Effects of iron chelating agent on Schistosoma mansoni infected murine model.

Schistosomiasis is one of the major health problems in many tropical and developing countries. Infection takes place once cerceriae penetrate human skin, then it changed into schistosomules. The schistosomules takes iron in the form of heme from host's haemoglobin, ferritin and transferrin. Iron is a vital element not only for growth and sexual maturity of schistosomules to adults but also for oogenesis. Since the trapped eggs are the pathological causative agent for most of pathogenesis and complications, the current work was designed to study the effects of early deprivation of schistosomules from iron in the host (in vivo) by chelating it with deferoxamine (DFO). The iron chelation has effects on growth, maturity and egg deposition, as well as it has ameliorative effects on liver pathology such as hepatic fibrosis. Mice were classified into four groups, normal control, DFO treated only, Schistosoma mansoni (S. mansoni) infected DFO untreated and S. mansoni infected DFO treated. The infected DFO treated mice showed significant reduction in fecal egg excretion with increased percentage of dead eggs and this was accompanied with a significant reduction of both total worm burden and hepatic egg load and increased dead egg percentage compared to the infected DFO untreated group. There was also a significant reduction in both serum and hepatic tissue ferritin concentrations in the infected DFO treated mice in comparison to the infected DFO untreated group. Additionally, a significant decrease in number and size of granulomas with subsequent improvement of liver fibrosis was recorded in the infected DFO treated group. This immunopathology was also associated with significant up regulation of Interlukine12 (IL12), Interferon gamma (IFN γ) and significant down regulation in interleukin4 (IL4), interleukin10 (IL10) in both serum and hepatic tissue in the infected DFO treated compared to other groups. Entirely, DFO succeeded in diminishing the growth, maturity and fecundity of S. mansoni with a subsequent improvement of hepatic pathology. As a result of the above findings, it can be concluded that DFO could be considered as a useful treatment against schistosomal infection.

Quantitative assessment of the effects of sulfamethoxazole on Toxoplasma gondii loads in susceptible WT C57BL/6 mice as an immunocompetent host model.

The abundance of Toxoplasma gondii with or without sulfamethoxazole (SMX) treatment was evaluated with quantitative competitive polymerase chain reaction in various organs of wild-type C57BL/6 mice, a susceptible immunocompetent host, after peroral infection with a cyst-forming Fukaya strain of T. gondii. SMX affected different organs in three ways: T. gondii was reduced independently of SMX (skin and kidney); T. gondii was not eradicated with continuous treatment (brain, heart, and lung); and T. gondii was eradicated with continuous treatment (tongue, skeletal muscle, and small intestine). The SMX concentrations in the brains, hearts, and lungs were higher in infected mice than in uninfected mice. These results indicate that even in an immunocompetent host, chemotherapy is necessary to reduce the parasite load and thus reduce the risk of recurrent disease.

Human wound myiasis caused by Phormia regina and Sarcophaga haemorrhoidalis in Minia Governorate, Egypt.

Myiasis is the parasitic infestation of human by the larvae (maggots) of dipterous fly that grow within the host while feeding on its tissue. Cutaneous myiasis is the most considerably encountered clinical form. Moreover, wound (traumatic) myiasis is the main clinical manifestation of cutaneous myiasis. In this research, we aimed to study the type of infesting larvae that are responsible for wound myiasis in the patients in Minia city, Egypt. Three cases of wound myiasis have been noticed among 280 patients with wounds at different parts of bodies. Two of them were diabetic patients. The third one had a history of hypertension with right side hemiplegia 2 years ago. All of them were elderly. The larvae removed from cases 1 and 3 were identified macroscopically and microscopically as the third-stage larvae of Sarcophaga haemorrhoidalis. The larvae removed from case 2 were the third-stage larvae of Phormia regina, which is very rare worldwide. In addition to the open and obsolete wound, diabetes mellitus and low socio-economic circumstances were shown to be attributed as important predisposing risk factors that led to the occurrence of myiasis in these patients.

Polymerase Chain Reaction: A Better Method for Diagnosing Chronic Schistosoma mansoni Infections.

For more effective diagnosis of the acute and chronic stages of Schistosoma mansoni infection in humans, the polymerase chain reaction (PCR) technique was compared with the Kato-Katz method. A total of 150 stool samples were collected from inpatient and outpatient clinics at the Department of Tropical Medicine, Minia University Hospital, Egypt. Three groups of patients, 50 with acute intestinal schistosomiasis, 70 with chronic intestinal schistosomiasis and 30 normal healthy controls were studied. Stool samples were analyzed by PCR and the Kato-Katz method. The mean number of eggs per gram of feces was 4.6 when estimated by the Kato-Katz method in positive stool samples from acute schistosomiasis cases but only 1.7 in chronic cases. In acute intestinal schistosomiasis, 15 and 45 out of 50 cases were positive by Kato-Katz and PCR, respectively. In the chronic intestinal schistosomiasis cases, 6 and 68 out of 70 cases were positive by the Kato-Katz and PCR methods, respectively. We conclude that PCR appears to be an effective diagnostic technique for S. mansoni infection, especially where a low worm burden exists, such as in chronic cases.

Human Cystic Echinococcosis in the Nalut District of Western Libya: A Clinico-epidemiological Study.

Human cystic echinococcosis (CE) is an endemic disease in the Mediterranean area that has not yet been fully documented in western Libya. The present study describes the clinico-epidemiologic profile of CE in western Libya's Nalut district. From April 2008 to July 2011, 36 cases of CE were confirmed following surgical removal of cysts. The cysts were most frequently found in the liver (61.1%), followed by the lungs (19.4%), kidneys (11.1%), peritoneal cavity (11.1%), and spleen (5.6%). Among the 36 patients, 6 possessed plural cysts and 3 had cysts in 2 organs. Blood samples from this group were examined for the presence of serum anti-hydatid IgG antibodies, which revealed positivity in 25 patients (69.4%). An additional 300 blood samples were collected randomly from the inpatient and outpatient clinics at Nalut Hospital. Twenty-seven samples (9%) were found to be positive for the anti-hydatid IgG antibody among which the prevalence of infection tended to be higher in men (12%) than in women (6%). This study demonstrates that CE is a major parasitic infectious disease of public health significance in Libya, notably in the western part of the country, and that disease awareness needs to be raised nationwide.

Cutaneous leishmaniasis in the Nalut District, Libyan Arab Jamahiriya: a clinico-epidemiologic study and Leishmania species identification.

Cutaneous leishmaniasis (CL), an endemic disease in the littoral zones of the Mediterranean area, the Middle East, East Africa, and especially in Libya, has not been fully documented. The present study clarifies the clinico-epidemiologic profile of CL and the molecular genotyping of the Leishmania spp. in the Nalut district, Libya. Two hundred and twenty-three CL patients were examined at the out-patient clinics of Nalut Hospital from March 2006 to February 2007. CL was diagnosed by clinical, microscopic, culture, polymerase chain reaction (PCR), and PCR-restriction fragment length polymorphism (RFLP) analyses. The disease was observed year-round, with the highest prevalence between November and February. Fifty-nine percent of patients were younger than 20 yr. Nodulo-ulcerative lesions, indurated ulcers, papulo-ulcerative lesions, and subcutaneous nodular lesions were observed in 170, 25, 15, and 13 patients, respectively. Two hundred patients (89.7%) had dry type of lesions, whereas 23 patients (10.3%) presented a wet type of lesion. One hundred and fifty-nine (71.3%) of 223 patients were confirmed positive for CL by the presence of the amastigote form of Leishmania by stained Giemsa smear, and 170 (76.2%) were positive according to the presence of the promastigote form of Leishmania by culture in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS). PCR confirmed 203 (91.0%) positive cases. Genotyping of Leishmania spp. by RFLP analysis revealed that L. tropica was the most common species at all ages, and L . infantum was second under 20 yr of age. In summary, CL is endemic in the Nalut district, Libya; PCR was the most sensitive parasite diagnostic test, and L. tropica was the most common species.

Clinical varieties of Toxocariasis canis in Children's Hospital, Mansoura University: is it an underestimated problem?

Human toxocariasis is a worldwide parasitic disease. Children are more frequently infected because of the closer contact with contaminated soil and relatively frequent geophagia. Toxocariasis in children has variable modes of presentation but clinical diagnosis is difficult. Various clinical phenotypes of toxocariasis in symptomatic children attending Children's Hospital Mansoura University were studied. A total of 480 children were included in the study with mean age 7.24 +/- 4.22 years, 61.9% were boys and 200 age-sex-matched healthy controls. Patients were examined clinically, and the anti-Toxocara antibodies in the blood of children were performed by ELISA using T. canis larval excretory-secretory products as antigen. Eosinophils level in peripheral blood was measured. Sero-positive cases were 12 % of patients and only 3.5% of controls. Statistical analysis showed a significant association between infection and male sex (P <0.001). Sero-positive children were older than the sero-negative (P <0.001). Eosinophilia was detected in 86.2% of sero-positive children. Sero-positivity and degree of eosinophilia were more frequently detected among patients with allergy (bronchial asthma and urticaria). Degree of eosinophilia was found to be positively correlated to the optical density (OD) ELISA of anti-Toxocara IgG.

Evaluation of ELISA and IHAT in serological diagnosis of proven cases of human hydatidosis.

Cystic hydatid disease (Hydatidosis) is the most serious tape-worm infection prevalent in the cattle and sheep raising area of the world. Hydatidosis in man (as an accidental host) is caused by infection with the ova containing larval stage of Echinococcus granulosus. In the last decade, different techniques have been employed for sero-diagnosis of hydatidosis; as IHA, IFA, ELISA, CCLE (Counter Current Immuno-electrophoresis). This paper evaluated the validity of ELISA and IHA. Since whole hydatid cyst fluid was used as a source of antigen for serodiagnosis. Thirty surgical and pathological hydatidosis proven patients were examined. The sensitivity and specificity of ELISA were 96.7% & 97.5% respectively, and that of IHA were 86.7%, &95% respectively.

Evaluation of the effects of sulfamethoxazole on Toxoplasma gondii loads and stage conversion in IFN-gamma knockout mice using QC-PCR.

Toxoplasma gondii abundance with or without sulfamethoxazole treatment was evaluated by quantitative competitive polymerase chain reaction (QC-PCR) assay in various organs of IFN-gamma knockout BALB/c (B/c) mice after peroral infection with the cyst-forming Fukaya strain. T. gondii infection was observed in the brain, skin, tongue, heart, and skeletal muscle of the mice treated with sulfamethoxazole, although the parasite was not observed during the treatment in the mesenteric lymph node, spleen, small intestine or kidney. After discontinuing the therapy, T. gondii reappeared within five days in all organs. Reverse transcriptase (RT)-PCR showed that sulfamethoxazole treatment accelerated the stage conversion of T. gondii from tachyzoites into bradyzoites in the brain, lung, and heart. In contrast, after discontinuing sulfamethoxazole treatment, T. gondii underwent stage conversion from bradyzoites into tachyzoites in these organs. These results indicate that we successfully established an animal model for evaluating chemotherapy regimens in immunocompromised hosts infected with T. gondii.

Induction of protective immunity by primed B-1 cells in Toxoplasma gondii -infected B cell-deficient mice.

We examined the role of B-1 cells in protection against Toxoplasma gondii infection using B cell-deficient mice (muMT mice). We found that primed but not naïve B-1 cells from wild-type C57BL/6 mice protected B cell-deficient recipients from challenge infection. All muMT mice transferred with primed B-1 cells survived more than 5 months after T. gondii infection, whereas 100% of muMT mice transferred with naïve B-1 cells succumbed by 18 days after infection. Additionally, high expression of both T help (Th) 1- and Th2-type cytokines and a high level of nitric oxide production were observed in T. gondii-infected muMT mice transferred with primed B-1 cells. Thus, it was clearly demonstrated that B-1 cells play an important role in host protection against T. gondii infection in muMT mice.

Induction of protective immunity by DNA vaccination with Toxoplasma gondii HSP70, HSP30 and SAG1 genes.

The vaccination with Toxoplasma gondii heat shock protein 70 (T.g.HSP70) gene (a virulent tachyzoite-specific) induced the most prominent reduction in T. gondii loads in various organs of B6 and BALB/c mice at the acute and chronic phases of toxoplasmosis compared with T.g.HSP30 (a bradyzoite-specific) and SAG1 (a tachyzoite-specific) genes. A single gene gun vaccination with 2 microg of T.g.HSP70 gene induced a significant reduction in the number of T. gondii organisms compared with 50 microg of T.g.HSP70 gene vaccination by intramuscular (i.m.) or intraperitoneal (i.p.) injection. The vaccine effects of T.g.HSP70 gene persisted for more than 3 months.