Molecular prevalence of Cryptosporidium isolates among Egyptian children with cancer.

Immunocompromised individuals especially children with cancer are at risk for acquiring cryptosporidiosis, which can result in severe morbidity and mortality. This work was conducted to evaluate the prevalence of Cryptosporidium parasite and its genotypes in children with cancer. Stool specimens were collected from 145 children in the Oncology unit of Pediatric Department, Zagazig University Hospital, Sharqiyah province, Egypt. Cryptosporidium infection was evaluated using modified Ziehl-Neelsen (MZN) staining and nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. The prevalence of Cryptosporidium infection in oncological children was 29.7% using microscopy and 25.5% using nested PCR. Genotypic characterization showed that 23 (62.2%) had C. hominis, 11 (29.7%) C. parvum, and 3 specimens (8.1%) were mixed infection of both genotypes. Cryptosporidiosis was significantly associated with diarrhea. However, no statistically significant difference was detected between the age, gender, residency, animal contact and malignancy type concerning to Cryptosporidium infection. This study concluded that Cryptosporidium is a prevailing opportunistic parasite among children with cancer. It should be considered in oncological patients especially those suffering from diarrhea which requires proper management to reduce its complications.

Effect of nanoparticles on the therapeutic efficacy of praziquantel against Schistosoma mansoni infection in murine models.

Praziquantel (PZQ) is the main treatment of Schistosomiasis mansoni. However, resistance to it was described. So, there is a necessity to develop novel drugs or to enhance the present drugs. This work aimed to assess the efficacy of PZQ alone and when loaded on liposomes in treatment of S. mansoni infection by parasitological and histopathological studies in experimental murine models. 112 male laboratories bred Swiss Albino mice were used in this work. They were divided into four groups: Group 1: control group; Group 2: infected then treated by PZQ (500 mg/kg) at 7, 30 and 45 days post infection; Group 3: infected then treated by liposome encapsulated PZQ (lip.PZQ) (500 mg/kg) at 7, 30 and 45 days post infection; Group 4: infected then treated by free liposomes at 7, 30 and 45 days post infection. The results showed that G3 caused the highest significant reduction of the total worm count, eggs/gram liver tissue and intestine (97.2%, 99.3%, 99.5%) respectively. Followed by G2 (85.1%, 97.6%, 89.8%) respectively. Regarding the histopathological studies, G3 showed the highest significant reduction in number and diameter of hepatic granuloma (97.6% and 98.1%), followed by G2 (77.2% and 75%) when compared to other groups. In conclusion, lip.PZQ is more effective than free PZQ from all aspects especially when administered 45 days PI.

House dust mites among allergic patients at the Allergy and Immunology Unit, Zagazig University: an immunologic and serologic study.

In the twenty-first century, the occurrence of allergic diseases has increased. Prevention and control of house dust mites (HDMs) are required as they play a major role in allergic conditions. The present work aimed to detect HDM allergy (Dermatophagoides pteronyssinus and Dermatophagoides farinae) among allergic patients attending the Allergy and Immunology Unit, Zagazig University. Ninety-six patients with a history of allergic diseases were included in this study. They were examined for allergy to D. pteronyssinus and D. farinae using different diagnostic tools: the skin prick test (SPT) and measurement of specific IgE antibodies to HDM allergen extracts. Ninety-six allergic patients were recruited in this study [60 females (62.5%) and 36 males (37.5%) aged between 5-60 years]. SPT (81.2 and 79.2%) and IgE (70.9 and 75%) gave positive results for both D. pteronyssinus and D. pteronyssinus, respectively. The common risk factors were use of cotton bedding > 10 years old, older homes > 20 years, crowded homes, family history, home dampness and homes at the ground floor. It was concluded that allergies to D. pteronyssinus and D. farinae contribute to allergic diseases in Zagazig City. Use of the SPT and IgE level is a promising diagnostic tool in the diagnosis of D. pteronyssinus and D. farinae.

The impact of anthelminthic therapeutics on serological and tissues apoptotic changes induced by experimental trichinosis.

Trichinosis is a sharable parasitic disease caused by Trichinella spp., the disease occurred on eating inappropriate cooked pork infected by the parasite encysted larvae. This study aimed to evaluate experimentally the impact of treatment by thiabendazole, praziquantel (PZQ) and prednisone on T. spiralis induced parasitological, serological and apoptotic changes. Forty albino rats were infected orally each by ± 1000 larvae, divided into four groups each of 10 rats, group (A) infected control, group (B) thiabendazole tested, group (C) PZQ tested and group (D) prednisone tested. On the seventh and 40th days post-infection, all groups were evaluated parasitologically by the number of the intestinal worms and the muscular encysted larvae, while IFN-γ and TNF-α were estimated by ELISA, histopathological and histochemical assessment of the tissue changes during both phases were performed by different stains. In conclusion, thiabendazole was a potent and curable drug, it showed nearly 100% efficacy on intestinal worms, highly significant variations in cytokines levels during both the intestinal and muscular phases, while it induced moderate effects on encysted muscular larvae number, In addition it ameliorated myocytes apoptotic changes induced by trichinosis.

Therapeutic potential effect of bone marrow-derived mesenchymal stem cells on chronic liver disease in murine Schistosomiasis Mansoni.

Some reports have shown that mesenchymal stem cells (MSCs) therapy could ameliorate chemically-induced hepatic fibrosis. This research assesses the therapeutic action of bone marrow mesenchymal stem cells (BM-MSCs) on chronic diseased liver in Schistosoma mansoni infected mice. All infected female mice divided into three groups, one group (15 mice) treated with oral praziquantel (PZQ), second group (15 mice) received intravenous injection of BM-MSCs and third group (15 mice) treated with both MSCs + PZQ. Two control groups (15 mice each) subdivided into one infected and second healthy one. BM-MSCs were obtained from bones of both femur and tibia of male mice (30 mice), then cultured and characterized morphologically by detection of CD105 by flow cytometer. Liver tissues for all groups were examined histopathologically. Measuring of the collagen 1 gene expression was done by real-time PCR and immunohistochemical study to detect stem cells differentiation for detection of MSCs engraftments in liver tissue. MSCs treatment caused marked improvement and regression of fibrosis, and prevents deposition of collagen and reduced the expression of collagen 1 gene in infected mice on their liver tissues, especially when used with PZQ in mice treatment. It can be concluded that, MSCs is a good therapeutic method for liver fibrosis caused by S. mansoni infection.

Assessment of spiramycin-loaded chitosan nanoparticles treatment on acute and chronic toxoplasmosis in mice.

Toxoplasmosis is a zoonotic parasitic disease with worldwide distribution. Chitosan is a natural polymer which is commonly used in the production of nanomedicines. It is known to enable higher drug permeation, being biocompatible and has very low toxicity, besides its antimicrobial effects. Our study aimed to assess the effect of spiramycin-loaded chitosan nanoparticles (SLCNs) in treatment of acute and chronic toxoplasmosis in mice. 200 male Swiss albino mice were included in our study, divided to two main groups; Toxoplasma gondii RH strain infected group and ME49 strain infected group, each main group was subdivided into four subgroups; subgroup I: infected control, subgroup II: infected and received chitosan nanoparticles (CS NPs); 20 µg of CS NPs in 100 µl of PBS/mouse/dose, subgroup III: infected and treated with spiramycin (Rovamycin); 100 mg/kg/day, subgroup IV: infected and treated with 100 mg/kg/day spiramycin-loaded chitosan nanoparticles. Effect of treatment was assessed parasitologically and histopathologically. It was noticed that SLCNs significantly decreased the mortality rate of infected mice with both strains compared to high mortality rate of mice in the infected control subgroups. Moreover, there was a significant decrease in the number of organisms of SLCNs treated subgroup as compared to the other subgroups. Histopathological studies showed a marked improvement of the pathological pictures of brain, liver, spleen and eye in the subgroup received SLCNs as opposed to other groups. In conclusion, the present study revealed that loading of spiramycin on chitosan nanoparticles increased its antiparasitic effect on acute and chronic T. gondii infection.

Molecular approach for detecting early prepatent Schistosoma mansoni infection in Biomphalaria alexandrina snail host.

The present study aimed to evaluate a polymerase chain reaction (PCR) assay used for detection of Schistosoma mansoni infection in Biomphalaria alexandrina snails in early prepatent period and to compare between it and the ordinary detection methods (shedding and crushing). Biomphalaria alexandrina snails are best known for their role as intermediate hosts of S. mansoni. DNA was extracted from infected snails in addition to non-infected "negative control" (to optimized the efficiency of PCR reaction) and subjected to PCR using primers specific to a partial sequence of S. mansoni fructose-1,6-bus phosphate aldolase (SMALDO). SMALDO gene was detected in the infected laboratory snails with 70, 85, and 100 % positivity at the 1st, 3rd, and 7th day of infection, respectively. In contrast, the ordinary method was not sensitive enough in detection of early prepatent infection even after 7 days of infection which showed only 25 % positivity. By comparing the sensitivity of the three methods, it was found that the average sensitivity of shedding method compared to PCR was 23.8 % and the average sensitivity of crushing method compared to PCR was 46.4 % while the sensitivity of PCR was 100 %. We conclude that PCR is superior to the conventional methods and can detect positive cases that were negative when examined by shedding or crushing methods. This can help in detection of the areas and times of high transmission which in turn will be very beneficial in planning of the exact timing of the proper control strategy.

The relation between the prevalence of soil transmitted parasites in the soil and among school children in Zagazig district, Sharkyia Governorate, Egypt.

Soil transmitted helminths are highly prevalent throughout the developing countries of the world and Egypt is categorized as one of the developing countries. This study aimed to detect the relation between the presence and prevalence of STH in the soil and their reflection on the prevalence among school-children living in rural and urban areas in Zagazig district, Egypt. The present study was carried out on (859) school-children aged 6-13 years in Zagazig district all over the year 2013. They were subjected to stool examination. Out of them 100 suspected children were serotested for anti-TES IgG antibodies Toxocara spp. Soil samples also were collected from their places and regions. The results showed overall prevalence of parasitic infections was 21.07 % for helminthic infection and 39.46 % for protozoa. The prevalence of parasitic infections was more among children in rural areas (28, 49.2, 11.1 %) than those who live in urban areas (6.4, 18, 2.2 %) for helminths, protozoa and Toxocara as the most prevalent helminth respectively. Highly significant association was found between soil-transmitted helminthic infections detected by stool examination of the studied children group and the prevalence of geohelminthes in the examined soil samples, taken from residency areas of those children. This association was missed regarding protozoal infections. According to the results, low level of education and consequently poor socio-economic and hygienic conditions of families appear to be powerful determinants of infection. Thus, improving environmental sanitation is imperative for the control of soil-transmitted helminths.

Geohelminths distribution as affected by soil properties, physicochemical factors and climate in Sharkyia governorate Egypt.

Soil-transmitted helminths are mainly a group of parasitic nematodes causing human infection through contact with parasite eggs or larvae; they survive in the warm and moist soil of the tropical and subtropical countries. This study was carried out in Sharkyia governorate from October, 2011 to October, 2013, to correlate between the prevalence and distribution of these parasites in the soil and the physicochemical factors affecting the examined samples of the soil. One hundred and twenty samples of different types of soil (clay, silt, sand) from different localities were collected and examined. Diagnosis of geohelminths was confirmed by the recovery of their eggs and larvae with other protozoa by different parasitological methods. The modified baermann method was found to be more efficient in detection of geohelminths larvae than charcoal culture method. Among the examined sites geohelminths were much more numerous in the soil of rural areas especially in the spring and summer seasons, while the contamination of canal banks by geohelminths was the worst (80 %). An insignificant correlation was reported between the soil texture and the number of positive samples in the examined areas while the relationship was directly proportional among (moisture, PH, organic). It appeared that the most common geohelminthic stage was Toxocara spp. eggs besides other types of protozoa especially Balantidium coli cysts. This suggests that factors other than soil texture are important in the prevalence of geohelminths in the soil e.g. temperature, moisture, PH and organic matter. So, to change some of these factors in a trial to control geoparasites transmission but with keeping the environment should be tried. These results also open the way to further studies to highlight the mutual affection between inhabitants of these sites and the prevalence of these geoparasites.

Studies on the role of tumor necrosis factor- alpha (TNF-α) in hepatocytes induced apoptosis in vaccinated, Schistosoma mansoni-challenged mice.

Tumour Necrosis Factor-alpha (TNF-α) plays a complex role in pathophysiological changes caused by schistosomiasis in the liver cells as induced apoptosis. So, The highlighted experimentally the role of TNF-α in hepatocytes apoptosis, using that as an assessment of the efficacy of antischistosomal vaccination by mixed crude antigens preparations [Cercarial antigen preparation (CAP) + soluble worm antigen preparation (SWAP) + soluble egg antigen(SEA)] by parasitological, histo-pathological and histochemical studies using Feulgen stain of hepatoytes DNA, a serological study also of serum TNF-α level by ELISA. Fifty two laboratory bred Albino male mice, were used in this study. They were classified into four groups (13 mice in each group), G1: normal control, G2 as infected control while G3 supported by Freund's Adjuvant (F. Adj) then infected and G4 vaccinated with combined antigens (CAP, SWAP and SEA) + F. Adj, then infected. Mice were sacrificed by cervical dislocation 9 weeks post infection, parasitological (Kato-Katz thick smear for egg count), histopathologial {haematoxylin and eosin (H&E) staining of hepatic sections}, histochemical (feulgen staining of hepatocytes DNA) and ELISA to estimate serum TNF-α level were performed. The data showed that vaccination with combined antigens showed protective effect on vaccinated then Schistosoma challenged mice, hepatocytes induced apoptosis was directly proportional with the TNF-α serum level, and the protection degree of potential combined vaccine was inversely proportional with serum TNF-α level and induced apoptosis.