Remarks on Eimeria spp. (Apicomplexa: Eimeriidae) from Kobus spp. (Bovidae: Reduncini), with supplementary morphological data of Eimeria congolensis Ricci-Bitti et al., 1973 from a new host subspecies, the common waterbuck Kobus ellipsiprymnus ellipsiprymnus (Ogilbyi, 1833).

Reduncin bovids of Kobus spp. (Bovidae: Reduncini) are natively distributed in sub-Saharan Africa, although some populations have been introduced into parks and zoos around the world. The majority of the species has declining populations, being categorized as threatened by the International Union for Conservation of Nature and Natural Resources; therefore, protective measures for the conservation of Kobus spp. are necessary, including the study of their parasites, such as the eimeriid coccidians (Apicomplexa: Eimeriidae). In this context, the aim of the current study was to brings together the taxonomic data from the descriptions and reports of Eimeria spp. from reduncin bovids, based on the detailed morphological identification of Eimeria congolensis Ricci-Bitti, Pampiglione & Kabala, 1973 from a new host subspecies, the common waterbuck Kobus ellipsiprymnus ellipsiprymnus (Ogilbyi, 1833), in a safari park of Portugal. Five Eimeria spp. are recorded from reduncin bovids, in addition to six more reports identified generically as Eimeria sp., which were compared and taxonomically rearranged. The oocysts identified as E. congolensis in the current study were compatible with the original description and were supplemented for some taxonomic characters not originally included, such as: Stieda body flattened to nipplelike, sub-Stieda body rounded to trapezoidal, sporocyst residuum granular and membrane-bound, in addition to greater details of the micropyle, among others. Finally, the current study highlights the importance of studying the coccidians of reduncin bovids for the conservation of Kobus spp. due to the possibility of these Eimeria spp. are extra-intestinal parasites, which can potentially cause severe coccidiosis associated with increased morbidity and mortality in certain threatened populations of Kobus spp.

Isospora pichororei n. sp. (Chromista: Apicomplexa: Eimeriidae) from rufous-capped spinetails Synallaxis ruficapilla Vieillot, 1819 (Passeriformes: Furnariidae: Synallaxiinae) in South America.

Spinetails are a suboscine passerines of the genus Synallaxis Vieillot, 1818 which have great interest for ornithology, given the wide diversity of 37 species that are distributed throughout the Neotropical region. Despite this wide diversity and distribution, Synallaxis spp. have never been recorded as hosts of coccidian parasites. In this context, the current study describes a new species of Isospora Schneider, 1881 from rufous-capped spinetails Synallaxis ruficapilla Vieillot, 1819 captured in the Itatiaia National Park, which is a federal conservation unit in Southeastern Brazil. The oocysts of Isospora pichororei Genovez-Oliveira & Berto n. sp. are subspheroidal to ovoidal, measuring on average 25 by 21 µm. Micropyle is present, but discrete. Oocyst residuum absent, but one or two polar granules are present. Sporocysts are ellipsoidal with slightly pointed posterior end, measuring on average 17 by 10 µm. Stieda and sub-Stieda bodies are present. Sporocyst residuum is clustered among the vermiform sporozoites, which have striations, refractile bodies and nucleus. This morphology was different from the other Isospora spp. recorded in the host family Furnariidae. Molecular identification was targeted by the amplification and sequencing of a locus of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. This sequence had the highest similarity of 99.5% with a sequence deposited for Isospora oliveirai Ortúzar-Ferreira & Berto, 2020, which is a coccidian species that parasitizes suboscine tityrids Schiffornis virescens (Lafresnaye, 1838), also in the Itatiaia National Park. Phylogenetic analysis grouped some species in subclades, including I. pichororei with I. oliveirai; however, it was inconclusive in an expectation of parasite-host coevolution. Finally, I. pichororei is established as new to science, being the first description from Synallaxinae and the third description from Furnariidae. Furthermore, this is the first Isospora sp. from the host family Furnariidae to have a molecular supplementation by sequencing a locus of the cox1 gene of the mitochondrial genome.

POPULATION DYNAMICS OF ENTERIC PARASITES IN THE ENDANGERED VANCOUVER ISLAND MARMOT (MARMOTA VANCOUVERENSIS).

Enteric parasites can have wide-ranging effects throughout an ecosystem, often driving coevolutionary and ecological processes. Parasites have long been overlooked in conservation efforts because of the negative impact inflicted on their hosts; however, parasites make up a significant component of Earth's biodiversity and host conservation efforts need to be parasite inclusive. The Vancouver Island marmot (VIM), Marmota vancouverensis, is an endangered alpine rodent endemic to Vancouver Island, British Columbia, Canada. Captive-bred VIMs are released to augment the wild population, but their susceptibility to parasites is unknown. The objectives of this study were to describe the diversity, prevalence, severity, and temporal variation of VIM enteric parasites. Noninvasive fecal samples were collected from wild and captive marmots and analyzed using a modified McMaster fecal egg floatation technique to indicate parasite prevalence and relative mean abundance. We identified oocysts and ova from 3 parasite taxa including a protozoan coccidium not previously described in the VIM (prevalence 68%), an ascarid nematode Baylisascaris laevis (prevalence 82%), and an anoplocephalid cestode Diandrya vancouverensis (prevalence 8%). Depending on the species, comparisons revealed variation in parasite infection by sex, by colony, and between wild and captive VIMs, but not among age classes or by female reproductive status. Finally, captive VIMs displayed significant monthly variation in parasite prevalence and mean egg abundance, suggesting a seasonal influence on parasite egg shedding. This information is critically important for future research investigating the influences of these trends on the health, ecology, and conservation of VIMs and their parasites.

Toxoplasma gondii seroprevalence and risk factors in cats in Rio de Janeiro.

Seroprevalence studies on cats are essential for monitoring the occurrence of Toxoplasma gondii infection. The present research investigated anti-T. gondii antibodies, risk factors, clinical signs, hematology and serum biochemistry in cats from different regions of Rio de Janeiro. An overall 18.7% (17/91) of the cats were seroreactive, and age was associated with increased chances of seroprevalence of anti-T. gondii antibodies. Clinical signs, hematology and serum biochemistry parameters did not help achieve an antemortem diagnosis of cat toxoplasmosis. The parasite circulates in cats from three major regions of Rio de Janeiro, and the present data set will contribute to future epidemiological studies in this endemic state of Brazil.

Prevalence and risk factors for gastrointestinal parasitosis in small ruminant farms in the Sahelian zone of Niger.

The general aim of this study is to analyse the risk factors for gastrointestinal parasitosis in small ruminants in order to contribute to the emergence of targeted treatment methods, at herd and agro-climatic zone levels, for the integrated and sustainable management of parasitic diseases in Sahelian livestock systems. The methodology was based on a questionnaire survey conducted in 37 villages and coprological analysis using the McMaster method on faecal samples from 968 small ruminants, including 555 goats and 413 sheep. Multiple logistic regression was used to highlight the risk factors associated with each type of parasitosis encountered. The results showed that the most widespread farming system remained 100% traditional, with feeding based essentially on natural grazing. Coprological results showed the prevalence of nematodosis (70.2%), Cestodosis (4.1%) and Coccidiosis (79.9%), with an average prevalence of coinfection of 56.9%. These parasite loads were significantly higher during the rainy season and in the more arid northern Sahelian zone, with a marked reduction at the end of the season. Average parasitic egg excretions were 1089 EPG of nematodes and 6864 EPG of coccidia. Parasite loads were higher in the wetter southern strip and varied significantly by breed. Of the five breeds of small ruminants studied, the ara-ara sheep had the highest parasitic loads and prevalences for nematodosis (78.6%), coccidiosis (89,3%) and coinfection (70.9%), appears to be the most susceptible to parasitosis. As for risk factors for severe parasite pressure, animals at the end of the rainy season, older animals and those with poor body condition were at risk of nematodiasis or coinfection. On the other hand, animals at the beginning of the rainy season, farms located in less arid southern Sahelian zones and male subjects were the groups at significant risk of coccidiosis. In these extensive Sahelian farming conditions, the control of these parasitoses by selective treatment of animals could be developed, targeting in particular the risk groups highlighted in this study.

Coccidia of Guinea fowls: Validity of recorded Eimeria spp. (Apicomplexa: Eimeriidae) and first molecular identification of Eimeria grenieri Yvoré & Aycardi, 1967.

Guinea fowls, Numida meleagris (L., 1758), are galliform birds native to sub-Saharan Africa, but introduced in several countries around the world for domestic breeding and/or animal production. This species is considered more resistant to disease by Eimeria spp. than other domestic galliform birds. Here we review the Eimeria spp. known to infect species of Numididae and provide the first molecular identification of an Eimeria sp. from Guinea fowls. There are currently 3 named eimerians from Guinea fowls; Eimeria numidae Pellerdy, 1962; Eimeria grenieri Yvoré and Aycardi, 1967; and Eimeria gorakhpuri Bhatia & Pande, 1967. We reviewed each of these species descriptions and documented their taxonomic shortcomings. From that, we suggest that E. gorakhpuri is a junior synonym of E. numidae. In conclusion, we have morphologically redescribed in detail E. grenieri from N. meleagris from Rio de Janeiro and provided molecular supplementation through sequencing of three non-overlapping loci in cox1 and cox3 genes and fragments of small and large subunit mitochondrial rDNA.

A New Isosporan (Apicomplexa: Eimeriidae) from the Pacific Blue-Tailed Skink, Emoia caeruleocauda (Sauria: Scincidae: Eugongylinae), from Guam, U.S. Territory.

PURPOSE: Nothing is known about coccidians (Apicomplexa: Eimeriidae) from the Pacific blue-tailed skink, Emoia caeruleocauda. Here, we report mensural and morphometric data on a new species of Isospora from E. caeruleocauda from Guam, US Territory. METHODS: Feces from four E. caeruleocauda collected by hand in November 2023 were placed in individual vials containing 2.5% potassium dichromate. They were examined for sporulated oocysts after flotation in Sheather's sugar solution, measured, and photographed. RESULTS: A single (25%) E. caeruleocauda was found to be passing oocysts representing a new species of Isospora. Oocysts of Isospora guamensis n. sp. are ellipsoidal to ovoidal with a bi-layered wall, measure (L × W) 16.5 × 11.8 µm, and have a length/width (L/W) ratio of 1.4; a micropyle and an oocyst residuum were absent but a polar granule was present. Sporocysts are ovoidal and measure 9.4 × 6.5 µm, L/W 1.4; Stieda and sub-Stieda bodies were present but a para-Stieda body was absent. The sporocyst residuum is composed various-sized granules in a compact rounded or irregular mass, sometimes dispersed between the sporozoites. The new species can be differentiated from all other isosporans from skinks by possessing the smallest oocysts known from this host family. CONCLUSION: This is the first time an isosporan coccidian has been reported from E. caeruleocauda as well as the first report of a coccidian from a Guam-inhabiting skink.

Effects of Dietary Inclusion of a Proprietary Combination of Quillaja saponaria and Yucca schidigera on Intestinal Permeability and Immune Response in Broiler Chickens during a Coccidia Challenge.

This study assessed the impact of Magni-Phi Ultra (MPU) inclusion on intestinal integrity and immunity in broiler chickens challenged with coccidia during peak and recovery phases. A total of 128 male Ross 708 broiler chicks were randomly allotted to one of four treatment groups (four chicks/cage). Treatments included an uninfected control (UUC); a coccidial challenge (CC) infected control (IUC); a CC fed salinomycin at 66 ppm (SAL); and a CC fed Magni-Phi Ultra at 0.11 g/kg of diet (MPU). At 16 days post-hatch, all birds in the CC groups were orally gavaged with a 3× dose of a live coccidia vaccine. At 5 dpi, the birds fed MPU and SAL showed decreased plasma FITC-d, oocyte shedding, and lesion scores and higher BWG compared to the IUC birds (p < 0.05). Jejunum IL-17, IL-10, and IFN-ϒ mRNA expression was higher in the IUC compared to the UUC (p < 0.05) group at 5 dpi. At 12 dpi, the birds fed MPU or SAL had lower plasma FITC-d and jejunum IFN-ϒ and IL-10 mRNA expression compared to the IUC birds (p < 0.05). This study indicates that MPU supports intestinal integrity and mucosal immune responses during the peak and recovery phases of infection, which may lead to improved health and performance.

Cassia alata's dual role in modulating MUC2 expression in Eimeria papillata-infected jejunum and assessing its anti-inflammatory effects.

Coccidiosis poses significant hazards to animals, particularly in terms of compromised health, reduced productivity, and economic losses in livestock farming. The conventional treatments for coccidiosis often involve synthetic drugs, contributing to concerns about drug resistance and environmental impact. The pressing need for eco-friendly alternatives is highlighted in this study, emphasizing the importance of exploring medicinal plants like Cassia alata leaf extracts (CAE) against Eimeria papillata-induced infection in mice. The CAE exhibited significant phenolic (2.17 ± 0.03 g/100 g) and flavonoid (0.14 ± 0.01 g/100 g) content and demonstrated notable antioxidant activity. In infected mice, the CAE treatment led to a substantial reduction in oocyst output (~6 fold), ameliorating necrotic enteritis and inflammatory changes in the jejunum. Additionally, CAE treatment increased goblet cell numbers (9.3 ± 0.1 / villus) and decreased macrophage infiltration in the intestinal villi. Molecular analyses revealed CAE's positive modulation of MUC2 gene and notably reduced the levels of pro-inflammatory cytokines (specifically IL-1ß, IL-10, and IFN-γ) when contrasted with the infected cohort. Furthermore, CAE treatment significantly reduced nitric oxide levels (44.03 ± 2.4 µmol/mg), showcasing its anti-inflammatory properties. The findings of this study not only contribute to the understanding of CAE's therapeutic potential but also underscore the importance of seeking eco-friendly alternatives in the face of coccidiosis challenges, addressing both the well-being of animals and the sustainability of agricultural practices. RESEARCH HIGHLIGHTS: Cassia alata extract (CAE) exhibited significant phenolic and flavonoid content, displaying notable antioxidant activity. In infected mice, CAE treatment led to a substantial reduction in oocyst output, ameliorating necrotic enteritis and inflammatory changes in the jejunum. CAE treatment increased goblet cell numbers and decreased macrophage infiltration in the intestinal villi, while molecular analyses revealed its positive modulation of the MUC2 gene and notable reduction in pro-inflammatory cytokine levels. Additionally, CAE treatment significantly reduced nitric oxide levels, showcasing its anti-inflammatory properties.

Morphology, morphometry, and phylogeny of the protozoan parasite, Eimeria labbeana-like (Apicomplexa, Eimeriidae), infecting Columba livia domestica.

Introduction: Eimeria spp. are intracellular protozoan parasites of the phylum Apicomplexa causing economic losses to various wild and domestic animals. An eimerian species infecting Columba livia domestica was identified in this study. Methods: A total of 15 faecal samples were examined by floatation technique, a prevalence rate of 60% was reported. Eimerian oocysts were sporulated in 2.5% potassium dichromate solution then identified using morphological and molecular (DNA amplification of the 18S rRNA and ITS-1 genes) diagnostic techniques. Results: Sporulated oocysts were identified as Eimeria labbeana-like, after morphometry with typical bi-layered wall with spherical to subspherical oocysts morphology. A polar granule is present, but no micropyle or oocyst residuum. Sporocysts are elongated ovoidal with stieda body. Sporocyst residuum with many granules and sporozoites with refractile bodies and nucleus. Both 18S rRNA and ITS-1 sequences have been deposited in GenBank database. DNA sequences from the partial 18S rRNA generated from the oocysts were found to be related to eimerian and isosporan parasites found in domestic pigeons. For the first time, ITS-1 sequences for E. labbeana-like were provided. Conclusion: The necessity of using molecular techniques to describe pigeon intestinal coccidian parasites in conjunction with traditional morphology-based tools was emphasized in this work in order to understand the biology of such parasites.

Assessing the efficacy of the ovicidal fungus Mucor circinelloides in reducing coccidia parasitism in peacocks.

The biological control of gastrointestinal (GI) parasites using predatory fungi has been recently proposed as an accurate and sustainable approach in birds. The current study aimed to assess for the first time the efficacy of using the native ovicidal fungus Mucor circinelloides (FMV-FR1) in reducing coccidia parasitism in peacocks. For this purpose, an in vivo trial was designed in the resident peacock collection (n = 58 birds) of the São Jorge Castle, at Lisbon, Portugal. These animals presented an initial severe infection by coccidia of the genus Eimeria (20106 ± 8034 oocysts per gram of feces, OPG), and thus received commercial feed enriched with a M. circinelloides suspension (1.01 × 108 spores/kg feed), thrice-weekly. Fresh feces were collected every 15 days to calculate the coccidia shedding, using the Mini-FLOTAC technique. The same bird flock served simultaneously as control (t0 days) and test groups (t15-t90 days). The average Eimeria sp. shedding in peacocks decreased up to 92% following fungal administrations, with significant reduction efficacies of 78% (p = 0.004) and 92% (p = 0.012) after 45 and 60 days, respectively. Results from this study suggest that the administration of M. circinelloides spores to birds is an accurate solution to reduce their coccidia parasitism.

Oral delivery of Eimeria acervulina transfected sequentially with two copies of the VP2 gene induces immunity against infectious bursal disease virus in chickens.

Chicken coccidiosis caused by Eimeria spp. can occur on almost all poultry farms, causing huge economic losses to the industry. Genetically manipulated Eimeria parasites as a vaccine vector to deliver viral antigens have been reported. In our preliminary study, transgenic E. acervulina expressing a VP2 gene (Ea-VP2) of the infectious bursal disease virus (IBDV) demonstrated partial protection against IBDV infection. To enhance immune responses, we aimed to increase the VP2 gene copy number in transgenic E. acervulina. In this study, we used a novel plasmid vector carrying a VP2 gene fused with three flag tags and a red fluorescent reporter gene (mCherry). The vector was introduced into Ea-VP2 sporozoites through nucleofection, leading to the generation of Ea-2VP2. Subsequent analysis revealed a notable escalation in the fluorescent rate, increasing from 0.11 to 95.1% following four consecutive passages facilitated by fluorescent-activated cell sorting. Verification via PCR, Western blot, and immunofluorescence confirmed the successful construction of the Ea-2VP2 population. Despite lower fecundity compared to wild-type E. acervulina, Ea-2VP2 maintained immunogenicity. Our research effectively created a transgenic E. acervulina strain transfected sequentially with two copies of the VP2 gene from IBDV. This modification resulted in an increased humoral immune response after primary immunization in chickens. Additionally, it demonstrated a degree of protection within the bursa against IBDV infection. Future studies will focus on further enhancing immune response levels.

Prevalence of coccidia in lagomorphs in China between 1981 and 2023: A systematic review and meta-analysis.

Lagomorpha coccidiosis, caused by coccidia, is a prevalent disease affecting rabbits, hares and pikas. This meta-analysis aimed to estimate the pooled prevalence of coccidia infection in lagomorphs and identify potential risk factors. A systematic search of six databases yielded 102 studies published between 1981 and 2023. The pooled prevalence of Eimeriidae, Sarcocystidae and Cryptosporidiidae in lagomorphs was 76.4%, 6.2% and 3.9%, respectively. Rabbits had the highest prevalence of Eimeriidae (76.8%) and Sarcocystidae (7.4%), while pikas had the highest prevalence of Cryptosporidiidae (6.2%). Juvenile rabbits exhibited the highest prevalence of Eimeriidae (84.6%) and Cryptosporidiidae (9.9%). Northwest China had the highest prevalence of Eimeriidae (87.8%). Over time, the prevalence of Eimeriidae declined (Coefficient: -0.0062; P<0.05), but remained high (65.0%) in the past five years. Our findings highlight the prevalence of Eimeriidae infection in lagomorphs and the need for further research on Sarcocystidae and Cryptosporidiidae infections. We emphasize the importance of developing lagomorpha coccidia vaccines and implementing vaccination schedules for juvenile rabbits to mitigate coccidia infections.

Identification and genetic characterization of a novel species of Choleoeimeria Schneider, 1875 from a captive-bred bilby (Thylacomyidae; Macrotis lagotis) (Reid, 1837) in Western Australia.

A novel Eimeria sp. from a captive-bred bilby (Macrotis lagotis Reid, 1837) has been identified in Western Australia. The bilby was bred at the Kanyana Wildlife Rehabilitation Centre, Perth, as part of the National Bilby Recovery Plan. Oocysts (n = 31) irregular blunt ellipsoidal, 17-18 × 11-12 (17.2 × 11.3); length/width (L/W) ratio 1.4-1.5 (1.5). Wall bi-layered, 0.8-1.0 (0.9) thick, outer layer smooth, c.2/3 of total thickness. Micropyle barely discernible. Oocyst residuum is absent, but 2-3 small polar granules are present. Sporocysts (n = 31) ovoidal, 7-8 × 5-6 (7.8 × 5.7); L/W ratio 1.3-1.4 (1.4). Stieda, sub-Stieda and para-Stieda bodies absent or indiscernible; sporocyst residuum present, usually as an irregular body consisting of numerous granules that appear to be membrane-bound or sometimes diffuse among sporozoites. Sporozoites vermiform with a robust refractile body. Further molecular characterization was conducted on the sporulated oocysts. At the 18S locus, it sat in a large clade of the phylogenetic tree with two isolates of Eimeria angustus from quendas (Isoodon obesulus Shaw, 1797) and the Choleoeimeria spp. It shared the highest identity with E. angustus (KU248093) at 98.84%; at the COI gene locus, it was unique and most closely related to Choleoeimeria taggarti, which is hosted by another species of marsupial, the yellow-footed antechinus (Antechinus flavipes flavipes), with 90.58% genetic similarity. Based on morphological and molecular data, this isolate is a new species and named as Choleoeimeria yangi n. sp.

Factors associated with strongylida infections in sheep on farms in peri-urban Nairobi, Kenya.

Gastrointestinal parasite infections are among the major limitations to production in sheep in many parts of the world. It is important to continually assess their levels of infection in order to institute control measures and reduce the impact. This study determined the factors associated with the strongylida egg counts in sheep on selected farms in peri-urban Nairobi, Kenya. This was a cross-sectional study in which farm and animal-level data, including faecal samples, were collected from 1640 sheep from 30 purposively selected farms in Ruai, and Kamulu wards in Kasarani sub-county and Utawala and Mihango wards in Embakasi East Sub-County, in Nairobi County Kenya. The faecal samples were subjected to coprological examination using a modified McMaster technique to determine counts of strongylida eggs and coccidia oocysts with a detection level of 100 egg or oocyst per gram (EPG or OPG) of faeces. The positive faecal samples for strongylida eggs were pooled per farm and cultured for morphological identification of larval stage three. Descriptive statistics and multilevel mixed-effect logistic regression analyses were used to determine factors associated with strongylida egg count ≥600 EPG (p < 0.05). The receiver operating characteristics curve was used to assess the overall diagnostic performance in the final model. Strongylida eggs were detected in 45.5% (746/1640) of the sheep, and the mean EPG was 486.0± 858.9 with a median of 200 and a range of 0-16,700. The coccidia oocysts were detected in 49.4% (810/1640) of the sheep with a mean OPG was 341.7± 1782.4, a median of 0 and a range of 0-60,000. In the coprocultures, the nematode genera identified (% differential count of L3) were Haemonchus (90%), Trichostrongylus (5%) and Oesophagostomum (5%). In the final multivariable regression model, the odds of detecting EPG ≥ 600 was 1.44 times higher for sheep shedding coccidia oocysts than those that did not. The odds for detecting EPG ≥ 600 was 4.01 times for sheep in Ruai ward compared with those in the combined Kamulu, Utawala and Mihango wards. The receiver operating characteristic curve area was 73.1%, suggestive of good model performance. The results suggest that gastrointestinal strongylida and coccidia infections are common in sheep and farmers should be educated on the importance of appropriate control measures.

Mixed Eimeria and Cryptosporidium infection and its effects on pathology and clinical outcomes in juvenile Asian seabass (Lates calcarifer) cultured in Thailand.

Coccidiosis is an important disease in juvenile fish because of severe intestinal injury during infection. We first reported the mixed infection of intestinal coccidia and its association with health status and pathological findings in juvenile Asian seabass (Lates calcarifer) cultured in Thailand. Two groups of Asian seabass, 60-day fish and 90-day fish, were sampled to investigate prevalence and coccidian infection intensity using morphological characterization and PCR. Phylogenetic analysis of 18S rRNA gene amplified from the intestines revealed Eimeria sp. and Cryptosporidium sp. infection. The prevalence of Eimeria sp. and Cryptosporidium sp. in sampled fish was 100%. Clinical outcomes assessed, using health assessment index (HAI) scoring and semi-quantitative grading of intestinal lesions and inflammation, demonstrated that all fish developed variety of pathology and clinical illness; however, infection intensity in 60-day fish was significantly higher (p < .05) than 90-day fish. The HAI score of 60-day fish was poorer than 90-day fish, which correlated to a high infection intensity (r = .397), analysed by Pearson correlation coefficient. Overproduction of intestinal oxidants contributing to mucosal injury was examined by nitrotyrosine expression. The high production of reactive nitrogen species indicated severe inflammatory response, and intestinal injuries occurred mainly in the 60-day fish.

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.

IFN-γ inhibitory molecules derived from Eimeria maxima inhibit IL-12 secretion by modulating MAPK pathways in chicken macrophages.

IFN-γ plays a crucial role in resisting intracellular parasitic protozoa, such as Eimeria species. In our previous study, we identified 4 molecules derived from Eimeria maxima (E. maxima) that significantly inhibited IFN-γ production. However, the mechanism underlying this inhibitory effect remains unknown. In this study, we first investigated the effects of these 4 IFN-γ inhibitory molecules on the expression levels of chicken Toll-like receptors (chTLRs), IL-12, IL-10, TGF-ß, and TNF-α in chicken macrophage HD11 and bone marrow-derived dendritic cells (BMDCs). The results demonstrated that these 4 inhibitory molecules significantly downregulated the mRNA levels of chTLR-2, chTLR-4, chTLR-21, and both mRNA and protein levels of IL-12. Subsequently, to clarify the effects of these 4 inhibitory molecules on the IL-12 secretion-related signaling pathways in chicken macrophages, qRT-PCR and Western blot were used to detect the changes of key molecules involved in the signaling pathways of IL-12 secretion (NF-κB, ERK1/2, p38, JNK, STAT3) following coincubation with these inhibitory molecules. Finally, RNAi was employed to verify the function of key molecules in the signaling pathway. The results revealed a significant upregulation in the expression of ERK1/2 phosphorylated protein induced by the 4 inhibitory molecules. Knockdown of the ERK1/2 gene significantly reduced the inhibitory effect of the 4 E. maxima inhibitory molecules on IL-12. These findings indicate that the 4 inhibitory molecules can inhibit the secretion of IL-12 by upregulating the expression of ERK1/2 phosphorylated protein, which is a key molecule in the ERK-MAPK pathway. Our study may contribute to elucidating the mechanisms underlying immune evasion during E. maxima infections, thereby providing new insights for the control of chicken coccidiosis.

A NEW EIMERIA SPECIES (APICOMPLEXA: EIMERIIDAE) PARASITIZING COMMERCIAL CHUKAR PARTRIDGES (ALECTORIS CHUKAR) IN SOUTHERN ONTARIO.

A commercial producer hatching and rearing chukar partridges (Alectoris chukar) in Ontario, Canada had flocks experiencing coccidiosis. Microscopic analysis of Eimeria species isolated from a field sample indicated the presence of 2 distinct oocyst morphotypes; the most abundant species was determined to be Eimeria chapmani, based on oocyst morphology and sequence-based genotyping, and the less abundant, second Eimeria sp. was an undescribed parasite. Oocysts of the unknown Eimeria sp. were large and oval-shaped; dimensions averaged 27.9 µm by 17.0 µm (shape index = 1.65 µm). Oocysts contained at least 1 polar granule and 4 almond-shaped sporocysts with average dimensions measuring 12.5 µm by 6.9 µm (shape index = 1.83). Each sporocyst featured a Stieda body, sub-Stieda body, and sporocyst residuum; a sporocyst contained 2 sporozoites that each possessed a small anterior refractile body and a larger posterior refractile body. Virtually all oocysts sporulated after 24 hr when suspended in potassium dichromate at room temperature (22 C) on a rotary platform. Experimental infections with various doses of oocysts demonstrated elevated parasite shedding from birds gavaged with higher challenge doses; fecundity generally decreased in heavier infections. The approximate prepatent period of the parasite was 4-5 days (unsporulated oocysts observed histologically at 90 hr postinfection and in feces by day 5) and patency lasted until day 12 postinfection. To characterize the endogenous development of the Eimeria sp., tissues were collected at 8 regions along the intestinal tract (including the ceca and rectum) every 6 hr throughout the estimated prepatent period. Parasites were observed to infect the descending and ascending duodenum, midjejunum, proximal and distal ileum, and the ceca. The endogenous stages identified included intracellular sporozoites, 3 generations of merogony, and gametogonic stages. Sequences of the mitochondrial genome (GenBank MW934555) and nuclear 18S ribosomal DNA (GenBank MW934259) were obtained using polymerase chain reaction amplification for Sanger sequencing, and these were unique from all published sequences on GenBank. Molecular data, in conjunction with the unique biology of the Eimeria sp. isolated from the chukar partridge flock, support that this coccidium is new to science.

RNA-Seq of Phenotypically Distinct Eimeria maxima Strains Reveals Coordinated and Contrasting Maturation and Shared Sporogonic Biomarkers with Eimeria acervulina.

Strains of Eimeria maxima, an enteric parasite of poultry, vary in virulence. Here, we performed microscopy and RNA sequencing on oocysts of strains APU-1 (which exhibits more virulence) and APU-2. Although each underwent parallel development, APU-1 initially approached maturation more slowly. Each strain sporulated by hour 36; their gene expression diverged somewhat thereafter. Candidate biomarkers of viability included 58 genes contributing at least 1000 Transcripts Per Million throughout sporulation, such as cation-transporting ATPases and zinc finger domain-containing proteins. Many genes resemble constitutively expressed genes also important to Eimeria acervulina. Throughout sporulation, the expression of only a few genes differed between strains; these included cyclophilin A, EF-1α, and surface antigens (SAGs). Mature and immature oocysts uniquely differentially express certain genes, such as an X-Pro dipeptidyl-peptidase domain-containing protein in immature oocysts and a profilin in mature oocysts. The immature oocysts of each strain expressed more phosphoserine aminotransferase and the mature oocysts expressed more SAGs and microneme proteins. These data illuminate processes influencing sporulation in Eimeria and related genera, such as Cyclospora, and identify biological processes which may differentiate them. Drivers of development and senescence may provide tools to assess the viability of oocysts, which would greatly benefit the poultry industry and food safety applications.