Giardia intestinalis assemblages among Egyptian symptomatic/asymptomatic cases in Cairo

Giardia intestinalis is frequent enteric protozoa, affecting humans worldwide. Human infections aremainly caused by two genetically different assemblages call A & B. This cross-sectional study characterizedthe Giardia genotypes isolated from the stool of symptomatic and asymptomatic Egyptians inCairo and correlated these genotypes with the demographic and clinical data of the cases. Stool sampleswere collected from 389 individuals (245 complaining of gastrointestinal (GIT) symptoms and144 apparently healthy asymptomatic individuals), and microscopically examined. Positive Samplesfor G. intestinalis were molecularly characterized by Copro-nPCR targeting beta-giardin gene, andthen analyzed by RFLP for assemblage identification. Giardia was detected in 62 samples (15.9%); 53samples in symptomatic cases (21.6%) and 9 samples in asymptomatic individuals (6.25%). DNA ofpositive samples was amplified by nPCR-RFLP assays. There was a significant predominance of assemblageB among symptomatic (82.7%) and asymptomatic (77.8%) groups, while the rest of sampleshad assemblage A. Among the clinical data, only flatulence was significantly associated with Giardiainfection with assemblage B. Assemblage B is the predominant genotype found in Egypt in symptomaticand asymptomatic patients suggesting an anthropologic transmission cycle.Key Words: Giardia; genotyping; Assemblage; nPCR-RFLP; beta-giardin

Stool Microbiota Diversity Analysis of Blastocystis-Positive and Blastocystis-Negative Individuals

Blastocystis is a unicellular eukaryote found in the gastrointestinal tract of both human and other animal hosts. The clinical significance of colonic Blastocystis colonization remains obscure. In this study, we used metabarcoding and bioinformatics analyses to identify differences in stool microbiota diversity between Blastocystis-positive and Blastocystis-negative individuals (n = 1285). Alpha diversity was significantly higher in Blastocystis carriers. At phylum level, Firmicutes and Bacteroidetes were enriched in carriers, while Proteobacteria were enriched in non-carriers. The genera Prevotella, Faecalibacterium, Flavonifracter, Clostridium, Succinivibrio, and Oscillibacter were enriched in carriers, whereas Escherichia, Bacteroides, Klebsiella, and Pseudomonas were enriched in non-carriers. No difference in beta diversity was observed. Individuals with Blastocystis-positive stools appear to have gut microbiomes associated with eubiosis unlike those with Blastocystis-negative stools, whose gut microbiomes are similar to those associated with dysbiosis. The role of Blastocystis as an indicator organism and potential modulator of the gut microbiota warrants further scrutiny.publishedVersio

Ginger (Zingiber Officinale)-derived nanoparticles in Schistosoma mansoni infected mice: Hepatoprotective and enhancer of etiological treatment.

BackgroundNanotechnology has been manufactured from medicinal plants to develop safe, and effective antischistosmal alternatives to replace today's therapies. The aim of the study is to evaluate the prophylactic effect of ginger-derived nanoparticles (GNPs), and the therapeutic effect of ginger aqueous extract, and GNPs on Schistosoma mansoni (S. mansoni) infected mice compared to praziquantel (PZQ), and mefloquine (MFQ).Methodology/principal findingsEighty four mice, divided into nine different groups, were sacrificed at 6th, 8th, and 10th week post-infection (PI), with assessment of parasitological, histopathological, and oxidative stress parameters, and scanning the worms by electron microscope. As a prophylactic drug, GNPs showed slight reduction in worm burden, egg density, and granuloma size and number. As a therapeutic drug, GNPs significantly reduced worm burden (59.9%), tissue egg load (64.9%), granuloma size, and number at 10th week PI, and altered adult worm tegumental architecture, added to antioxidant effect. Interestingly, combination of GNPs with PZQ or MFQ gave almost similar or sometimes better curative effects as obtained with each drug separately. The highest therapeutic effect was obtained when ½ dose GNPs combined with ½ dose MFQ which achieved 100% reduction in both the total worm burden, and ova tissue density as early as the 6th week PI, with absence of detected eggs or tissue granuloma, and preservation of liver architecture.Conclusions/significanceGNPs have a schistosomicidal, antioxidant, and hepatoprotective role. GNPs have a strong synergistic effect when combined with etiological treatments (PZQ or MFQ), and significantly reduced therapeutic doses by 50%, which may mitigate side effects and resistance to etiological drugs, a hypothesis requiring further research. We recommend extending this study to humans

Mapping gut parasitism patterns in a cohort of Egyptians

Abstract In developing countries, the prevalence of intestinal parasitic infection is still significant, particularly due to geographical and socioeconomic variables. The objective of this study was to map the distribution pattern of intestinal parasitic infection in a cohort of the Egyptian population, as well as to assess associated risk factors. A cross-sectional hospital-based study was conducted on 386 patients. A single fecal specimen was collected from the study individual and examined microscopically for the detection of parasitic infection. DNA was extracted from all samples and utilized to amplify Entamoeba histolytica complex species, Cryptosporidium species, Giardia intestinalis assemblages, and Blastocystis species using PCRs. Typing of Cryptosporidium species and Giardia intestinalis assemblages was performed using restriction enzymes RasI and HaeIII respectively. While Blastocystis spp. subtypes (ST) were identified through sequencing of PCR products and phylogenetic analysis. 59.6% (230/386) of the study patients were infected with one or more intestinal parasites, 87.4%; 201/230 of patients had mono-parasitic infections, and 12.6%; 29/230 had multiple-parasitic infections (P < 0.0001). The predominant protozoa were Blastocystis, followed by Entamoeba histolytica complex, and Giardia intestinalis both as mono-parasites and as part of multiple parasites. Molecular assays showed that Blastocystis ST3, Entamoeba dispar, Giardia intestinalis assemblage B, and Cryptosporidium hominis were the most prevalent species. Intestinal parasitic infection was significantly associated with age, gender, residence, and water source. Multi-parasitism showed that residency in a rural area was a risk factor (OR 4.49; 95% CI 1.51–13.37; P = 0.007). Egyptians residing in rural areas have a high prevalence of intestinal multi-parasitism. Therefore, to lessen the prevalence and effects of these infections in this group, effective and sustainable control methods, providing health education focusing on good personal hygiene habits, and providing a safe drinking water supply should be implemented

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. Keywords: Archived microscopic slides, Cutaneous leishmaniasis, ITS1-PCR, kDNA-PC

Blastocystis SPP. infection prevalence and associated patient characteristics as predictors among a cohort of symptomatic and asymptomatic Egyptians

Blastocystis species (spp.) is a large unicellular intestinal protozoan parasite that has a worldwide distribution. It has unclear pathogenicity and is linked to many clinical disorders. The study purpose was to determine the prevalence of Blastocystis spp. molecularly in a cohort of symptomatic and asymptomatic individuals and to assess the association of Blastocystis spp. with the patient characteristics as possible predictors of the occurrence of Blastocystis spp. Fecal specimens were collected from 139 Egyptians of both sexes, aged from 5 months to 74 years. All fecal specimens were examined coproscopically for detection of gut parasites and cultured on Modified Jones’ medium for detection of Blastocystis spp. Molecular assay using nested PCR (nPCR) was performed for cultured Blastocystis. The association between detection of Blastocystis spp. and patient demographics and clinical data was determined. Prevalence of parasitic infections was 62 (44.6%) using coproscopy. Blastocystis spp. was the most prevalent parasite (21.6%) in both symptomatic and asymptomatic individuals, followed by E. histolytica complex (13.7%) and Giardia intestinalis (10.8%). Cryptosporidium spp. (2.2%) and E. coli (2.1%) were the least detected parasites. Among studied patient characteristics, only age showed statistical significance in association with detection of Blastocysitis spp.

Molecular prevalence of Entamoeba species in a cohort of Egyptians: Entamoeba dispar predominance

Background and objective: Intestinal and extraintestinal amebiasis is caused by the protozoan parasite Entamoeba (E.) histolytica, it is of considerable morbidity and mortality in developing countries. E.histolytica complex species includes E.histolytica, E. moshkovskii and E. dispar are morphologically indistinctable. The current study goal was to use molecular assays to detect the true prevalence of E. histolytica complex species among a cohort of Egyptians. Methods:  A single stool specimen was collected from 133 patients, examined coproscopically before and after concentration. DNA was extracted from microscopically positive stool specimens for E. histolytica complex species were molecularly idenitified using multiplex PCR. Results: The  coroscopic prevalence of intestinal parasites was 51.1% (68/133) of them 30 cases had E.histolytica complex (22.6%; 30/133). Among coproscopically positive samples, E. dispar was the most common parasite (63.3%;  19/30),  followed by  E.  histolytica  (23.4%;  7/30) and E. moshkovskii  (13.3%;  4/30). There was statistical significance association between sociodemographic characteristics and Entamoeba species in asymptomatic individuals, while in the symptomatic individual, only age groups were stastically significant. Conclusion: E.dispar is the predominant Entamoeba species among studied individuals. There is a need for molecular diagnosis of Entamoeba to determine the true prevalence of E. histolytica and avoid overmedication

Molecular identification of Leishmania major species in phlebotomine sand flies from Al Ahsa, Eastern KSA

الملخص: أهداف البحث: يعتبر داء الليشمانيات الجلدي من المخاطر التي تهدد الصحة العامة في المملكة العربية السعودية. العديد من المناطق السعودية مستوطنة للغاية بما في ذلك مدينة الأحساء، شرق المملكة العربية السعودية. لا يوجد توثيق جزيئي سابق لطفيل الليشمانيا في النواقل في الأحساء. هدفت هذه الدراسة إلى التعرف الجزيئي على أنواع الليشمانيا من ذباب الرمل الذي تم جمعه من الأحساء. طريقة البحث: أجريت الدراسة المقطعية الحالية في الأحساء من يوليو 2020 إلى مايو 2021. تم جمع ذبابة الرمل من 3 مناطق بها أعلى نسبة إصابة بداء الليشمانيات الجلدي وذباب الرمل وفقا لبيانات مركز الوقاية من الأمراض المنقولة بالنواقل في الأحساء. تم استخدام مصيدة الضوء المصغرة سي دي سي والمصائد اللاصقة لجمع ذباب الرمل. تم فرز إناث ذباب الرمل الفليبوتومين وفصلها لمزيد من التحليل. تم استخدام أحواض من 3-10 إناث من ذباب الرمل لاستخراج الحمض النووي الجيني. تم تضخيم الحمض النووي المستخرج باستخدام بروتوكول يستهدف جنس الليشمانيا والبادئات الخاصة بالأنواع. بالنسبة لانتواع الليشمانيا، تم استخدام طريقة تعدد الأشكال بعد تفاعل البوليميراز المتسلسل. النتائج: تم تضمين 113 بركة من ذباب الرمل في الدراسة. من بين 113 مجموعة من العينات، كانت 10 منها إيجابية لجنس الليشمانيا بعد بروتوكولنا التجريبي. أدى توصيف أنواع الليشمانيا عن طريق تعدد الأشكال في تفاعل البوليميراز المتسلسل، وتعدد الأشكال، إلى جعل الليشمانيا الرئيسية هي النوع الوحيد الموجود في ذباب الرمل الذي تم جمعه. الاستنتاجات: هذا هو أول توثيق جزيئي لأنواع الليشمانيا في ذباب الرمل الفليبوتومين في منطقة الأحساء. كانت الليشمانيا الكبرى هي النوع الوحيد الموجود في دراستنا. مزيد من البحث الشامل حول الناقل والخزانات أمر حيوي لإنشاء ديناميكيات انتقال الليشمانيا في الأحساء. تساعد هذه المعلومات في التخطيط الاستراتيجي لأساليب الوقاية والسيطرة على أنواع الليشمانيا في المنطقة. Abstract: Objectives: Cutaneous leishmaniasis (CL) is considered an overlooked public health threat in KSA. CL is endemic to several Saudi regions, including Al Ahsa City, Eastern KSA. To our knowledge, no prior molecular identification of Leishmania parasites in vectors in Al Ahsa has been published. The aim of this study was to perform molecular identification of Leishmania species in sand flies collected from Al Ahsa. Methods: This cross-sectional study was conducted in Al Ahsa from July 2020 to May 2021. Sand flies were collected from the three areas with the highest rates of CL cases and sand flies, according to data from the Vector Borne Diseases Prevention Center in Al Ahsa. CDC miniature light traps and sticky traps were used to collect sand flies. Pools of 3–10 female sand flies were subjected to genomic DNA extraction. The extracted DNA was then amplified with a protocol targeting the Leishmania genus and using species-specific primers. For Leishmania species identification, a PCR-restriction fragment length polymorphism (PCR-RFLP) method was used. Results: Ten of 113 pools of samples tested positive for the Leishmania genus, according to our experimental protocol. Characterization of Leishmania species by PCR-RFLP established Leishmania major as the only species found in the collected sand flies. Conclusion: This is the first documentation of molecular identification of Leishmania species in phlebotomine sand flies in the Al Ahsa region. L. major was the only species identified in our study. Further comprehensive research investigating the vectors and reservoirs will be crucial to establish the dynamics of transmission of Leishmania in Al Ahsa