The impact of chlorine, ultraviolet-C, and microwave treatment on the survivability of Blastocystis sp. cysts.

Blastocystis sp. is a common widely distributed gut protozoan, with water transmission identified as one of its transmission routes. This study aimed to investigate the effect of chlorine, ultraviolet (UV)-C, and microwave (MW) treatments on the in vitro viability of cysts of Blastocystis sp. Purified Blastocystis sp. cysts were molecularly subtyped. Viable cysts were subjected to different free chlorine concentrations (1, 2, and 4 ppm), different doses of UV-C (5.13, 10.26, 20.52, and 40.47 mJ/cm2), and MW irradiation times (10, 15, 30, and 45 s). Viability reduction percentage, log10 inactivation, and micrometre-based optical microscopy examined cyst number and appearance after each disinfection trial. The three disinfectants' efficacy and application conditions were assessed. The analysed isolates of Blastocystis cysts were subtype 3, possessed varying sizes and shapes, but two identical genomes. The cysts of Blastocystis sp. were resistant to chlorine at all doses and exposure durations tested. UV-C at a dose of 40 mJ/cm2 and MW treatment for 15 s were able to completely disinfect the cysts. The MW was the most effective disinfectant against Blastocystis cysts based on all evaluated factors. MW irradiation is the most efficient water treatment method for eradicating Blastocystis cysts in an easy and safe manner.

Anti-Cryptosporidium parvum activity of Artemisia judaica L. and its fractions: in vitro and in vivo assays.

Background: This study investigates the toxic activity of Artemisia judaica ethanolic extract (ArEx) as well as its phenolic fraction (ArPh), and terpenoid fraction (ArT) against Cryptosporidium parvum (C. parvum) oocysts. Methods: Over a 4 months period, estimation of the total phenolic (TPC), total flavonoids (TFC), and total terpenoids contents (TTC) in ArEx; investigation of the in vitro antioxidant activity of ArEx, ArPh, and ArT; evaluation of ArEx, ArPh, and ArT toxic activity against C. parvum oocysts using MTT assay; parasitological analysis on ArPh-treated C. parvum oocysts and comet assay were performed both in vitro and in vivo (infectivity). Results: The ArEx TPC, TFC, and TTC was 52.6 ± 3.1 mgGAE/g, 64.5 ± 3.1 mg QE/g, and 9.5 ± 1.1 mg Linol/g, respectively. Regarding the phytochemical in vitro antioxidant activity, the ArPh exhibited the highest antioxidant activity compared to the ArEx and ArT. The ArPh showed promising free radical scavenging activity of DPPH and ABTS•+ with IC50 values of 47.27 ± 1.86 µg/mL and 66.89 ± 1.94 µg/mL, respectively. Moreover, the FRAP of ArPh was 2.97 ± 0.65 mMol Fe+2/g while its TAC was 46.23 ± 3.15 mg GAE/g. The ArPh demonstrated toxic activity against C. parvum oocysts with a potent IC50 value of 31.6 µg/mL compared to ArT (promising) and ArEx (non-effective). ArPh parasitological analysis demonstrated MIC90 at 1000 µg/ml and effective oocysts destruction on count and morphology. ArPh fragmented oocysts nuclear DNA in comet assay. Beginning at 200 µg/mL, ArPh-treated oocysts did not infect mice. Conclusion: To combat C. parvum infection, the phenolic fraction of A. judaica L. shows promise as an adjuvant therapy or as a source of potentially useful lead structures for drug discovery.

Gastrointestinal parasites in Africa: A review.

Data on human gastrointestinal parasites (GIP) infections in the african sub-regions and countries are mainly lacking in terms of prevalence and population stratification by afflicted age group, symptomatology, multi-parasitism, and diagnostic methods. This study aims to describe the GIP reported in african countries and discuss the extent of the burden in the african context. Only 68.42% (39/57) of african countries reported human cases of GIP with helminths (45%, CI: 40-50%, I2: 99.79%) as the predominant parasitic group infecting the african population. On a regional scale, Central Africa had the highest pooled prevalence for GIP (43%, CI: 32-54%, I2: 99.74%), while the Central African Republic led all countries with a pooled prevalence of 90% (CI: 89-92%, I2: 99.96%). The vulnerable population (patients who are minorities, children, old, poor, underfunded, or have particular medical conditions) was the most affected (50%, CI: 37-62%, I2: 99.33%), with the predominance of GIP in the 6 to <20 years age group (48%, CI: 43-54%, I2: 99.68%). Reports on multi-parasitism (44%, CI: 40-48%, I2: 99.73%) were almost double the reports of single infections (43%, CI: 27-59%, I2: 99.77%) with combined molecular and non-molecular techniques demonstrating the best performance for GIP identification. The current review spans more than 40 years of GIP reports from the african continent. Geographical characteristics, environmental factors, habits of its inhabitants, and their health status play a crucial role in GIP modulation and behaviour in its captive hosts. Strategies for regular and enhanced surveillance, policy formation, and high-level community awareness are necessary to identify the true incidence in Africa and the transmission of the pathogens via water and food.

Subtypes and phylogenetic analysis of Blastocystis sp. isolates from West Ismailia, Egypt.

In Egypt, Blastocystis sp. is not yet on the diagnostic list of parasitology reports, and information about its subtypes (STs) is scarce. This study investigated its prevalence and its STs/alleles, performed phylogenetic analysis, and considered the distribution of risk factors associated with Blastocystis sp. infections in West Ismailia, Ismailia governorate. Sociodemographic data, exposure factors, and previous parasitic infection status were recorded for symptomatic and asymptomatic individuals. Microscopy, polymerase chain reaction, sequencing, and phylogenetic analysis for Blastocystis sp. isolated from fecal samples were performed. Eighty Blastocystis sp.-infected individuals (15.3%) were examined. The age of the individuals ranged between 0.60 and 85.0 (mean 17.10 ± 15.70), the male/female ratio was 33/47, and the asymptomatic/symptomatic ratio was 55/25. The findings demonstrate clear evidence of direct contact with animals, poor water quality, and previous parasitic infections. Eleven samples yielded three Blastocystis STs (ST1: allele 4, ST2: alleles 9 and 12, and ST3: allele 34), with ST3 (45.5%) representing the most common subtype. Phylogenetic analysis with a robust bootstrap revealed three distinct clades for isolates of each subtype. This study updates the epidemiological knowledge of the distribution of Blastocystis sp. STs in Egypt and expands the current understanding of the prevalence, risk factor frequencies, and genetic diversity of this protist in the studied area.

Cryptosporidium and Cryptosporidiosis: The Perspective from the Gulf Countries.

The present review discusses the burden of cryptosporidiosis in the Gulf Cooperation Council (GCC), which is underreported and underestimated. It emphasizes that the Cryptosporidium parasite is infecting inhabitants and expatriates in the Gulf countries. Children under 5 years are a vulnerable group that is particularly affected by this parasitic disease and can act as carriers, who contribute to the epidemiology of the disease most probably via recreational swimming pools. Various risk factors for cryptosporidiosis in the GCC countries are present, including expatriates, predisposing populations to the infection. Water contamination, imported food, animal contact, and air transmission are also discussed in detail, to address their significant role as a source of infection and, thus, their impact on disease epidemiology in the Gulf countries' populations.

Inhibitory activity of chitosan nanoparticles against Cryptosporidium parvum oocysts.

Cryptosporidium is a ubiquitous harsh protozoan parasite that resists many disinfectants. It remains viable and infective for a long time in water and food causing global outbreaks. Chitosan (the deacetylated chitin molecule) was used in its nanosuspension form to evaluate its effect against Cryptosporidium parvum. The experiments were performed in vitro in serial concentrations and confirmed in mice in vivo infectivity assay. Chitosan nanoparticles (Cs NPs) were toxic to Cryptosporidium oocysts. The effect appeared to decrease the number of Cryptosporidium oocysts and altered their content. The destruction rate of oocysts was dependent on the dose of chitosan and the time of exposure (P < 0.05). Higher doses of Cs NPs over a prolonged period exhibited a significantly higher destruction rate. Using staining and light microscopy, remarkable destructive changes were observed in the oocysts' morphology. The minimal lethal dose for > 90% of oocysts was 3000 µg/ml, no mice infections in vivo were observed. The results in this study elucidate Cs NPs as an effective anti-cryptosporidial agent.

Correction to: Strongyloides stercoralis in Patients on Corticosteroids Therapy Using Enzyme­Linked Immunosorbent Assay and Gelatin Particles Indirect Agglutination Tests: A Diagnostic Approach.

In the printed version of the article, the name of the third author was misspelled. The correct name is as follows: Amal A. El-Moamly.

Strongyloides stercoralis in Patients on Corticosteroids Therapy Using Enzyme-Linked Immunosorbent Assay and Gelatin Particles Indirect Agglutination Tests: A Diagnostic Approach.

INTRODUCTION: Patients on corticosteroids therapy (POCT) are more likely to flare up concealed Strongyloides infection and develop Strongyloides hyperinfection syndrome and/or dissemination. Such critical complications can lead to high mortality rates. Rapid detection methods are, therefore, necessary to detect Strongyloides infection in POCT with the advantage of being applicable in a developing country. METHODS: Two hundred POCT have been enrolled in this study to determine the rate of infection with Strongyloides. Three different groups of POCT (Strongyloides infected, non-infected, infected with other parasites) were used to evaluate the antibodies detection capability of two serological techniques (enzyme-linked immunosorbent assay (ELISA) and gelatin particles indirect agglutination (GPIAT) against the results of the gold standard agar plate culture (APC). RESULTS: With APC, the infection rate of Strongyloides stercoralis in POCT was 9.5% (19/200). POCT with Strongyloides infection displayed related risk factors (job, rural settlements, and soil contact) for infection combined with the subtropical nature of Ismailia Governorate. With regard to serology, ELISA detection results were poor compared to APC with sensitivity and specificity of 42.1% and 82.6%, respectively, and positive and negative predictive values of 72% and 30%. GPIAT appeared to be closely related to APC with sensitivity and specificity of 89.4% and 81.8%, respectively, and positive and negative predictive values of 80.9% and 96.7%. Statistical moderate correlation was detected between GPIAT and ELISA. CONCLUSION: The GPIAT technique is more convenient, easier, cheaper and faster to rule out the infection of Strongyloides in POCT. It might be the test of choice for routine immunodiagnosis of human strongyloidiasis.

Anti-Blastocystis Activity In Vitro of Egyptian Herbal Extracts (Family: Asteraceae) with Emphasis on Artemisia judaica.

Achillea fragrantissima (Forssk.) Sch. Bip. (known as Qaysoom), Echinops spinosus L. (known as Shoak Elgamal) and Artemisia judaica L.(known Shih Baladi) are members of the Asteraceae family known for their traditional medical use in Egypt. The ethanol extracts of these plants were evaluated for their efficacy against a protozoan parasite (Blastocystis). Two different molecular subtypes of Blastocystis were used (ST1 and ST3). Significant growth inhibition of Blastocystis was observed when exposed to both A. judaica (99.3%) and A. fragrantissima (95.6%) with minimal inhibitory concentration (MIC90) at 2000 µg/mL. Under the effect of the extracts, changes in Blastocystis morphology were noted, with the complete destruction of Blastocystis forms after 72 h with the dose of 4000 µg/mL. Different subtypes displayed different responses to the herbal extracts tested. ST1 exhibited significantly different responses to the herbal extracts compared to ST3. A. judaica was selected as the herb of choice considering all of its variables and because of its effective action against Blastocystis. It was then exposed to further fractionation and observation of its effect on ST1 and ST3. Solvent portioned fractions (dichloromethane (DCM), ethyl acetate (EtOAc) and n-hexane) in A. judaica were found to be the potent active fractions against both of the Blastocystis subtypes used.

The impact of water crises and climate changes on the transmission of protozoan parasites in Africa.

Waterborne protozoa (WBP) are important cause of several outbreaks all over the world. The report system of WBP in Africa is weak. More than one third of African countries (21/54) reported WBP with absent reports in the remaining countries (33/54). The top reported WBP were Cryptosporidium, Giardia, FLA and Entamoeba contaminating different African water resources. Other protozoa were less documented even though it is abundant and robust. More than one protozoa were detected in contaminated African water including drinking sources, a prediction index to popular epidemics and real presence of undocumented WBP outbreaks. Risk factors in Africa were observed to be abundant and multi-factorial 'socioeconomic, governmental, pathogen in water and climate change. Climate change is an important factor impacting Africa. Increasing droughts in Africa with other extreme weather events will lead to water crises. Incidence and transmission of WBP will change, with new manifested strains/species. Recognizing future consequences of water crises in Africa are important. Governments and population unity will be needed to protect against expected raise and spread of WBP diseases and water shortages.

Antiprotozoal activity of magnesium oxide (MgO) nanoparticles against Cyclospora cayetanensis oocysts.

Outbreaks of Cyclospora cayetanensis infection have been linked to consumption of food and water contaminated by oocysts that can survive both physical and chemical disinfectants. Magnesium oxide (MgO) nanoparticles (NPs) can be potentially used in food as bactericides. In this study, C. cayetanensis pre- and post-sporulated oocysts were exposed to MgO NPs with different doses ranging from 1.25-25 mg/ml. With comparison to control, the antiprotozoal activity of MgO NPs was evaluated by identifying the median effective concentration dose (EC50), lethal concentration dose (LC90), microscopically changes on treated oocysts and rates of sporulation. Among pre- and post-sporulated oocysts, MgO NPs ≥ EC50 was observed after 24 h at concentrations 10 and 12.5 mg/ml, respectively, while ≥ LC90 was observed after 24 h, 48 h and 72 h at concentrations 15, 12.5 and 10 mg/ml, respectively. MgO NPs treated oocysts showed abnormal morphological changes such as an increase in size, wall injury, deposition of vacuolated homogenous particles in the cytoplasm, evacuation of oocyst's contents, and collapse. Sporocysts of treated oocysts were noticed to be peripherally shifted. Sporulation failure of treated oocysts achieved ≥90% after 24 h and 72 h of incubation with 15 and 12.5 mg/ml, respectively, while it was 10.1% among non-treated. All the differences were statistically significant. Our results demonstrated that MgO NPs has a significant anti-Cyclospora effect on both unsporulated and sporulated oocysts, especially considering that it could be biologically synthesized, that way it can be used safely as a preventive agent in food and water disinfectant treatment.

Comparison of current methods used to detect Cryptosporidium oocysts in stools.

In this review all of the methods that are currently in use for the investigation of Cryptosporidium in stool material are highlighted and critically discussed. It appears that more qualifications and background knowledge in this field regarding the diagnosis of the Cryptosporidium parasite is required. Furthermore, there is no standardization for the protocols that are commonly used to either detect oocysts in faeces or to diagnose the Cryptosporidium infection. It is therefore necessary to initiate further education and research that will assist in improving the accuracy of the diagnosis of Cryptosporidium oocysts in the faecal micro-cosmos. Where ambient concentrations of oocysts are low in stool material, detection becomes a formidable task. Procedures for ring tests and the standardization of multi-laboratory testing are recommended. It is also necessary to enhance the routine surveillance capacity of cryptosporidiosis and to improve the safety against it, considering the fact that this disease is under diagnosed and under reported. This review is intended to stimulate research that could lead to future improvements and further developments in monitoring the diagnostic methodologies that will assist in harmonizing Cryptosporidium oocysts in stool diagnosis.

An overview of methods/techniques for the detection of Cryptosporidium in food samples.

Cryptosporidium is one of the most important parasitic protozoa of concern within the food production industry, worldwide. This review describes the evolution and its development, and it monitors the methodology that has been used for Cryptosporidium in food material since 1984, when the first publication appeared regarding the detection of Cryptosporidium parvum in food materials. The methods that are currently being used for the detection of Cryptosporidium oocysts in food material (mainly vegetables) and all of the other available published methods are discussed in this review. Generating more consistent and reliable data should lead to a better understanding of the occurrence, transport and fate of the oocysts in food material. Improvements in monitoring and developing effective methodology, along with food security, offer more practical possibilities for both the developed and developing worlds.

Predominance of Giardia lamblia assemblage A among iron deficiency anaemic pre-school Egyptian children.

Intestinal parasites and nutritional deficiency can coexist and influence each other. This study aimed to clarify the association between Giardia genotypes and presence of iron deficiency anaemia (IDA) among pre-school Egyptian children. Two groups (IDA and non-anaemic) of giardiasis children (44/group) were selected according to their recovery response after treatment of giardiasis. Each group included 24 and 20 gastrointestinal symptomatic and asymptomatic, respectively. Giardia human genotypes were performed by intergenic spacer (IGS) gene based polymerase chain reaction (PCR) with high-resolution melting curve (HRM). PCR/HRM proved that Tms of assemblage A and B ranged from 79.31 ± 0.29 to 84.77 ± 0.31. In IDA patients, assemblages A and B were found among 40/44 (90.9 %) and 4/44 (9.1 %), respectively, while in non-anaemic patients, assemblages A and B were found in 10/44 (22.7 %) and 32/44 (72.7 %), respectively, beside two (4.6 %) cases had mixed infection. The difference was statistically significant. No significant relation was found between symptomatic or asymptomatic assemblages and IDA as assemblage A was found in 21/24 (87.5 %) and 19/20 (95 %) of symptomatic and asymptomatic, respectively, while 3/24 (12.5 %) and 1/20 (5 %) of assemblage B were symptomatic was asymptomatic, respectively. A significant relation was found between assemblage A subtypes distribution among IDA patients as AI and AII were detected on 23 (52.3 %) and 16 (36.4 %) of patients, respectively, while one case (2.3 %) had mixed infection. In conclusion, assemblage A is predominant among IDA giardiasis children suggesting its role in enhancing the occurrence of IDA while B has a protective role.