Evaluation of Effectiveness between Two Different Facilities for Drinking Water Having Different Water Sources for Removal of Free-Living Amoeba in Qalyubia Governorate, Egypt

Background: Conventional drinking water treatment plants (CDWTPs) and Ground water (GWTP) are the main 2 types of drinking water treatment using freshwater as a source for drinking water in Egypt. Objective: The Egyptian standards for drinking water denied the presence of any type of living protozoa in drinking water produced for human use. Martials and methods: 48 water bodies were selected from Benha and Kaha districts in Qalyubia Governorate, Egypt. Mean turbidity, pH, conductivity, temperature, ammonia, nitrite, iron, manganese, magnesium and residual chlorine were recorded in each water body from two sites. Centrifuged samples were cultured on non-nutrient agar plates with Escherichia coli. Positive sample isolates were subjected to DNA extraction and polymerase chain reaction using genus and speciesspecific primers targeting the internal transcribed spacer region (ITS) and Mp2CL5 gene. Results: The prevalence of Naegleria species, N. fowleri and Acanthamoeba in the study area were 54.1% and 0% (N. fowleri and Acanthamoeba) of all sample examined. The removal of free-living amoebae from drinking water, it was shown that conventional DWTP (Benha) could get rid of 91% of FLAs present in the raw untreated water, while ground DWTP removed only 55.6% of these organisms. Conclusions: The conventional drinking water treatment system for surface water was better than that of only chlorine disinfection for ground water in removing free-living amoeba (FLAs). In general, the persistence of FLAs in drinking tap water unfortunately exerts public health hazards

Mapping gut parasitism patterns in a cohort of Egyptians

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

In vitro and in vivo anthelmintic and chemical studies of Cyperus rotundus L. extracts

Abstract Background Trichinellosis, a zoonosis caused by the genus Trichinella, is a widespread foodborne disease. Albendazole, one of the benzimidazole derivatives, is used for treating human trichinellosis, but with limited efficacy in killing the encysted larvae and numerous adverse effects. Cyperus rotundus L. is a herbal plant with a wide range of medicinal uses, including antiparasitic, and is frequently used in traditional medicine to treat various illnesses. Methods LC-ESI-MS was used to identify the active phytoconstituents in the methanol extract (MeOH ext.) of the aerial parts of C. rotundus and its derivate fractions ethyl acetate (EtOAc fr.), petroleum ether (pet-ether fr.), and normal butanol (n-BuOH fr.). The in vivo therapeutic effects of C. rotundus fractions of the extracts were evaluated using the fraction that showed the most promising effect after detecting their in vitro anti-Trichinella spiralis potential. Results C. rotundus extracts are rich in different phytochemicals, and the LC-ESI-MS of the 90% methanol extract identified 26 phenolic compounds classified as phenolic acids, flavonoids, and organic acids. The in vitro studies showed that C. rotundus extracts had a lethal effect on T. spiralis adults, and the LC50 were 156.12 µg/ml, 294.67 µg/ml, 82.09 µg/ml, and 73.16 µg/ml in 90% MeOH ext., EtOAc fr., pet-ether fr. and n-BuOH fr., respectively. The n-BuOH fr. was shown to have the most promising effects in the in vitro studies, which was confirmed by scanning electron microscopy. The in vivo effects of n-BuOH fr. alone and in combination with albendazole using a mouse model were evaluated by counting adults in the small intestine and larvae in the muscles, in addition to the histopathological changes in the small intestine and the muscles. In the treated groups, there was a significant decrease in the number of adults and larvae compared to the control group. Histopathologically, treated groups showed a remarkable improvement in the small intestine and muscle changes. Remarkably, maximal therapeutic effects were detected in the combination therapy compared to each monotherapy. Conclusion Accordingly, C. rotundus extracts may have anti-T. spiralis potential, particularly when combined with albendazole, and they may be used as synergistic to anti-T. spiralis medication therapy

Polyphenolic Profile of Herniaria hemistemon Aerial Parts Extract and Assessment of Its Anti-Cryptosporidiosis in a Murine Model: In Silico Supported In Vivo Study

Herniaria hemistemon J.Gay is widely used in folk medicine to treat hernia. The present study aimed to annotate the phytoconstituents of H. hemistemon aerial-part extract and investigate its in vivo anticryptosporidial activity. The chemical characterization was achieved via the LC&ndash;ESI&ndash;MS/MS technique resulting in the annotation of 37 phytocompounds comprising flavonoids and phenolic acids. Regarding the anticryptosporidial activity, fifty dexamethasone-immunosuppressed mice were separated into five groups: GI, un-infected (normal control); GII, infected but not treated (model); GIII, infected and received NTZ, the reference drug; GIV, infected and received H. hemistemon extract (100 mg/kg); and GV, infected and received H. hemistemon extract (200 mg/kg). When GIII, GIV, and GV were compared to GII, parasitological analyses displayed highly significant differences in the mean numbers of Cryptosporidium&nbsp;parvum oocysts in the stool between the different groups. GV demonstrated the highest efficacy of 79%. Histopathological analyses displayed improvement in the small intestine and liver pathology in the treated groups (GIII, IV, and V) related to the model (GII), with GV showing the highest efficacy. Moreover, the docking-based study tentatively highlighted the potential of benzoic acid derivatives as lactate dehydrogenase inhibitors. The docked compounds showed the same binding interactions as oxamic acid, where they established H-bond interactions with ARG-109, ASN-140, ASP-168, ARG-171, and HIS-195. To sum up, H. hemistemon is a promising natural therapeutic agent for cryptosporidiosis

Polyphenolic Profile of <i>Herniaria hemistemon</i> Aerial Parts Extract and Assessment of Its Anti-Cryptosporidiosis in a Murine Model: In Silico Supported In Vivo Study

Herniaria hemistemon J.Gay is widely used in folk medicine to treat hernia. The present study aimed to annotate the phytoconstituents of H. hemistemon aerial-part extract and investigate its in vivo anticryptosporidial activity. The chemical characterization was achieved via the LC–ESI–MS/MS technique resulting in the annotation of 37 phytocompounds comprising flavonoids and phenolic acids. Regarding the anticryptosporidial activity, fifty dexamethasone-immunosuppressed mice were separated into five groups: GI, un-infected (normal control); GII, infected but not treated (model); GIII, infected and received NTZ, the reference drug; GIV, infected and received H. hemistemon extract (100 mg/kg); and GV, infected and received H. hemistemon extract (200 mg/kg). When GIII, GIV, and GV were compared to GII, parasitological analyses displayed highly significant differences in the mean numbers of Cryptosporidium parvum oocysts in the stool between the different groups. GV demonstrated the highest efficacy of 79%. Histopathological analyses displayed improvement in the small intestine and liver pathology in the treated groups (GIII, IV, and V) related to the model (GII), with GV showing the highest efficacy. Moreover, the docking-based study tentatively highlighted the potential of benzoic acid derivatives as lactate dehydrogenase inhibitors. The docked compounds showed the same binding interactions as oxamic acid, where they established H-bond interactions with ARG-109, ASN-140, ASP-168, ARG-171, and HIS-195. To sum up, H. hemistemon is a promising natural therapeutic agent for cryptosporidiosis

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

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

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

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

<i>Annona muricata</i> 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 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