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The Apicomplexa parasitic phylum rhoptry neck protein 2 (RON2) plays a key role in the process of invading host cells. Eimeria tenella, an intracellular protozoan shares a similar conserved invasion pattern. However, whether E. tenella RON2 participates in the process of invading the host intestinal epithelium is poorly understood. In this study, the sequence of EtRON2 was analyzed and expressed. The expression of the truncated extracellular N-terminal fragment of EtRON2 (403-700 aa, designated EtRON2 403-700 ) with a molecular mass of 38.3 kDa. EtRON2 in the sporozoite protein was detected at 151.4 kDa by rabbit anti-rEtRON2 403-700 antibody. Immunofluorescence results showed that EtRON2 was mainly localized to the nucleus and apex of the E. tenella sporozoite. qPCR results showed that the highest expression level of EtRON2 was detected in sporulated oocysts compared with other developmental stages of E. tenella. In vitro invasion inhibition assays showed that the capacity of sporozoites to invade DF-1 cells was significantly inhibited after pretreatment with the rabbit anti-rEtRON2 403-700 antibody. Silencing the EtRON2 gene by RNA interference (RNAi) significantly inhibited EtRON2 expression and significantly reduced the invasion of DF-1 cells by sporozoites. In vivo experiments revealed a significant decrease parasite burden and oocyst outputs in chicks after infection with EtRON2 gene-silenced sporozoites by cloacal inoculation. Recombinant EtRON2 403-700 (rEtRON2 403-700 ) immunizes chicks effectively against E. tenella infection by inducing humoral immunity and upregulating IFN-γ and CD8 + T lymphocytes. Furthermore, chicks exhibited increased relative weight gain rates, lower cecum lesion scores, and reduced oocyst outputs during the E. tenella challenge. H&E staining showed that the cecum tissue of chicks immunized with rEtRON2 403-700 showed relatively mild histopathological changes. In conclusion, the results of this study demonstrated that EtRON2 plays a key role in E. tenella invasion of the host intestinal epithelium and provides a potential target for vaccines against E. tenella infection., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Eimeria is an intracellular obligate apicomplexan parasite that parasitizes the intestinal epithelial cells of livestock and poultry, exhibiting strong host and tissue tropism. Parasite-host interactions involve complex networks and vary as the parasites develop in the host. However, understanding the underlying mechanisms remains a challenge. Acknowledging the lack of studies on Eimeria invasion mechanism, we described the possible invasion process through comparative analysis with other apicomplexan parasites and explored the fact that parasite-host interactions serve as a prerequisite for successful recognition, penetration of the intestinal mechanical barrier, and completion of the invasion. Although it is recognized that microbiota can enhance the host immune capacity to resist Eimeria invasion, changes in the microenvironment can, in turn, contribute to Eimeria invasion and may be associated with reduced immune capacity. We also discuss the immune evasion strategies of Eimeria, emphasizing that the host employs sophisticated immune regulatory mechanisms to suppress immune evasion by parasites, thereby sustaining a balanced immune response. This review aims to deepen our understanding of Eimeria-host interactions, providing a theoretical basis for the study of the pathogenicity of Eimeria and the development of novel anticoccidial drugs., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Weng et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Coccidiosis, an intestinal disease caused by Eimeria parasites, is responsible for major losses in the poultry industry by impacting chicken health. The gut microbiota is associated with health factors, such as nutrient exchange and immune system modulation, requiring understanding on the effects of Eimeria infection on the gut microbiota. This study aimed to determine the effects of Eimeria acervulina infection on the luminal and mucosal microbiota of the cecum (CeL and CeM) and ileum (IlL and IlM) at multiple time points (days 3, 5, 7, 10, and 14) post-infection. E. acervulina infection decreased evenness in CeL microbiota at day 10, increased richness in CeM microbiota at day 3 before decreasing richness at day 14, and decreased richness in IlL microbiota from day 3 to 10. CeL, CeM, and IlL microbiota differed between infected and control birds based on beta diversity at varying time points. Infection reduced relative abundance of bacterial taxa and some predicted metabolic pathways known for short-chain fatty acid production in CeL, CeM, and IlL microbiota, but further understanding of metabolic function is required. Despite E. acervulina primarily targeting the duodenum, our findings demonstrate the infection can impact bacterial diversity and abundance in the cecal and ileal microbiota., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

The purpose of this pilot study was to evaluate and determine the concentration of prostaglandin GF2α (PGF2α) and isoprostane 8-iso-PGF2α in plasma and intestine of specific pathogen-free (SPF) Leghorn chickens challenged with Eimeria maxima, with or without dietary supplementation of curcumin using solid-phase microextraction and ultra-performance liquid chromatography/tandem mass spectrometry. Eighty 1-day-old male SPF chickens were randomly allocated to one of four groups with four replicates (n = 5 chickens/replicate). Groups consisted of: (1) Control (no challenge), (2) Curcumin (no challenge), (3) Eimeria maxima (challenge), and (4) Eimeria maxima (challenge) + curcumin. At day 28 of age, all chickens in the challenge groups were orally gavaged with 40,000 sporulated E. maxima oocysts. No significant differences (P > 0.05) were observed in the groups regardless of the treatment or challenge with E. maxima. Enteric levels of both isoprostane 8-iso-PGF2α and PGF2α at 7 days and 9 days post-challenge were significantly increased (P < 0.01) compared to the non-challenge control chickens. Interestingly, the enteric levels of both isoprostane 8-iso-PGF2α and PGF2α at 7 days post-challenge were significantly reduced in chickens fed curcumin, compared to control chickens challenge with E. maxima. At 9 days post-challenge, only levels of isoprostane 8-iso-PGF2α in the enteric samples were significantly reduced in chickens challenged with E. maxima supplemented with curcumin, compared with E. maxima challenge chickens. No differences of isoprostane 8-iso-PGF2α or PGF2α were observed in plasma at both days of evaluation. Similarly, no significant differences were observed between the challenge control or chickens challenge with E. maxima and supplemented with curcumin at both times of evaluation. The results of this pilot study suggests that the antioxidant anti-inflammatory properties of curcumin reduced the oxidative damage and subsequent intestinal mucosal over-production of lipid oxidation products. Further studies to confirm and extend these results in broiler chickens are required.

Coccidiosis is one of the most prevalent diseases seen in the poultry industry leading to excessive economic losses. The aim of this study was to investigate the effect of butyric acid glycerol esters (BE) on the ileal and cecal microbiota in birds challenged with Eimeria maxima (EM). Ross 708 male broilers were fed a diet supplemented with 0 (control) or 0.25% BE from day 1. On day 21, half of the birds were infected with 10 3  EM oocysts. For determing microbiota, ileal and cecal contents and epithelial scrapings were collected at 7 and 10 D postinfection (PI). Alpha diversity of bacterial communities was mostly affected (P < 0.05) by time PI and EM infection. The richness of luminal bacterial populations in the ileum and ceca was affected (P < 0.05) by addition of BE and by time PI × EM × BE interaction, respectively. In the ileal and cecal luminal and mucosal bacterial communities, permutational multivariate analysis of variance (PERMANOVA, unweighted UniFrac) showed significant (P < 0.05) differences because of time PI and interaction between time PI, EM, and BE. Significant (P < 0.05) differences in taxonomic composition at the family level were observed in microbiota of luminal and mucosal populations of the ileum and ceca owing to time PI, EM, BE, and their interactions. The bacterial community present in the cecal lumen was characterized by the lowest number of differential bacteria, whereas the cecal mucosal community was characterized by the highest number of differentially abundant bacteria. In conclusion, our results show that EM infection and time PI has the biggest impact on microbial diversity in the chicken gut. The presence of BE in the diet had a limited effect on gut microbiota., (Copyright © 2020. Published by Elsevier Inc.)

Matrix metalloproteinases (MMPs) play an important role in intestinal extracellular matrix homeostasis. An overexpression of MMPs results in tissue destruction and local inflammation and has been associated with multiple inflammatory diseases. These host proteases might also be important in tissue damage caused by infectious agents, such as in intestinal damage in Clostridium perfringens-induced avian necrotic enteritis (NE). The aim of the present study was to elucidate the effect of a C. perfringens infection on the MMP activity in the small intestine of birds with a pre-disposing coccidial infection to obtain a more thorough understanding of the pathogenesis of NE. For this purpose, the gelatinolytic activity present in jejunal tissue of Eimeria infected birds which were challenged with either a pathogenic C. perfringens type G strain or a commensal C. perfringens type A strain was analyzed using substrate zymography. The results show that infection of broilers with Eimeria and different C. perfringens strains, independent of their pathogenicity, decreases the expression of a 40-45 kDa host collagenase in the jejunum, as compared to the expression in Eimeria-infected control birds. It was also shown that the expression of 2 MMPs with molecular weights of approximately 50-60 and 60-70 kDa was significantly lower in necrotic tissue as compared to the activity in macroscopically healthy tissue adjacent to the lesion. These results indicate that host collagenases are not elicited by the C. perfringens infection for permeabilizing the host mucosa to allow penetration of the NetB toxin in Eimeria infected broilers.

Coccidiosis is caused by protozoa of the genus Eimeria. These are intracellular parasites of enterocytes that rupture the host cell, causing lesions in the intestinal mucosa. The lesions caused by Eimeria reduce nutrient absorption capacity, negatively affecting productive gains in broilers, and representing a gateway for other enteropathogens. The objective of this study was to analyze the correlation between lesions caused by Eimeria and the prevalence of coccidiosis and other alterations found in the gastrointestinal tracts of broilers produced in Brazil from 2017 to 2018. Intestinal health evaluations were performed in 32 integrations (farm) of broilers in Brazil, totaling 726 birds analyzed between the ages of 22 and 40 days. Necropsied chickens were collected at three different points, with at least three birds per shed. We analyzed the following changes in the gastrointestinal tract: presence of cellular desquamation, fluid and mucus excess, ingestion of bedding, small and large intestine lesion, food passage, altered tone, "Turkish towel" lesions, worm infection, enteritis and gizzard erosion. The definition of macroscopic lesion scores caused by Eimeria acervulina, Eimeria maxima, Eimeria tenella followed a specific methodology. Mucosal oocyst counts for E. maxima (E. maxima micro) was performed using an optical microscope with a magnification of 100×. We found that the species E. acervulina had the highest prevalence (5.5%). With respect to E. acervulina, a positive correlation was observed with cellular desquamation, bedding ingestion and passage of food. The second highest prevalence was E. maxima (average of 4%), showing positive correlations with cellular desquamation, fluid excess, bed ingestion, feed passage and E. acervulina. E. tenella represented the lowest prevalence (0.8%) among the species of Eimeria analyzed, showing a positive correlation with altered intestinal tone. On microscopic evaluation, E. maxima was present in 45% of mucosa scrapings, representing subclinical coccidiosis of 1125% (11.25-fold) greater than the rate of clinical coccidiosis. Regarding other alterations that were visualized in the gastrointestinal tract, we have recorded the incidence of altered intestinal tone (0.1%), worm infection (0.4%), small intestine (0.8%), enteritis (1%), duodenitis (1.5%), "Turkish towel" lesions (3.3%), excess fluid (4.5%), bed ingestion (6.9%), excess mucus (8.4%), food passage (10.3%), cellular desquamation (11%) and gizzard erosion (13.4%). We conclude that monitoring is of paramount importance to understand the intestinal health status of poultry lots. Microscopic E. maxima is present in 45%. We identified factors that correlate with reduction in intestinal health, impairing zoo-economic performance., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Up to now, four coccidian species have been identified in Rallidae (Aves: Gruiformes): Eimeria mongolica, E. alakuli, E. paludosa and E. porphyrulae. Here, we described an Eimeria species, E. paludosa, from a common gallinule (Gallinula galeata) in Mexico. Oocysts were ovoid and wall pitted single-layered. A prominent micropyle was present, the oocyst residuum absent and the polar granule was present. On histological examination of tissues, endogenous stages (meronts, microgametocytes and macrogametocytes) were seen in the epithelial cells of the small intestine (upper and lower intestine). In addition to a new locality, this is the first description of E. paludosa from G. galeata and is the third description of a coccidian infecting Rallidae in the Americas.

Three-week-old turkey poults were infected with pure lines of three species of Eimeria ( E. adenoeides , E. gallopavonis , and E. meleagrimitis ) recently isolated from commercial turkey farms. The lines had been propagated from a single oocyst and identified by species-specific PCR amplification of the mitochondrial cytochrome c oxidase subunit I (COI) gene. Five to six days after infection their intestines were removed and examined for the presence of intestinal lesions. A description and review of the pathology caused by these parasites is provided, and a scoring system developed by which the severity of the lesions can be evaluated. The system is similar to that described by Johnson, J. and Reid, W. M. [1970. Anticoccidial drugs: lesion scoring techniques in battery and floor-pen experiments with chickens. Experimental Parasitology , 28, 30-36] for chickens in which a score of zero to four is assigned to lesions of increasing severity. The intestinal lesions observed here, and their assigned scores, are supported by representative illustrations. It is hoped that they may prove a useful tool for evaluating the pathology caused by E. adenoeides , E. gallopavonis , and E. meleagrimitis in the turkey. RESEARCH HIGHLIGHTS A scoring system has been developed for intestinal lesions caused by three species of Eimeria that infect the turkey.The lesions attributable to these species are illustrated.

Eimeria species are intestinal protozoan parasites that cause lack of production, malabsorption and mortality in floor raised chickens. Administering an oral antibody to interleukin 10 (aIL-10) reduces the symptoms of coccidiosis in broilers, indicating interleukin 10 (IL-10) is key to Eimeria pathology. IL-10 is an anti-inflammatory cytokine and acts as a stand down signal to reduce inflammation and host pathology during disease. Related protozoan parasites exploit IL-10 to reduce pathogen-damaging host inflammatory responses. We hypothesize that IL-10 is increased during Eimeria infection through an unknown host-pathogen interaction, and by feeding aIL-10 to neutralize excess IL-10 the bird is allowed to mount an effective immune response to Eimeria. To determine the effects of aIL-10 during the intestinal immune response, intestinal pathology and the relationship between IL-10, interferon gamma (IFNγ) and Eimeria infection were evaluated in this study. In both experiments, broilers were administered either a 10x dose of Advent® Eimeria vaccine or saline. Duodenum, jejunum and cecum samples were collected, processed, stained and examined under a microscope. Evaluation of intestinal histomorphology during aIL-10 administration showed minimal differences in birds fed aIL-10 during infection compared to animals fed a control antibody during Eimeria infection. To further evaluate aIL-10's positive effect during infection, immunofluorescent histochemistry was performed on chicken intestines days 3-7 post Eimeria infection for IL-10 and IFNγ presence in intestinal mucosa in control and infected birds, in regions with and without visible Eimeria burden. IL-10 and IFNγ had significant changes between days 4.5-7 post-infection in birds fed aIL-10 compared to animals fed a control antibody. Overall we found that the duodenum had increased IL-10 presence and increased IFNγ presence, and the jejunum and cecum had decreased IL-10 presence and decreased IFNγ presence. These differences in spatial regulation of IL-10 and IFNγ may indicate Eimeria species induce slightly different cytokine responses., (Published by Elsevier B.V.)

Ovine Eimeria spp. infections cause increased mortality, reduced welfare and substantial economic losses, and anticocccidials are important for their control. Recent reports of anticoccidial resistance against ovine Eimeria spp. necessitate the development of in vitro methods for the detection of reduced anticoccidial efficacy, especially since the in vivo methods are both expensive, time consuming and requires the use of otherwise healthy animals. The aim of the present study was therefore to approach a preliminary standardization of in vitro assays for evaluation of the efficacy of the most commonly used anticoccidials in ruminants. For this purpose, apart from the evaluation of inhibition of oocyst sporulation, most effort was concentrated on assessment of the capacity of the different anticoccidials to inhibit both the invasion and further development (up to the first schizogony) of E. ninakohlyakimovae sporozoites in bovine colonic epithelial cells (BCEC). For this purpose, infected cultures were monitored 1, 8 and 15 days post infection to determine the infection rate, number of immature schizonts and number, size and appearance of mature schizonts, respectively. No clear inhibitory effect was found with any of the anticoccidial formulations tested, and we could not identify why there were no measurable effects from the different anticoccidials. Despite the lack of positive results, further investigations should be encouraged, as this could decrease the need for animal experiments and could be used in the initial assessment of anticoccidial efficacy of new drugs., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Differentiating the Eimeria species causing cecal coccidiosis in turkeys is challenging. To obtain benchmark biological data for Eimeria gallopavonis Hawkins 1952 and Eimeria meleagridis Tyzzer 1929 and to support the stability of the species concept for each, genetically typed, single oocyst-derived lines of E. gallopavonis Weybridge strain and E. meleagridis USAR97-01 were used to redescribe the biological, pathological, and morphological features of these parasites. Oocysts of E. meleagridis and E. gallopavonis overlap in dimensions, but oocysts of the former have a single polar granule compared with multiple in the latter. Mature first-generation meronts of E. gallopavonis were observed histologically as early as 48 h post-inoculation alongside the villi in jejunum (before and after Meckel's diverticulum), ileum, cecal neck and rectum, but not cecal pouches. Three asexual cycles were observed suggesting that early workers apparently overlooked one asexual cycle. Examination of endogenous development of a culture labeled "Eimeria adenoeides Weybridge strain" suggested that this strain (found in a number of publications as a large oocyst strain of "Eimeria adenoeides") matched the species description of E. gallopavonis and so has been renamed herein. Macroscopic lesions induced by E. gallopavonis consisted of caseous material distally from posterior of the yolk stalk through the remaining intestinal tract, excluding the cecal pouches. For E. meleagridis, only the first asexual generation was observed outside of the cecal pouches within the jejunum around the yolk stalk. Second- and 3rd-generation asexual stages developed almost exclusively in the cecal pouches (but not cecal necks). Macroscopic lesions described for E. meleagridis were similar to those of E. adenoeides. Marked corrugation of the cecal serosal surface was observed. Cecal pouches contained creamy colored, caseous material varying from loose material to granular. Distinguishing features of the Eimeria species infecting the lower part of the small intestine are summarized in the present study, and new type specimens were designated for E. gallopavonis and E. meleagridis to provide a stable reference for future work with these parasites.

The endogenous life cycle of Eimeria caliginosa was studied in experimentally infected dusky rice rats, Melanomys caliginosus. All endogenous stages were located in the epithelial cells of villi in the small intestine. Both Giemsa-stained mucosal scrapings and histological sections were studied for the diagnosis of all the life-cycle stages. Eimeria caliginosa has 3 generations of meronts (M) that differ by size, shape, and number of merozoites (m), which also differ in their size, shape, and location of their nuclei within the cytoplasm of the meronts. The 3 meront types, M 1 -M 3 , respectively, had 20-33 (m 1 ), 5-9 (m 2 ), and 13-16 (m 3 ) merozoites. Macrogametocytes and microgametocytes, as well as macrogametes and microgametes, completed the sexual cycle, which concludes with the formation of unsporulated oocysts. This parasite's endogenous development produced severe intestinal lesions in the experimentally infected dusky rice rats. In our ongoing work to understand the biodiversity present in plants and animals of the protected Reserva Biológica Alberto Manuel Brenes (ReBAMB) field station in Costa Rica, we now have discovered 3 new Eimeria species, and this is the second complete life cycle in which we document both the asexual and sexual stages.

Intracellular generation of nitric oxide (NO) and superoxide anion (SOA) can result in the formation of 3'-nitrotyrosine proteins (NTp). Nitrated proteins usually are associated with significant perturbation in protein function, apoptosis, autophagy, and cell death. We undertook the present study to establish the temporal dynamics of NTp generation in cytokeratin-18-positive epithelial cells (ETCs) of broiler chickens in response to infection with Eimeria acervulina. Duodenal tissue was harvested from noninfected (NOI) and infected (INF) broilers on days (d) 1, 3, 6, 7, and 10 postinfection (PI) and fixed, embedded, and sectioned for quantitative image analysis, immunohistochemistry with antibodies specific to NTp and the SOA-generating enzyme xanthine oxidase (XO). The pixel density characteristics for NTp and XO representative of ETCs demonstrated that NTp and XO increased in intestinal villi as early as d1 PI (P < 0.05 vs. NOI). Progressive increases in NTp were evident in ETCs through d6 PI. For XO, increases in cell content increased only through d3. On d6 and d7 PI, high levels of NTp were present in immune infiltrating cells (IIC) where no XO was detected. The increases in ETC NTp occurred in a defined pattern, significant by villus-to-crypt location for day of infection, initiating in the distal villus and progressing down into the crypts. Two NTp patterns were observed for ETCs: a high level associated with ETCs harboring parasites and a low-level increase in ETCs not containing Eimeria but in proximity to such. The data suggest that NTp and XO responses may mediate some of the processes through which ETCs respond to Eimeria to limit the extent of infection by this pathogen.

Rabbit coccidiosis is caused by infection with one or usually several Eimeria species, parasitizing in hepatobiliary ducts or intestinal epithelium of rabbits. To date, 11 species of rabbit coccidia have been well documented. Here we report a new species of Eimeria from rabbits. Sporulated oocysts were ellipsoidal to slightly ovoidal, 37.4 (32.6-41.2) μm in length, 23.5 (20.9-25.5) μm in width, with a shape index (length/width) 1.6 (1.43-1.91) and smooth, bilayered, homogeneously thick wall. The micropyle was obvious and with an inner diameter of 6.2 (5.0-7.5) μm. Both oocyst residuum and polar granule were absent. Sporocysts were ellipsoidal to elongate, 17.2 (13.2-20.0) μm long and 8.4 (7.5-9.1) μm wide, with a shape index (length/width) of 2.1 (1.74-2.21) and the presence of Stieda body and sporocyst residuum. The prepatent period was 132h. Phylogenetic analysis showed that 18S rDNA sequence of the new species clustered together with the 11 rabbit Eimeria species into a clade. However, ITS-1 sequence of the new species shared low similarities (27.1%-30%) with those of 11 rabbit Eimeria species. As the data above supported the erection of a new species, we named it as Eimeria kongi n. sp., in honor of Fanyao Kong, a Chinese parasitologist. The finding of the new species has important implications for the diagnosis and prevention of rabbit coccidiosis., (Copyright © 2017. Published by Elsevier B.V.)

Eimeria tenella, one of the most important parasitic protozoa in the genus Eimeria, is responsible for chicken caecal coccidiosis resulting in huge economic losses to poultry industry. The present study investigated the changes in caecal microflora of E. tenella-infected chickens and the regulating effect of coated sodium butyrate, a potential alternative to antibiotics. Using high-throughput sequencing of 16S rRNA V3-V4 region of bacteria we found significant changes in caecal microflora of E. tenella-infected chickens indicated by an increase of Firmicutes (mainly Ruminococcaceae, Lachnospiraceae and vadin BB60) and Proteobacteria (mainly Enterobacteriaceae) and a decrease of Bacteroidetes (predominantly Bacteroidaceae). Inclusion of coated sodium butyrate in the diet of chickens per se had no significant effect on caecal microflora of normal healthy chickens but significantly prevented the increase in Firmicute abundance and decrease of Bacteroidetes abundance in E. tenella-infected birds. No significant changes to caecal microflora were observed at the phylum level between control and E. tenella-infected birds given coated sodium butyrate. In conclusion, our results show that coated sodium butyrate can balance the disorders of cecal microflora caused by E. tenella; thus, it can be a useful supplement for the control of avian coccidiosis., (Copyright © 2017 Elsevier Inc. All rights reserved.)

This study used morphological techniques to describe and name four new species of coccidia from the brown kiwi (Apteryx mantelli). Four distinct eimerian oocyst species were recovered that we describe as new species. The largest of these, Eimeria paraurii n. sp. measured 32.2 × 19.8 μm and is morphologically similar to gametocytes previously described histologically in colorectal polyps (Morgan et al. in Parasitol Res 111(4):1689-1699, 2012). Eimeria apteryxii n. sp. measured 23.9 × 14.9 μm and is similar to renal oocysts described histologically in brown, rowi (A. rowii) and Haast tokoeka kiwi (A. australis "Haast") (Morgan et al. in Avian Pathol 42(2):137-146, 2013). Eimeria kiwii n. sp. measured 14.8 × 13.9 μm and resembled gametocytes described previously in kiwi intestinal epithelial cells in brown kiwi (Morgan et al. in Parasitol Res 111(4):1689-1699, 2012). Eimeria mantellii n. sp. measured 17.8 × 10.7 μm and did not appear similar to any coccidia previously described in histological studies in kiwi. These are the first species of Eimeria to be described and named from brown kiwi. Because the morphological descriptions in the present study were determined from a limited number of kiwi droppings from two geographical locations, it is likely that these represent only a portion of Eimeria species present in other populations of both brown kiwi and other Apteryx species from around New Zealand.

The present study aimed to investigate the dynamic expression patterns of death-receptor adapter proteins TNF-receptor-associated death-domain protein (TRADD) and Fas-associated death-domain protein (FADD) in E. tenella-induced host-cell apoptosis. Culture techniques for primary chick embryo cecum epithelial cells, ELISA, hematoxylin-eosin staining, fluorescence quantitative PCR techniques, and Hoechst-Annexin V-PI apoptosis staining were used to detect the apoptosis rates and dynamic expression patterns of TRADD and FADD in E. tenella host cells at 4, 24, 48, 72, 96, and 120 h. The rates of early apoptosis, late apoptosis, and necrosis of E. tenella-infected group (group T0) were significantly lower (P < 0.05) or highly significantly lower (P < 0.01) than those of control group (group C) at 4 h, but higher (P < 0.05 or P < 0.01) at varying degrees than those of the same group at 24 to 120 h. Compared with group C, both the mRNA and protein expression levels of TRADD in the group T0 cells increased highly significantly (P < 0.01) at 4 and 24 h, and significantly (P < 0.05) at 48, 72, and 120 h. Compared with the mRNA expression in the group C cells, that of TRADD in the group T0 cells increased significantly (P < 0.05) at 96 h. Both the mRNA and protein expression of FADD in the group T0 cells displayed no significant difference (P > 0.05) from those in the group C cells at 4 h. However, FADD expression in the group T0 cells were significantly (P < 0.05) or highly significantly higher (P < 0.01) than those in the group C cells at 24 to 120 h. These observations indicate that in the early developmental stages of E. tenella, the host-cell apoptosis rate decreased, and TRADD expression increased. In the middle and later developmental stages of E. tenella, the host-cell apoptosis rate increased and expression of TRADD and FADD increased. The variation trends of TRADD and FADD expression were significantly positively correlated with the change rule of the host-cell apoptosis rate, respectively. These results indicate that TRADD and FADD may play an important role in E. tenella-induced host-cell apoptosis., (© 2017 Poultry Science Association Inc.)

For this study, threonine (Thr) deficiency was hypothesised to exacerbate the intestinal damage induced by feed withdrawal with coccidial infection because of its high obligatory requirement by the gut; two dietary Thr treatments (0·49 and 0·90 %) were applied to chicks from 0 to 21 d of age. At 13 d of age, feed was withdrawn for 24 h from one-half of birds of each dietary treatment with subsequent gavage of a 25× dose of coccidial vaccine. Overall, there were four treatments with eight replicate cages per treatment. Under combined challenge, birds fed the Thr-deficient diet had 38 % lower 13-21-d body weight gain (P≤0·05) compared with birds fed the Thr-control diet. At 21 d, the challenged group fed low Thr had higher number of oocysts (+40 %, P=0·03) and lower crypt depth (-31 %, P0·05). Overall, Thr deficiency worsened the detrimental effects of combined feed withdrawal and coccidial infection on growth performance and oocyst shedding by impairing intestinal morphology, barrier function, lymphocyte profiles and their cytokine expressions.

Cystoisospora belli in patients with the acquired immunodeficiency syndrome (AIDS) has been described as cause of chronic diarrhea and disseminated cystoisosporosis. Diagnosis of intestinal cystoisosporosis can be achieved at the tissue level in the villus epithelium of the small bowel. Disseminated cystoisosporosis is diagnosed by microscopy identification of unizoite tissue cysts in the lamina propria of the intestine. We report a case of disseminated cystoisosporosis in a human immunodeficiency virus (HIV)-infected patient with detection of parasitemia. We studied a 39-year old patient with AIDS and chronic diarrhea by analysis of stool and duodenal biopsy samples. Blood samples were also collected and examined by light microscopy and molecular techniques for C. belli DNA detection. The unizoite tissue cyst stages were present in the lamina propria, with unsporulated oocysts in feces. Zoites were present in blood smears and DNA of C. belli was detected in blood samples. Our study identified a new stage in the life cycle of C. belli. Detection of parasitemia is a novel and noninvasive tool for diagnosis of disseminated cystoisosporosis.

Four experiments were conceived in order to test the efficacy of artemisinin, a sesquiterpene lactone derived from Artemisia annua, in single experimental infection of broiler chickens with Eimeria acervulina (1 × 10(5) oocysts), Eimeria maxima (5 × 10(4) oocysts) or Eimeria tenella (1 × 10(4) oocysts), and mixed infection with all 3 species (3.2 × 10(4) Eimeria spp. oocysts). For each experiment, three different dosages of artemisinin (5, 50 and 500 ppm) were compared with a negative control (uninfected, unmedicated), a positive control (infected, unmedicated) and a classical anticoccidial (monensin). The weight gain (WG), feed conversion ratio (FCR), oocysts shedded per gram of feces (OPG), lesion score, oocysts sporulation rates and mortality rate were recorded in all groups. The dosage of 5 ppm of artemisinin improved the WG and FCR for the chickens infected with E. acervulina. The OPG was significantly decreased in all the groups medicated with artemisinin and challenged with a mixed infection (p ≤ 0.01). The lesion score of the chickens challenged with Eimeria was reduced by different concentrations of artemisinin, depending on the species involved, but this compound did not have a positive effect on the lesions caused by E. acervulina. Histopathological analysis revealed superficial erosions of the intestinal mucosa, mixt. mononuclear and heterophilic inflammatory infiltrate in the lamina propria and intralesional presence of various developmental stages of parasite in groups infected with Eimeria spp.The sporulation rate of E. acervulina and E. maxima oocysts was significantly affected by 500 ppm of artemisinin, whilst the dosage of 5 ppm affected the sporulation of E. tenella oocysts. These data suggest that artemisinin is not effective against single eimerian infections but could be used as an alternative in mixed coccidiosis, especially if its effect on the oocysts sporulation would be fully investigated., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Metabolic and morphometric alterations of the duodenal villi caused by parasitism of chickens by Eimeria maxima were evaluated, using 100 male Cobb birds, randomly distributed into two groups (control and infected). The infected group was inoculated with 0.5 ml of a solution containing 5 × 10³ sporulated oocysts of Eimeria maxima. Ten birds per sample were sacrificed on the 6th, 11th, 22nd and 41st days post-infection (dpi). In order to evaluate the alterations, samples of duodenum, jejunum and ileum fragments were collected after necropsy for histological analysis. Villus biometry was determined by means of a slide graduated in microns that was attached to a binocular microscope. To evaluate the biochemical data, 5 ml of blood were sampled from the birds before sacrifice. The statistical analyses were performed using the GraphPad 5 statistical software for Windows. Tukey's multiple comparison test (p <0.05) was performed for the different dpi's and the unpaired t test for the difference between the groups. Infection by E. maxima causes both qualitative and quantitative alterations to the structure of the intestinal villi, thereby interfering with the absorption of nutrients such as calcium, phosphorus, magnesium, protein and lipids, with consequent reductions in the birds' weights.

The molecular and cellular mechanisms leading to immune protection against coccidiosis are complex and include multiple aspects of innate and adaptive immunities. Innate immunity is mediated by various subpopulations of immune cells that recognize pathogen associated molecular patterns (PAMPs) through their pattern recognition receptors (PRRs) leading to the secretion of soluble factors with diverse functions. Adaptive immunity, which is important in conferring protection against subsequent reinfections, involves subtypes of T and B lymphocytes that mediate antigen-specific immune responses. Recently, global gene expression microarray analysis has been used in an attempt to dissect this complex network of immune cells and molecules during avian coccidiosis. These new studies emphasized the uniqueness of the innate immune response to Eimeria infection, and directly led to the discovery of previously uncharacterized host genes and proteins whose expression levels were modulated following parasite infection. Among these is the IL-17 family of cytokines. This review highlights recent progress in IL-17 research in the context of host immunity to avian coccidiosis., (Copyright © 2013. Published by Elsevier Ltd.)

Enteric coccidiosis may cause significant morbidity and mortality in juvenile brown kiwi (Apteryx mantelli). Morphology of sporulated oocysts indicates that at least two Eimeria species are able to infect the brown kiwi. A histological study of the endogenous stages of coccidia was undertaken in the intestinal tracts of ten naturally infected young kiwi. Sequential sectioning of the entire intestinal tract allowed identification and recording of the distribution of the various coccidial life stages. Macromeronts measuring 268 × 162 μm when mature were found mainly within the lamina propria of the proximal one third of the small intestine. A smaller form of lamina propria meront was also identified (8.7 × 6.4 μm) with a similar distribution to the macromeronts. Small meronts (4.4 × 3.8 μm) were also identified in mucosal epithelial cells, with the overall peak in distribution within the intestinal tract being distal to the lamina propria meronts. Three morphologically distinctive gametocytes were identified. Type A gametocytes contained within epithelial cells shared the same distribution as the epithelial meronts. Polyps containing large numbers of type B gametocytes within the distal intestinal tract were found in two cases, and type C gametocytes were identified throughout the entire intestinal tract in one case only. The observational nature of this study precludes complete knowledge of the parasite life cycles using histology alone. However, it is likely that each of the three morphologically distinct gametocytes represents a separate species of enteric coccidia.

This is the first report of an intestinal Eimeria infection in Asian seabass (Lates calcarifer) at the histopathological and ultrastructural levels. The Eimeria infection was often associated with severe pathology and significant mortality in the absence of other pathogens. This showed that it is an important disease of juvenile L. calcarifer in small scale nurseries in Vietnam. Heavy infection and high prevalence levels of the Eimeria infection are suspected to be linked to the low daily water exchange rates practised in these nurseries. Although systemic iridovirus infection was concurrently observed in some of the fish examined, it was not as consistently present in diseased fish as the Eimeria infection., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Innate immune reactions conducted by macrophages may affect the outcome of primary infections and are crucial for the transition to adaptive immune responses. In bovine coccidiosis little is known on early monocyte/macrophage-mediated responses. We therefore investigated in vivo, in vitro and ex vivo reactions of monocytes and macrophages against Eimeria bovis, one of the most pathogenic Eimeria species in cattle. Macrophages significantly infiltrate the gut mucosa of E. bovis-infected calves, particularly after challenge infection. Furthermore, peripheral monocytes of infected animals, as precursor cells of macrophages, exhibited enhanced ex vivo phagocytic and oxidative burst activities. Enhanced levels of both activities were found early after infection and towards the end of first merogony. In vitro exposure of macrophages to sporozoites led to phagocytosis of the pathogen, whilst monocytes failed to do so. Phagocytosis occurred independently of the viability of the sporozoites, indicating that active invasion by the parasites was negligible. Phagocytosis occurred in the absence of immune serum, but could clearly be enhanced by addition of immune serum, suggesting macrophage-derived antibody-dependent cytotoxicity. Furthermore, co-culture of macrophages with sporozoites and stimulation with merozoite I antigen induced distinct levels of cytokine and chemokine gene transcription. Thus, the transcription of genes encoding for IFN-gamma, IL-12, TNF-alpha, IL-6, CXCL1, CXCL8, CXCL10 and COX-2 was upregulated after sporozoite encounter. In contrast, soluble merozoite I antigen only induced the gene transcription of IL-6 and IL-12 and failed to upregulate IFN-gamma and TNF-alpha gene transcripts. In monocytes, IFN-gamma and CXCL10 were found upregulated, all other immunoregulatory molecules tested were not affected. In summary, our results strongly suggest that macrophage-mediated, innate immune reactions play an important role in the early immune response to E. bovis infections in calves.

Although chronic intestinal helminth infections may suppress allergen-induced airway pathology by inducing a combination of modified T-helper (Th) 2 and immunosuppressive cytokines, a similar capacity of natural acute intestinal infections has remained untested, despite their global prevalence. Here, we show that allergic airway phenotypes including eosinophilia, eotaxin mRNA, and Th2 cytokines are significantly suppressed in animals that were infected by and that have cleared the intestinal parasite Eimeria vermiformis. Unlike in helminth-infected animals, regulation requires temporal coincidence of infection with sensitization; depends on interferon-gamma; and is not associated with an enhanced antigen-specific immunoglobulin G1 response. Moreover, regulation was effective following allergen sensitization in different anatomical sites, and in young and adult mice. These data highlight a transient anatomical dissemination of "functional immunologic dominance" following infection of the gut mucosa. They strongly support the hypothesis that airway allergies are naturally suppressed by both acute and chronic mucosal pathogens, but by different mechanisms.

A case of coccidiosis in a young, red male squirrel (Sciurus vulgaris L.) has been described in this report. The squirrel was found dead and presented to the department of pathology for necropsy. A traumatic lesion was observed on the face that could have caused death. At necropsy the large and small intestines were swollen due to fluid and gas. During the examination of gut content numerous coccidia oocysts were observed. After sporulation, the oocysts were identified as those of Eimeria lancasterensis. In the histopathological examination numerous coccidia developmental stages were observed in the epithelium of small intestine. This is the first report of Eimeria lancasterensis identification in squirrels in Turkey.

The aim of this study was to evaluate a nasal vaccine using antigens derived from sporozoites of Eimeria tenella incorporated into Iscom to protect broiler chicks. Forty-five one-day-old chickens (Cobb), unvaccinated against coccidiosis, were used in this experiment. The birds were maintained in separated battery cages and divided into three groups: G1 (n=15), G2 (n=15), and G3 (n=15). G1 received 50 microg of sporozoites+Iscom vaccine, G2 received Iscom without antigens, and G3 received only PBS. The treatments were administered by nasal route on days 0, 7, and 21 of the experiment. On the 28th day, all birds were challenged with 105 sporulated oocysts of E. tenella. On the challenge day, three birds from each group were euthanized to evaluate lymphocyte proliferation. Lesion scores were obtained from five birds from each group, 7 days after challenge. The remaining animals were euthanized on the 50th day. The mean lymphocyte proliferation responses were significantly different (P=0.03); G1 was 2.3-2.6 times more elevated than G2, and G3 (P<0.001). 83% of the birds from G1 showed an IgY antibody reaction by ELISA at challenge. The means for oocysts shedding were 16,890+/-20,511, 48,080+/-50,047, and 65,020+/-74,461, for G1, G2 and G3 birds, respectively. There was no significant difference (P>0.17) in oocysts shedding between groups. However, the G1 and G2 chicks demonstrated reduction in percentage of oocyst shedding when compared to control birds (G3) by 74.02% and 26.05%, respectively. The average lesion scores were G1=0.4, G2=1, and G3=2. This study demonstrated that the lowest lesion score and oocyst shedding were observed in the birds from the group that received antigens derived from sporozoite with an Iscom adjuvant (G1). These results suggest that this vaccine can induce protection against avian coccidiosis.

To investigate the influence of genetic differences in the MHC on susceptibility to avian coccidiosis, M5.1 and M15.2 B-haplotype-disparate Fayoumi chickens were orally infected with live Eimeria maxima oocysts, and BW gain, fecal oocyst production, and expression of 14 immune-related genes were determined as parameters of protective immunity. Weight loss was reduced and fecal parasite numbers were lower in birds of the M5.1 line compared with M15.2 line birds. Intestinal intraepithelial lymphocytes from M5.1 chickens expressed greater levels of transcripts encoding interferon-gamma (IFN-gamma), interleukin-1beta (IL-1beta), IL-6, IL-8, IL-12, IL-15, IL-17A, inducible nitric oxide synthase, and lipopolysaccharide-induced tumor necrosis factor-alpha factor and lower levels of mRNA for IFN-alpha, IL-10, IL-17D, NK-lysin, and tumor necrosis factor superfamily 15 compared with the M15.2 line. In the spleen, E. maxima infection was associated with greater expression levels of IFN-gamma, IL-15, and IL-8 and lower levels of IL-6, IL-17D, and IL-12 in M5.1 vs. M15.2 birds. These results suggest that genetic determinants within the chicken MHC influence resistance to E. maxima infection by controlling the local and systemic expression of immune-related cytokine and chemokine genes.

The progression of coccidiosis provoked by Eimeria acervulina was followed in chicks fed on OTA-contaminated as well as on OTA-free diets. More heavy progress of duodenal coccidiosis, including mortality, occurred in OTA-treated chicks as can be seen from the higher value of lesion (3.50) and oocyst (31.65) indices. A stronger decrease of serum total protein was found in OTA-treated chicks (22.80 g/l) than in chicks infected with E. acervulina(24.20 g/l), but that decrease was strongest in chicks treated with OTA and simultaneously infected with E. acervulina (19.71 g/l). The serum concentration of uric acid was significantly increased in all chicks exposed to OTA, most notably in those additionally infected with E. acervulina (1020.6 (micro mol/L), whereas the serum enzyme activity of AST was increased only in chicks infected with E. acervulina and highest in those fed OTA contaminated diet (122.2 U/L). OTA induced degenerative changes in kidneys, liver and heart as well as a depletion of lymphoid tissue in the lymphoid organs and a decrease of body weight. Coccidiosis induced only a slight growth depression and duodenal hemorrhages in addition to characteristic duodenal damages. The impairment of kidney function, histopathological changes and general growth depression were stronger when chicks infected with E. acervulina were also given OTA.

The expression levels of mRNA encoding a panel of 28 chicken cytokines and chemokines were quantified in intestinal lymphocytes following Eimeria acervulina and Eimeria tenella primary and secondary infections. Compared with uninfected controls, transcripts of the pro-inflammatory cytokines IFN-alpha, IL-1beta, IL-6, and IL-17 were increased up to 2020-fold following primary infection. By contrast, following secondary infection by either microorganism, pro-inflammatory mRNAs levels were relatively unchanged (< or = 20-fold). Transcripts encoding the Th1 and Th1 regulatory cytokines IFN-gamma, IL-2, IL-10, IL-12, IL-15, IL-16, and IL-18 were uniformly increased 14-2471-fold after E. acervulina primary infection, but either unchanged (IL-15, IL-16, IL-18), increased (IFN-gamma, IL-10, IL-12), or decreased (IL-2) following E. tenella primary infection. Following secondary infections, Th1 cytokine mRNA levels were relatively unchanged, with the exception of IL-12 which was increased 1.5 x 10(5)-fold after E. acervulina and decreased 5.1 x 10(4)-fold after E. tenella infection. Transcripts for the Th2 or Th2 regulatory cytokines IL-3 and GM-CSF were increased up to 327-fold following primary or secondary infection with both parasites, while IL-4 and IL-13 mRNAs were decreased 25- to 2 x 10(5)-fold after primary or secondary infection. The dynamics of chicken chemokine expression revealed modest changes (<100-fold) following primary or secondary infection except for lymphotactin. When lymphocyte subpopulations were similarly analyzed, IFN-gamma, IL-2, IL-3, IL-15, and MIF were most highly increased in TCR2(+) cells following E. acervulina infection, while TCR1(+) cells only expressed high levels of IL-16 following E. tenella infection. In contrast, CD4(+) cells only expressed highest levels of IL-10 after E. acervulina infection, whereas these cells produced abundant transcripts for IFN-gamma, IL-3, IL-15, and MIF after E. tenella infection. We conclude that coccidiosis induces a diverse and robust primary cytokine/chemokine response, but a more subdued secondary response.

Coccidiosis, a major intestinal parasitic disease of poultry, induces a cell-mediated immune response against the etiologic agent of the disease, Eimeria. In the current study, the expression levels of gene transcripts encoding pro-inflammatory, Th1, and Th2 cytokines, as well as chemokines were measured in intestinal intraepithelial lymphocytes (IELs) after Eimeria maxima infection. In addition, changes in IEL numbers were quantified following E. maxima infection. Transcripts of the pro-inflammatory and Th1 cytokines IFN-gamma, IL-1beta, IL-6, IL-12, IL-15, IL-17, and IL-18 were increased 66- to 8 x 10(7)-fold following primary parasite infection. Similarly, mRNA levels of the Th2 cytokines IL-3, IL-10, IL-13, and GM-CSF were up-regulated 34- to 8800-fold, and the chemokines IL-8, lymphotactin, MIF, and K203 were increased 42- to 1756-fold. In contrast, IFN-alpha, TGF-beta4, and K60 transcripts showed no increased expression, and only the level of the Th2 cytokine IL-13 was increased following secondary E. maxima infection. Increases in intestinal T cell subpopulations following E. maxima infection also were detected. CD3(+), CD4(+), and CD8(+) cells were significantly increased at days 8, 6, and 7 post-primary infection, respectively, but only CD4(+) cells remained elevated following secondary infection. TCR1(+) cells exhibited a biphasic pattern following primary infection, whereas TCR2(+) cells displayed a single peak in levels. Taken together, these data indicate a global chicken intestinal immune response is produced following experimental Eimeria infection involving multiple cytokines, chemokines, and T cell subsets.

Specific pathogen-free (SPF) rabbits were inoculated with oocysts of an original strain (OS) of Eimeria piriformis and the first newly developed oocysts recovered from the intestine were used for infection of other rabbits. The prepatent period (PP) was shortened after 12 passages from 194 to 170 hours and remained stable after 5 passages without any selection pressure. Oocysts of the precocious line (PL) exhibited peculiar morphology. Besides refractile bodies (RB) within sporozoites, one huge RB joined with a residual body was present inside each sporocyst. The parasite developed in the proximal colon and, to a lesser extent, in other parts of the large intestine. All stages were seen in the epithelium of crypts. In OS, four asexual generations preceded gamogony and, like in other rabbit coccidia, two types of meronts were observed: meronts of type A that develop into polynucleate merozoite, in which endomerogony takes place, and meronts of the type B that form uninucleate merozoites. The endogenous development of PL was identical with that of OS except for the last merogony which was absent. This resulted in earlier appearance of gamogony and shortening of PP. These observations of the life cycle of E. piriformis substantially improve on its description made 50-60 years ago.

The SPF rabbits were inoculated with oocysts of Eimeria flavescens and the first newly developed oocysts were recovered. They were used for inoculation of other rabbits which consequently excreted oocysts sooner than in the previous passage. By repeated use of this method, the prepatent period was shortened after 18 passages by more than 60 h. The endogenous development of this precocious line (PL) differed from that of the original strain (OS). Compared to OS, two asexual generations, second (or third) and fourth, were absent in PL. The first merogony took place in the jejunum and ileum in OS and, in contrast, in the large intestine in PL. Like in other rabbit coccidia, two types of meronts (A and B) were seen in each generation. However, the ratio of B: A meronts in the last (fifth) asexual generation as well as ratio of microgamonts:macrogamonts differs in OS and PL.

Intraepithelial lymphocytes (IELs) play a critical role in protective immune response to intestinal pathogens such as Eimeria, the etiologic agent of avian coccidiosis. A list of genes expressed by intestinal IELs of Eimeria-infected chickens was compiled using the expressed sequence tag (EST) strategy. The 14,409 ESTs consisted of 1851 clusters and 7595 singletons, which revealed 9446 unique genes in the data set. Comparison of the sequence data with chicken DNA sequences in GenBank identified 125 novel clones. This EST library will provide a valuable resource for profiling global gene expression in normal and pathogen-infected chickens and identifying additional unique immune-related genes.

The initial phases of invasion of mammalian coccidia of the genus Eimeria into host tissue are still poorly known. This process, including the passage of oocysts through the intestinal lumen, excystation of sporozoites, their penetration into epithelial cells and migration to the target site was studied in both naive and immune mice infected with Eimeria falciformis. After oral infection, the intact oocysts were transported with enteral contents to the large intestine, where the excystation of sporozoites and their penetration into superficial epithelium took place. The sporozoites subsequently migrated into the epithelium of crypts, which is the specific site of asexual multiplication. The immune status of the hosts did not affect the passage of oocysts, excystation and penetration of sporozoites. However, the migration of sporozoites towards their target site (crypts) was impeded in immune mice and sporozoites tended to remain in superficial mucosa rather than migrate to the crypts.

To study the pathophysiology of diarrhoea in coccidial infections, Na+ and Cl- fluxes (J(Na), J(Cl)), short circuit current (I(sc)) and tissue conductance (g(t)) were determined in stripped gut epithelia of Eimeria separata infected rats employing the Ussing chamber technique. E. separata invades enterocytes of the caecum and proximal colon. Na+ absorption was generally reduced in infected tissues, Cl- absorption only in the caecum. I(sc) values were increased in the caecum and reduced in the proximal colon. Tissue conductance was not affected. Values tended to normal with time after infection. Theophylline caused markedly increased I(sc) and g(t) values in the caecum epithelia of infected rats. In the epithelia of the distal colon, i.e. the non-infested part of the large intestine, g(t) values remained unaffected but I(sc) was fourfold increased. This I(sc) increase was strongly sensitive to amiloride, suggesting a compensatory activation of Na+ channels in the distal colon of infected rats. Accordingly, serum levels of aldosterone, which activates Na+ channels in the distal colon, were increased to eightfold levels in infected animals. Thus compensatory Na+ absorption was under endocrine control.

The endogenous cycle of Eimeria flavescens was studied in specific pathogen-free rabbits by means of histology and transmission electron microscopy. In total, five asexual generations were observed and two types of meronts and merozoites were found in each generation. Type A gave rise to a smaller number of thick polynucleate merozoites in which daughter merozoites were formed by endomerogony, while in the type B meronts slender uninucleate merozoites arose from ectomerogony. The first generation meronts were found in the crypts and proximal part of the villi of the duodenum and jejunum, whereas the three following generations developed in the superficial epithelium of the large intestine (cecum, vermiform appendix and colon). The last merogony as well as gamogony took place in crypts of the large intestine.

Coccidia of the genus Eimeria are present in most pigs raised on dirt in the United States. They are generally considered nonpathogenic in weaned pigs. Oocysts of Eimeria spinosa Henry, 1931 were observed in tissue sections and intestinal contents of a weaned male pig that died suddenly on a farm in Iowa. Microscopically, necrotizing enteritis associated with many thick-walled coccidial oocysts was present in intestinal sections. Examination of intestinal contents demonstrated oocysts that were thick-walled and had small projections on the surface of the oocyst wall, characteristic of E. spinasa Henry, 1931 of swine. Twenty-live oocysts in intestinal contents measured 20.4 by 14.2 microm. No pathogenic bacteria were detected in the pig by culture methods, but lesions suggestive of salmonellosis were observed in some tissues. The specific cause of death was not determined; however, E. spinosa infection was considered to have contributed to the death of this pig. The results suggest that E. spinosa may be pathogenic for pigs.

1. Poultry granulocytes are not clearly distinguished from each other with haematoxylin-eosin (HE) stain; thus, histochemical techniques must be used. Three experiments were carried out using 4-week-old Leghorn chickens. 2. Three, 80-chicken groups were orally infected with (1) 10(8) colony forming units (CFUs) Salmonella enteritidis, or (2) 10(4) Eimeria tenella oocysts, or (3) 10(8) CFUs S. enteritidis + 10(4) E. tenella oocysts. Ten chickens from each group were euthanased and caecum samples obtained. Caecum samples were fixed in 10% formalin (buffered, pH 7.4) at 4, 8, 12 h, 1, 3, 5, 7, and 14 d post-inoculation (PI). 3. Samples were stained using three different staining techniques: HE for the identification of heterophils and eosinophils, Ziehl-Neelsen for mast cells, and p-phenilenediamine dihydrochloride plus pyrocatechol (PPD + PC) for eosinophils. 4. Birds from Experiment 1 showed no changes in the numbers of granulocytes. Birds from Experiments 2 and 3 showed higher numbers of heterophils in caecal mucosa and submucosa separately, on d 5 and 7. In Experiment 3, a decrease was observed in submucosal mast cells on d 3. Chickens from Experiments 2 and 3 showed increased numbers of mucosal mast cells between d 7 and 14. 5. PPD + PC positively stained eosinophils, but not heterophils. 6. Numbers of heterophils and mast cells were increased during the acute inflammatory process caused by E. tenella. Therefore, mast cells could play a role as primary inflammatory cells. Eosinophils seem not to be part of the inflammatory process caused by E. tenella.

Intestinal coccidiosis was diagnosed histologically in the small intestine of a spinner dolphin (Stenella longirostris). Numerous intralesional coccidia were present in mucosal epithelial cells. Schizonts, gamonts, and unsporulated oocysts were seen. Schizonts were up to 30 x 20 microm and contained up to 16 merozoites, which measured 10-12 x 2 microm. Unsporulated oocysts were about 9-12 x 8-10 microm. This is the first report of intestinal coccidiosis in a spinner dolphin.

The development and application of a standardized model for testing of anticoccidial disinfectants are described. Due to its economic impact, tenacity of oocysts, and reproducibility of the course of infection Eimeria tenella has been chosen as test organism. Oocysts of the Houghton strain were more susceptible to disinfection with 4% TP4 (Preventol) than oocysts of a field isolate (FI 292/1) as determined by sporulation inhibition and lysis. Scoring of intestinal lesions and of oocyst numbers in mucosal scrapings in chicken infected with various doses of oocysts were found unsuitable for assessment of disinfectants. Because strain differences were observed only Houghton strain oocysts were applied for further testing. Guidelines for standardized in vivo testing of disinfectants have been stipulated by the German Veterinary Society (DVG) on the basis of these studies. When applied for testing of Neopredisan (NP) in two separate laboratories similar results were obtained. Inhibitory activity (IA; proportion of inactivated oocysts) of 92.9 and 90.6% were calculated for 3% NP and of 95.2 and 96.8% for 4% NP after treatment with the disinfectant over 120 min. According to the guidelines IA of at least 95% is required for certification of sufficient disinfecting efficacy by the DVG.

An electron microscopic study of the endogenous development of Eimeria mulardi Chauve, Reynaud and Gounel, 1994 was carried out in mule ducks which are hybrids of the domestic duck (Anas platyrhynchos) and the muscovy duck (Cairina moschata). All of the endogenous stages were seen within the nucleus of the host cell. Merozoites arose from ectomerogony and three mutually similar merogonies were noted. The asexual stages were found in leukocyte-like cells in the lamina propria of the jejunum, ileum and caecum, while the gamonts developed in glandular epithelial cells in the same part of the intestine.

The Cathepsin-D proteinase was demonstrated immunohistochemically in cases of intestinal coccidiosis in chickens; its expression concerned almost the cyto-invasive stages of reproductive cycle of several strains of coccidia (merozoites of the II-nd generation and also microgametes). The authors consider Cathepsin-D as a potentional "penetration/spreading factor", being significant for intracellular invasion and spreading of coccidia into host cells. In diagnostic practice, an immunohistochemical demonstration of Cathepsin-D is a relatively simple and sensitive method, which brings information on the extent of parasitic lesions, and on participation of particular evolutive forms of coccidial life cycle; it may also be useful in proving isolated coccidia in the intestinal epithelium, poorly recognized in routine HE preparation. Some Cathepsin-D protease reactivity in intestinal Cryptosporidiosis is also briefly mentioned. The examined cases of cerebral toxoplasmosis and primary acanthamoebic meningoencephalitis (PAME) showed in above mentioned methods negative results.

The phenotype of cells transporting sporozoites of Eimeria necatrix during a primary infection was determined using a panel of six monoclonal antibodies to various chicken lymphocyte surface markers. Sporozoites and cells harboring them were examined at 8, 12 and 18 h postinfection using two-color immunoflorescence and confocal microscopy. The majority of parasites observed within lymphocytes were found in CD 8 + (15%) or CD 3+ (13-22%) cells at all time periods examined. Smaller numbers were found within deltagamma TCR+ (5%) and alphabeta TCR+ (5%) lymphocytes. No sporozoites were found within CD 4+ or IgM+ lymphocytes at any of the time periods.

This study aimed to characterise the local (intestinal) immune response of rats after primary and challenge infections with Eimeria separata. Naive rats and rats which had been immunised by two moderate infections were exposed to a heavy infection with 100000 oocysts per animal. Necropsies were performed 0, 24 and 48 h after infection and lymphocyte subpopulations were microscopically quantified in the caecum mucosa after marking by immunohistological techniques. There was no difference between naive and immune rats concerning the number of CD45R(+) (B) cells, whereas significantly more CD3(+) (T) cells were found in the caecum wall of the immune rats. CD4(+) T cells predominated in animals after primary infection, whereas CD8(+) T cells represented the major T-cell subset in challenged rats. The proportion of TCRgammadelta(+) T cells did not differ in the mucosa between the groups examined, whereas challenged rats showed significantly increased numbers of TCRalphabeta(+) T cells in the caecum wall when compared with animals after a primary infection. Thus, CD4(+) T cells may be particularly involved in the immune response to a primary infection of rats with E. separata whereas immunity to a challenge infection seems to be mediated predominantly by CD8(+) and TCRalphabeta(+) T cells.

1. Coccidiosis, caused by different Eimeria species, is believed to be a more prominent problem in loose-housed layers kept on litter than in battery cages. In this study, the impact and development of Eimeria infections were investigated in layers kept in litter-based, high stocking density systems for loose-housed hens. 2. Layers from 57 flocks on 26 farms were followed by necropsy of a representative sample of birds that died or had to be culled. Coccidiosis was diagnosed in 11 flocks (19.3%) from 9 (31%) of the farms. The outbreaks occurred when the birds were 19 to 32 weeks old. E. maxima was identified in 6 and E. tenella in 3 of the outbreaks. 3. Sixteen of the flocks were also monitored with faecal and litter samples collected at regular intervals. Oocysts were detected in samples from all these flocks. The pattern of oocyst excretion was similar in most of the flocks, with maximum counts at 4 to 8 weeks after introduction to the laying house. There was no significant correlation between the levels of oocysts in faeces and clinical coccidiosis. 4. Raising pullets without any coccidiostat, to increase their chance to develop immunity against coccidia, was not found to decrease the risk of coccidiosis during the production period when compared to the practice of giving amprolium and ethopabate during the rearing period.

We examined exogenous and endogenous development of Eimeria procera in experimentally infected grey partridges (Perdix perdix). Our examination included data on morphology, localization, duration of schizogony and gametogony and morphology of sporulated oocysts. The endogenous stages of E. procera developed in large numbers within the epithelial cells of caecal crypts. The asexual development comprised three generations of schizonts. The first fully developed macrogametes and microgamonts were observed on Day 5 post-infection (p.i.) in histologic section. The patent period began on Day 6 p.i. and ended on Day 11 p.i. with peak production of oocysts on Days 7 and 8. Long oval oocysts of E. procera measured 25.78-28.13 microm in length and 14.06-15.24 microm in width, sporulation time ranged from 18 to 24h at 25 degrees C and from 36 to 48h at 20 degrees C.

Ten poultry farms (broiler breeder pullets, layer pullets, and broilers) in the provinces of Entre Rios and Buenos Aires in Argentina were examined for presence of Eimeria spp. Litter samples obtained from flocks 7-11 wk old were taken to the laboratory for oocyst counting and sporulation, then concentrated for inoculation into coccidia-free chickens. Species were identified by prepatent period, oocyst size, location and appearance of lesions in the intestine, microscopic examination of mucosal smears, and histology (to confirm Eimeria brunetti). On this basis, Eimeria praecox was found in two samples, Eimeria mitis in two, Eimeria acervulina in nine, Eimeria maxima in seven, Eimeria necatrix in three, Eimeria tenella in seven, and E. brunetti in four. These results confirm the presence of all seven recognized species of Eimeria in chickens in the Republic of Argentina.