Thabet Sakran

Endogenous stages of Choleoeimeria baltrocki (Daszak et Ball, 1991) n. comb. infecting the gall bladder of gold skink, Eumeces schneiderii Daudin, 1802 from Egypt

Examination of 98 gold skink, Eumeces schneiderii Daudin, 1802 from Alexandria and Marsa-matroh Governorates, Egypt revealed oocysts of Choleoeimeria baltrocki (Daszak et Ball, 1991) n. comb. in the gall bladder and faeces. The prevalence of the infection was 40.8%. Sporulated oocysts were found in the faeces and the gall bladder contents. Sporulated oocysts were tetrasporocystic, cylindrical, reaching a mean of 38.7 (36–42) × 19.9 (17–25) μm with bilayered smooth and colourless wall. The dizoic sporocysts were subspherical and measuring 10.8 (9.5–13) × 9.3 (8–10.5) μm with a large globular sporocyst residuum. Sporocyst wall was bilayered joined by meridional suture. Sporozoites were crescent-shaped, blunt at one end and slightly tapered at the other. Endogenous stages (meronts, gamonts, gametes and young oocysts) were confined to the gall bladder epithelium and the infected cells were hypertrophied. Based on the morphological features of the exogenous stages and endogenous development of the present parasite, its generic affiliation was revised and placed into the genus Choleoeimeria.

Lambs Infected with UV-Attenuated Sporocysts of Sarcocystis ovicanis Produced Abnormal Sarcocysts and Induced Protective Immunity against a Challenge Infection

The present study surveyed the prevalence of natural infection of the sheep esophagus muscle with sarcocysts of Sarcocystis ovicanis and examined induction of protective immunity using UV-attenuated sporocysts. The overall prevalence of natural infection of the sheep was 95%. Infectivity of the collected sarcocysts was confirmed by shedding of sporulated oocysts after feeding infected esophageal tissues to dogs. To induce protective immunity, lambs were immunized 3 times (once a week) with 1.5 x 10(4) sporocysts exposed to UV-light for 30 min (UV-30 group) or 60 (UV-60 group) min and then challenged with 1.5 x 10(4) normal sporocysts at the 3rd week post the 1st vaccination. These lambs showed high survival and less clinical signs of sarcocystosis than normal infected lambs. The attenuated sporocysts produced abnormal cysts; small in size and detached from the muscle fiber. These abnormalities were more obvious in UV-60 group than UV-30 group. Also, the IFN-gamma level and lymphocyte percentage were increased while the total leukocyte count was decreased in the UV-60 group compared with other groups. The high level of IFN-gamma may be an evidence for the induction of Th1 responses which may have protective effect against a challenge infection.

A new record of Myxobolus brachysporus and M. israelensis in the tilapia (Oreochromis niloticus) collected from the Nile River, Egypt.

The present study was carried out as part of an ongoing general survey for myxosporean parasites infecting tilapias in the River Nile, Egypt. In the present study, 77 Nile tilapia (Oreochromis niloticus) were collected from boat landing sites at Beni-Suef governorate, Egypt and examined for the myxosporean infection. The infection was encountered as a huge number of free spores in the kidney and the spleen. The infection showed a prevalence of 51.9% (40/77) for Myxobolus brachysporus while it was 25.9% (20/77) for Myxobolus israelensis. Mature spores of M. brachysporus were ellipsoidal and measured 8.6 × 13.2 μm. The polar capsules were subcircular with 5–6 filament turns and measured 4.7 × 3.6 μm. Spores of M. israelensis were ellipsoidal in the frontal view and fusiform in the lateral view. Spore measurements were 13.4 μm long and 8.7 μm wide. The polar capsules were elongated with 6–7 filament coils and measured 8.6 × 3.1 μm. The findings presented here proved that tilapia fishes in the Nile River are still suffering from infections with Myxobolus species. Therefore, further studies should be carried out to survey the Myxobolus infection among tilapias under culture conditions to clarify the pathological impacts of this parasite in tilapias aquaculture.

Diagnostic potential of Fasciola gigantica -derived 14.5 kDa fatty acid binding protein in the immunodiagnosis of bubaline fascioliasis

The 14.5 kDa fatty acid binding protein (FABP) was isolated from the crude extract of adult Fasciola gigantica worms. Polyclonal anti-FABP IgG was generated in rabbits immunized with prepared FABP antigen. Sandwich enzyme-linked immunosorbent assay (ELISA) was applied to detect coproantigen in stools and circulating Fasciola antigen (CA) in sera of 126 water buffaloes by using purified and horseradish peroxidase (HRP)-conjugated anti-FABP IgG. Sandwich ELISA sensitivity was 96.97% and 94.95%; while specificity was 94.12% and 82.35% for coproantigen and CA detection, respectively. However, sensitivity and specificity of the Kato-Katz technique was 73.74% and 100%, respectively. The diagnostic efficacy of sandwich ELISA was 96.55% and 93.1% for coproantigen and CA detection, respectively. In contrast, the diagnostic efficacy of the Kato-Katz technique was 77.59%. In conclusion, these results demonstrate that the purified 14.5 kDa FABP provides a more suitable antigen for immunodiagnosis of early and current bubaline fascioliasis by using sandwich ELISA.

Immunomodulatory effect of diethylcarbamazine citrate plus filarial excretory-secretory product on rat hepatocarcinogenesis.

Diethylcarbamazine citrate (DEC) had a significance in anti-filarial chemotherapy, while excretory-secretory product (ES) is released from adult filarial females. The target of the current study was to examine the immunomodulatory effect of DEC, Setaria equina ES or a combination of them on rat hepatocellular carcinoma (HCC) induced by diethylnitrosamine (DEN). In vitro effect of combined DEC and ES or ES alone on lipopolysaccharide (LPS)-stimulated rat peripheral blood mononuclear cells (PBMCs) was tested through IFN-γ assay in culture supernatants. In addition, single or repeated doses of DEC, ES or DEC+ES have been applied in white albino rats to test the effect on HCC. Levels of IFN-γ and anti-ES IgG antibodies in rat serum were assayed using ELISA. Hemolytic complement activity (CH50) was determined in serum while the concentration of nitric oxide (NO) was assayed in liver tissue. The infiltration of NK cells as well as the expression of MHC Iproliferating cell nuclear antigen (PCNA), inducible NO synthase (iNOS), Bcl2 and p53 were determined using immunohistochemistry. There was a dose-dependent increase in IFN-γ after in vitro exposure to DEC+ES. Repeated ES doses increased NO concentration (p<0.05) and expression of iNOS but reduced CH50 (p<0.001), while repeated DEC+ES doses could increase anti-ES IgG (p<0.01), IFN-γ level (p<0.05) and NK cell infiltration. The same treatments could also reduce the expression of MHC I expression, PCNA, Bcl2 and p53. This study has shown immunomodulatory and protective effects of DEC+ES repeated doses on rat HCC.

Immunoprotective responses against murine sarcocystosis by β - Irradiated sporocysts

This study aimed to induce protective immunity against infection with Sarcocystis muris in experimental mice using β-irradiated sporocysts. Mice were vaccinated with 50 sporocysts of S. muris which were exposed to 1.84 μSv β-irradiation for 2, 4 and 8 h. After challenge infection, different samples were taken for evaluation. Serum and intestinal wash were assayed for IFN-γ and IgA, respectively. Mesenteric lymph nodes (MLNs) and spleen were investigated for CD4+ and CD8+ T cells using immunohistochemistry. For liver, the morphological changes in parasitic stages and the count of infiltrated CD8+ T, NK1.1+ and FasL+ cells were also investigated. Real time (RT) - PCR was used for detection of liver MHC I, CD1d, IFN-γ, perforin and FasL as well as the parasite 18S ribosomal(r) RNA in liver and muscle tissues. Alterations of liver parasitic stages as well as a decrease in the infection with the parasite in both of liver and muscle tissues were dependent on radiation exposure time. An investigation for the mechanism of immunoprotection showed an increase in liver NK1.1+ & FasL+ cells, serum IFN-γ and intestinal IgA, while CD4+ and CD8+ T showed a remarkable increase in MLNs and spleen. FasL expression increased in the liver dependently on radiation exposure time, while perforin, MHC I and CD1d were not. β-irradiated sporocysts with 1.84 μSv for 8 h s could induce the highest protection against infection with Sarcocystis. This could be largely relied on the increased infiltration of NK cells and associated higher expression of FasL in the liver.

Detection rates of waterborne protozoa in water sources from Fayoum Governorate

Background: Waterborne protozoal infections are common health problems in many parts of the world especially in developing countries. Water is a major vehicle for transmission of protozoa such as G. lamblia, Cryptosporidium spp. as well as pathogenic and opportunistic free living amoeba (FLA).Objective: This study aims to detect the presence of protozoal agents in tap water and storage water tanks at Fayoum Governorate, Egypt.Material and Methods: A total of ninety five water samples were collected from different water sources, taps (65) and tanks (30), from 6 Fayoum districts. The samples were processed to detect the presence of G. lamblia cysts by Lugol’s iodine stain, Cryptosporidium oocysts by modified Ziehl-Neelsen stain and FLA by cultivation. After cultivation, Acanthamoeba spp. were identified according to their morphological features and flagellation test was performed to detect amoeboflagellates.Results: All water samples collected from tanks (100%) were contaminated by protozoa of medical importance, while only 6 (9.2%) of the tap water samples were pathogen free. The majority of water samples were contaminated with mixed protozoal infections. The overall detection rates of contaminants in water sources were 86.3%, 52.6%, 13.7% by FLA, Cryptosporidium spp. and G. lamblia, respectively.Conclusions: The recorded detection rates of waterborne protozoa present a hazard to the community resulting in silent morbidities and mortalities. It is strongly recommended to adopt proper water safety measures.

Detection for cross-reactive proteins in filarial worm Setaria equina, MCF-7 human breast cancer, and Huh-7 hepatoma cells

ABSTRACT This study aimed to detect the cross-reactive proteins in filarial parasite adult worm Setaria equina and two different tumor cell lines (MCF-7 human breast cancer and Huh-7 hepatoma cells). This was performed using rabbit anti-S. equina extract (SeqE) or DEC (Diethylcarbamazine citrate) polyclonal IgG antibodies by indirect ELISA and western blotting. The results indicated cross-reactive bands at 70 and 75 kDa in all extracts by anti-DEC and SeqE antibodies, respectively. In addition, the expression of 70 kDa protein was only reduced in filarial worms and Huh-7 after in vitro DEC treatment compared to the control.