Impact of latent toxoplasmosis on pneumonic and non-pneumonic COVID-19 patients with estimation of relevant oxidative stress biomarkers.

Susceptibility to COVID-19, the most devastating global pandemic, appears to vary widely across different population groups. Exposure to toxoplasmosis has been proposed as a theory to explain the diversity of these populations. The aim of the present study was to investigate the possible association between latent toxoplasmosis and COVID-19 and its probable correlation with markers of oxidative stress, C-reactive protein (CRP) and ferritin. In a case-control study, blood samples were collected from 91 confirmed (48 non-pneumonic; NP, and 43 pneumonic; P) COVID-19 patients and 45 healthy controls. All participants were tested for IgG anti-Toxoplasma gondii antibodies and oxidative stress markers (nitric oxide [NO], superoxide dismutase [SOD] and reduced glutathione [GSH]), and CRP and serum ferritin levels were determined. In COVID-19 patients, IgG anti-T. gondii antibodies were found in 54% compared to 7% in the control group, with the difference being statistically significant (P ˂ 0.001). However, no significant correlation was found between the severity of COVID-19 and latent T. gondii infection. Latent toxoplasmosis had a strong influence on the risk of COVID-19. NO and SOD levels were significantly increased in COVID-19 patients, while GSH levels decreased significantly in them compared to control subjects (P ˂ 0.001 for both values). CRP and ferritin levels were also significantly elevated in P COVID-19 patients infected with toxoplasmosis. This is the first study to look at the importance of oxidative stress indicators in co-infection between COVID-19 and T. gondii. The high prevalence of latent toxoplasmosis in COVID-19 suggests that T. gondii infection can be considered a strong indicator of the high risk of COVID-19.

Green synthesis of zinc oxide/Allium sativum nano-composite and its efficacy against murine cryptosporidiosis.

Cryptosporidiosis is a global health problem threats life of immunocompromised patients. Allium sativum (A. sativum) is one of the therapeutic options for cryptosporidiosis. This study develops green synthesized ZnO-NPs based on A. sativum extract, and assesses its therapeutic application in treating experimental cryptosporidiosis in immunosuppressed mice. FTIR, scanning electron microscopy, and zeta analyzer were used for characterization of bio ZnO-NPs. The morphology of prepared materials appeared as sponge with many pores on the whole surface that allows the feasibility of bio ZnO-NPs for different biological activities. Its structural analysis was highly stabilized with negative charge surface which indicated for well distribution into the parasite matrix. Twenty-five immunosuppressed Cryptosporidium parvum infected mice, classified into 5 groups were sacrificed at 21th day after infection with evaluation of parasitological, histopathological, oxidative, and proinflammatory biomarkers. Treated mice groups with 50 and 100 mg/kg of AS/ZnO-NPs showed a highly significant decline (79.9% and 83.23%, respectively) in the total number of expelled oocysts. Both doses revealed actual amelioration of the intestinal, hepatic, and pulmonary histopathological lesions. They also significantly produced an increase in GSH values and improved the changes in NO and MDA levels, and showed high anti-inflammatory properties. This study is the first to report green synthesis of ZnO/A. sativum nano-composite as an effective therapy in treating cryptosporidiosis which gave better results than using A. sativum alone. It provides an economical and environment-friendly approach towards novel delivery synthesis for antiparasitic applications. RESEARCH HIGHLIGHTS: Green synthesis of ZnO-NPs was developed using A. sativum extract. The morphology of prepared ZnO-NPs appeared as sponge with many pores on SEM The study evaluates its therapeutic efficacy against murine cryptosporidiosis The green synthesized ZnO-NPs significantly reduced percent of oocyst shedding, improved the pathological changes, and showed high antioxidant and anti-inflammatory potentials.

Newly fabricated zinc oxide nanoparticles loaded materials for therapeutic nano delivery in experimental cryptosporidiosis.

Cryptosporidiosis is a global health problem that threatens the lives of immunocompromised patients. This study targets to fabricate and investigate the efficiency of zinc oxide nanoparticles (ZnO-NPs), nitazoxanide (NTZ)-loaded ZnO-NPs, and Allium sativum (A. sativum)-loaded ZnO-NPs in treating cryptosporidiosis. Further FTIR, SEM, XRD, and zeta analysis were used for the characterization of ZnO-NPs and loaded materials. The morphology of loaded materials for ZnO-NPs changed into wrapped layers and well-distributed homogenous particles, which had a direct effect on the oocyst wall. The charge surface of all particles had a negative sign, which indicated well distribution into the parasite matrix. For anti-cryptosporidiosis efficiency, thirty immunosuppressed Cryptosporidium parvum-infected mice, classified into six groups, were sacrificed on the 21st day after infection with an evaluation of parasitological, histopathological, and oxidative markers. It was detected that the highest reduction percent of Cryptosporidium oocyst shedding was (81.5%) in NTZ, followed by (71.1%) in A. sativum-loaded ZnO-NPs-treated groups. Also, treatment with A. sativum and NTZ-loaded ZnO-NPs revealed remarkable amelioration of the intestinal, hepatic, and pulmonary histopathological lesions. Furthermore, they significantly produced an increase in GSH values and improved the changes in NO and MDA levels. In conclusion, this study is the first to report ZnO-NPs as an effective therapy for treating cryptosporidiosis, especially when combined with other treatments that enhance their antioxidant activity. It provides an economical and environment-friendly approach to novel delivery synthesis for antiparasitic applications.

Toxocariasis: potential association with bronchial asthma, and pneumonia among pediatric children.

Toxocariasis is an underestimated geohelminthic infection which shows respiratory changes concurrent with larval migration. The purpose of the present study was to detect Toxocara seropositivity in asthmatic and pneumonic children, and in turn to evaluate its association with the children clinical manifestations, laboratory test results, and sociodemographic risk factors. A total of 50 asthmatic, 50 pneumonic children and 50 healthy controls were subjected to stool analysis by direct wet mount and concentration techniques to exclude possible cross reactivity. Blood samples were collected for complete blood count and assessment of eosinophil count. Sera were examined for anti-Toxocara IgG antibodies, and measurement of total IgE level. Anti Toxocara IgG was detected in 27.3% (41/150) of the studied children. It was significantly higher in asthmatic group compared to controls (26%. p value = 0.033) and significantly highest in pneumonia group compared to both bronchial asthma group (46%. p value = 0.030) and control group (10%. p value = 0.001). There was a significant association between anti Toxocara IgG seroprevalence and each of eosinophilia, total IgE and both combined. Anti Toxocara IgG showed significant higher percentage in asthmatic children who recorded history of soil contact and pets contact as compared to control and pneumonic groups. Toxocara IgG seropositivity was highly associated with fever, cough, wheezes and dyspnea with statistical significance. Toxocara seropositivity has to be considered as a vital associated factor for asthmatic and pneumonic children, and eventually better to be considered in differential diagnosis by pediatricians. Further studies are still needed to explore the correlation between toxocariasis and different patient categories.

Detection of unusual Cryptosporidium parvum subtype in patients with gastrointestinal cancer in Egypt.

While the importance of cryptosporidiosis in immunocompromised persons is well known, the prevalence of Cryptosporidium spp. in cancer patients is not clear. The current study was designed to assess the occurrence and genetic characteristics of Cryptosporidium spp. in patients with gastrointestinal (GI) cancer in Egypt. Stool samples were collected from 100 patients with GI malignancies and 20 healthy individuals without any GI manifestations (control group). They were screened by microscopy and the immunochromatographic RIDA®QUICK Cryptosporidium kit. Subtyping of Cryptosporidium spp. was conducted by sequence analysis of the glycoprotein 60 (gp60) locus. Sociodemographic, environmental data and information on GI symptoms, cancer types, and clinical treatment were obtained via a questionnaire. By microscopy and RIDA®QUICK, only 7% (7/100) of GI cancer patients were positive for Cryptosporidium, compared with 40% (40/100) by gp60 nPCR. No positives were obtained from the control group. Male sex (P = 0.02) and younger age (P = 0.004) were major Cryptosporidium risk factors for infection. The occurrence of Cryptosporidium was also significantly more frequent (P = 0.003) in watery stool samples. Sequence analysis of the gp60 amplicons (~ 400 bp) identified a novel C. parvum subtype with nine TCA repeats and eleven ACATCA repeats. A formal subtype designation could not be made due to the short sequence length. More studies should be conducted to verify the common occurrence of this unusual C. parvum subtype and establish its genetic identity.

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

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

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

BACKGROUND: Nanotechnology has been manufactured from medicinal plants to develop safe, and effective antischistosmal alternatives to replace today's therapies. The aim of the study is to evaluate the prophylactic effect of ginger-derived nanoparticles (GNPs), and the therapeutic effect of ginger aqueous extract, and GNPs on Schistosoma mansoni (S. mansoni) infected mice compared to praziquantel (PZQ), and mefloquine (MFQ). METHODOLOGY/PRINCIPAL FINDINGS: Eighty four mice, divided into nine different groups, were sacrificed at 6th, 8th, and 10th week post-infection (PI), with assessment of parasitological, histopathological, and oxidative stress parameters, and scanning the worms by electron microscope. As a prophylactic drug, GNPs showed slight reduction in worm burden, egg density, and granuloma size and number. As a therapeutic drug, GNPs significantly reduced worm burden (59.9%), tissue egg load (64.9%), granuloma size, and number at 10th week PI, and altered adult worm tegumental architecture, added to antioxidant effect. Interestingly, combination of GNPs with PZQ or MFQ gave almost similar or sometimes better curative effects as obtained with each drug separately. The highest therapeutic effect was obtained when ½ dose GNPs combined with ½ dose MFQ which achieved 100% reduction in both the total worm burden, and ova tissue density as early as the 6th week PI, with absence of detected eggs or tissue granuloma, and preservation of liver architecture. CONCLUSIONS/SIGNIFICANCE: GNPs have a schistosomicidal, antioxidant, and hepatoprotective role. GNPs have a strong synergistic effect when combined with etiological treatments (PZQ or MFQ), and significantly reduced therapeutic doses by 50%, which may mitigate side effects and resistance to etiological drugs, a hypothesis requiring further research. We recommend extending this study to humans.

Blastocystis spp. and Giardia intestinalis co-infection profile in children suffering from acute diarrhea.

Giardia intestinalis and Blastocystis spp. are two common zoonotic intestinal parasites responsible for the majority of diarrheic cases, especially in children. The study was planned to determine the impact role of both Giardia and Blastocystis co-infection in children up to 12 years old with acute diarrhea in Beni-Suef, Egypt. Also, to estimate the general prevalence rates and risk factors associated with intestinal parasitic infections. Stool samples collected from 125 children, within a period of 7 months, were examined using direct wet mount and trichrome stain in comparison to in vitro culture on Jones medium (for detecting Blastocystis spp.) and Immunochromatography techniques (for detecting Giardia). At least one type of intestinal parasites was harbored in 72% of the studied children, single infection constituted 37.6% while, 34.4% were polyparasitized children. The predominant parasites involved were Blastocystis spp.(53.6%), followed by Giardia intestinalis (27.2%). Co-infection between both parasites was recorded in 20% of cases. Two risk factors were significantly associated with higher risk of Giardia-Blastocystis co-infection including age category (5-7 years) and family size (≥ 6 members) by univariate analysis (P = 0.004, 0.009 respectively) and also by the multivariate model (OR = 0.526, 0.336). Abdominal pain was the only gastrointestinal manifestation associated with the co-infection using the univariate analysis (P = 0.0427). The present study detects a high prevalence of intestinal parasitic infection; in particular, Giardia-Blastocystis co-infection in diarrheic young children (up to 12 years) in Beni-Suef Governorate, Egypt and recommends further subtypes identification of both parasites.

Human intestinal myiasis caused by Clogmia albipunctata larvae (Diptera: Psychodidae): First report in Egypt.

This is the first case report in Egypt and Middle East region of human intestinal myiasis caused by Clogmia albipunctata larvae. A 36-year-old male patient presented to the Outpatient Clinic of Internal Medicine in Beni-Suef University Hospital complaining of abdominal pain, bloating, and diarrhea intermittent with constipation. Patient's stool sample was referred to the Department of Medical Parasitology for examination of passed larvae and treatment prescription. Examination of larvae by light and scanning electron microscopy confirmed the morphological identification of fourth instar larvae of C. albipunctata. Human intestinal myiasis is an unusual cause of gastrointestinal manifestations.

Assessment of Giardia and Cryptosporidium Assemblages/Species and Their Viability in Potable Tap Water in Beni-Suef, Egypt Using Nested PCR/RFLP and Staining.

BACKGROUND: The protozoan Giardia and Cryptosporidium are responsible for most water-borne diseases all over the world. The extent and number of outbreaks of waterborne diseases suggests a significant risk of their potential transmission via drinking water. This study aimed to document the prevalence and viability of Giardia and Cryptosporidium (oo) cysts in tap water samples in Beni-Suef Governorate, Egypt and to detect the predominant Giardia and Cryptosporidium assemblages/species using nested PCR/ Restriction Fragment Length Polymorphism (RFLP) confirmed by further sequencing of positive samples. METHODS: A total of 80 tap water samples were collected throughout a year from four big centers and filtered using the membrane filtration method. Samples were stained by Lugol's iodine, Modified Zeihl-Neelsen (MZN) (to detect prevalence) and trypan blue stain (to detect viability). Nested PCR-RFLP and sequencing were used for molecular characterizations and genotyping of the detected Giardia and Cryptosporidium. RESULTS: Giardia and Cryptosporidium DNA was detected in 20 (25%) and 29 (36.3%) samples respectively, with predominance of Giardia assemblage B (85%) and C. hominis (75.9%). The prevalence and viability of both parasites (oo) cysts showed seasonality which peaked in summer and were greater in Beba center and in rural areas. CONCLUSION: To our knowledge, no studies have been done in these areas before. The anthroponotic transmission has an important role in giardiasis and cryptosporidiosis epidemiology in this studied area.

Toxoplasma gondii in cancer patients receiving chemotherapy: seroprevalence and interferon gamma level.

Toxoplasma gondii is an opportunistic parasite causing life-threatening diseases in immune-compromised patients. The purpose of the study is to determine the seroprevalence of Toxoplasma gondii in chemotherapy receiving cancer patients in relation to different types of malignancies, and to estimate the level of interferon gamma in Toxoplasma seropositive and seronegative cancer patients and healthy controls. Anti-Toxoplasma IgG and IgM antibodies, and interferon gamma were analyzed in 120 cancer patients receiving chemotherapy (60 having hematological malignancies and 60 with solid organ tumors) and 60 healthy controls using ELISA method. Toxoplasma (IgG and IgM) were determined in (66.7% and 9.2%) of the cancer group compared to (33.3% and 6.7%) of the control group with statistical significance only in IgG seropositivity (p < 0.001, OR = 4). Patients with hematological malignancies had higher IgG seropositivity than solid organ tumors (40% vs 26.7%). The difference between the groups was statistically significant (p = 0.002, OR = 3.5). Median level of interferon gamma was in the same range between cancer patients and control group. However, it was highly elevated in Toxoplasma seropositive (76 pg/ml) than seronegative (44.5 pg/ml) cases with statistical significance (p < 0.001). T. gondii infection remains a major threat to cancer patients and still needs proper screening, diagnosis and treatment.

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

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

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

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

Blastocystis subtypes isolated from irritable bowel syndrome patients and co-infection with Helicobacter pylori.

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.

Case Report of Human Urinary Myiasis Caused by Clogmia albipunctata (Diptera: Psychodidae) with Morphological Description of Larva and Pupa.

BACKGROUND: Urinary myiasis is a form of myiasis caused mainly by larvae of Fannia scalaris, Musca, Sarcophaga, Lucilia, Wohlfahrtia, Calliphora, and rarely by Eristalis and Clogmia albipunctata. METHODS: This report presents a case of female patient complaining of dysuria and frequency of micturition associated with intermittent passage of small, motile, dark-colored worm-like organisms in urine. She was a married housewife aged 24 years old referred from the Tropical Outpatient Clinic of Beni-Suef University Hospital, Egypt. The patient was subjected to a full questionnaire sheet and investigations such as CBC, stool and urine analysis and urinary ultrasonography. Collected larvae and pupae from urine samples were examined macroscopically and microscopically. RESULTS: The examined larvae and pupae belonged to C. albipunctata. Ivermectin was prescribed to the patient with complaint withdrawal and complete disappearance of the larvae from urine. CONCLUSION: This study reports the first case of urinary myiasis caused by C. albipunctata in Beni-Suef Governorate, the second in Egypt and third case worldwide. The study throws some light on the medical importance and management of urinary myiasis.