean species composition showed little variation during the 10-day sampling period in November, remained similar in April, and varied slightly in June. Twelve Eimeria spp. were identified in soil samples in proportions similar to those seen in fecal samples. The results indicate that clinically normal weanling beef heifers are likely to be infected with a diverse, but relatively stable, community of Eimeria spp.     

Coproparasitological examinations and molecular determination of Eimeria species in Madura cattle reared on Madura Island,Indonesia

Madura cattle, which are native to Indonesia and mainly kept on Madura Island, East Java, are expected to contribute to improving the regional meat self-sufficiency. Eimeria spp. are the most pathogenic protozoans among gastrointestinal parasites in livestock but no molecular surveys of Eimeria spp. in Madura cattle have been conducted to date. In this study, a total of 183 fecal samples were collected from Madura cattle and 60 (32.8%) were positive for parasites of protozoans and nematodes by the sugar floatation method. Among the samples with parasites, Eimeria spp. oocysts were detected in 50 samples (27.3%) with an average OPG value of 1686.1. Eimeria spp. were successfully identified to the species level in 26 samples with Eimeria bovis being the most prevalent, followed by E. zuernii and E. aubrunensis. A total of 21 samples showed mixed infection of more than two species of Eimeria. E. bovis and E. zuernii have been recognized as having high virulency and, thus, these parasites are potential sources of severe coccidiosis and the cause of infections in other cattle. Although additional studies are warranted, these results can be helpful for improving the management and productivity of Madura cattle.     

Eimeria species (Apicomplexa: Eimeriidae) from arctic ground squirrels (Spermophilus parryii) and red squirrels (Tamiasciurus hudsonicus) in Alaska and in Siberia, Russia

Fecal samples from arctic ground squirrels (Spermophilus parryii) collected in Alaska (n = 90) and Russia (n = 46) and from red squirrels (Tamiasciurus hudsonicus) in Alaska (n = 35) were examined for the presence of Eimeria spp. (Apicomplexa: Eimeriidae). Four species were recovered from arctic ground squirrels, including Eimeria callospermophili (prevalence = 18%), Eimeria cynomysis (23.5%), Eimeria lateralis (19%), and Eimeria morainensis (77%). A single species, Eimeria tamiasciuri (91%), was recovered from red squirrels. Eimerians recovered from arctic ground squirrels represent new host records, and the single species from red squirrels is a new geographic record. Alaskan arctic ground squirrel prevalence was higher for E. callospermophili (Alaska = 22% vs. Russia = 9%), E. cynomysis (34% vs. 2%), and E. lateralis (27% vs. 4%), but not E. morainensis (78% vs. 76%).     

Eimeria spp. (Apicomplexa: Eimeriidae) from the eastern chipmunk (Tamias striatus) in Pennsylvania with a description of one new species

Intestinal contents of 41 eastern chipmunks (Tamias striatus) from Armstrong County, Pennsylvania, were examined for the presence of Eimeria spp. Three previously named species were identified: E. lateralis (prevalence = 9%), E. ovata (3%), and E. vilasi (74%); 1 new species, Eimeria tamiensis n. sp. (74%), is described here. This report extends the geographic ranges of the named species into Pennsylvania. Sporulated oocysts of E. tamiensis n. sp. are ovoid and 18.6 x 14.5 (16-23 x 12-17) microm, with no micropyle or residuum. Sporocysts are ellipsoid and 8.6 x 5.4 (7-10 x 4-8) microm. with a Stieda body and granular residuum. Prevalences of E. lateralis and E. vilasi were similar to those reported previously. The differences in prevalences may be due to different life-history strategies of high- and low-prevalence Eimeria species.     

Phylogenetic position of Eimeria antrozoi, a bat coccidium (Apicomplexa: Eimeriidae) and its relationship to morphologically similar Eimeria spp. from bats and rodents based on nuclear 18S and plastid 23S rDNA sequences.

Partial plastid 23S and nuclear 18S rDNA genes were amplified and sequenced from 2 morphologically similar Eimeria species. E. antrozoi from a bat (Antrozous pallidus) and E. arizonensis from deer mice (Peromyscus spp.), as well as some other Eimeria species from bats and rodents. The phylogenetic trees clearly separated E. antrozoi from E. arizonensis. The phylogenies based on plastid 23S rDNA data and combined data of both plastid and nuclear genes grouped 2 bat Eimeria and 3 morphologically similar Eimeria species from rodents into 2 separate clades with high bootstrap support (100%, 3 rodent Eimeria species; 72-97%, 2 bat Eimeria species), which supports E. antrozoi as a valid species. The rodent Eimeria species did not form a monophyletic group. The 2 bat Eimeria species formed a clade with the 3 morphologically similar rodent Eimeria species (E. arizonensis, E. albigulae, E. onychomysis, all from cricetid rodents) with 100% bootstrap support, whereas 2 other rodent Eimeria species (E. nieschulzi, E. falciformis, from murid rodents) formed a separate clade with 100% bootstrap support. This suggests that the 2 Eimeria species from bats might be derived from rodent Eimeria species and may have arisen as a result of lateral host transfer between rodent and bat hosts.     

Comparison of Helicobacter spp. genetic sequences in wild and captive seals, and gulls

Helicobacter species are widely distributed in the gastrointestinal system of humans and many animal taxa. Investigations of natural infections are essential to elucidating their role within the host. The feces of fur seals Arctocephalus pusillus doriferus and sea lions Neophoca cinerea from 3 separate captive populations, as well as a wild colony from Kangaroo Island, Australia, were examined for the occurrence of Helicobacter spp. The feces from several wild silver gulls Larus novahollandiae were also investigated. As detected by PCR, 18 of 21 samples from captive and 12 of 16 samples from wild seals were positive for Helicobacter spp. Three species were identified in these animals. Whilst one possibly novel type was identified from wild fur seals, the majority of wild and captive individuals had the same species. This species also occurred in more than 1 seal type and in silver gulls, and shared a 98.1 to 100% identity to other Helicobacter spp. from harp seals and sea otters. A similar sequence type to species identified from cetaceans was also detected in several captive seals. This study reports for the first time the presence of Helicobacter spp. in wild and captive seals and demonstrates the diversity and broad-host range of these organisms in the marine host.     

The species of Eimeria in rabbits for meat production in Britain

Eight species of coccidia were recognized in 596 faeces samples from 3 commercial rabbitries in South East England. The level of infection was related to management methods and at one site it was reduced by an outbreak of mucoid enteritis and/or its treatment with oxytetracycline. In samples from rabbits managed conventionally in the wire cages over droppings-pits, 96% contained oocysts and of these, 60% had 1000-10000 oocysts/g. Mixed infections were common, 67% of the animals carrying 2-4 different species. Eimeria, media, E. magna and E. perforans occurred most frequently; E. coecicola, E. irresidua and E. flavescens were less common and E. intestinalis and E. piriformis were relatively rare. E. stiedai was not recorded.     

Molecular identification of Eimeria species in Spanish bats

This is the first study reporting the detection and molecular characterization of Eimeria in bats in Spain, specifically in 12 of 32 chiropteran species described in the Iberian Peninsula. A total of 76 faecal samples were collected from different bat roosting sites across Spanish territory. The DNA was extracted from the samples and sequenced by targeting the Eimeria SSU-rRNA gene. Two Eimeria species were detected in 29 of the 76 faecal samples (38%), and the bat-specific Eimeria rioarribaensis and rodent-specific Eimeria jerfinica were detected in 4 (5%) and 25 (33%) of the samples, respectively. This is the first report of E. rioarribaensis in the bats Rhinolophus euryale, Myotis myotis and Nyctalus lasiopterus, extending the host and geographical ranges for this bat coccidian parasite. The identification of the rodent-specific parasite species E. jerfinica in bats indicates the occurrence of this species in Spain, although its presence has not previously been reported in wild rodents in this country. Considering that most of the Eimeria spp. reported in bats were described only on the basis of morphometric data, molecular studies are required to determined which Eimeria species occur in bats, to complete the identification of these species and to clarify the phylogeny.     

Characterization of the complete mitochondrial genomes of five Eimeria species from domestic chickens

Chicken coccidiosis caused by members of the genus Eimeria causes significant economic losses worldwide. In the present study we sequenced the complete mitochondrial DNA (mtDNA) sequences of six Eimeria species and analyzed features of their gene contents and genome organizations. The complete mt genomes of E. acervulina, E. brunetti, E. maxima, E. necatrix, E. tenella and E. praecox were 6179bp, 6148bp, 6169bp, 6214bp, 6213bp and 6174bp in size, respectively. All of the mt genomes consist of 3 genes for proteins (cox1, cox3, and cytb), 12 gene fragments for the large subunit (LSU) rRNA, and 7 gene fragments for the small subunit (SSU) rRNA, but no transfer RNA genes. The organization of the mt genomes is similar to that of Plasmodium, but distinct from Babesia and Theileria. The putative direction of translation for 3 genes (cox1, cox3, and cytb) was the same in all six Eimeria species. The contents of A+T of the mt genomes were 65.35% for E. acervulina, 65.43% for E. brunetti, 64.53% for E. maxima, 65.04% for E. necatrix, 64.98% for E. tenella and 65.59% for E. praecox. The AT bias has a significant effect on both the codon usage pattern and amino acid composition of proteins. Phylogenetic analyses using concatenated nucleotide sequences of the 2 protein-coding genes (cytb and cox1), with three different computational algorithms (Bayesian analysis, maximum parsimony and maximum likelihood), all revealed distinct groups with high statistical support, indicating that the six Eimeria spp. represent six distinct but closely-related species. These data provide novel mtDNA markers for studying the molecular epidemiology and population genetics of the six Eimeria spp., and should have implications for the molecular diagnosis, prevention and control of coccidiosis in domestic chickens.     

Molecular characterization and phylogenetic analysis of Eimeria from turkeys and gamebirds: implications for evolutionary relationships in Galliform birds

In order to determine the evolutionary relationships among Eimeria species that parasitize birds of the Galliformes, the 18s rDNA gene and a portion of the cytochrome oxidase subunit 1 (cox-1) were amplified from Eimeria species isolated from turkeys, chukars, and pheasants. The phylogenetic analysis of these sequences suggests that species infecting chickens are polyphyletic and, therefore, do not all share a direct common ancestor. Both the 18s rDNA and the cox-1 sequences indicate that Eimeria tenella and Eimeria necatrix are more closely related to Eimeria of turkeys and pheasants than to other species that infect the chicken. It is, therefore, likely that the chicken Eimeria spp. represent 2 separate ancestral colonizations of the gut, one of which comprises E. tenella and E. necatrix that infect the ceca, while the other includes Eimeria acervulina, Eimeria brunetti, Eimeria maxima, and Eimeria mitis, which infect the upper regions of the intestine.     

Identification of a novel species of Eimeria Schneider, 1875 from the woylie,Bettongia penicillata Gray (Diprotodontia: Potoroidae) and the genetic characterisation of three Eimeria spp. from other potoroid marsupials

Faecal samples (n = 1,093) collected from the woylie Bettongia penicillata Gray, in south-western Australia were examined for the presence of coccidian parasites. Eimeria sp. oöcysts were detected in 15.2% of samples. Faecal samples obtained from the eastern bettong Bettongia gaimardi (Desmarest) (n = 4) and long-nosed potoroo Potorous tridactylus (Kerr) (n = 12) in Tasmania, were also screened for the presence of Eimeria spp. (prevalence 50% and 41.7%, respectively). Morphological and genetic comparison with other known species of Eimeria indicates that the material identified in woylies is novel. This study aimed to (i) morphologically describe and genetically characterise Eimeria woyliei n. sp. found in woylies; and (ii) genetically characterise Eimeria gaimardi Barker, O’Callaghan & Beveridge, 1988, Eimeria potoroi Barker, O’Callaghan & Beveridge, 1988, and Eimeria mundayi Barker, O’Callaghan & Beveridge, 1988, from other potoroid marsupials. Molecular phylogenetic analyses conducted at the 18S rDNA and mitochondrial cytochrome c oxidase subunit 1 (cox1) loci revealed that E. woyliei n. sp. was most closely related to Eimeria setonicis Barker, O’Callaghan & Beveridge, 1988, at the 18S rDNA locus, and Eimeria trichosuri O’Callaghan & O’Donoghue, 2001, at the cox1 locus. Eimeria woyliei n. sp. is the sixth species of Eimeria to be formally described from potoroid marsupials.