Cystoisospora spp. infection at a dog breeding facility in the Madrid region: Infection rate and clinical management based on toltrazuril metaphylaxis.

Canine coccidiosis caused by Cystoisospora canis and Cystoisospora ohioensis-complex is common in kennels. While often underestimated, coccidiosis may cause severe clinical signs in puppies and sometimes even lead to death, so preventative measures are important. This study examines Cystoisospora spp. infection at a Labrador retriever breeding facility in Madrid, Spain. To identify environmental factors associated with infection, dams were examined throughout a reproductive cycle (from oestrus to 60 days postpartum) and their puppies during their first 60 days of life. Also assessed was the efficacy of combined treatment with emodepside (0.9 mg/ml) and toltrazuril (18 mg/ml) at a dose of 0.5 ml/kg of weight, equivalent to 0.45 mg/kg and 9 mg/kg, respectively, in puppies on day 35 of life. Oocyst shedding was detected in 4.6-18.6% of 45 dams examined and in 2.2-9.1% of their litters (315 puppies). In both cases, peak opg elimination was recorded on day 30 postpartum/of life. The species of Cystoisospora detected were C. canis (91.3%) and C. ohioensis-complex (8.7%). While in both dams and puppies opg counts were higher in autumn when rainfall was at its highest, correlation between opg and rainfall emerged as significant only in puppies (p = 0.031). The treatment of 35 day-old puppies with toltrazuril was 100% effective in controlling this infection in the kennel. Our findings therefore suggest the need for a strict hygiene regime and the use of toltrazuril as blanket treatment to reduce Cystoisospora transmission in dog breeding facilities.

Parasitological and molecular investigation of consequences of raw meat feeding (BARF) in dogs and cats: implications for other pets living nearby.

Considering the original nature of carnivorism among dogs and cats, nowadays these pets are increasingly fed BARF (Biologically Appropriate Raw Food/Bone And Raw Food) diet to improve their health conditions. However, this diet may also carry health risks, such as vitamin or mineral deficiencies, and infection with pathogens including parasites. In our study, fecal samples of 89 pet animals kept on BARF diet were subjected to coprological examination followed by molecular analyses. Six of them shed eggs of Dicrocoelium dendriticum. This result was confirmed with PCR and sequencing, and in one case, the DNA of Fasciola hepatica was also demonstrated. In addition, oocysts of Cystoisospora canis, a Cystoisospora ohioensis-like sp. and Eimeria stiedai, as well as sporocysts of a Sarcocystis sp. were also detected. All samples were negative for Neospora caninum and Toxoplasma gondii. In conclusion, no evidence was found for the infection of BARF-fed dogs and cats with parasites that are usually associated with this diet and considered as clinico-pathological risk factors for these pets themselves (e.g., N. caninum, T. gondii). However, fluke eggs (probably originating from ruminants) and oocysts of E. stiedai (from rabbit liver in the food) were demonstrated as pseudoparasites. These species are usually not considered among parasite-associated risks of BARF-feeding, implying that other animals living near BARF-fed pets are neglected in this context. However, where intermediate hosts of D. dendriticum occur in urban areas, BARF-feeding may indirectly affect later other dogs and cats. It was also shown here that BARF-feeding may contribute to the contamination of the environment with E. stiedai oocysts, thus increasing the risks of biliary coccidiosis in nearby living pet rabbits that would otherwise not have access to oocysts of E. stiedai.

On the efficacy of preventive toltrazuril treatments and the diagnosis of Cystoisospora suis infections in intensively raised piglets in farms from southeast Spain.

This study investigated the presence and abundance of Cystoisospora suis oocysts in faecal samples from 131 one- to three-week-old pig litters belonging to eight intensively raised, indoor herds in Spain. Seven herds used preventive anticoccidial toltrazuril treatments administered orally or by intramuscular injection, and one did not use preventive anticoccidial treatments. The diagnosis was performed using two oocyst flotation-concentration methods, Bailenger's method in every herd and the more recent Joachim's method in four herds. Oocysts were detected in every farm, the proportion of oocyst-positive samples was higher with Bailenger's technique, and the estimated overall prevalence (95% confidence interval) was 40 (32-49)%, including 47 (29-65)% in non-medicated litters, 52 (38-67)% in orally medicated litters and 28 (16-40)% in intramuscularly medicated litters (p < 0.05). However, mixed logistic regression models indicated that the risk of infection was not significantly associated with preventive anticoccidial treatments (p > 0.05), and increased with age, was higher in herds with partially compared to fully slatted dung floors in farrowing pens and in litters with pigs with diarrhoea (p < 0.05). The median (range) oocysts per gram of faeces (OpG) in infected litters by Bailenger's method was 623 (35-49048) and mixed negative binomial models revealed no significant association between infection intensity in positive litters and pen's floor type and piglets age, faecal consistency and treatment status (p > 0.05). The apparent low efficacy of Toltrazuril suggests treatment administration failures, reduced residual efficacy or low susceptibility of C. suis strains in study farms and needs further investigation.

Isospora juruviarae n. sp. (Apicomplexa: Eimeriidae) from chivi vireos Vireo chivi (Vieillot, 1817) (Passeriformes: Vireonidae) in South America.

Chivi vireos Vireo chivi (Vieillot, 1817) are passerine birds widely distributed throughout Brazil, but mainly observed in the Atlantic Forest of the South and Southeast regions of the country. In this context, the current study identifies a new species of Isospora Schneider, 1881 from V. chivi captured in the Marambaia Island, on the coast of the State of Rio de Janeiro, Southeastern Brazil. The oocysts of Isospora juruviarae Andrade & Berto n. sp. are subspheroidal to ovoidal, measuring on average 26 by 24 µm. Micropyle is absent or inconspicuous. Oocyst residuum absent, but polar granules are present. Sporocysts are ellipsoidal with pointed posterior end, measuring on average 17 by × 11 µm. Stieda and Sub-Stieda bodies are present. Sporocyst residuum is present among the vermiform sporozoites, which have refractile bodies and nucleus. This morphology was different from the other Isospora spp. recorded in the same family, superfamily and parvorder as the host. Molecular identification was targeted by the amplification and sequencing of two different loci of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and one locus of the 18S small subunit ribosomal RNA (18S) gene. Phylogenetic analyses were not very efficient in forming monophyletic groups associated with host taxon, zoogeographical region or taxonomic character; however, they confirmed the identification as a new species through comparison with sequences from Isospora spp. of wild passerines. Finally, based on the morphological and molecular analyses of the oocysts recovered from the chivi vireo V. chivi in the current work, I. juruviarae is considered new to science, being the second species recorded in the host family Vireonidae and the first to have a supplementation by molecular identification.

Morphological and molecular biology identification of Cystoisospora sp. in the blue fox, Alopex lagopus (Linnaeus, 1758).

To investigate the prevalence and molecular characteristics of Cystoisospora sp. in blue fox (Alopex lagopus), Sheather's sugar floatation method was conducted to detect coccidia in 423 fresh fecal samples randomly collected from blue fox farms from three cities in China. The overall prevalence of coccidia was 1.4% (6/423), and three Cystoisospora sp. (Cystoisospora fennechi, Cystoisospora sp. I and Cystoisospora vulpina) were identified by their morphological characteristics. The 18S ribosomal RNA (rRNA) and cytochrome c oxidase subunit I (COI) locus sequences were sequenced for molecular biological identification, homology comparison, and phylogenetic analysis of Cystoisospora sp. by single-oocyst selection technology and multi-locus-nested PCR amplification. At the 18S rRNA and COI loci, C. vulpina had 99.48% and 99.59% homology, respectively, with Cystoisospora canis and Cystoisospora ohioensis from canines. Phylogenetic analysis indicated that C. vulpina was clustered in a clade with Cystoisospora sp. from Canidae, which the relatives are consistent with the hosts. To our knowledge, this is the first report on molecular identification and evolutionary analysis of C. vulpina at two different loci.

SARCOCYSTIS AND OTHER PARASITES IN FECES OF BOBCATS (LYNX RUFUS) FROM MISSISSIPPI.

Rectal contents of 56 adult bobcats (Lynx rufus) in 2014 and 2017 from remote areas of Mississippi were examined microscopically for parasite stages after the sugar flotation method. Among the helminths, eggs/larvae found were: Paragonimus sp. in 12, Toxocara cati-like in 16, trichurid-capillarid-like in 3, hookworms in 27, and lungworms in 28. Among the protozoa, oocysts/cysts found were: Cystoisospora felis-like in 2, Cystoisospora rivolta-like in 4, Cryptosporidium sp. in 1, and Giardia sp. in 1. Additionally, numerous Sarcocystis sporocysts were detected in the feces of 12 bobcats; sporocysts were described morphologically. The status of C. felis derived from the bobcat and other wild felids is reviewed and compared with C. felis from the domestic cat. It is the first record of C. rivolta from the bobcat. The presence of eggs of Paragonimus sp. and T. cati in feces of 21.4% and 28.5%, respectively, suggests a role for the bobcat in the dissemination of these zoonotic helminths in the environment in the wild. Taxonomy of coccidia of wild Felidae is discussed and Isospora lyncisLevine and Ivens, 1981 from the Lynx is now regarded as a species inquirenda.

THE PHARMACOKINETICS AND PHARMACODYNAMICS OF ORAL PONAZURIL IN THE TREATMENT OF SYSTEMIC ISOSPOROSIS IN PASSERINE BIRDS.

Systemic isosporosis, previously atoxoplasmosis, is a significant cause of mortality in juvenile passerine birds. Recommended treatment regimens are empiric and vary in efficacy. The goal of this study was to determine the pharmacokinetics and pharmacodynamics of ponazuril for treatment of systemic isosporosis. Ponazuril, diluted with water to create an oral suspension (50 mg/ml), was administered (100 mg/kg) to 72 European starlings (Sturnus vulgaris) by a single dose via direct oral gavage (n = 24), a single dose injected into superworm larvae (Zophobas morio; n = 24), or a daily dose mixed with commercial dog food to top-dress feed for 5 d (n = 24). Peak plasma concentrations were 5.84, 2.46, and 9.13 µg/ml for the direct gavage, injected larvae, and top-dressed feed groups, respectively. With repeated dosing, mean plasma concentrations from the top-dressed feed group were maintained between 8.12 to 13.11 µg/ml. Results suggested ponazuril at a dosage of 100 mg/kg administered via direct gavage or top-dressed feed, but not via injected larvae, would exceed the concentrations needed to inhibit merogony of other apicomplexan parasites in cell culture (5 µg/ml). To assess the pharmacodynamics of this dose, seven passerine birds, red-vented bulbuls (Pycnonotus cafer; n = 2), blue-grey tanager (Thraupis episcopus; n = 1), and red-capped cardinals (Paroaria gularis; n = 4), were identified as shedders of systemic Isospora spp. via fecal qPCR. Birds were then treated with ponazuril (100 mg/kg) daily on top-dressed feed for 14 d. Fecal shedding was assessed via qPCR for 6 wk from the initiation of treatment. Treatment was associated with reduction in proportions of fecal shedding during the treatment period and the week following treatment, but shedding resumed in all birds by the end of sampling. Results support that treatment of breeding birds with 100 mg/kg ponazuril could reduce the shedding of active oocysts and decrease risk of clinical infection in susceptible juveniles.

Brief report: Isospora gilvusi n. sp. (Apicomplexa: Eimeriidae) from the warbling vireo Vireo gilvus Vieillot (Passeriformes: Vireonidae) in Morelia, Mexico.

A new coccidian species, Isospora gilvusi n. sp. (Apicomplexa: Eimeriidae) collected from the warbling vireo Vireo gilvus, is reported from Morelia, Michoacán State, Mexico. Sporulated oöcysts of the new species are spherical to subspherical, 27-31 × 27-29 (30.1 × 28.4) µm, with a length/width (L/W) ratio of 1.1; one or two polar granules are present, but micropyle and oöcyst residuum are absent. Sporocyts are ovoid to drop-shaped, 16-17 × 11-12 (17.0 × 11.5) µm, with a L/W ratio of 1.7; Stieda and sub-Stieda bodies are both present, but para-Stieda body is absent; sporocyst residuum diffuse. At the histological study, endogenous stages were observed in the epithelial cells of the duodenum. This is the second species of Isospora recorded infecting a bird of the family Vireonidae in the New World.

Isospora speciosae n. sp. (Apicomplexa: Eimeriidae) from the black-polled yellowthroat Geothlypis speciosa Sclater, 1859 (Passeriformes: Parulidae) in Ciénegas del Lerma, Mexico.

A new coccidian species, Isospora speciosae n. sp. (Apicomplexa: Eimeriidae) collected from the black-polled yellowthroat Geothlypis speciosa Sclater, is reported from the marsh Ciénegas del Lerma Natural Protected Area, Mexico. Sporulated oöcysts of the new species are subespherical to ovoidal, 24-26 × 21-23 (25.7 × 22.2) µm, with a length/width (L/W) ratio of 1.1; one or two polar granules are present, but micropyle and oöcyst residuum are absent. Sporocyts are ovoidal, 17-19 × 9-11 (18.7 × 10.2) µm, with a L/W ratio of 1.8; Stieda and sub-Stieda bodies are both present, but para-Stieda body is absent; sporocyst residuum compact. The new species is the sixth species of Isospora recorded in a bird of the family Parulidae in the New World.

Isospora bertoi n. sp. of the saffron finch, Sicalis flaveola (Aves: Passeriformes) from Brazil1.

A new species of coccidia (Protozoa: Apicomplexa: Eimeriidae) is described from the saffron finch, Sicalis flaveola, is reported from Brazil. Sporulated oocysts of Isospora bertoi n. sp. are spherical to subspherical; 23.6 (21.1-26.5) x 22.0 (19.4-24.6) µm; shape Index (L/W ratio) 1.1 (1.0-1.2) µm; with bilayer smooth walls, ~1.1 µm. Micropyle and oocyst residuum are absent, but polar granules are present. Sporocysts are elongated ellipsoidal, 16.2 (13.6-17.9) x 10.1 (8.9-12.4) µm. Stieda body is button-shaped and Sub-Stieda and Para-Stieda body are absent. Sporocyst residuum is compact and composed of hundreds of granules scattered among the sporozoites. The sporozoite is claviform with an elongated posterior refractile body and nucleus.

Integrative taxonomy for the traditional coccidians (Chromista: Miozoa: Eimeriidae) from island canaries (Aves: Passeriformes: Fringillidae): Worldwide distribution, morphological and molecular characterization, revaluations and establishment of junior synonyms.

Island canaries Serinus canaria (Linnaeus) are finches native to the North Atlantic Islands, however, they have a worldwide distribution in captivity due to their relevance as a pet bird. Coccidians are the most reported parasites of passerines worldwide, both in the wild and in captivity, being frequently associated with disease in passerines kept in rehabilitation centers and commercial breeders. This study aimed to identify coccidians from island canaries kept in captivity in Brazil. Three hundred and fifteen genomic DNA extracted from fecal samples of island canaries from different breeders from Southern and Southeastern Brazil were used to perform a nested PCR assay to amplify a partial fragment of the 28S small subunit ribosomal RNA gene (28S) of Isospora spp. Microscopic screening and morphological identification of Isospora oocysts was performed in fecal samples corresponding to PCR positive DNA samples. Fecal samples have been formalin-stored for approximately four years. Positivity rate for both microscopy and PCR was 10.5% (33/315). Posteriorly, Isospora serini (Aragão, 1933) Box, 1975 and Isospora canaria Box, 1975 were morphologically identified from fresh fecal samples of island canaries maintained by a breeder in the State of São Paulo, Southeastern Brazil, providing a genotypic characterization via sequencing of the mitochondrial cytochrome c oxidase subunit 1 (COI) and 28S genes. The 28S and COI sequences referring to the morphological identification of I. canaria was, respectively, 100% and 99% similar to sequences deposited as Isospora serinuse Yang, Brice, Elliot & Ryan, 2015 from island canaries kept in a rehabilitation center in Australia. The COI sequence referring to the morphological identification of I. serini was 100% similar to a sequence of an extraintestinal Isospora, corroborating this identification/sequencing since I. serini is the first isosporan with an extra-intestinal cycle demonstrated. The comparison of morphological and molecular data from I. canaria and I. serini from this study with published data of Isospora spp. from canaries worldwide, allowed the specific identification from preliminary generic identifications, correction of misidentifications, as well as the establishment of junior synonyms. Finally, this study provides morphological and molecular data that ensure the correct identification of the two Isospora spp. from island canaries in future studies worldwide.

A new isosporan (Apicomplexa: Eimeriidae) from barn swallow, Hirundo rustica (Aves: Passeriformes: Hirundinidae: Hirundininae), in Oklahoma, USA.

Compared to other commonly examined passerine birds for coccidian (Apicomplexa) parasites, little is available on the coccidia of birds of the family Hirundinidae, including barn swallows (Hirundo rustica). Feces was collected in May 2022 from two nesting H. rustica in McCurtain County, Oklahoma, USA; the samples contained oöcysts representing a new species of Isospora. Oöcysts of Isospora zimmermani n. sp. are subspheroidal to ovoidal with a smooth bi-layered wall, measure (L × W) 22.8 × 20.8 µm, and have a length/width (L/W) ratio of 1.1; a micropyle and oöcyst residuum were absent but polar granule(s) are present. Sporocysts are ellipsoidal to ovoidal and measure 15.2 × 9.6 µm, L/W 1.6; a knob-like Stieda body and a rounded sub-Stieda body are present. The sporocyst residuum is composed of compact spheroid with a dense, irregular mass of finer granules lying between and dispersed among the sporozoites. The new species represents only the second isosporan reported from H. rustica but the first from the USA, and fourth species documented from the avian family Hirundinidae. This article was registered in the Official Register of Zoological Nomenclature (ZooBank) as urn:lsid:zoobank.org:pub:1F1E71C3-56E8-4EA7-A9B0-1E585FAA9DE0.

A New Isospora (Apicomplexa: Eimeriidae) from the Dark-Eyed Junco, Junco hyemalis (L.) (Passeriformes: Passerellidae), in West-Central Arkansas, USA.

PURPOSE: Many passeriform birds are excellent hosts of coccidian parasites but little is known of the coccidians infecting members of the family Passerellidae, particularly from the dark-eyed junco, Junco hyemalis (Linnaeus, 1758). Here, we report from morphometric and mensural data a new species of Isospora from J. hyemalis in Arkansas, USA. METHODS: During November 2020, seven adult J. hyemalis were taken with a mist net from Polk County, Arkansas, USA, and their faecal samples examined for coccidian parasites. Samples were placed in individual vials containing aqueous potassium dichromate. They were examined for coccidia after flotation in Sheather's sugar solution, measured, and photographed. RESULTS: A single J. hyemalis (14%) was found to be passing a new species of Isospora. Oöcysts of Isospora pittmani n. sp. were spheroidal to subspheroidal with a smooth bi-layered wall, measured (L × W) 24.8 × 23.8 µm, and had a length/width (L/W) ratio of 1.04; a micropyle and oöcyst residuum was absent but typically one to up to four polar granule(s) was present. Sporocysts are ovoidal to ellipsoidal and measured 15.0 × 10.0 µm, L/W 1.5; a flattened Stieda body was present as well as a barely discernible sub-Stieda body but para-Stieda bodies were absent. The sporocyst residuum was composed of various-sized granules dispersed between and across the sporozoites. A spheroidal to ellipsoidal posterior refractile body and spheroidal anterior refractile body occasionally seen, but a nucleus was not observed. Only two previous isosporans have been reported from passerellid birds and the new species can readily be distinguished from them. CONCLUSION: This is the first coccidian reported from J. hyemalis and, more importantly, the first isosporan known from any member of the family Passerellidae in North America.

Redescription of Isospora vanriperorum Levine, 1982 (Apicomplexa: Eimeriidae) from the Northern Cardinal, Cardinalis cardinalis (L.) (Passeriformes: Cardinalidae), in North Texas, USA.

Little is known about the coccidian parasites of North American birds of the family Cardinalidae. A single isosporan, Isospora vanriperorum was previously described over 40 years ago from the northern cardinal, Cardinalis cardinalis from Hawaii. Since then, it was also reported in the green-winged saltator, Saltator similis from Brazil but the identification was invalidated recently. Feces were collected in July 2022 from two C. cardinalis found dead in Lamar County, Texas, USA. One sample contained oöcysts representing I. vanriperorum. Oöcysts are spheroidal to subspheroidal with a smooth uni-layered wall, measure (L × W) 23.8 × 22.4 µm and have a length/width (L/W) ratio of 1.04; a micropyle and oöcyst residuum are absent but polar granule(s) are present. Sporocysts are ovoidal and measure 15.5 × 10.5 µm, L/W 1.5; a knob-like Stieda body and a rounded sub-Stieda body are present. The sporocyst residuum is more often composed of various-sized granules dispersed between and across the sporozoites; it is less often in a compact rounded mass. This isosporan has now been reported from the northern cardinal for the second time but most importantly, from the mainland USA for the first time.

First detection of coccidia species in blue peafowl (Pavo cristatus) in China via both microscopic and molecular methods.

Coccidia are protozoan parasites in the class Conoidasida. To determine the prevalence and molecular characteristics of coccidia species in blue peafowl (Pavo cristatus) in Henan, China, 240 fecal specimens were collected and screened for the presence of Eimeria spp. and Isospora spp. The overall prevalence was 65.0% (156/240), and seven different coccidia species were identified: E. pavonis (51.3%, 123/240), E. arabic (40.0%, 96/240), E. riyadhae (37.1%, 89/240), E. mandali (22.9%, 55/240), E. mayurai (14.2%, 34/240), I. mayuir (10.9%, 26/240), and I. lacazei (8.5%, 21/240). E. arabic and E. riyadhae were detected for the first time in China. Additionally, we provide molecular data of the seven different coccidia species at the 18S and 28S ribosomal RNA and the COI loci. Sequence homology percentages among the five species of Eimeria at the 18S rRNA, 28S rRNA, and COI loci were 96.0%-98.6%, 90.7%-98.2%, and 85.0%-94.9%, respectively, whereas for two species of Isospora the sequence homology percentages were 98.8%, 99.1%, and 95.4% at three corresponding loci. This is the first report of the molecular data of the seven coccidia species in blue peafowl in China.

Cystoisospora suis infection in suckling piglets in Brazil: Prevalence and associated factors.

Piglet coccidiosis is a parasitic disease caused by the protozoan Cystoisospora suis, which is regarded as the most prevalent gastrointestinal parasite in intensive pig farms. Despite the availability of highly effective chemo-metaphylaxis (toltrazuril), coccidiosis is still prevalent in European and other countries. We conducted a cross-sectional study on swine farms in Brazil, to assess the prevalence of Cystoisospora suis in fecal samples and determined the associated factors. In total, 666 litters from 50 farms were sampled twice within one week between samplings (mean age at sampling: 10.75 and 17.7 days). Of 666 litters, 225 (33.8%) were positive at least once, and the expected within farm prevalence of C. suis oocysts was 32.9% (25.4-41.3%; 95% confidence interval). Oocysts were more prevalent in fecal samples collected from farms with diarrhea (odds ratio = 6.75). The room temperature was also positively associated with oocyst detection; a one-degree increase in room temperature increased the chance of a litter being positive by 23.2%. Up-to-date, this is the most comprehensive technical evidence of factors associated with C. suis infection in Brazilian industrial piglet farms.

Molecular identification of Isospora coerebae Berto, Flausino, Luz, Ferreira amp; Lopes, 2010 (Chromista: Miozoa: Eimeriidae) from the bananaquit Coereba flaveola (Linnaeus, 1758) (Passeriformes: Thraupidae: Coerebinae) from Brazil.

Isospora coerebae Berto, Flausino, Luz, Ferreira Lopes, 2010 is a coccidian protozoan described from bananaquits Coereba flaveola (Linnaeus, 1758), on Marambaia Island, which is located on the southeastern Brazilian coast. In this current work, I. coerebae is identified from C. flaveola in a protected area close to Marambaia Island, but on the mainland, establishing a new location of parasitism, in addition to providing a preliminary genotypic characterization via sequencing of two regions of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene. Its oocysts are sub-spherical, 24.4 22.9 m, with smooth, bilayered wall, ~1.7 m thick. Micropyle, polar granules and oocyst residuum are absent. Sporocysts are elongate ovoidal, 17.6 10.5 m. Stieda body prominent and rounded and sub-Stieda body short and wide. Sporocyst residuum is composed of scattered granules of different sizes. Sporozoites are vermiform with a prominent posterior refractile body. The oocysts of the current work are morphologically equivalent to the original description of I. coerebae, which have a typical and easily identifiable morphology, mainly in the Stieda and Sub-Stieda bodies. The two sequenced gene regions of the COI gene approximated I. coerebae to Isospora spp. from Southeastern Brazil, but also from Isospora spp. from passerines of North America, Europe and Asia. Although there is a small overlap between the two genic regions sequenced in the current work, it is estimated that the longer COI sequence, which was recently designed and still not widespread, should show better phylogenetic results in the future.

The First Isospora Species (Apicomplexa: Eimeriidae) Described from the Northern Yellow-Shafted Flicker (Colaptes auratus luteus) in Ontario, Canada.

PURPOSE: Members of the genus Isospora commonly infect a wide variety of wild birds. By combining morphometric and genetic data, we are able to differentiate and describe a new apicomplexan parasite, Isospora picoflavae n. sp., from the Northern Yellow-shafted Flicker (Colaptes auratus luteus) in Ontario, Canada. METHODS: Unsporulated oocysts were observed in intestinal contents obtained during necropsy; these oocysts completed exogenous sporulation within 7 days at room temperature. Complete nuclear 18S rDNA and complete mitochondrial genome sequences were obtained from this previously undescribed Isospora sp. to compare with related coccidia. RESULTS: Sporulated oocysts were subspherical in shape measuring an average of 22.7 × 21.7 µm (n = 53; mean shape index 1.05) containing a near-spherical polar granule. Sporocysts were ovoidal to ellipsoidal in shape, measuring an average size of 17.0 × 10.5 µm (n = 9; mean shape index 1.62). Sporocyst residuum was concentrated in an irregular, spherical mass in the middle of each sporocyst. The isolated oocysts differed morphologically from other Isospora species reported to infect members of the family Picidae. Based on phylogenetic analyses using either combined nuclear 18S rDNA and mitochondrial cytochrome c oxidase I (COI) sequences or complete mitochondrial genome sequences, this Isospora sp. n. isolated from the Northern Flicker grouped within a clade containing Isospora species described from various passeriform hosts. CONCLUSION: Based on combined morphological and molecular data, the oocysts found in the gastrointestinal contents of Colaptes auratus luteus represent a new species of Isospora named herein as Isospora picoflavae n. sp.

Isospora leptopogoni n. sp. (Apicomplexa: Eimeriidae) from the sepia-capped flycatcher Leptopogon amaurocephalus Tschudi, 1846 (Passeriformes: Rhynchocyclidae) in South America.

Coccidian protozoan species recorded from flycatchers are few, but they have been described with a certain frequency in recent years. In this context, the present study describes a new Isospora sp. from sepia-capped flycatchers Leptopogon amaurocephalus Tschudi, 1846 captured in the Itatiaia National Park and in a reforestation area which is about 60 km away from the park boundaries, in addition to providing a molecular identification via sequencing of the mitochondrial cytochrome c oxidase subunit 1 gene. Isospora leptopogoni n. sp. has oöcysts that are subspheroidal to ovoidal, measuring on average 22.0 × 19.7 µm, with a smooth, bi-layered wall, c.1.7 µm thick. The micropyle is delicate or inconspicuous. Oöcyst residuum is absent, but one to three polar granules are present. Sporocysts are lemon-shaped, measuring on average 14.7 × 9.3 µm, with a knob-like Stieda body and a rectangular to rounded sub-Stieda body. Sporocyst residuum is present, consisting of compactly bounded granules. Sporozoites are vermiform, with refractile bodies and nucleus. Isospora leptopogoni is different from other Isospora spp. mainly due to its lemon-shaped sporocysts, the presence of micropyle and details of Stieda and sub-Stieda bodies. Phylogenetic analysis placed I. leptopogoni close to other Isospora spp. recorded from phylogenetically related hosts and from the same biogeographic region. Finally, the recurrent finding of this coccidian species in the same L. amaurocephalus specimen in a specific locality in the Itatiaia National Park suggests that the dispersion of I. leptopogoni needs continuous transmissions between susceptible passerines as the area of movement of each L. amaurocephalus specimen appears to be quite small.