Microscopy detection and molecular characterisation of Giardia duodenalis infection in outpatients seeking medical care in Egypt.

Introduction: Giardiosis remains one of the most prevalent enteric parasitic infections globally. Earlier molecular-based studies conducted in Egypt have primarily focused on paediatric clinical populations and most were based on single genotyping markers. As a result, there is limited information on the frequency and genetic diversity of G. duodenalis infections in individuals of all age groups. Methods: Individual stool samples (n = 460) from outpatients seeking medical care were collected during January-December 2021 in Kafr El-Sheikh governorate, northern Egypt. Initial screening for the presence of G. duodenalis was conducted by coprological examination. Microscopy-positive samples were further confirmed by real-time PCR. A multilocus sequence typing approach targeted amplification of the glutamate dehydrogenase (gdh), beta-giardin (bg), and triose phosphate isomerase (tpi) genes was used for genotyping purposes. A standardised epidemiological questionnaire was used to gather basic sociodemographic and clinical features of the recruited patients. Results: Giardia duodenalis cysts were observed in 5.4% (25/460, 95% CI: 3.6-7.9) of the stool samples examined by conventional microscopy. The infection was more frequent in children under the age of 10 years and in individuals presenting with diarrhoea but without reaching statistical significance. Stool samples collected during the winter period were more likely to harbour G. duodenalis. All 25 microscopy-positive samples were confirmed by real-time PCR, but genotyping data was only available for 56.0% (14/25) of the isolates. Sequence analyses revealed the presence of assemblages A (78.6%, 11/14) and B (21.4%, 3/14). All assemblage A isolates were identified as sub-assemblage AII, whereas the three assemblage B sequences belonged to the sub-assemblage BIII. Patients with giardiosis presenting with diarrhoea were more frequently infected by the assemblage A of the parasite. Conclusion: This is one of the largest epidemiological studies evaluating G. duodenalis infection in individuals of all age groups in Egypt. Our molecular data suggest that G. duodenalis infections in the surveyed population are primarily of anthropic origin. However, because assemblages A and B are zoonotic, some of the infections identified can have an animal origin. Additional investigations targeting animal (domestic and free-living) and environmental (water) samples are warranted to better understand the epidemiology of giardiosis in Egypt.

Annona muricata Leaf as an Anti-Cryptosporidial Agent: An In Silico Molecular Docking Analysis and In Vivo Studies.

Cryptosporidiosis is a serious parasitic diarrheal disease linked to the occurrence of colorectal cancer in immunocompromised patients. The FDA-approved drug nitazoxanide (NTZ) achieved a temporary effect, and relapses occur. Annona muricata leaf is widely used in traditional medicine to treat a wide range of disorders, including antiparasitic and anticancer effects. So, this study aimed to investigate Annona muricata leaf antiparasitic and anticancer properties compared to NTZ in Cryptosporidium parvum (C. parvum) acutely and chronically infected immunosuppressed mice. A molecular docking analysis was performed to evaluate the effectiveness of some biologically active compounds that represented the pharmacological properties of Annona muricata leaf-rich extract toward C. parvum lactate dehydrogenase compared to NTZ. For the in vivo study, eighty immunosuppressed albino mice were classified into four groups as follows: group I: infected and treated with A. muricata; group II: infected and treated with nitazoxanide; group III: infected and received no treatment; and group IV: were neither infected nor treated. Furthermore, half of the mice in groups I and II received the drugs on the 10th day post-infection (dpi), and the other half received treatment on the 90th day post-infection. Parasitological, histopathological, and immunohistochemical evaluations were performed. The docking analysis showed that the lowest estimated free energy of binding of annonacin, casuarine, L-epigallocatechin, P-coumaric acid, and ellagic acid toward C. parvum LDH, were -6.11, -6.32, -7.51, -7.81, and -9.64 kcal/mol, respectively, while NTZ was -7.03 kcal/mol. Parasitological examination displayed a significantly high difference in C. parvum oocyst mean counts in groups I and II compared to group III (p-value < 0.001), with group I demonstrating the highest efficacy. The analyses of histopathological and immunohistochemical results revealed that group I showed restoration of the normal villous pattern without evidence of dysplasia or malignancy. A. muricata leaf has proved to be a reliable agent for Cryptosporidium treatment. This paper argues for its promising use as an antiparasitic agent and for the prevention of neoplastic sequels of Cryptosporidium infection.

Prevalence and influencing factors of self-medication during the COVID-19 pandemic in the Arab region: a multinational cross-sectional study.

BACKGROUND: The novel coronavirus pandemic (COVID-19) has begun with a wave of misinformation and fear of infection. This may have led people to self-medicate inappropriately. The World Health Organization describes self-medication (SM) as utilizing medicines to relieve symptoms or health conditions without consulting a physician. Inappropriate drug use is a burden on both health resources and patient health in the Arab region. This study aimed to detect the prevalence and influencing factors of self-medication among the general Arab population during the COVID-19 pandemic. METHODS: A multinational cross-sectional study was conducted among the general population of ten Arab countries from early August to late October 2021. Participants aged 18 years or older could join the study via social media platforms. A convenience sampling technique was used. A developed and validated web-based questionnaire was used to collect data on self-medication practice, associated influencing factors, information sources, commonly used medications, and commonly treated conditions. Descriptive, univariate, and multivariate regression analyses were applied using IBM SPSS v 26 and R v 4.0.0 software. RESULTS: A total of 8163 participants completed the questionnaire, and 518 participants were excluded from the analysis due to inconsistencies in their data. Almost two-thirds (62.7%) of participants reported practicing self-medication during the COVID-19 pandemic. At the country level, Egypt had the highest prevalence of self-medication practice (72.1%), while Palestine had the lowest prevalence (40.4%). The most commonly used drugs were analgesics, antipyretics, and vitamins (86, 65.1, and 57.1%, respectively), while antitussives and antibiotics scored 47.6 and 43.3%, respectively. Experience with similar health conditions (74.6%) and urgency of the problem (47.2%) were the most frequent factors that led to self-medication. Additionally, 38.2% of the self-medicated participants (SMPs) used drugs as prophylaxis against COVID-19. Pharmacist consultation was the most common source of information about self-medication (66.7%). Multivariate analysis showed that predictors of self-medication were older age (p = 0.008), presence of chronic illness (p = 0.015), and having monthly income or medical insurance that does not cover the treatment cost (p = 0.001, p < 0.001, respectively). CONCLUSION: Self-medication is considered a common practice across the Arab population. It is necessary to regulate policies and raise awareness among the public about self-medication.

Genotype diversity, phylogenetic analysis and seasonality of isolates of Acanthamoeba spp. in swimming pools in Kafrelsheikh, Egypt.

Species of Acanthamoeba Volkonsky, 1931 are the commonest among free-living amoebae that are widespread in different water resources but with lacking phylogenetic data. This study aims at detecting molecular prevalence and genetic diversity of Acanthamoeba isolates in Kafrelsheikh Governorate, Egypt. Forty-eight water samples were collected from 12 swimming pools; four samples during each season over one year. Samples were filtered, cultivated on non-nutrient agar plates and examined microscopically. Polymerase chain reaction (PCR) and sequence analysis of positive samples targeting diagnostic fragment 3 (DF3) of the small subunit rRNA gene were done. Cultivation succeeded to detect 14 (29%) positive samples while PCR missed three positive samples. The obtained sequences were phylogenetically analysed. The phylogenetic tree was constructed for them with sequences of reference species from the NCBI database. The identified species were Acanthamoeba castellanii Douglas, 1930 (T4), A. astronyxis (Ray et Hayes, 1954) (T9) and A. hatchetti Sawyer, Visvesvara et Harke, 1977 (T11). The prevalence of species of Acanthamoeba was higher during summer and fall. Therefore, the control of the presence of Acanthmoeba spp. in swimming pools needs immediate, effective and practical measures to prevent and control infection with species of Acanthamoeba.

Multiattribute Analysis of Trichomonas vaginalis Diagnostics and Its Correlation with Clinical Complaints and Contraceptive Methods in a Symptomatic Egyptian Cohort.

Background: Trichomonas vaginalis (T. vaginalis) infection has been long considered among the sexually transmitted diseases that possesses a clear effect on women's health especially in the childbearing period. Methods: A 234 females of age range 18-45 years old attending the Gynecology and Obstetrics Outpatient Clinic of Kasr El Aini Hospitals were enrolled in a cross-sectional study. The taken vaginal swabs were subjected to wet mount microscopy, Giemsa stain, modified Diamond's culture, and nested polymerase chain reaction (nPCR) amplification. Multiattribute and analytical hierarchy processes were conducted to detect laboratory utility. Univariate and multivariate analyses were done to detect the multiple risk factors that may be associated with Trichomonas infection. Results: Based on nPCR, the prevalence of trichomoniasis was 26.9%. Wet mount, Giemsa stain, and culture showed 100% specificity but of low sensitivity (28.57%, 28.57%, and 57.14%, respectively). On the multivariate analysis, nPCR showed the highest rank for diagnostic performance and culture had the lowest rank. For univariate analysis, there was a significant correlation between T. vaginalis infection and vaginal discharge, burning sensation, dyspareunia, and the use of intrauterine device (IUD) (P value < 0.05). Conclusion: The routine screening of trichomoniasis using nPCR was reliable, sensitive, and specific. Also, it could financially be considered a more suitable option in batch screening. Significant higher rates of infection were reported among IUD users compared to condom or hormonal-based methods.

Laboratory utility of coproscopy, copro immunoassays and copro nPCR assay targeting Hsp90 gene for detection of Cryptosporidium in children, Cairo, Egypt.

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.