Ayman A. El-Badry

Molecular characterization of cutaneous leishmaniasis in Al-Madinah Al-Munawarah province, western Saudi Arabia.

BACKGROUND Leishmaniasis is a parasitic disease affecting a large number of people worldwide. In this study we carried out the molecular characterization of cutaneous leishmaniasis (CL) in Al-Madinah Al-Munawarah Province, Saudi Arabia, confirming Leishmania major and Leishmania tropica as the prevalent species using molecular techniques. METHODS One hundred and five patients with suspected CL were identified from four different localities in Al-Madinah Al-Munawarah Province and Al-Miqat Hospital, Al-Madinah, Saudi Arabia. Thirty-four of the 105 patients were selected at random for molecular investigation. RESULTS Characterization of CL species by internal transcribed spacer 1 (ITS1) PCR-restriction fragment length polymorphism (RFLP) and kinetoplast DNA (kDNA) PCR established L. major and L. tropica as the causative organisms. kDNA PCR had a sensitivity of 90.7%, whereas ITS1 PCR had a sensitivity of 70.1%, thus facilitating the diagnosis and species identification. Parasite culture alone detected 39.2% and smear alone 55.3% of the positive samples. With the exception of kDNA PCR, all other assays were 100% specific. CONCLUSIONS This study provides the first findings for the comprehensive molecular characterization of CL in Saudi Arabia.

Molecular copro-prevalence of Cryptosporidium in Egyptian children and evaluation of three diagnostic methods

ObjectiveTo determine molecular prevalence of Cryptosporidium in a cohort of Egyptian children and compare three diagnostic tests.MethodsStool samples from children with diarrhea (n=150) and from apparently healthy children (n=100) were examined for Cryptosporidium using microscopy, enzyme linked immuosorbant assay (ELISA) and nested polymerase chain reaction (nPCR).ResultsnPCR detected Cryptosporidium in 22.4% of children. Acid-fast stain and ELISA showed false negativity but 100% specificity with nPCR as gold standard.ConclusionCryptosporidium is a common cause of diarrhea in children in Egypt.

Molecular characterization of Leishmania infection in sand flies from Al-madinah Al-munawarah province, western Saudi Arabia

Cutaneous leishmaniasis (CL) is caused by various species of the genus Leishmania. The disease is considered a major health problem in different areas of Saudi Arabia including Al-madinah Al-munawarah province. We aimed to identify Leishmania species isolated from sand fly vectors by molecular analysis. Sand fly sampling was carried out from May 2010 to October 2010 in province of Al-madinah Al-munawarah from four different localities. Female sand flies collected were subjected to DNA extraction followed by molecular analysis using the semi-nested PCR and conventional PCR protocols, respectively, against minicircle kDNA and ribosomal internal transcribed spacer 1 (ITS1-rDNA). The PCR positive specimens against ITS1-rDNA locus were digested for further confirmation of species identification. A total of 2910 sand flies were collected. Phlebotomus papatasi accounted for 93.8% (1673 males and 1057 females), however, the number of Phlebotomus sergenti was only 180 (109 males and 71 females). Sixty-two out of 250 (23.7%) female P. papatasi tested for Leishmania parasite were positive for Leishmania major using the semi-nested PCR method against kDNA. All of the 62 positive specimens produced a band size 650 bp. A 31% of female P. sergenti were positive against kDNA of Leishmania tropica and produced a 720 bp band. These positive P. sergenti for L. tropica DNA produced ITS1-PCR-RFLP profile showed two bands of ∼200 bp and 57 bp which are specific for L. tropica, confirming the presence of L. tropica in P. sergenti. However, the ITS1-PCR-RFLP profile showed two bands of ∼203 bp and 132 bp which are specific for L. major in P. papatasi. We concluded that, the semi-nested PCR method against kDNA and the ITS1-PCR-RFLP analysis are useful tools for molecular identification of both L. major and L. tropica. A multicenter study is necessary in order to evaluate the extent of the disease and functional analysis of new Leishmania genes.

Serum Malondialdehyde Levels as a Biomarker of Cellular Injury In Human Fascioliasis

Abstract Macrophages, neutrophils and other phagocytic cells are key components of the antiparasitic, antimicrobial and tumoricidal immune responses. These cells are capable of generating large amounts of reactive oxygen species (ROS) and reactive nitrogen species (RNS). ROS and RNS have a possible role in the pathogenesis of parasitic disease. Lipid peroxidation is a well-established mechanism of cellular injury and is used as an indicator of oxidative stress in cells and tissues. To examine oxidant status and lipid peroxidation in fascioliasis patients, the malondialdehyde (MDA) (an end-product of lipid peroxidation) has been studied. Serum MDA level was measured in 34 patients infected with Fasciola gigantica and their age and gender were matched to 36 healthy controls. The difference between MDA levels of patients infected with Fasciola gigantica and the control group was statistically significant both in females ( P P F. gigantica infection.

Infectious Nosocomial Diarrhea in the Surgical Wards: Role of Parasites and Microbes Imply Stool Analysis

Abstract Objective This is the first study to document the etiology of non-outbreak infectious nosocomial diarrhea in surgical wards, in Al-Madinah Al-Munawarh hospitals, Saudi Arabia. Methods Fecal samples were collected from all hospitalized patients in surgical wards having diarrheic stool 72 hours after hospitalization with no co-morbid diarrhea or intestinal disturbances on hospitalization during the period from September, 2007 to March, 2008. They were analyzed by copro-scopical, fecal culture, and copro-ELISA methods. Results Enteropathogens were revealed in 51.9% of cases; about third of them had combined infections. C.difficile toxin AB Cryptosporidium parvum (6.6%), B.hominis (6.6%), G.lamblia (3.5%) and E.histolytica (3.1%). Heavy yeast colonization was isolated from 3.5% of cases and Rotavirus coproantigens in 1.2%. Conclusion Stool specimen analysis 72 h after admission would be of value for parasites and microbes in hospitalized patients. Patients who had prolonged duration of hospitalization, complicated surgical procedures, pre-existing co-morbidity, taking medications especially antibiotics, or at extremes of ages were high risk groups. The numbers of fecal specimens examined, fecal concentration, and fecal ELISA, increase the recovery rate of stool examination. In addition, the morbidity was greater than reported, which implicate the request for stool examination and the role of parasites and microbes as an etiology of nosocomial diarrhea in surgical wards.

Molecular prevalence and estimated risk of cutaneous leishmaniasis in Libya.

BACKGROUND/PURPOSE Cutanoeus leishmaniasis (CL) is an endemic disease in the Mediterranean area including Libya. The aim of the present study is to detect the prevalent Leishmania species obtained from smeared cutaneous lesions in addition to studying the diverse sociodemographic risk factors of the reported cases from different provinces of Libya. METHODS A total of 250 archived microscopic slides from clinically suspected cases of CL attending the leishmaniasis clinic in the Dermatology Department, Tripoli Central Hospital, Tripoli, Libya, were microscopically examined. Leishmania-DNA was amplified using combined polymerase chain reaction (PCR) targeting kinetoplast-DNA (kDNA) and ribosomal internal transcribed spacer 1 (ITS1)-DNA with restriction fragment length polymorphism analysis for direct Leishmania species identification. RESULTS Using kDNA and ITS1-PCR, 22.5% and 20% of cases were positive, respectively. Only 14.4% of cases were positive using microscopy. Nominating ITS1-PCR as the reference standard, kDNA-PCR assay was highly sensitive while microscopy was 100% specific but of limited sensitivity (72%) with a substantial agreement and an overall accuracy of 94.4%. Leishmania major and Leishmania tropica were the predominant species reported from the north-western provinces including Tripoli, Zintan, and Gharyan with their related subprovinces; Asabaa, Mizdan, Alkawasem, and Alorban. CL prevailed more among men and residents of rural areas. House wives and students were the most affected professions. Children were the least affected, while the middle-aged were the most affected age group. CONCLUSION L. major and L. tropica are the predominant species in the north-western regions of Libya. ITS1-PCR-restriction fragment length polymorphism assay offered a sensitive, specific, and faster diagnostic method especially with negative parasitologic examination.

Blastocystis subtypes isolated from irritable bowel syndrome patients and co-infection with Helicobacter pylori

Irritable bowel syndrome (IBS) is a chronic functional gastrointestinal disease presenting clinically by abdominal pain with alteration of bowel habits. Although IBS has uncertain etiology, chronic gut inflammation due to persistent exposure to an infectious agent including Blastocystis sp. was proposed. The aim of this study was to detect the prevalence of Blastocystis sp. subtype (ST) isolated from stool of IBS patients and to assess Blastocystis sp. and H. pylori co-infection in IBS patients from Beni-Suef Governorate, Egypt. Stool samples were collected from 115 IBS patients, following Rome III criteria. All stool samples were microscopically examined by wet mount and permanent trichrome stain, cultured on Jones’ medium with further sequencing of positive Blastocystis isolates and screened for detection of H. pylori coproantigen. Blastocystis sp. was the predominant parasite in IBS patients; it had statistical significant association with both rural residence (OR = 10) and flatulence (OR = 8.2). There was a predominance of Blastocystis sp. ST3 (81%) followed by ST1 (19%). Blastocystis culture results (19.1%) were superior than microscopy (16.5%). The majority of Blastocystis-positive IBS patients (72.7%) were co-infected with H. pylori with statistical significance; however, H. pylori was higher in Blastocystis-negative IBS patients (47/64) than in Blastocystis-positive IBS patients (17/64). Interestingly, IBS is usually associated with gut dysbiosis, while the most prevalent parasite in our IBS patients was Blastocystis sp., which is frequently found in asymptomatic individuals. Whether Blastocystis sp. is a cause or a consequence of IBS still needs further investigation, with a particular focus on correlation of IBS with different Blastocystis sp. subtypes and gut microbiomes.

Wuchereria bancrofti microfilariae and quantitative circulating antigen detection in selected endemic areas in Egypt

Back ground and objective Wuchereria bancrofti is responsible for 90% of cases of lymphatic filariasis throughout the tropics and in some subtropical areas worldwide, including Egypt. To combat this disease, the WHO has launched a program aiming to eliminate lymphatic filariasis by the year 2020 in all the endemic countries using mass drug administration (MDA) to interrupt the disease′s transmission. The aim of the present work was to study W. bancrofti infection in selected endemic areas in Egypt by performing parasitological examination and enzyme-linked immunosorbent assay (ELISA) antigen detection test, and to analyze the demographic, clinical, and MDA data of the study population in relation to W. bancrofti infection. Patients and methods A total of 300 blood samples were collected from residents in endemic areas in five governorates. Parasitological examination and Og4C3 ELISA test were performed to identify W. bancrofti infection. Results Microfilariae were identified in one individual while circulating filarial antigens (CFAs) were detected in 10 individuals. Statistical analysis of the collected data showed that CFAs were significantly higher in the male population than in the female population, whereas analysis regarding other demographic, clinical, and MDA data showed no statistical significance. Conclusion The study results showed that CFAs are still detected in endemic communities in Egypt, and that the prevalence is higher in the male population than in the female population. Although the Og4C3 ELISA test is a useful research tool for the study of W. bancrofti infections, its cost and format hinder its wide use in endemic areas.

Molecular detection of Capillaria philippinensis: An emerging zoonosis in Egypt

Human infection with Capillaria philippinensis is accidental; however, it may end fatally if not diagnosed and treated in the proper time. The first case was detected in the Philippines in 1963, but later reported in other countries around the world, including Egypt. In this report, molecular diagnosis using a specific nested PCR for detection of C. philippinensis in faeces is described based on the amplification of small ribosomal subunit. The test showed sensitivity and specificity, as it detected all the positive cases and gave no cross-reaction with human DNA and DNA of other tested parasites. This method can be very useful not only for improvement of diagnosis, but also to understand the different environmental routes of transmission by detection of C. philippinensis DNA-stages in the possible fish intermediate hosts and reservoir animal host, helping to improve strategies for surveillance and prevention of human disease.

First identification of Naegleria species and Vahlkampfia ciguana in Nile water, Cairo, Egypt: Seasonal morphology and phylogenetic analysis

BACKGROUND/PURPOSE In Egypt, there is a scarcity of data concerning Naegleria (N.) family, with a shortage of phylogenetic studies. This studys aim was molecular detection, sequencing and phylogenetic analysis of morphologically identified Nagleria and to determine natural seasonal distribution of Nagleria species in water sources of Greater Cairo, Egypt. METHODS A total of 120 water samples were collected during each season over a year. Every water sample was filtrated and cultured on non-nutrient agar (NNA). Morphologically positive Nagleria-like isolates were subjected to Nagleria genus and species-specific PCR targeting rDNA gene, PCR products were sequenced and obtained sequences were phylogenetic analyzed. RESULTS Nile River water was the only source found to contained Naegleria. For the first time in Egypt, Vahlkampfia ciguana and the Naegleria species N.australiensis, N.philippinensis and N.neojejuensis were identified from the Nile water. The pathogenic Naegleria fowleri, previously reported in Egypt, was however not detected in this study. CONCLUSION Interestingly, there were no seasonal variations in prevalence of Naegleria spp.; yet, there was seasonal diversity in the water samples of the same site. These newly discovered Vahlkampfiidae in Egyptian aquatic environments indicate the need for further phylogenetic investigations using bigger sample sizes in order to determine their potential risk for human health.