Detection of Cryptosporidium spp. in Diarrheic Immunocompetent Patients in Beni-Suef, Egypt: Insight into Epidemiology and Diagnosis.

Cryptosporidium species is an important cause of gastrointestinal infections globally. This study aimed to shed light on its role in diarrheic immunocompetent patients in Beni-Suef, Egypt and to compare three diagnostic methods. Two hundred diarrheic patients, 37±16.8 year old, were enrolled. Stool samples were examined by light microscopy, using modified Ziehl-Neelsen stain (MZN) for Cryptosporidium spp. oocysts. Coproantigens were detected by sandwich ELISA. DNA molecular diagnosis was done by nested PCR. PCR yielded the highest detection rates (21.0%), compared to ELISA (12.5%) and MZN staining method (9.5%). The higher infection rates were in 20-40 year-old group, followed by 40-60 year-old. Association between epidemiologic factors was statistically not significant; positivity and gender, clinical manifestations, residence, source or water, or contact with animals. Cryptosporidiosis is an important enteric parasitic infection in Beni-Suef and PCR remains the gold standard for diagnosis.

Molecular seasonality of Giardia lamblia in a cohort of Egyptian children: a circannual pattern.

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.

Electron Microscopic Alterations in Pediculus humanus capitis Exposed to Some Pediculicidal Plant Extracts.

Head lice, Pediculus humanus capitis, infestation is an important public health problem in Egypt. Inadequate application of topical pediculicides and the increasing resistance to the commonly used pediculicides made the urgent need for the development of new agents able to induce irreversible changes in the exposed lice leading to their mortality. The aim of the present work is to evaluate pediculicidal efficacy of some natural products such as olive oil, tea tree oil, lemon juice, and ivermectin separately in comparison with tetramethrin-piperonyl butoxide (licid), as a standard pediculicide commonly used in Egypt. The effects of these products were evaluated by direct observation using dissecting and scanning electron microscopes (SEM). Results showed that after 1 hr exposure time in vitro, absolute (100%) mortalities were recorded after exposure to 1% ivermectin and fresh concentrate lemon juice. The mortalities were decreased to 96.7% after exposure to tea tree oil. Very low percentage of mortality (23.3%) was recorded after 1 hr of exposure to extra virgin olive oil. On the other hand, the reference pediculicide (licid) revealed only mortality rate of 93.3%. On the contrary, no mortalities were recorded in the control group exposed to distilled water. By SEM examination, control lice preserved outer smooth architecture, eyes, antenna, respiratory spiracles, sensory hairs, and legs with hook-like claws. In contrast, dead lice which had been exposed to pediculicidal products showed damage of outer smooth architecture, sensory hairs, respiratory spiracles and/or clinching claws according to pediculicidal products used.

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.

The potential curative and hepatoprotective effects of platelet rich plasma on liver fibrosis in Schistosoma mansoni experimentally infected mice.

Trapped Schistosoma mansoni eggs trigger fibrotic liver disease that can continue to liver cirrhosis and failure. This work evaluates the outcome of platelet rich plasma (PRP) on S. mansoni-induced liver fibrosis by intraperitoneal (IP) and intrahepatic (IH) routes with/without Praziquantel (PZQ) treatment. Swiss albino mice (n = 162) were divided into non-infected (n = 66) and infected (n = 96) groups, then subdivided into non-treated and treated subgroups with PRP(IP), PRP(IH) 6th and 10th weeks post-infection, PZQ, PZQ + PRP(IP) and PZQ + PRP(IH) 6th and 10th weeks post-infection. Effects of treatments were evaluated by parasitological, histopathological and Immunohistochemical assessments. In the early assessment (12th week post-infection) of infected-treated groups, the mean granuloma number showed significant reduction in groups treated with PZQ + PRP (IH) 10th week, PRP (IP), PZQ + PRP (IP) and PZQ + PRP (IH) 6th week (33.33%, 33%, 27.77% and 27.22%, respectively). Furthermore, the mean granuloma diameter showed significant reduction in groups treated with PRP (IH) 10th week and PZQ + PRP (IP) (24.17% and 15.5%, respectively). Also, the fibrotic index showed significant reduction in groups treated with PZQ + PRP (IP), PRP (IP) and PZQ + PRP (IH) 6th week (48.18%, 46.81% and 41.36%, respectively). Transforming growth factor ß1(TGF-ß1) expression was in correlation with parasitological and histopathological results. Diminished TGF-ß1 expression was mostly in infected groups treated with PZQ + PRP (IP), PZQ + PRP (IH) 6th week and PRP (IP) (88.63%, 88.63% and 77.27%, respectively). In the late assessment (14th week post-infection) of infected treated groups, TGF-ß1expression was reduced in groups treated with PZQ, PRP (IH) 10th weeks, PRP (IP) (83.33%, 66.66%, 33.33% respectively). PRP showed promising anti-fibrotic effects on S. mansoni-induced liver fibrosis.

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.

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.

In vitro amoebicidal effect of Aloe vera ethanol extract and honey against Acanthamoeba spp. cysts.

This study evaluated in vitro effect of different concentrations of Aloe vera (A. vera) ethanol extract and honey against Acanthamoeba spp. cysts in comparison with chlorhexidine (the drug of choice for treatment of Acanthamoeba infection) at different incubation periods. Four different concentrations of the tested agents were used, 100, 200, 400, and 600 µg/ml for A. vera ethanol extract and 25, 50, 100, and 200 µg/ml for honey. Isolated Acanthamoeba spp. cysts from keratitis patients were incubated with different concentrations of the tested agents as well as chlorhexidine 0.02% (drug control) for different incubation periods (24, 48, 72 h). After each incubation period, the effect of A. vera extract and honey against Acanthamoeba cysts was assessed by counting the number of viable cysts, determining the inhibitory percentage and detecting the morphological alternations of treated cysts compared to non-treated and drug controls. Both A. vera ethanol extract and honey showed a concentration and time-dependent effect on the viability of Acanthamoeba cysts. In comparison with chlorhexidine (the drug control), A. vera ethanol extract possessed a potent cysticidal activity at all tested concentrations throughout different incubation periods, except for concentration 100 µg/ml which recorded the lower inhibitory effect. With increasing the dose of A. vera ethanol extract to 200, 400, 600 µg/ml, the recorded inhibitory percentages of Acanthamoeba cysts viability were 82.3%, 92.9% and 97.9% respectively, after 72 h compared to 76.3% of chlorhexidine. Similarly, honey at concentrations of 50-100 µg/ml gave higher inhibitory effect of 59% and 76.7%, respectively compared to chlorhexidine which showed an inhibitory percentage of 55.7% after 24 h. Meanwhile, the lowest tested concentration of honey (25 µg/ml) gave an inhibitory effect by 47.7-67% which was less than that of chlorhexidine throughout different incubation periods. With increasing the dose of honey to 200 µg/ml, the inhibitory effect was 98.9% after 72 h higher than that of chlorhexidine (76.9%). Using a scanning electron microscope, Acanthamoeba cysts treated by A. vera ethanol extract showed alternations in their shapes with flattening, collapsing, and laceration of their walls. Also, treated cysts by honey were highly distorted and difficult to identify because most of them were shrinkage and collapsed to a tiny size. On the other hand, chlorhexidine showed less structural and morphological changes of Acanthamoeba cysts. A. vera ethanol extract and honey had considerable cysticidal effects on Acanthamoeba cysts. They may give promising results for treatment of Acanthamoeba keratitis.

GIARDIA INTESTINALIS AND HELICOBACTER PYLORI CO-INFECTION: ESTIMATED RISKS AND PREDICTIVE FACTORS IN EGYPT.

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.

PROTOZOA CAUSING FOOD POISONING.

Food poisoning also called foodbome illness, or illness caused by eating contaminated food is a term used to cover an unpleasant range of illnesses. Food poisoning symptoms vary with the source of contamination. Most types of food poisoning cause one or more of the following signs and symptoms: nausea, vomiting, watery diarrhea, abdominal pain and cramps and fever Signs and symptoms may start within hours after eating the contaminated food, or they may begin days or even weeks later. Sickness caused by food poisoning generally lasts from a few hours to several days.

THE ANTIGENIC RELATIONSHIP BETWEEN SCHISTOSOMA MANSONI AND ITS INTERMEDIATE SNAIL HOST.

Schistosomiasis is a public health problem, in many developing: countries including Egypt, Determination of the antigenic relationship between S mansoni and its intermediate snail host (IMH) Biomphalaria alexandrina can open a new field for diagnosis and control of the dis- ease. In the present study infected and non-infected B. alexandrina foot and visceral hump tissue as well as S. mansion crude Ag (SWAg) were fractionated using SDS-PAGE. It's specific and cross reacted protein fractions were determine using EITB versus experimentally prepared mice hyper immune sera (HIS) versus each antigen. After treatment of fractionated S.mansoni crude worm antigens (SWAg) versus HIS produced after vaccination of mice by the same Ag, 8 kda protein fractions ranged from 35-140 kda were reacted specifically. Treatment of fractionated B.alexandrina infected and non-infected foot and visceral hump Ag versus previous HIS revealed presence of common polypeptides bands between SWAg and non-infected snail antigens. The fraction at 135 kda, 68 kda, were detected in all cases, while that at 40-42 kda and that at 35 kda was diagnosed in SWAg and that of infected snails only. The fraction at 68 kda was reacted specifically between SWAg and all tested fractionated snail antigens either that of foot or visceral hump when they treated separately by HIS of mice vaccinated by each snail A eparately. The fraction at 135 kda was common between SWAg and snail (infected and non-infected) visceral hump antigen. The fraction at the level of 110 kda was diagnosed inSWAg, in non-infected foot antigen and visceral hump Ag. The fraction at the level of 46-48 (da are common between SWAg and snail foot and visceral hump Ag after treated by HIS of mice vaccinated by foot Ag, Presence of common antigenic fractions between snail tissues and Schistosoma species can prefer an easily source of antigen valuable for diaguosis or vaccination as well as can be considered as new tool for determination to the snail IMH of new discovered trematode Darasites.

TICK-BORNE INFECTIOUS DISEASES WITH REFERENCE TO EGYPT.

Ticks are hematophagous ectoparasites of man, domestic and street animals, and birds of health and economic hazard worldwide. They transmit many micro-organisms and protozoan infectious diseases and their toxins cause different signs and symptoms that may be fatal according to the infesting tick saliva protein. In Egypt where there are many rural and urban areas, so many genera and species of ticks are encountered. The Egyptian veterinary and agricultural authorities deal with ticks from econom- ic point of view. But from medical point of view, ticks have specific role in transmission of zo- onotic infectious diseases as well as their saliva causes tick paralysis. When dealing with chil- dren from tick infested areas, tick paralysis should be considered in differential diagnosis of the clinically confused diseases as poliomyelitis, myasthenia gravis; Guillain-Barre; paralytic rabies botulism; transverse myelitis and /or the diphtheritic polyneuropathy. Tick-borne relapsing fever is a zoonosis and is endemic in many countries. The two main Borrelia spp. involved in. North America are B. hermsii (in the mountainous West) and B. turicatae (in the southwest). Crimean Congo hemorrhagic fever (CCHF), babesiosis and infantile tick paralysis were identified as emerging diseases disaster. Also, Lyme disease is the commonest worldwide and encountered in Egypt since a long tilne. And now what about other tick-borne diseases, taking into consideration the tick populations is endemic riot only in Egypt but worldwide vectors.

EGYPTIAN EOSINOPHILIC AND INFECTIOUS MENINGOENCEPHA- LITIS AND THEIR IMPACT ON PSYCHOLOGICAL ASPECTS.

Meningoencephalitis is an acute inflammation of the brain and spinal cord & their covering protective membranes. Meningitis can be life-threatening because of the inflammation's proximity to the brain and spinal cord; therefore, the condition is classified as a medical emergency. The commonest symptoms of meningitis are headache and neck stiffness associated with fever, confusion or altered consciousness, vomiting, and an inability to tolerate light (photophobia) or loud noises (phonophobia). Children often exhibit only nonspecific symptoms, such as irritability and drowsiness. If a rash is present, it may indicate a particular cause of meningitis; for instance, meningitis caused by meningococcal bacteria may be accompanied by a characteristic rash. A broad variety of allergic, infectious, neoplastic, and idiopathic diseases are associated with increased blood and/or tissue eosinophilia and range in severity from self-limited conditions to life-threatening disorders. Although accepted upper limits of normal blood eosinophil numbers vary somewhat, a value above 600 eosinophils /microL of blood is abnormal in the vast majority of cases. Generally speaking, there are several possible causes of eosinophils in the CSF; undoubtedly parasitic infection is one of the main causes.

COMPARATIVE STUDY ON IMMUNOBLOT VERSUS PCR IN DIAGNOSIS OF SCHISTOSOMIASIS MANSONI IN EXPERIMENTAL INFECTED MICE.

This study compared PCR and Western blot techniques in diagnosis of schistosomiasis mansoni. Forty Swiss albino mice were used, thirty two mice were infected with cercariae of S. mansoni and eight mice were kept uninfected which were used as a control. Blood was obtained from four infected mice weekly beginning from the 1st week to the 8th week post infection. The study found that PCR was positive from the first week post infection, while Western blot technique was positive from the second week post infection. Thus, PCR diagnosed schistosomiasis mansoni earlier than Western blot technique, but both were able to diagnose.

Toxocara canis and chronic urticaria in Egyptian patients.

Toxocariasis is a common parasitic condition present all over the world with high seroprevalence rates even among asymptomatic individuals. Recent diagnostic techniques have revealed a wider scope of clinical syndromes due to toxocariasis including dermatological disorders and particularly urticaria. The patients with chronic urticaria were enrolled for serological toxocariasis investigations. The results pointed to a possible role of T. canis in-ection in chronic urticaria patients, especially those exposed to an increased risk of environmental exposure to toxocariasis.

The red fox, Vulpes v. aegyptiaca, a new host of leishmania major in Sinai Peninsula.

A leishmanial isolate was obtained from the ear of one red fox or Nile fox (Vulpes v. aegyptiaca) out of eight from North Sinai Governorate. The isolate was typed by the enzymatic variant profiles of the four enzymes, GPI, G6PD. 6PGD and PGM against the three Old World reference strains, L. major, L. tropica and L. donovani and proved to be Leishmania major. This is the second time that L. major has been shown to occur in canine hosts. The list of the mammalian hosts of L. major was reviewed and discussed.

A scanning electron microscopy study of Musca domestica larvae.

The micro-morphology of the house fly, Musca domestica larvae was studied by scanning electron microscopy (SEM). The ultra-structure of the mouth-hooks, oral ridges, spinulation of body segments and posterior spiracles, was photographed and described.