A total of 450 stool samples were collected from inpatient and outpatient clinics of Pediatric Department, Minia University Hospital, Minia District, Egypt. Two groups of patients were studied, including 200 immunosuppressed and 250 immunocompetent children. Stool samples were subjected to wet saline and iodine mounts. A concentration technique (formol-ether sedimentation method) was carried out for stool samples diagnosed negative by wet saline and iodine mounts. Samples were stained by 2 different methods; acid fast stain (modified Ziehl-Neelsen stain) and Giemsa stain. Total 188 cases (94%) were diagnosed positive for parasitic infections among immunosuppressed children, whereas 150 cases (60%) were positive in immunocompetent children (P<0.0001). The most common protozoan infection in immunosuppressed group was Cryptosporidium parvum (60.2%), followed by Blastocystis hominis (12.1%), Isospora belli (9.7%), and Cyclospora caytenensis (7.8%). On the other hand, Entamoeba histolytica (24.6%) and Giardia lamblia (17.6%) were more common than other protozoans in immunocompetent children.
Schistosomiasis is an endemic disease in 74 countries causing more than 250,000 deaths every year. Accordingly, the development of an effective drug for eradication of schistosomiasis is an open research field. The current chemotherapy for control is praziquantel (PZQ). However, PZQ does not improve liver fibrosis. Therefore, the aim of this study is to evaluate the combined effect of alpha lipoic acid (ALA) with PZQ on the liver fibrosis induced by Schistosoma mansoni challenged mice. Evaluation was based on the worm burden count, ova load, granuloma size, and histopathology of the liver. Reduced glutathione (GSH) was measured in the tissue as a biomarker for impaired antioxidant function. Malondialdehyde (MDA) was also measured in the tissue as a biomarker for oxidative stress. The serum level of matrix metalloproteinase 1 was measured as a biomarker for fibrotic status of the liver. Liver function enzymes such as ALT, AST, and GGT were also measured. Four groups of ten mice each were used in this study. The first group was infected with 50 ± 10 S. mansoni cercariae. The second group was also infected and was treated with PZQ 9 weeks post-infection (PI). The third group was treated with PZQ and ALA 9 weeks PI. The fourth group was used a healthy control. The present study revealed remarkable improvement in all parameters measured (parasitological and biochemical) as well as significant improvement of hepatic pathology in the third group which was treated with PZQ and ALA. The treatment of mice with PZQ and ALA results in reduction in the worm burden, egg count, and granuloma size. Furthermore, this combined treatment increased the tissue level of the antioxidant (GSH) and decreased the tissue level of MDA in this group.
Controversy surrounding the pathogenic role of Blastocystis spp. in humans and lack of well-established diagnostic criteria led to debates concerning the treatment for that organism. Furthermore, some strains develop resistance against the recommended drugs. Thus, using natural medicine has many positive aspects to address these points. In an earlier study, we addressed in vitro effect of garlic and ginger on Blastocystis spp. isolates as an alternative treatment. Accordingly, this study was conducted to evaluate in vivo activities of these two herbs on mice infected with Blastocystis spp. Antiprotozoan activities were determined by monitoring Blastocystis shedding in stools and histopathological changes of the intestine of infected mice. Additionally, assessment of the antioxidant effect (via measuring the level of malondialdehyde (MDA) production) of these herbs on the treated groups of mice was done. Also, their effects on nitric oxide (NO) production were assessed. In this work, treatment of infected mice with garlic, ginger, and nitazoxanide (NTZ) reduced the shedding of cysts significantly compared to the infected untreated group, P value ≤0.001, 0.0001, and 0.0003, respectively. As well, histopathological examination revealed that Blastocystis was frequently observed within the lumen, at the tip of the epithelium, and/ or infiltrated in an enterocyte in the infected group without treatment compared to that of the infected treated ones. Furthermore, mice infected with Blastocystis exhibited increased levels of NO (440.09 ± 3.7 vs. 276.66 ± 0.8, P ≤ 0.001) and MDA production (106.19 ± 0.43 vs. 63.06 ± 0.45, P ≤ 0.0004) compared to that of the uninfected controls. Treatment of infected mice with garlic, ginger, and NTZ reduced NO levels to 54.41 ± 1.2, 47.70 ± 1.2, and 37.43 ± 0.98 and MDA levels to 22.38 ± 0.17, 63.34 ± 3.89, and 66.76 ± 9.1, respectively. We conclude that using ginger and garlic for treatment of blastocystosis is beneficial.
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.
Four cases of gastric or intestinal myiasis are reported. The cases contain 2 males (1 child 10 years old, and 1 adult 40 years old) and 2 females (1 girl 18 years old, and 1 adult 50 years old) from Minia Governorate, Southern Egypt. Three of them, including cases no. 1, 3, and 4, were gastric myiasis, and complained of offensive hematemesis of bright red blood. Minute moving worms, larvae of the fly, were found in the vomitus. On the other hand, case no. 2 had intestinal myiasis, and complained of abdominal distention, nausea, vomiting, and diarrhea. The stool of case 2 was mixed with blood, and minute moving worms were observed in the stool. Endoscopy was performed to explore any pathological changes in the stomach of the patients. The larvae were collected and studied macroscopically, microscopically, and us-ing a scanning electron microscope (SEM) to identify their species. Three different types of larvae were identified. The larvae isolated from case 1 were diagnosed as the second stage larvae of Sarcophaga species, and the larvae isolated from case 2 were the third stage larvae of Sarcophaga species. On the other hand, the larvae isolated from cases 3 and 4 were diagnosed as the third stage larvae of Oestrus species.
This study investigated the effect of breast-feeding in protection against protozoan infection in infants with persistent diarrhea. Infants were classified into 2 groups; 161 breast-fed infants and the same number of non-breast-fed infants. Microscopic examinations of stool were done for detection of parasites and measuring the intensity of infection. Moreover, serum levels of IgE and TNF-α were measured by ELISA. Cryptosporidium spp., Entamoeba histolytica/Entamoeba dispar, Giardia lamblia, and Blastocystis sp. were demonstrated in infants with persistent diarrhea. The percentage of protozoan infections was significantly lower in breast-fed infants than that in the non-breast-fed infants. The levels of IgE and TNF-α were significantly lower in the breast-fed group than in the non-breast-fed group. There were significant positive associations between the serum levels of IgE and TNF-α and the intensity of parasite infection in the breast-fed group. It is suggested that breast-feeding has an attenuating effect on the rate and intensity of parasite infection.
Experimental vaccination with radiation-attenuated cercariae (RAC) confers possible practical levels of resistance to challenge infection by humoral and by cellular mechanism. Here, we aimed to identify possible vaccine antigens by using specific IgG antibody from RAC vaccinated miniature pig. Two milligrams of soluble egg antigen (SEA) or schistosomal worm antigen preparation (SWAP) was fractionated using two dimensional liquid chromatography From the positive fractions we chose 17 of SEA and 10 of SWAP that had no reactivity with normal cercariae infected (NCI) sera and had single peak of 214 nm; and automated N-terminal amino acid sequence based on in situ Edman Reaction was conducted. Four sequences were obtained and applied to the homology search in NCBI database. A total of eight candidate genes were listed up and their cDNA clones from schistosomula stage were obtained. Two of the recombinant proteins (AAW27472.1 and AXX25883.1) showed strong reactivity with the RAC vaccinated serum but marginal with NCI serum. This protocol using proteome PF2D could be applicable in identifying immunoreactive proteins from crude extract for the development of vaccines or for diagnostics.
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