Background:Fasciolosis is an important zoonotic disease affecting the productive performance of farm animals in Egypt.Aim:The aim of the present study was comparing the ovicidal effect of different extracts as an alcoholic (Methanolic and Ethanolic) and aqueous Moringa oleifera leaf extracts on Fasciola gigantica non-embryonated and developed eggs.Materials and Methods:Tested concentrations of extracts ranged from 12.5 to 800 mg/ml. Nitroxynil was used as reference drug with a dose of 100 mg/ml.Results:M. oleifera alcoholic and aqueous extracts showed a concentration-dependent ovicidal effect on F. gigantica non-embryonated and developed eggs. Based on LC50 values, water extract showed the highest ovicidal activity since it registered the lowest values of 2.6 mg/ml on non-embryonated eggs. Non-embryonated eggs were more susceptible to aqueous extract than developed eggs. On the other hand, the developed eggs were more susceptible to ethanolic extract than non-embryonated eggs even the lowest LC50 (12.38 mg/ml).Conclusion:M. oleifera leaf extracts especially aqueous extract could be a promising step in the field of controlling fascioliasis. Further, in vivo studies are needed to enlighten the therapeutic potential of M. oleifera extracts in treating F. gigantica infection.

Toxoplasmosis is an important worldwide foodborne zoonotic disease. Infected cattle meats are considered a serous cause of human toxoplasmosis. Here, this study assesses the infection with Toxoplasma gonddi (T. gondii) in cattle using meat juice samples from diaphragmatic muscles collected at the slaughter. An in house indirect enzyme linked immunosorbent assay (ELISA), and commercial latex agglutination test (LAT) followed by immunoblotting were developed on the meat juice (fluids) using tachyzoites of locally isolated T. gondii strain. The comparative analysis of the results of the tested juice samples showed an excellent agreement between the in-house indirect ELISA and LAT test in the positive and negative of meat juice. Relative sensitivity was higher for ELISA on diaphragms fluids random samples 80.39%, for the LAT test was 68.6%. Immune-reactive bands of T. gondii local strain Ag with naturally infected meat juice were 116, 83, 65, 30 and 23 KDa. The obtained results concluded that the development of an effective ELISA test to be used in for detection of toxoplasmosis infection of slaughtered cattle in large-scale would be exactly valuable, since the important role that beef plays in epidemiology of T. gondii, in particular the hazard of transmission to human and food safety.

Background and Aim: Cryptosporidiosis is a leading cause of diarrheal disease worldwide and is an animal and public health burden. This study aimed to evaluate the protective potential of affinity-purified Cryptosporidium parvum oocyst antigen as a vaccine candidate according to fecal oocyst shedding, humoral and cellular immune responses, histopathological changes, and the number of parasite developmental stages in ileal and hepatic tissues. Materials and Methods: We isolated oocysts from naturally infected buffalo calves and identified them molecularly as C. parvum isolates (GenBank: ON730707 and ON730708) by targeting the Cryptosporidium oocyst wall protein gene. We propagated the C. parvum oocysts in mice. In addition, we prepared crude antigen from the isolated oocysts by purification using cyanogen bromide-activated Sepharose-4B affinity chromatography coupled with rabbit hyperimmune serum. Then, we divided 81 parasite-free mice into three groups: (1) non-vaccinated non-infected mice, (2) mice orally infected with 1 × 105 C. parvum oocysts on week 4 of the experiment, and (3) mice immunized twice with 40 μg/kg of the purified fraction at 2-week intervals. Then, we challenged the vaccinated group with C. parvum oocysts after 2 weeks, and the positive control group was infected at the same time. Results: We observed a prolonged prepatent period and decreased oocyst shedding in the vaccinated infected mice compared with the non-vaccinated infected mice (t < 0.001). The vaccinated mice had significantly higher immunoglobulin G levels than those in the other two groups at all examined weeks. In addition, the production of cytokines interferon-gamma, interleukin (IL)-10, IL-12, and IL-15 was activated post-vaccination. After the challenge, all tested cytokines were significantly increased (p < 0.001) in the two infected groups compared with the non-vaccinated non-infected group, with the highest levels in the vaccinated infected group. Vaccinated infected mice exhibited significantly fewer pathological lesions in the ileum and liver than non-vaccinated infected mice, which showed prominent histopathological lesions. Endogenous developmental stages of C. parvum indicated that the ileum was more parasitized than the liver and that vaccination resulted in a lower number of oocysts in ileal and hepatic tissues (p < 0.05). Conclusion: Our prepared affinity-purified vaccine candidate could be promising in protecting against cryptosporidiosis.

In the current study, cross-reaction between two important zoonotic parasites; extracellular helminthes Fasciola gigantica and intracellular protozoa Toxoplasma gondii was proved by enzyme linked immunosorbent assay. Five antigens were used to identify and compare the cross-binding activities in the prepared antisera. Two F. gigantica antigens; adult flukes (FgA) and eggs (FgEA) were used to detect IgG in T. gondii naturally infected human sera (TgIHS) and experimentally infected sera of sheep (TgISS), mice (TgIMS) and rats (TgIRS). Three types of T. gondii antigens; RH (TgRHA), local sheep isolate (TgLA) and ME49 isolate (TgMEA) were used to detect cross binding activities in F. gigantica experimental infected rabbit sera (FgIRS) and F. gigantica naturally infected bovine sera (FgIBS). The cross-binding activities in the prepared antisera were strongly directed towards FgA and TgLA rather than the other antigens. The characterization of the five antigens using SDS-PAGE showed 4 common bands of FgA and TgLA; 165, 97, 76, and 65 kDa. While two common bands were observed between TgRHA, TgMEA and FgA; 165, and 65 kDa. Whereas, two common bands found between three types of T. gondii antigens and FgEA were identified; 165 and 65 kDa. The immunogenic cross-reactive bands between FgA and TgLA with F. gigantica infected bovine sera were identified by immunoblot. In FgA, the common immunogenic bands were 165, 65 and 14 kDa. While in TgLA, common immunogenic bands were 165 and 65 kDa. Whereas, the common immunogenic band between FgA and TgLA identified with T. gondii experimentally infected sheep sera was 65 kDa. The current research proves cross reaction between F. gigantica and T. gondii. One common band of 65 kDa showed broad immunogenic cross-reactivity with the developed antisera raising the prospect of being putative common immunodiagnostic candidate of both infections.

Background: Cystic echinococcosis (CE), a zoonotic disease that affects animal and human health, is of increasing economic importance due to high morbidity rates and high economic losses in the livestock industry. Aim: The present study was conducted to purify the antigen from hydatid cyst fluid (HCF) with high diagnostic efficacy of camel hydatidosis using indirect enzyme-linked immunosorbent assay (ELISA). Materials and Methods: The HCF antigen was purified using Sephacryl S-300 column chromatography. Characterization of fractions was performed using reducing and non-reducing sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. Further, antibodies against Echinococcus granulosus cysts in camel serum were detected using indirect ELISA. Results: The purification process resulted in three fractions of antigens: FI, FII, and FIII. Indirect ELISA showed that higher diagnostic efficacy was observed in FI than in FII and FIII. Indirect ELISA, in which FI was utilized, showed 88% sensitivity and 91.7% specificity. Non-reducing SDS-PAGE showed that FI had two bands of molecular weights 120 and 60 kDa. Western blot analysis of FI demonstrated that 60, 38, and 22 kDa were antigenic bands when reacted with naturally infected camel sera with E. granulosus cysts. Using indirect ELISA, F1 recorded an infection percentage of 81.7% in randomly collected camel serum samples. Conclusion: FI is a promising antigen for accurate diagnosis of camel CE using indirect ELISA.