Hepatic coccidiosis in Serrasalmus rhombeus Linnaeus, 1766 from the Amazon basin: morphological and histopathological aspects

Calyptospora species are coccids that commonly cause liver infections in fish all around the world. This paper describes the morphology and histopathological characteristics of liver infection caused by Calyptospora sp. in black piranha Serrasalmus rhombeus, from the Capim River, in the municipality of Ipixuna do Pará, state of Pará (Brazil). Specimens were collected, analyzed and necropsied and tissue fragments containing parasites were prepared for histology and scanning electron microscopy. Parasitism was detected in 33.0% of the analyzed specimens, which had spherical oocysts in the liver, with four pyriform sporocysts presenting sporozoites internally. A histological examination revealed oocysts positioned close to blood vessels, causing necrosis and degeneration of hepatic parenchyma, while the presence of mononuclear cell infiltrate and melanomacrophages indicated the onset of an inflammatory process. This is the first record of the genus Calyptospora in fish from the Capim River.

Expression Analysis and Serodiagnostic Potential of Microneme Proteins 1 and 3 in Eimeria stiedai

Eimeria stiedai is an apicomplexan protozoan parasite that invades the liver and bile duct epithelial cells in rabbits and causes severe hepatic coccidiosis, resulting in significant economic losses in the domestic rabbit industry. Hepatic coccidiosis lacks the typical clinical symptoms and there is a lack of effective premortem tools to timely diagnose this disease. Therefore, in the present study we cloned and expressed the two microneme proteins i.e., microneme protein 1 (EsMIC1) and microneme protein 3 (EsMIC3) from E. stiedai and used them as recombinant antigens to develop a serodiagnostic method for an effective diagnosis of hepatic coccidiosis. The cDNAs encoding EsMIC1 and EsMIC3 were cloned and the mRNA expression levels of these two genes at different developmental stages of E. stiedai were determined by quantitative real-time PCR analysis (qRT-PCR). The immunoreactivity of recombinant EsMIC1 (rEsMIC1) and EsMIC3 (rEsMIC3) proteins were detected by Western blotting, and indirect enzyme-linked immunosorbent assays (ELISAs) based on these two recombinant antigens were established to evaluate their serodiagnostic potential. Our results showed that the proteins encoded by the ORFs of EsMIC1 (711 bp) and EsMIC3 (891 bp) were approximately 25.89 and 32.39 kDa in predicted molecular weight, respectively. Both EsMIC1 and EsMIC3 showed the highest mRNA expression levels in the merozoites stage of E. stiedai. Western blotting analysis revealed that both recombinant proteins were recognized by E. stiedai positive sera, and the indirect ELISAs using rEsMIC1 and rEsMIC3 were developed based on their good immunoreactivity, with 100% (48/48) sensitivity and 97.9% (47/48) specificity for rEsMIC1 with 100% (48/48) sensitivity and 100% (48/48) specificity for rEsMIC3, respectively. Moreover, rEsMIC1- and rEsMIC3-based indirect ELISA were able to detect corresponding antibodies in sera at days 6, 8, and 10 post E. stiedai infection, with the highest positive diagnostic rate (62.5% (30/48) for rEsMIC1 and 66.7% (32/48) for rEsMIC3) observed at day 10 post infection. Therefore, both EsMIC1 and EsMIC3 can be used as potential serodiagnostic candidate antigens for hepatic coccidiosis caused by E. stiedai.

Clinico-pathologic observations of spontaneous hepatic coccidiosis in broiler rabbits maintained in Bannerghatta biological park in Karnataka state of India

The present study reports the clinico-pathologic changes observed in spontaneous hepatic coccidiosis caused by Eimeria stiedae in rabbits at Bannerghatta Biological Park (BBP) in Karnataka state of India. The clinical signs observed in the affected rabbits included sudden death, stunted growth, anorexia, weight loss, diarrhoea and abdominal pain. Increased liver enzymes and bilirubin was noticed upon biochemical examination. At necropsy, lesions included grossly enlarged liver (hepatomegaly) with presence of discrete yellowish-white nodules ranging from 1mm to 5mm size throughout the parenchyma containing a thick creamy white fluid. Faecal sample examination demonstrated the presence of Eimeria oocyts. The impression smears from the liver and intestinal mucosa also showed Eimeria oocysts, degenerative changes, necrotic cells and mononuclear cell aggregation to form oocyst granuloma. Histological observation revealed distension of bile duct, hyperplasia of epithelium along with numerous coccidian oocysts at the centre. Granulomatous tissues were observed to have encircled the bile duct with infiltration of inflammatory cells. Microscopic examination of exudates from liver also revealed numerous oocysts of E. stiedae. Multiple areas of coagulative necrosis of hepatic cells surrounded with inflammatory cells were also found.