Comparison of three immunodiagnostic tests for experimental Heterophyes heterophyes infection in dogs

The aim of this study was to compare the performance of three in-house diagnostic tests, i.e. counter current immunoelectrophoresis (CCIE), intradermal (ID) and indirect fluorescent immunoassay (IFI), for the diagnosis of Heterophyes infection. One hundred and twenty puppies were randomly divided into eight groups (n=15/group). Heterophyes heterophyes infections were established in these puppies by administering a dose of 50 H. heterophyes encysted metacercariae/puppy by gavage. Forty puppies of similar age and sex were divided into eight groups, of five puppies each and were used as negative controls. Sera pooled from separate infected and uninfected groups were tested against H. heterophyes antigens, weekly for 8 weeks post-infection (PI). The ID assay detected infected puppies sooner than any of the serological tests. Sero-conversion was first detected 2 weeks PI by ID assay and 1 week later by CCIE and IFI assays. ID test performed well till the end of the experiment (sensitivity and specificity: 100% and 90%, respectively). Both IFI and CCIE assays had a sensitivity of 40% and 20%, respectively for early detection of antibody, which was less sensitive than ID but both assays were more specific (100%) than the ID assay. The lowest agreement was between ID and IFI tests (40.3%), whilst the highest was observed between CCIE and IFI tests (67.2%). Of the three evaluated methods, the ID test could be recommended for scientific and laboratory diagnosis of heterophyosis in naturally infected animals. However, since none of the investigated method are optimal (i.e, offers 100% specificity and sensitivity), the choice of test employed must depend on the aim of the study.     

Host-dependent variations in the seasonal prevalence and intensity of heterophyid encysted metacercariae (Digenea: Heterophyidea) in brackish water fish in Egypt

The prevalence of heterophyid (Digenea: Heterophyidae) encysted metacercariae (EMC) in its second intermediate host, the fish Mugil spp. and Tilapia spp. was studied in a subtropical permanent Lake in northeastern Egypt. Seasonal changes in the occurrence of the EMC in different fish hosts were monitored in a longitudinal field survey lasting 12 months from June 2006 to May 2007. This study tested two hypotheses; (i) prevalence and intensity of heterophyid EMC fluctuate seasonally throughout the year and (ii) variation in the prevalence and intensity of heterophyid EMC is host-dependent. A total of 832 fish specimens comprising 5 species collected from Manzala Lake, northeastern Egypt were examined by artificial gastric juice digestion for EMC. All five species of brackish water fish examined were found to harbor the EMC of the family Heterophyidae in their muscles. The overall infection prevalence of EMC over 12 months was 23.2%. The adult flukes recovered from puppies experimentally infected with morphologically different metacercariae from different fish species were compatible with six species belong to five genera of Heterophyidae, namely, Heterophyes heterophyes, Heterophyes aequalis, Pygidiopsis genata, Phagicola sp., Haplorchis sp. and Stictodora sp. EMC of H. heterophyes were most abundant, detected in 56% of the total fish examined. P. genata was ranked second, followed by Phagicola sp., H. aequalis, Haplorchis sp., and Stictodora sp., respectively. Seasonal differences in infection were observed for all heterophyid species studied in all fish species examined. Heterophyid infections reached peak prevalences during the summer season 38.2% followed by spring 26.6% and autumn 19.3% seasons, whereas the lowest prevalence was recorded in the winter 8.7%. Intensity of heterophyid EMC followed the same seasonal pattern, being high during summer months and low in winter months. All fish species were infected with all the heterophyid digeneans, but with different prevalence. Possible reasons for these findings are discussed with reference to host, climatic and biotic factors.     

Euclinostomum heterostomum and E. ardeolae in tilapia species of Aswan Governorate,Egypt: morphological,molecular, and histopathological characterization

Euclinostomum heterostomum and Euclinostomum ardeolae, both encysted metacercariae (EMC), were found to infect farmed Oreochromis niloticus in the Sahary fish hatchery and wild O. niloticus, Sarotherodon galilaeus, and Tilapia zillii in Lake Nasser, Aswan Governorate, Egypt, at a prevalence of 25.25% and an infection intensity of 1–14 EMC/fish. Macroscopic and microscopic examinations were used to identify them morphologically. PCR amplification, sequencing of the 28S large ribosomal RNA region, and phylogenetic analysis were used for molecular characterization. E. heterostomum and E. ardeolae were isolated from farmed O. niloticus kidneys, peritoneum, abdominal cavity, and under the gills attached to the head bone at prevalence of 13.5% and 6%, respectively, and from wild O. niloticus, S. galilaeus, and T. zillii at a prevalence of 15% and 4.5%, 18% and 10%, and 22.5% and 11.5% respectively. The molecular analyses of rRNA-28S marker revealed that the two Euclinostomum species examined are related yet distinct. This is the first molecular and phylogenetic evidence linking E. ardeolae to Euclinostomum. The present study added two new sequences to the GenBank databases for the genus Eulinostomum from Egypt, with accession numbers MW604803 and MW604806. Euclinostomum spp. have been shown to have detrimental and destructive effects on the kidneys of infected fish; these effects may eventually result in the parasites’ cysts replacing the kidney’s tissues. The kidneys of tilapia spp. infected with Euclinostomum spp. displayed degenerative alterations, with dilated renal tubules associated with migratory tunnels composed primarily of leucocytic infiltrations. Euclinostomum spp. exhibited clear hemophagic characteristics.