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

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.

Mapping gut parasitism patterns in a cohort of Egyptians.

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.

Diagnostic yield of stool culture and probable predictive factors.

BACKGROUND AND AIM: Stool analysis is commonly performed to diagnose certain gastrointestinal diseases. The diagnostic yield of stool culture,  a method of stool analysis, is variable worldwide and is unclear in the Kingdom of Saudi Arabia. This study was conducted to determine the diagnostic yield of stool culture from the year 2008 to 2020 and to determine the predictors for a positive stool culture. Furthermore, antibiotic susceptibility patterns of the detected copro-pathogens in the same time period were collected and studied. METHODS: This is a retrospective case-control study in which patients' data was collected from the hospital's electronic health record. The results of all stool analyses performed from 2008 to 2020 and associated patients' characteristics were collected.  Characteristics of cases with a positive stool culture were compared to the characteristics of those without to identify the predictors for positive stool cultures. RESULTS: Copro-pathogens were detected in 89.4% of cultured stool samples. Salmonella spp (1590/1775, 89.6%) was the most common organism followed by Shigella spp. (84/1775, 4.7%) and Campylobacter spp (45/1775, 2.5%). Male sex, the 1-5 age group, positive fecal occult blood test results, and positive stool leukocyte test results were associated with a positive stool culture result. Cultured copro-pathogens were highly sensitive to Trimethoprim/Sulfamethoxazole and Ampicillin. CONCLUSIONS: Stool analysis was found to be a test of high diagnostic yield. However, there is still a need for more studies on this subject with a focus on possible predictive factors for specific organisms.

Genotypic and phylogenic analyses of cutaneous leishmaniasis in Al Ahsa, Eastern Saudi Arabia during the coronavirus disease 2019 pandemic: First cases of Leishmania tropica with the predominance of Leishmania major.

During the coronavirus disease 2019 lockdown period, a surge in sandflies and cutaneous leishmaniasis (CL) cases was observed in Al-Ahsa, Saudi Arabia. Skin punch biopsies were obtained from 100 patients clinically diagnosed with CL in Al-Ahsa who had no travel history in the last 6 months. Impression smears were used following a three-step polymerase chain reaction (PCR) protocol using genus-specific primers targeting kDNA and ITS1. Leishmania speciation was determined by ITS1 PCR/nested PCR-restriction fragment length polymorphism and sequencing. A phylogenetic tree was constructed. The associated patient characteristics were analyzed. Using internal transcribed spacer one (ITS1)-PCR/nested PCR, 98 cases were considered true-positive CL. Leishmania major was the predominant species, and Leishmania tropica was identified in three cases. Microscopy had poor sensitivity and perfect specificity. Direct ITS1-PCR missed nine cases. Sex, residence, and treatment outcome were significantly associated with the occurrence of Leishmania; distribution of skin lesion(s) and treatment outcome were significantly associated with Leishmania genotype. This is the first time that L. tropica was identified as a cause of CL in human in Al-Ahsa, in addition to the predominant zoonotic species, L. major. We recommend using ITS1-nested PCR for negative cases by ITS1-PCR. Further exploration of Leishmania transmission dynamics in vectors and reservoir animals is essential for designing effective preventive measures.

Anti-blastocystosis activity of antioxidant coated ZIF-8 combined with mesoporous silicas MCM-41 and KIT-6.

The biocompatible hybrid Zeolitic imidazolate framework-8 (ZIF-8)/structured silica nanocomposite can be loaded with antioxidants such as curcumin and resveratrol to offer multiple advantages of drug functionalization and structural stability. blastocystosis, an enteric parasite, has various outcomes and its treatment includes drugs which have side effects and do not result in a full cure. We aimed to design novel biocompatible nanocomposites containing natural antioxidant, resveratrol or curcumin and ZIF-8/mesoporous silica. We also assessed their anti-blastocystosis activities as bioactive novel nanocomposites. The nano-silica (MCM-41 and KIT-6) was synthesized using a hydrothermal technique and made composite with ZIF-8 using an ultrasonic technique. The antioxidants, curcumin and resveratrol, were loaded over ZIF-8/MCM-41 and ZIF-8/KIT-6 using a rotary evaporator technique to form novel nanocomposites with bioactive properties. The formulated nanocomposites were characterized. To test their biological activity, suspension of cultured blastocystosis cysts (subtype 3) were exposed to increasing concentrations of nanocomposites and the minimal lethal concentration of each nanocomposite was calculated. The bioactive nanocomposites (ZIF-8/KIT-6, ZIF-8/KIT-6/Resveratrol and ZIF-8/MCM-41/Curcumin) were formulated. Anti-blastocystosis activity of the tested nanocomposites was both dose and time dependent. ZIF-8/KIT-6/Resveratol showed the maximum percentage of growth inhibition (~ 100%) at a concentration of 500 µg/ml after 5 h of exposure. More than 90% of blastocystosis cysts' growth was significantly inhibited at all concentrations of ZIF-8/MCM-41/Curcumin, with different times of exposure, while it occurred only at the highest concentration of ZIF-8/KIT-6 (800 µg/ml). Using cheap, simple, reproducible and scalable techniques, we nano-formulated innovative bioactive nanocomposites, by incorporating the bioactive ZIF-8 (Zn2+ with imidazole), structured mesosilica and natural antioxidant compounds, curcumin or resveratrol, to generate multifunctional modalities. These eco-friendly, naturally based, safe, economical, biocompatible, and bioavailable nanocomposites are potential nanotherapeutics. The anti-blastocystosis results of these three nanocomposites indicate their potentially promising innovative and safe use as alternative Blastocystosis therapies.

Susceptibility to Rubella Infection and Incidence of Congenital Rubella Infection: 6 Years Retrospective Study.

Introduction: Rubella is considered one of the most serious and most common congenital infections. Despite global efforts for elimination, rubella cases are still being reported in many parts of the world. The purpose of this study is to determine the level of immunity to rubella in the community and most importantly among women at childbearing age in the eastern province of Saudi Arabia and compare it with the target set by the World Health Organization (WHO) along with the incidence of acute rubella infection and the associated congenital rubella infection and congenital rubella syndrome. Methods: This is a retrospective cross-sectional study over the six years period (Jan 2014-Jun 2020) on all individuals tested for rubella IgM and IgG in a university teaching hospital. Results: Nighty one percent (15,894/17,469) of the population tested showed evidence of rubella immunity with 8.8% (1546/17,469) being susceptible. Among women at childbearing age, susceptibility to rubella was higher with 9.2% (1220/13,278) of women showing no evidence of immunity. In addition, acute rubella infection was reported for 0.17% (29/17,469) of the population tested and 0.15% (20/13,278) in women at childbearing age. No cases of congenital rubella infection were reported in the study period. Discussion: The level of Rubella immunity in the population is 91% and is less than the WHO target for rubella control therefore, risk of resurge of cases is present, indicating the need for continued national surveillance and more efforts to improve vaccination coverage in the kingdom.

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.

A decade of travel-associated malaria at King Fahad Hospital of the University in the Eastern Province of Saudi Arabia.

Travel-associated malaria is a health hazard, even in non-malaria endemic regions. This is a hospital-based retrospective study of 12,931 febrile patients who presented at King Fahad Hospital of the University (KFHU) from January 2009 to December 2019. Patients either returning from malaria endemic countries and/or for whom malaria was suspected, had blood films microscopically screened for malaria parasites. Malaria prevalence was very low in febrile patients attending KFHU. Out of the 12,931 febrile patients, 0.63% (n = 81) were malaria positive, all travel-related, except for one case of transfusion malaria. Indian nationals were the most infected (29.6%, n = 24), followed by Sudanese nationals (24.7%, n = 20). P. falciparum (47%, n = 38) and P. vivax (42%, n = 24) were the predominant species. The majority of P. falciparum (64.5%, n = 20) cases were from African nationals and the majority of P. vivax (72.7%, n = 24) cases were from Asia. The highest percentage of malaria patients were adult (90%, n = 73), males (85.2%, n = 69), ages ranged from 6 to 65, with a mean of 34.6 years. Most of the malaria cases presented at the emergency room (ER), only 3 required critical care. Only sex, hospitalized in-patient (IP) and attendance at ER were statistically associated with malaria. In the presence of a potential vector, travel-associated malaria in non-malaria endemic areas should be monitored to guide control strategies.Author summary: Malaria is a neglected potentially fatal tropical mosquito-born disease. Travel-associated malaria is a health hazard, even in non-malaria endemic regions. In spite of previous efforts to estimate malaria prevalence, morbidity and mortality in Saudi Arabia in the last decade, there have been no studies that determine the prevalence of malaria in Al-Khobar, Eastern Province of Saudi Arabia. Malaria prevalence was very low in febrile patients (81/12,931) attending King Fahad Hospital of the University over a decade. Cases were all travel-related, except for one case of transfusion malaria. Indian nationals were the most infected (29.6%), followed by Sudanese nationals (24.7%). P. falciparum (47%) and P. vivax (42%) were the predominant species. The majority of P. falciparum (64.5%) cases were from Africa and the majority of P. vivax (72.7%) cases were from Asia. No patient factors predicted malaria in febrile travelers. In non-malaria endemic areas, in the presence of a potential vector, patients with acute fever coming from endemic areas or having received blood transfusion, should be screened for travel-associated malaria to guide control strategies.

An Association Between Blastocystis Subtypes and Colorectal Cancer Patients: A Significant Different Profile from Non-cancer Individuals.

PURPOSE: Blastocystis is a common enteric human parasite of non-conclusive pathogenicity which may be determined by subtype (ST) variation. Colorectal cancer (CRC) is considered one of the primary causes of cancer mortality. Blastocystis ST7 has been shown to reduce beneficial intestinal microbiota and may exacerbate CRC. This study assessed the possible association between Blastocystis STs and CRC in comparison to non-cancer patients. MATERIAL AND METHODS: A total of 200 fecal samples were obtained from CRC (100) and non-CRC (100) individuals attending Beni-Suef University Hospital, Egypt. Blastocystis was searched for in all samples using microscopy and culturing. Positive subculture samples were genetically sequenced and subtyped using conventional polymerase chain reaction (PCR). Blastocystis STs were determined by sequencing and a phylogenetic tree was created. Related patient characteristics and tumor stages were analyzed for association with presence of Blastocystis. RESULTS: Blastocystis was identified in 52% and 42% of CRC and non-cancer individuals, respectively. ST1, 2, and 3 were isolated from both cancer and non-cancer individuals; however, for the first time, ST7 was only isolated from CRC stool samples with significant association. Associated patient characteristics were evaluated as predictors. CONCLUSION: Blastocystosis is highly prevalent in CRC patients, predominantly in the latest CRC grades and stages. To the best of our knowledge, this is the first study to report the identification of Blastocystis ST7 in CRC patients. To determine whether certain STs of Blastocystis are associated with CRC would require further research, including the role played by gut microbiota.

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

BACKGROUND: Nanotechnology 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 FINDINGS: Eighty 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/SIGNIFICANCE: GNPs 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.

Seropositivity of syphilis among individuals screened in a tertiary hospital in the Eastern Province of Saudi Arabia.

BACKGROUND: Sexually transmitted infections are a serious public health problem. Syphilis, a multistage, curable chronic disease caused by the spirochete Treponema pallidum, remains a major health problem worldwide. The disease re-emerged in the era of HIV in many countries despite the accessibility of curative therapy and continuing public health efforts to eliminate it. OBJECTIVE: Analyse the seropositivity for syphilis. DESIGN: Retrospective cross-sectional. SETTING: Tertiary hospital. PATIENTS AND METHODS: We retrospectively studied individuals who underwent screening tests for syphilis between January 2014 and December 2018. The samples that were positive by both screening and confirmatory tests were considered as confirmed positive for syphilis. MAIN OUTCOME MEASURES: Syphilis positivity identified by chemiluminescence immunoassay, the rapid plasma reagin test, and specific antibodies against Treponema pallidum. SAMPLE SIZE: 11 832. RESULTS: Of the 11 832, 54 (0.45%) were confirmed as seropositive for syphilis. Thirty-three (61.1%) were non-Saudi; 21 (38.9%) were Saudis. Thirty (55.6%) cases were males. Twenty-two (40.74%) were married and 29 (53.70%) were unmarried. Of the 54 diagnosed as syphilis positive, 28 (51.9%) were expatriate workers screened for pre-employment. The percentage of syphilis among Saudis was 0.36%. In an overall chi-square analysis, a P<.0001 indicated a difference among nationalities in the frequency of syphilis. A post-hoc analysis showed that Somalians (P=.004) and Sudanese (P=.005) differed significantly from other nationalities. CONCLUSION: The study showed that syphilis was low among the screened population. More than half of the syphilis positive cases in this study were household employees. Screening for syphilis assists in planning complementary services for target populations and improves syphilis control. LIMITATIONS: Retrospective design. Hospital-based findings may not be representative of the seroprevalence of syphilis in the general population. CONFLICT OF INTEREST: None.

Evaluation of Praziquantel Effectivenss After Decades of Prolonged Use in an Endemic Area in Egypt.

PURPOSE: Praziquantel (PZQ) is the preferred drug for schistomiasis treatment because of its safety. As PZQ is used for mass drug administration (MDA) in schistosomiasis endemic areas, the effectiveness of the drug, used solely for decades, should be continuously monitored to detect drug resistance. We aimed to assess the effectiveness of PZQ to cure Schistosoma mansoni infection and to reduce the intensity of infection in an endemic area by estimating the cure rate (CR), egg reduction rate (ERR), and comparing these estimates to the levels recommended by the World Health Organization (WHO). METHODS: A total of 342 children aged 5-15 years living in Kafr-El-Sheikh were screened for S. mansoni infection. Stool samples were examined microscopically using Kato-Katz (KK) technique. Among the screened children, 106 children had S. mansoni ova in stool, 100 of them received the first dose of PZQ (40 mg/kg). Four weeks later, 96 of 100 children received the second dose of PZQ. Stool samples, collected 4 weeks after each dose of PZQ, were examined using KK. The effectiveness of PZQ was assessed based on ERR and CR. RESULTS: CR after the first dose of PZQ was 66.7%, increased to 79.12% after second dose (X2 = 3.05, P = 0.08). Median egg count before treatment was 30.00 (6.00-744), that significantly decreased after two doses of PZQ to 0.00 (0.00-221.33) (Z = 8.29, P = 0.001). Children aged 10-15 years showed higher CR (91.3%) than those aged 5-9 years (OR = 5.25, CI 1.58-17.40). CONCLUSIONS: PZQ is still an effective agent against S. mansoni in endemic areas, achieving a high CR and ERR with predominantly low intensity of infection. Age is a main predictor of response to PZQ.

Giardiasis in symptomatic children from Sharkia, Egypt: genetic assemblages and associated risk factors.

Giardia intestinalis (G. intestinalis) is a common enteric protozoan parasite worldwide and in Egypt. Identification of true prevailing Giardia assemblages helps in identification of the sources of infection. The study's aim was to determine the true prevalence of Giardia assemblages in Egyptian children from Sharkia governorate presenting with gastrointestinal symptoms and to investigate their association with molecularly detected Giardia. A total of 617 stool specimens were collected from children presenting with gastrointestinal symptoms in Alquraeen, Sharkia governorate, Egypt for 17 months. All stool specimens were microscopically examined by wet mount smear before and after stool concentration to recover parasitic stages. Giardia copro-DNA was amplified from microscopically detected stool specimens using Copro-nPCR targeting the tpi gene for Giardia, followed by sequencing products of nPCR. The molecular prevalence of Giardia among symptomatic children was 9.88%, 83% of which were assemblage B and 17% were assemblages A. Giardia affected both sexes and all ages and was most prevalent in preschool children. Abdominal pain was the most common GIT symptom followed by diarrhoea. However, none of the patients' demographic variables (sex, age, weight and height) nor clinical symptoms showed significant association with molecular detection of Giardia. Giardia was common among symptomatic children from Sharkia, Egypt, with the predominance of assemblage B, which suggests the possibility of sharing common transmission source and route. Giardia had age, sex and clinical symptom distributions without statistical significance. The results necessitate further genomic studies targeting multiple gene targets for a better understanding of the ecology, dynamics of transmission, pathogenicity and clinical impact of Giardia infection, to improve its management and strategic control.

Risk-free point-of-care visceral leishmaniasis diagnostics: combining buffy coat microscopy and immunoassay in tertiary rural hospitals in Sudan.

Visceral Leishmaniasis (VL), a life-threating disease in Sudan and Eastern Africa, is usually diagnosed by a painful and invasive tissue aspirate microscopy. This study assessed the diagnostic effectiveness of buffy coat (BC) microscopy and the rK39 immunoassay test separately and combined as an easy non-invasive method applied to peripheral blood sample for field diagnosis of VL. 151 VL suspected patients were recruited from tertiary rural hospitals in Bazura, Gedaref state, from 2014-2015. All patients were tested for VL using rK39 ICT and microscopy smears from LN aspirate and BC in addition to PCR from BC as a reference standard test. Both BC and LN aspirate microscopy showed perfect specificity (100%) with false negative results, while the majority of true positives (81%) had a low-parasite burden. ICT showed almost perfect agreement but limited by its poor specificity. Each of these three tests is inadequate as a consistent single diagnostic tool and should be replaced by PCR in routine practice. Combining the results of risk-free BC microscopy and rk39 ICT, using peripheral blood sample, improved VL diagnosis with almost perfect agreement and 93.4% accuracy. Our findings indicate that combined BC microscopy and ICT are accurate, simple and easy point-of-care VL diagnostic tools in community and rural hospitals that can replace or reduce the use of invasive tissue aspirates microscopy, when PCR is unavailable. This is particularly of value in endemic rural areas, decreasing the delay in final diagnosis and preventing deaths caused by VL.

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 study's 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.

Pregnane glycoside from Huernia saudi-arabica as latent schistosomicidal mediator.

Development of a novel agent for control of schistosomiasis is a mandate. In-vitro anti-schistosomal activity of the aerial parts of Huernia saudi-arabica were examined. Chromatographic investigations of the ethanol extract (EE) were afforded three compounds. Pregnane glycoside (CI) 12-ß-p-hydroxy-benzoyl-20-O-acetyl-boucerin-3-O-ß-D-glucopyranosyl-(1→4)-ß-D-cymaropyranosyl-(1→4)-ß-D-cymaropyranoside, in addition to two flavonoids (CII) luteolin-4'-O-ß-D-neohesperidoside and (CIII)quercetin-3-rutinoside were recognized via spectral analysis. The schistosomicidal effects were evaluated using scanning electron microscope (SEM). In-vitro bioassays on the viability (mobility, morphological changes and mortality) of Schistosoma mansoni adults, cercariae, miracidia and eggs at different concentrations 2.5, 5, 12.5, 25 and 50 µg/ml of EE and 2.6, 5.2, 13, 26 and 52 µM of CI in incubation times 1,2,4,6,12hrs were carried out. EE and CI evidenced in-vitro anti-schistosomal activity with a dose and incubation time-dependent fashion. The effect of EE and CI was evident by the topography damage showed by SEM. EE proved moderate in-vitro cytotoxicity with IC50 of 8.48 µg/ml.

Intestinal parasitism in pediatric oncology children receiving chemotherapy: unexpected low prevalence.

BACKGROUND: Children with underlying malignancies and those on chemotherapy are at risk for having intestinal parasitic infections, which can lead to a severe course and death. This cross-sectional study was done to assess the copro-parasitological and copro-molecular prevalence of entero-parasites in children with malignancies and those on chemotherapy. PROCEDURE: Stool samples were collected from 137 Egyptian hospitalized cancerous children with different malignancies in the National Cancer Institute, and receiving chemotherapy.Faecal samples were examined microscopically. Genomic copro-DNA was extracted from fecal samples and amplified by 3 separate nPCR assays targeting Cryptosporidium, G. intestinalis and Entamoeba histolytica complex. RESULT: The overall prevalence of enteroparasites was 6.6 % (9 cases). Only Giardia copro-DNA was encountered in 2 (1.4%) faecal samples of patients. Coproscopy detected parasites in 7 cases: Blastocystis spp. in 5 cases (3.6%), Hymenolepis nana in 1 case (0.7%) and Ascaris lumbericoides in 1 case (0.7%). CONCLUSION: Low prevalence may be due to patient's use of prophylactic anti-parasitic and anti-fungal drugs, a standard protocol, basic hygienic practices and good nursing all of which are preventive against enteroparasites transmission. Among studied variables only diarrhoeic individuals who had a solid tumor, and soft/liquid stool with mucus and blood were predictors of intestinal parasitism.

Helicobacter pylori and enteric parasites co-infection among diarrheic and non-diarrheic Egyptian children: seasonality, estimated risks, and predictive factors.

Helicobacter pylori (H. pylori) and intestinal parasites are known for their high prevalence in children. Both of them infect the gastrointestinal tract with overlapping clinical pictures. This study was conducted to determine H. pylori prevalence and its association with intestinal parasites in children, moreover to estimate risk and predictive factors for their detection in stool samples. Single fecal samples were collected from 226 Egyptian pediatric patients (125 diarrheic and 101 non-diarrheic) attending gastroenterology outpatients' clinics, from February 2016 to June 2017. All stool specimens were microscopically examined to search for ova and parasites. Copro-DNAs detection of H. pylori and Cryptosporidium were performed using nested-PCR assays. H. pylori was detected molecularly in 36.8% of the total study population, with a higher prevalence in diarrheic than in non-diarrheic children. Intestinal parasites were detected in 27.4% of the total study populations, of these, 43.9% had co-existence with H. pylori colonized patients and was significantly associated with Cryptosporidium spp. and G. intestinalis. Estimated risk of the presence of H. pylori was in January. Our data provide a better understanding of the epidemiology of H. pylori infection when associated with intestinal parasites. H. pylori co-existence with G. intestinals and Cryptosporidium may suggest the association of H. pylori infection with markers of fecal exposure. Whether H. pylori provides favorable conditions for intestinal parasitosis or vice versa, still further investigations are needed with an emphasis upon determining correlation with gut microbiomes.

Molecular characterization and phylogenetic analysis of Acanthamoeba isolates in tap water of Beni-Suef, Egypt.

The genus Acanthamoeba is a free-living amoeba widely distributed in various aquatic environments. It is an etiologic cause of amoebic encephalitis and keratitis particularly for immunocompromised individuals. The purpose of the present study was to investigate Acanthamoeba species prevalence in household and hospital potable water in Beni-Suef governorate, Egypt, and to employ sequencing methods to identify positive Acanthamoeba species isolates and their potential health risks. Sixty tap water samples (30 household and 30 governmental and private hospital settings) collected from Beni-Suef governorate, Egypt were filtered, cultured on non-nutrient agar, identified by morphotyping keys after staining with Giemsa stain and then confirmed by PCR using Acanthamoeba specific primers. Twenty positive samples were successfully genetically characterized and phylogenetically analyzed to identify Acanthamoeba species. The total detection rate for Acanthamoeba was 48/60 (80%); Acanthamoeba contamination in water collected from domestic houses was higher than in hospitals; 27/30 (90%) versus 21/30 (70%) with statistical significant value (P value = 0.05). Sequencing of 20 positive isolates revealed Acanthamoeba T4 in 65% and T2 in 35%. To our knowledge, this is the first research that documents the occurrence and phylogeny of Acanthamoeba species in Beni-Suef, Egypt. The presence of a higher percentage of Acanthamoeba species in tap water, in particular T4, highlights the potential health hazards for immunocompromised individuals and emphasizes the urgent need for the implementation of effective filtration and disinfection measures.

Strongyloides stercoralis larvae found for the first time in tap water using a novel culture method.

Strongyloides stercoralis (S. stercoralis) is a soil-borne helminth but its prevalence in water samples is underestimated. The aim of the present study is to propose non-nutrient agar (NNA) as a novel culture method in the diagnosis of Strongyloides and to detect the prevalence rate of S. stercoralis larvae in tap water samples in Beni-Suef Governorate, Egypt. Eighty tap water samples were collected from Beni-Suef Governorate and each sample was filtered in three steps using three filters: one filter for microscopic examination of wet mount, while the remaining two filters were cultured on both NNA and nutrient agar plate (APC). S. stercoralis larvae, adults, and eggs were detected in two (2.5%) and four (5%) out of 80 samples after examination of wet mount preparation and cultures on both NNA and APC, respectively. NNA succeeded to maintain Strongyloides alive for up to 1 month. Our study is the first to detect Strongyloides larvae in tap water in Egypt and the first to use NNA as a novel culture media for Strongyloides.