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Reem A Al Dossary,1 Sarah Althuwaiqeb,2 Khaled R Alkharsah,1 Nahid HO Wanni,1 Basavaraja C Hunasemaranda,1 Obeid E Obeid,1 Zaheenul Islam Siddiqui,1 Ayman A El-Badry,1 Nourah Hasan Al Qahtani3 1Department of Microbiology, College of Medicine, Imam Abdulrahman bin Faisal University, Dammam, Saudi Arabia; 2Imam Abdulrahman bin Faisal University, Dammam, Saudi Arabia; 3Department of Obstetrics and Gynecology, College of Medicine, Imam Abdulrahman bin Faisal University, Dammam, Saudi ArabiaCorrespondence: Reem A Al Dossary, Email raaldossary@iau.edu.saIntroduction: 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.Keywords: rubella, immunity, susceptibility, congenital

This work assess serum levels of soluble Fas form (sFas) in patients with different stages of laryngeal squamous cell carcinoma(LSCC) to investigate its prognostic significance. We correlate its levels with the morphological changes of peripheral blood cells via buffy coat examinatin. The study population included 70 patients clinically diagnosed and pathohistologically confirmed LSCC in addition to 20 healthy controls. According to TNM classification 33(47.1%) patients were in stage I (group I) and 24(34.3%) in stage II (group II) , 13(18.5%) in stage III (group III). The results revealed that the mean serum level of sFas (pg/ml) in the control group was 51.2, in group I was 66.33, in group II was 81.33 and in group III was 112.45. Statistical analysis of the mean of sFas by ELISA test in the patients' groups in comparison to the control revealed a significant increase of both group II and III in comparison to the control group (P < 0.01) but there was no significant difference (P < 0.05) between group I in comparison to either the control or group II. There was a significant difference (P < 0.05) between group I in comparison to group III. LM examination revealed massive extent of the apoptotic cells in group III when compared to both group I and group II. EM examination of the buffy coat revealed apoptotic changes, mainly in the peripheral blood mononuclear cells (PBMNCs), represented by surface membrane ruffles and blebs with clumped nuclear chromatin and vacuolated cytoplasm. In conclusion, this study may help us to better understand one of the escape mechanisms in cancer larynx. This mechanism is represented by the significant increase in both the serum level of sFas and the morphological apoptotic changes that detected in PBMNCs. Soluble Fas may contribute to the progression of laryngeal cancer. It can be used as an attractive target for anticancer therapy and may be considered as a marker of disease progression and poor prognosis in laryngeal cancer. [ABSTRACT FROM AUTHOR]

Objectives To find a suitable DNA isolation system from whole blood; five different DNA extraction methods were evaluated, comparing manual, partially automated and fully automated separation. Methods Phenol-chloroform, simple extraction by microwaving and the Wizard Genomic DNA Purification system were compared in this study. In addition to these manual methods, the MagNA Pure platform was performed manually, partially automated and fully automated. The last technique was a combination between the Wizard SV 96 Genomic DNA Purification Kit and the MagNa Pure extraction in which the whole vacuum system for waste collection was replaced by the magnetic separation principals. Results Best results were observed with the MagNa Pure automatic extraction that obtained nucleic acid amount of 0.98 microg/microl with 2.91 purity ratio. The second best methods were the magnetic beads based techniques, either the original or the modified one which showed purity ratios, 2.45 and 2.36 respectively and the amount of the nucleic acids recovered were 0.79 microg/microl and 0.76 microg/microl respectively. Conclusions Magnetic DNA separation is recommended for its high-quality nucleic acid extraction in which the risk of cross-contamination is reduced. It requires minimal starting material and is both cost-effective, user friendly and can be optionally automated with no manual intervention steps. [ABSTRACT FROM AUTHOR]

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., (© 2023. The Author(s).)

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., (© 2023 [The Author/The Authors].)

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.

The epidemiology of herpes simplex virus (HSV) infections varies among populations depending on their demographic characteristics and exposure. This study describes the prevalence of HSV 1/2 IgG and IgM antibodies among individuals over a period of 5 years. A retrospective study was conducted to collect data on cases tested for HSV-1 and HSV-2 IgG and IgM antibodies for different medical conditions over five years between January 2014 and December 2018. 620 samples were tested for HSV 1/2 IgG and IgM during the study period. The total HSV seropositivity in the study population was 68% (422/620). The total seropositivity excluding children below 6 months of age was 65.3% (313/479). HSV-IgG seropositivity was significantly higher in married individuals (p<0.001, 95% CI 1.61-3.69). The HSV IgG seropositivity was significantly higher in children under the age of 6 months (N=109, 77.30%) than in children between 7 and 24 months old (27.6%) (Chi-square for linear trend, p<0.001), and it then tends to increase with age more than 24 months (Chi-square for linear trend, p=0.011). Eleven children showed laboratory evidence of recent HSV infection (6.2%) as indicated by HSV IgM antibodies and had diverse clinical conditions. HSV infection is highly prevalent in the Eastern Province of Saudi Arabia. Infection is most probably acquired during early childhood, and the tendency increases with age. However, a significant number of mothers are at risk of infection and transferring the infection to their fetuses., Competing Interests: The authors declare no conflict of interest., (©2022 JOURNAL of MEDICINE and LIFE.)

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., (© 2022. The Author(s).)

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., (© 2021. The Author(s) under exclusive licence to Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

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 (Zn 2+ 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., (© 2022. The Author(s).)

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.

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., (© 2022. The Author(s).)

Blastocystis is one of the less well-understood waterborne protozoa of medical and veterinary importance. The present study aimed to detect, identify, and subtype Blastocystis in cattle and humans in northern Egypt and to investigate the potential for zoonotic transmission. Fecal samples were collected from 136 humans (68 in contact with animals and 68 no animal contact) and 190 cattle from a rural area in Kafr El-Sheikh Province, Egypt. All samples were microscopically examined by direct wet mount technique and cultured in Jones' medium. Genomic DNA was extracted from positive cultures. A region of the Blastocystis small subunit ribosomal ribonucleic acid (SSU rRNA) gene was amplified using PCR. Blastocystis subtypes were identified by sequencing of the amplified SSU rRNA gene. The relatedness of sequence types obtained in this study and others submitted in GenBank was compared according to their genetic relatedness. Out of 136 human and 190 cattle samples, Blastocystis was detected in 53 (38.9%) and 37 (19.4%), respectively using either culture or direct wet mount. Blastocystis SSU rRNA gene was amplified in 55 samples (32 humans and 23 animals). Six subtypes were identified (STs 1-3 in humans and ST4, ST10 and ST14 in cattle) and were distinguishable with inter (up to 0.075) and intra (up to 0.024) subtype variations. A close phylogenetic relationship between human and animal isolated STs 1-4 was demonstrated. To the best of our knowledge, this is the first study to investigate the the potentiality for zoonotic transmission of certain Blastocystis subtypes in northern Egypt., Competing Interests: Conflict of interestNone., (© Indian Society for Parasitology 2021.)

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., Competing Interests: The authors have declared that no competing interests exist.

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 (X 2  = 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.

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.

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., Competing Interests: Conflict of interestAll the authors declare that they have no conflict of interest., (© Indian Society for Parasitology 2020.)

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., Competing Interests: Declaration of Competing Interests All the authors declare that they have no conflict of interest, (Copyright © 2020 Elsevier B.V. All rights reserved.)

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., (Copyright © 2018. Published by Elsevier B.V.)

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- β-p -hydroxy-benzoyl-20- O- acetyl-boucerin-3- O-β -D-glucopyranosyl-(1→4)- β -D-cymaropyranosyl-(1→4)- β- D-cymaropyranoside, in addition to two flavonoids (CII) luteolin-4'- O - β -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 IC 50 of 8.48 µg/ml.

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 ( 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., Competing Interests: Conflict of interestThe authors have declared that no competing interest exists.

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 (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.

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.

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., (Copyright © 2016. Published by Elsevier B.V.)

Increasing prevalence of Cryptosporidium raises the importance to explore different aspects of its infection. In the absence of reproducible in vitro culturing, animal model is the only experimental method to study Cryptosporidium . Our study evaluated Cryptosporidium infection using coproscopy, copro-antigen and copro-DNA for early detection of murine cryptosporidiosis. Hundred and forty albino mice (neonates and adult) were divided into two groups, control group received sterile PBS solution, and infected groups were inoculated with molecularly characterized Cryptosporidium parvum oocysts and further subdivided into three subgroups for infectious dose response detection. Mice fecal samples were collected every 4 h on the first day and then daily and examined for fecal oocysts, copro-antigen and copro-DNA. Four mice from each subgroup were killed at 12, 24 and 48 h post-infection (P-I), and their intestines were examined for cryptosporidial mucosal DNA. Cryptosporidium copro-antigen and copro-DNA were detected 4 and 8 h P-I in infected neonatal and adult mice, respectively, and intestinal mucosal DNA was detected after 12 h in both. Microscopy was able to detect oocysts 48 h P-I. Inoculated C. parvum oocysts were recovered in feces of infected mice without genotypic changes. Neonate mice showed higher susceptibility for cryptosporidial infection than adults without statistical differences for the given infectious doses. Both copro-immunoassay and copro-nPCR assays can early detect Cryptosporidium infection; however, nPCR was able to identify Cryptosporidium species, making nPCR a reliable biomarker for early detection in murine model.

Sources of heavy metals pollution in the Burullus Lagoon include phosphate fertilizers, sewage and oil spills from fishing boats. The benthic species identified in this lagoon are Adelosina carinata striata (Wiesner), Quinqueloculina bosciana (d'Orbigny), Quinqueloculina seminulum (Linnaeus), Ammonia tepida (Cushman), Elphidium excavatum (Terquem). A. tepida is the most dominant species in the lagoon. It constitutes more than 97% of the total benthic foraminiferal assemblages reflecting tolerance to the very low salinity in the Burullus Lagoon. The intensity of deformation was severe exhibiting a peculiar change in the coiling direction in A. tepida with increase in cadmium concentration. A. tepida exhibited a great morphological variability and the recorded morphological abnormalities show high spire giving the spiroconvex test, additional chamber, aberrant chamber shape and size, twisted tests with elongated axes of rotation and complex deformities, whereas E. excavatum showed additional calcite secretion (tumors)., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Giardia intestinalis (G. intestinalis) and Helicobacter pylori (Hpylori) are two intestinal pathogens sharing the same mode of infection. This study determines the prevalence of G. intestinalis and H pylori co-infection estimated risks and predictive factors for susceptibility to co-infection. Stool samples were collected from 801 patients suffering gastrointestinal symptoms and living in Greater Cairo. They were subjected to coproscopic examination for detection of intestinal parasites and copro PCR-restriction fragment length polymorphism (PCR-RFLP) and sequencing targeting the glutamate dehydrogenase (gdh) gene for Giardia. Positive samples for gidrdiasis were further subjected to copro- immunoassay to detect H pylori coprontigen. Among 63 cases of giardiasis by both microscopy and PCR (84.1 % as-semblage B and 15.9% AII), 52.5% were coinfected with H pylori. Co-infection was more frequent with assemblage B (50.9%) than assemblage A (40%). Among studied variables of assemblage type, gender, or harboring more than one parasite (polyparasitism), only school age children, was signifi-cantly associated (P value: 0.02) with Giardia and H pylori co-infection. Physicians in Egypt must consider G. intestinalis and H pylori as prevailing intestinal pathogens with predominance of Giardia assemblage B. Giardia and H pylori co-infection is common in school aged children and modulates gastrointestinal manifestations. Intestinal parasitism and H pylori association is complex and necessitates further genomic studies for a better understanding of the epidemiological and clinical impact of co-infection, as well as possible strategies for their treatment and control.

Lymphatic filariasis (LF) is a serious vector-borne health problem, and Wuchereria bancrofti (W.b) is the major cause of LF worldwide and is focally endemic in Egypt. Identification of filarial infection using traditional morphologic and immunological criteria can be difficult and lead to misdiagnosis. The aim of the present study was molecular detection of W.b in residents in endemic areas in Egypt, sequence variance analysis, and phylogenetic analysis of W.b DNA. Collected blood samples from residents in filariasis endemic areas in five governorates were subjected to semi-nested PCR targeting repeated DNA sequence, for detection of W.b DNA. PCR products were sequenced; subsequently, a phylogenetic analysis of the obtained sequences was performed. Out of 300 blood samples, W.b DNA was identified in 48 (16%). Sequencing analysis confirmed PCR results identifying only W.b species. Sequence alignment and phylogenetic analysis indicated genetically distinct clusters of W.b among the study population. Study results demonstrated that the semi-nested PCR proved to be an effective diagnostic tool for accurate and rapid detection of W.b infections in nano-epidemics and is applicable for samples collected in the daytime as well as the night time. PCR products sequencing and phylogenitic analysis revealed three different nucleotide sequences variants. Further genetic studies of W.b in Egypt and other endemic areas are needed to distinguish related strains and the various ecological as well as drug effects exerted on them to support W.b elimination.

Giardiasis is a prevailing intestinal disease in children. This study aimed to determine molecular prevalence of Giardia intestinalis in children attending Cairo University Pediatrics Hospitals, using copro-PCR assays, conventional methods and to evaluate diagnostic effectiveness of used tests. 229 fecal samples were collected from children suffering from gastrointestinal symptoms and examined for Giardia by microscopy, Immuno-chromatographic test (ICT), copro-DNA using two PCR assays targeting tpi [nested-PCR (nPCR)] and 18S [conventional-PCR (cPCR)] genes. Out of 229 samples assessed, Giardia was diagnosed in 13.9, 17, 17.9, 4.8 % of cases using microscopy, ICT, nPCR (tpi) and cPCR (18S), respectively. Nominating both PCR assays as composite reference standard, microscopy and ICT were of reliable specificity (100 and 96.9 %) and accuracy (95.6 and 93.6 %) but of limited sensitivity (78.6 and 76.2 %). Kappa agreement showed, there was substantial agreement of ICT (0.776) and almost perfect agreement of microscopy (0.839) with PCR assays. Giardia showed a molecular prevalence of 18.3 % (42/229). ICT assay for Giardia surpassed microscopy but both couldn't be used as a consistent single detection method due to their lowered sensitivities. nPCR targeting tpi is a reliable diagnostic test aiding to determine true prevalence of Giardia., Competing Interests: The authors declare that they have no reported conflicts of interest. Author contribution All manuscript authors contributed to every activity of it; idea of paper, study design, collection of materials, methodology, writing the paper and revising it.

Giardia lamblia (G. lamblia) is the most worldwide prevailing intestinal parasite, notorious for its broad range of seasonal and age-related prevalence. The potentially lethal nature of giardiasis makes it essential that the seasonality, the groups at risk, and other potential risk factors are identified. The present molecular epidemiological study was designed to determine the genetic diversity of G. lamblia infection, taking into account seasonal peaks, age distribution, and associated symptoms in a cohort of Egyptian diarrheic patients. Stool samples were collected from 1187 diarrheic patients attending outpatient clinics of Cairo University hospitals, of all age groups over a 12-month period. The patients were examined microscopically for fecal G. lamblia cysts, and/or trophozoites, and for copro-DNA detection using nested polymerase chain reaction (nPCR) assays targeting beta giardin gene. PCR-positive samples were characterized molecularly by nPCR restriction fragment length polymorphism (RFLP) to determine Giardia assemblages. The findings revealed circannual prevalence of Giardia, with a seasonal pattern peaking in mid-summer and late winter, with the summer peak preceded by a peak in temperature. Infection was prevailing in 224 (18.9 %) cases, mainly assemblage B (81.2 %) followed by assemblage A (18.8 %). There were statistically significant associations between the detection of Giardia and flatulence, persistent diarrhea, vomiting, and abdominal pain, while gender and intermittent diarrhea showed no association. The pre-school age group was the most vulnerable. This is the first study of molecular characterization of Giardia to determine its circannual prevalence in Egypt, a finding which carries promising potential for the diagnosis, treatment, and elimination of the disease.

Cryptosporidium is a significant cause of diarrhea worldwide especially in children. Infection may end fatally in immunocompromised patients. Multi-attribute analysis was used to determine the lab utility of 4 diagnostics; coproscopy of AF stained fecal smear, fecal immunoassays by ICT and ELISA and copro-nPCR assay targeting Hsp90 gene, for detection of Cryptosporidium in stool of 250 Egyptian children (150 diarrheic and 100 non-diarrhaeic children). Also, to determine Cryptosporidium molecular prevalence. Cryptosporidium was an important enteric pathogen among both diarrheic and non-diarrheic study children with a clearly high prevalence of 16.4 % (n = 41). Conventional methods had perfect specificity (100 %) but couldn`t be used as a consistent single detection method due to their lowered sensitivities. Multi-attribute analysis ranked nPCR the highest test for lab use. Being the test with the best diagnostic yield, nPCR is a reliable diagnostic test and is going to replace conventional methods for reliable detection of Cryptosporidium.

Giardiasis is a major health problem in both developed and developing world. A variety of methods for diagnosis of Giardia duodenalis cysts or trophozoites is available but still has certain limitations. 100 sample from diarrhoeal children who attending outpatient clinic in Abu El Rish hospital, .Kasr Al Ainy, Faculty of Medicine, Cairo University, Egypt. Giardiasis was diagnosed by direct wet mount, microscopy after formal- ethyl acetate concentration, Ridascreen ELISA assay and n-PCR targeting beta giardin (bg) gene. Using ELISA as reference standard, the methods' sensitivities, specificities, positive (PPV) and negative (NPV) predictive values and positive (LR+) and negative (LR-) likelihood ratios with 95% confidence interval (95% CI) were analyzed.The diagnostic methods were evaluated to determine their impact on the posttest probability using Fagan's nomogram. All the studied methods led to a LR+ higher than 10 indicating ability to ruling in giardiasis. n-PCR recorded LR- equal 0.00 and the probability of giardiasis would be 0% if the test was negative. The methods were also ranked on basis of Multiattribute utility theory and Analytical hierarchy process with ELISA ranked better than n-PCR.

Stool samples from 182 diarrhoeic (symptomatic) children and 100 apparently healthy (asymptomatic) children, matched for age, from Aboul-Reesh Cairo University Pediatrics Hospital were examined by ELISA and by nPCR (targeting COWP gene) for the detection of Cryptosporidium. The demographic and environmental data of the diarrhoeic group was recorded. The PCR amplified product of positive cases was then subjected to RFLP by digesting it with the restriction enzyme RsaI. The obtained fragments were resolved by electrophoresis and the bands were visualized and characterized versus a standard. ELISA results demonstrated a prevalence rate of 13.2% (24/182) among diarrhoeic group, and 8% (8/100) among non-diarrheic group, with overall detection rate of 11.3% (32/282). Higher rates of detection were obtained by nested PCR assay among diarrhoeic group 25.8% (47/182) and 16% (16/100) among non-diarrhoeic group with overall detection rate of 22.3% (63/282). Considering nPCR as the reference method, ELISA had a sensitivity of 47.6% and a specificity of 99.1%. RsaI digestion of nPCR product of COWP gene revealed the presence of 2 genotypes: genotype 1 with 4 bands (34, 106, 125 and 285) and genotype 2 in which 3 bands (34, 106 and 401). Among the 63 cases with cryptosporidiosis, 53 (88.3%) had genotype 1, and 7 (11.7%) had genotype 2. The higher prevalence of genotype 1 suggests a relatively greater risk of human source of infection than zoonosis.

Background: Aedes aegypti is the main vector of the yellow fever and dengue virus. This mosquito has become the major indirect cause of morbidity and mortality of the human worldwide. Dengue virus activity has been reported recently in the western areas of Saudi Arabia. There is no vaccine for dengue virus until now, and the control of the disease depends on the control of the vector., Objectives: The present study has aimed to perform phylogenetic analysis of Aedes aegypti based on mitochondrial NADH dehydrogenase subunit 4 ( ND4 ) gene at Almadinah, Saudi Arabia in order to get further insight into the epidemiology and transmission of this vector., Materials and Methods: Mitochondrial ND4 gene was sequenced in the eight isolated Aedes aegypti mosquitoes from Almadinah, Saudi Arabia, sequences were aligned, and phylogenetic analysis were performed and compared with 54 sequences of Aedes reported in the previous studies from Mexico, Thailand, Brazil, and Africa., Results: Our results suggest that increased gene flow among Aedes aegypti populations occurs between Africa and Saudi Arabia., Conclusions: Phylogenetic relationship analysis showed two genetically distinct Aedes aegypti in Saudi Arabia derived from dual African ancestor.

Lymphatic filariasis is a vector-borne health problem that has been focally endemic in Egypt for centuries. The chief vectors of transmission are Culicinae species. Control measures in the form of mass drug administration of DEC citrate treatment have been implemented in Nile delta for almost a decade. This study aimed to identify the prevalent mosquito species in endemic areas in Giza and Qualioubiya governorates and to monitor Wuchereria bancrofti infection by detecting the parasite DNA in collected mosquitoes. Adult mosquitoes were collected using light traps hung indoors. Microscopic examination was performed to identify and examine the morphologic characters of mosquitoes. Female Culex mosquitoes were subjected to semi-nested PCR to detect filarial DNA targeting repetitive DNA sequences (pWbl2 repetitive region) specific for W. bancrofti. The results revealed 3 species of mosquitoes Culex pipiens, Culex pusillus and Culex quinquefasciatus with the predominance of Culex pipiens (85.7%). Wuchereria bancrofti DNA was not detected in any of the collected mosquito pools. With progress of elimination programme in Nile Delta, follow up studies with larger sample size are recommended as the predominance of Culex pipiens the main lymphatic filariasis vector remains a risk of transmission in such areas.

Cryptosporidiosis is a worldwide gastrointestinal disease caused by the protozoan Cryptosporidium parasite. It has a broad range of seasonal and age-related prevalence. We aimed to study the molecular prevalence and seasonality of Cryptosporidium over a period of 1 year in a cohort of Egyptian diarrhoeic patients. Stool samples were collected from 865 diarrhoeic patients attending outpatient clinics of Cairo University hospitals, from all age groups over a 12-month period, examined microscopically for faecal Cryptosporidium oocysts by the acid-fast staining method and for copro-DNA detection using nested polymerase chain reaction (nPCR) assays. PCR-positive samples were characterised molecularly by nPCR-restriction fragment length polymorphism (RFLP) to determine Cryptosporidium genotypes. Cryptosporidium copro-DNA was detected in 19.5% of the collected samples throughout the year, with a major peak in summer (August) and a small rise in spring (April). Infection was mainly C. hominis (95.8%) followed by C. parvum (3.0%), affecting all age groups, with predominance in the pre-school age group, and decrease with age. There were statistically significant associations between the detection of Cryptosporidium and season, diarrhoea, patient age and drinking water, while gender, contact with animals and presence of mucus in stool showed no association. Cryptosporidium in diarrhoeic Egyptians was of distinct endemicity, with the bi-model mostly influenced by population dynamics, with a clear high prevalence in pre-school children and predominating anthroponotic (C. hominis) transmission throughout the year. The obtained results highlight Cryptosporidium as a water contaminant and an important cause of health problems in Egypt, necessitating further studies of the risk factors.

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., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Amoebiasis diagnosis is usually based on microscopy that cannot differentiate pathogenic E. histolytica from morphologically identical non-pathogenic species. 194 fecal samples were collected from diarrheic &/or dysenteric patients and examined for Entamoeba complex microscopically, E. histolytica/E. dispar coproantigen using ICT and E. histolytica coproantigen using Tech lab E. histolytica II ELISA test. Entamoeba complex trophozoites/cysts, E. histolytica/E. dispar coproantigen and E. histolytica coproantigen were detected in 22.2, 14.4 and 3.6 % of samples, respectively. Microscopy and ICT method had limited sensitivity with poor PPV (9.3 and 7.1 %, respectively) and both slightly agree with ELISA test. The prevalence of E. histolytica was low (3.6 %) in studied individuals and was 14 times lower than non-pathogenic amoebae. E. histolytica detection studied individuals was positively associated with mucoid and bloody stool, which makes them disease predictors. E. histolytica fecal ELISA assay for E. histolytica detection surpassed microscopy and E. histolytica/E. dispar ICT assay. This has highlighted the need for practical non-microscopic detection methods that can differentiate between amoeba infections to avoid unnecessary and possibly harmful therapies and to determine the true prevalence and epidemiology of E. histolytica.

Objective: To determine molecular prevalence of Cryptosporidium in a cohort of Egyptian children and compare three diagnostic tests., Methods: Stool 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)., Results: nPCR detected Cryptosporidium in 22.4% of children. Acid-fast stain and ELISA showed false negativity but 100% specificity with nPCR as gold standard., Conclusion: Cryptosporidium is a common cause of diarrhea in children in Egypt.

Labiatae family is represented in Saudi Arabia. The aim of the present study was to go insight to investigate the anticancer activity and antioxidative potentials of methanolic extracts of Mentha longifolia L. (ML) and Ocimum basilicum L. (OB) that grown in Madina province, western region, Saudi Arabia. OB exhibited the greater phenolic contents as mg gallic acid equivalent/g weight (mg GAE/g) for a value of 105 +/- 5.5 mg GAE/g. On the other hand, ML produced 29 +/- 3.12 mg GAE/g. The standard antioxidant vitamin E used in this experiment elicited a value of total phenolic contents equal 22 +/- 2.2 mg GAE/g. The percentage scavenging activity of against diphenylpicrylhydrazyl (DPPH) was 850 and 160% for OB and ML extracts, respectively. Vitamin E elicited% scavenging activity of against DPPH equal to 198%. Brine shrimp cytotoxic assay clearly indicated the cytotoxic effects of either ML or OB extract. The brine shrimp survival is inversely proportional to the concentration of either ML or OB extract used with LD50 191.23 and 235.50 ppm, respectively. Toxic effects on brine shrimps indicated the anticancer potential of ML or OB extract. The ML or OB extract was unable to produce pbluescript (pBS) plasmid DNA damage, while the plasmid DNA treated with EcoRI produced a single band as a result of DNA damage. Also, both ML and OB extract exhibited marked cytotoxic activity against MCF-7 cells at various concentrations (20, 40, 80, 160 and 320 microg mL(-1)). The 160 and 320 microg mL(-1) showed more cytotoxic effect against MCF-7 cells. Based on results achieved, we can concluded that, OB and ML extracts have the potency to act as powerful antioxidants and protect against DNA damage and have cytotoxic activity against MCF-7 cell line.

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., (Copyright © 2013 Elsevier Inc. All rights reserved.)

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., (Copyright © 2012 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

In order to overcome the false negative diagnosis of strongyloidiasis in the absence of rhabditiform larvae in stools, an ELISA-based Strongyloides stercoralis-specific coproantigen detection assay in stools of infected patients was evaluated. In a sandwich ELISA, a rabbit hyperimmune serum against S. stercoralis ES (excretory/secretory) adult antigen succeeded in capturing S. stercoralis coproantigen from infected patients and did not react with coproantigens prepared from the stool samples of patients infected with Schistosoma mansoni, Fasciola gigantica and Capillaria philippenensis. Coproantigen was able to detect anti-S. stercoralis IgG antibodies in sera of infected patients at the same OD level as produced with S. stercoralis E/S worm antigen using an indirect ELISA did not cross-react with sera from patients with S. mansoni, F. gigantica and C. philippenensis. S. stercoralis coproantigen detection proved a sensitive, simple, reliable and inexpensive ELISA-based, and an alternative to coproscopical methods in copropositive (with larvae in stool) and copronegative (without larvae in stool) stool samples. Fecal ELISA showed a positive relationship between copro-Ag and worm burdens, and considered a starting point for the development of species-specific copro-immunological diagnostic assays using monoclonal antibodies and dipstick technology.

By regulating matrix metalloproteinase (MMP) activity and controlling the breakdown of extracellular matrix components, tissue inhibitors of metalloproteinases (TIMPs) play an important role in the process of tumor invasion and metastasis. The present study was designed to clarify the role of TIMP-2 in nasopharyngeal carcinoma (NPC) patients and to evaluate its importance relative to clinicopathologic parameters. It was carried out in 30 patients with NPC and 20 controls. Tissue biopsies were studied and graded pathologically, and Western blot analysis was performed to assess TIMP-2 protein expression. Clinically, in accordance with TNM classification (T: tumor size, N: lymph node involvement, M: distant metastasis), 8 cases were diagnosed as stage II, 12 as stage III, and 10 cases as stage IV; however, pathologic typing with use of the World Health Organization (WHO) classification revealed the presence of 9 specimens of squamous cell carcinoma (WHO type 1), 6 cases of nonkeratinizing carcinoma (WHO type 2), and 15 cases of undifferentiated carcinoma (WHO type 3). The difference in percentage of TIMP-2 positivity between NPC patients (76.6%) and normal controls (30%) was statistically highly significant (P < .01). In addition, there was a significant positive correlation between TIMP-2 protein positivity and either the clinical staging or the histopathologic typing (P < .01) using Chi-square test (x(2)), suggesting that TIMP-2 can be used as a marker of the severity of NPC.Accordingly, we can assume that TIMP-2 may play a role in regional lymph node and/or distant metastasis and in progression of squamous cell carcinoma. Further studies are needed to investigate the role of TIMP-2 as a marker for tumor progression and to evaluate its potential value in the follow-up of patients.

In a trial at determining the most relevant immunoglobulin isotype that could reflect success of praziquantel treatment, an ELISA using soluble egg antigen (SEA) was applied on sera of Egyptian patients suffering from active intestinal schistosemiasis without hepatic complications, determining the levels of IgE, IgA, IgM, IgG1, IgG2, IgG3 and IgG4 raised against the SEA, both, pre- and early post-treatment. The positive results obtained to all anti-SEA immunoglobulin isotypes before treatment support the usefulness of this technique in the diagnosis of schistosomiasis. Except for IgG3 subclass, a statistically significant correlation was found between egg output-reflecting intensity of infection- and the different immunoglobulin levels, especially anti-SEA IgG4. When repeating the assay 5-6 months after treatment, the immunoglobulin levels showed either a rise (in case of IgE) or a drop (in case of IgA, IgM & IgG1-4), all of statistical significance, yet, IgG1-4 were still positive. So, ELISA could not give a definite indication of cure after anti-bilharzial treatment. IgE, IgG2 and IgG4 were revealed to be the most significant immunoglobulin isotypes at the post-treatment level, both statistically and due to their implications on resistance/ susceptibility to re-infection and also due to the correlation of IgG4 with the tendency to develop periportal fibrosis. Conclusively, although not having defined a particular Ig isotype as marker for cure, yet it exposed the urge for early post-treatment determination of IgE and IgG4 isotypes, which could serve as markers for picking up high risk patients susceptible to reinfection or liable to develop bilharzial periportal fibrosis, and who might benefit from a second course of specific treatment.

In order to overcome the false negative diagnosis of infection with C. philippinesis at time of absence of eggs in stool, coproantigen prepared from stools of infected patients was evaluated serologically. This antigen was able to detect anti-Capillaria antibodies in the sera of infected cases at the same OD level produced with Capillaria crude worm antigen using indirect ELISA technique C. philippenensis coproantigen did not cross-react with sera from patients with schistosomiasis mansoni, fascioliasis or strongyloidiasis at 1:00 serum dilution. Laboratory-prepared hyperimmune sera versus crude worm antigen of C. philippinensis succeeded in capturing Capillaria antigen prepared from the stools of infected patients and did not cross react with coproantigens prepared from stool samples of cases infected with S. mansoni or Fasciola using sandwich ELISA technique.

The effects of water extracts of Cynomorium coccineum and Withania somnifera on ovarian follicular development and serum levels of FSH and LH were studied in immature 17-day-old and 25-day-old-Wistar rats. Water extracts of the plants were given to the animals per os in a dose of 47 mg/100 g body weight for 6 days. Serum levels of FSH and LH were measured by ELISA. Folliculogenesis was studied with a light microscope. In 25-day-old rats, extracts of both plants elicited significant changes in gonadotrophin levels coupled with a significant increase in ovarian weight and profound folliculogenesis. Numerous primary, secondary, tertiary and antral follicles were present. A distinct zona pellucida was not seen and the oocyte was often detached. In 17-day-old animals there was a significant increase-in body weight but without significant changes in the ovarian weight and folliculogenesis., (Copyright 2000 John Wiley & Sons, Ltd.)