Assessment of the potential occurrence of Cryptosporidium species in various water sources in Sharqia Governorate, Egypt.

Cryptosporidium species are enteric apicomplexan parasites associated with diarrhoeal disease in humans and animals globally. Waterborne outbreaks resulting from contamination with the infective oocysts are common worldwide. Updated reports on waterborne protozoal infections are needed to identify emerging pathogens and susceptible populations. Therefore, this study aimed to assess the current profile of Cryptosporidium contamination of various water sources in Sharqia Governorate, Northeastern Egypt. For this purpose, eighty samples were collected from five different water types (canal, tap, tank, filtered, and groundwater), distributed in four major cities (El-Hessenia, Fakous, Zagazig, and Belbies) in Sharqia Governorate. All water samples were examined using conventional microscopy, ELISA, and real-time PCR (RT-PCR) techniques. Based on microscopic analysis, the Cryptosporidium protozoan was identified in 25% of the tested water samples. The RT-PCR assay has allowed for the quantification of Cryptosporidium oocysts in different types of water. Canal water exhibited the highest Cryptosporidium contamination levels (mean = 85.15 oocysts/L), followed by water tanks (mean = 12.031 oocysts/L). The study also provided a comparative evaluation of ELISA and RT-PCR for the diagnosis of Cryptosporidium infection. RT-PCR performed better than ELISA in terms of analytical accuracy (97.50% vs. 86.25%) and specificity (100% vs. 83.33%). However, ELISA showed a higher sensitivity (95.00% vs. 90.00%) for Cryptosporidium recovery. Our findings could serve as a platform for further investigations into the potential risks associated with water contamination in Sharqia Governorate. Supplementary Information: The online version contains supplementary material available at 10.1007/s12639-024-01675-1.

The Impact of Parasitic Infestation on Nutritional Status and Micronutrients among Children.

Background: Micronutrient deficiency is a great problem that is augmented by infection and poor nutrition. Iron, zinc, and selenium are trace elements needed for human growth. Objective: To investigate the impact of parasitic infection on nutritional status and serum iron, zinc, and selenium in children attending Pediatrics Outpatient Clinic of Zagazig University Hospitals. Subjects and Methods. A case-control study included 140 parasitic infected children and one hundred age- and sex-matched controls. Anthropometric measures were evaluated using specific Egyptian growth charts. Parasites were detected in stool specimens using standard microscopic methods. Atomic absorption spectrophotometer was used for the detection of serum iron, zinc, and selenium. To examine the statistical relationship between intestinal parasitic infection and the relevant variables (gender, residence, socioeconomic status, and age group), the nonparametric chi-square (χ2) test was used. Data were analyzed statistically using SPSS version 25. Results: Parasitic infected children showed a statistically significant low weight for age, height for age, and BMI. Serum iron, zinc, and selenium were significantly lower in parasitic infected children than controls. Serum iron, zinc, and selenium have significant positive correlations with weight, height, and BMI, respectively. Conclusion: Studied serum micronutrients especially zinc and iron and anthropometric indices were significantly lower in parasitically infected children.

Investigating the Occurrence of Soil-Transmitted Parasites Contaminating Soil, Vegetables, and Green Fodder in the East of Nile Delta, Egypt.

Background: Food-borne parasites are major sources of human and animal illness, posing severe health risks in places with contaminated soil, poor water quality, cleanliness, and poor sanitation. The usage of untreated organic fertilizers arising from the excreta of the parasites' definitive hosts either man or animal pollutes the agricultural soil and is reflected in its products of vegetables and green fodders causing serious health problems. Therefore, to the best of our knowledge, this will be the first study that investigated the combination of parasitic contamination of the agricultural soil and its products of raw eaten vegetables and green fodder in East Nile Delta, Egypt. Aim: The purpose of this study was to investigate the type and degree of contamination caused by parasites in regularly used raw vegetables, green fodder, and soil samples collected from open fields in Egypt's East Nile Delta. Study Procedures. A cross-sectional study comprised a simple random collection of 400 soil samples, 180 green fodder samples, and as well as 400 vegetable samples, including lettuce, radish, coriander, parsley, dill, watercress, tomatoes, green pepper, cucumber, and carrot, that were gathered throughout one year period from January to December 2021 to represent all seasons (winter, spring, summer, and autumn). The research locations were chosen from various open green fields and farming regions in Egypt's East Nile Delta producing ready-to-eat vegetables for human consumptions and planting green fodder for animal feeding. Concentrations, including sedimentation, and flotation, and staining techniques were used to recover the greatest number of parasitic life forms. The parasitic structures discovered were identified using biometric and imaging data and compared with known parasite morphology. Statistical analysis was performed with the SPSS software version 22 (IBM, Chicago, IL, USA). Data were presented in numbers and percentages. P-values equal to or less than 0.05 were considered to be statistically significant. The difference in parasitic contamination among the different categories was compared using the chi-square test. Results: In this investigation, 243 out of 400 soil samples (60.7%) confirmed positive for parasitic contamination (P < 0.05). Various parasitic life forms were significantly found in 249 out of 400 (62.25%) of the vegetable samples, with (65.1%) of them harboring one parasite species, whereas 9.2% significantly contained up to three parasites. Ascaris eggs, Trichuris eggs, and Giardia cysts were the most prevalent parasites, which were predominantly isolated from vegetables with uneven surfaces. 109 of 180 (60.0%) green fodder samples confirmed insignificantly positive for parasitic pollution. The proportion of parasite contamination in vegetable samples was insignificant although the highest was in spring (29.3%), followed by summer (27.7%), whereas it is significant in autumn (24.5%). The prevalence rate was the lowest in winter (20.1%). Conclusion and Recommendations. Our findings demonstrated a significant load of parasites notably the soil-transmitted parasitic infection in raw vegetables and green fodder cultivated in open fields as well as in their mother soil in the east of the Nile Delta, Egypt. These results confirm the urgent need to deploy strict control measures to the soil, especially during the pre-harvest period of raw eaten vegetables and green fodder, a critical step in reducing food-borne transmission of soil-transmitted parasites to man and animals.

Histopathological, Immunohistochemical and Biochemical Studies of Murine Hepatosplenic Tissues Affected by Chronic Toxoplasmosis.

Toxoplasmosis is a serious health problem in humans and animals resulting from obligatory intracellular invasion of reticuloendothelial tissue by Toxoplasma gondii. The profound pathologic effect of toxoplasmosis is confined to nervous tissue, but many other organs, including the liver and spleen, are insulted. Many molecules like caspase-3, CD3, and CD138 are implicated in the tissue immune response in a trial to alleviate hazardous toxoplasmosis impact. This study aimed to investigate the effect of chronic toxoplasmosis on the liver and spleen tissues of mice using biochemical and histopathological techniques and to detect the activity and level of expression of caspase-3, CD3, and CD138 in these tissues using immunohistochemical labeling. Compared with normal control, altered normal histological features accompanied by inflammatory reaction were recorded in hepatosplenic reticuloendothelial tissues in chronically infected mice. The biochemical profile of the liver has been changed in the form of increased liver enzymes, and oxidative stress has been evidenced by elevated nitric oxide (NO) concentration in liver homogenate. The levels of caspase3, CD3, and CD138 were markedly expressed in the liver and spleen of infected mice. Our findings revealed the persistent effect of latent toxoplasmosis on the host's histological architecture, metabolic, and immunological profile, creating a continued challenging host-parasite relationship.

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.

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.

Murine hepatocytes DNA changes as an assessment of the immunogenicity of potential anti-schistosomal vaccines experimentally.

Hepatic affection by granulomatous inflammation in schistosomiasis suggested that a potential anti-pathology vaccine could be generated based on limiting the presence of hazardous hepatocytes induced apoptosis and caused reduction of granulomas number and size . So, this work is concerned with experimental assessment of the efficacy of different Schistosoma mansoni antigens (SEA, SWAP and combined SEA and SWAP) on murine liver after challenge by Schistosoma infection, histopathological, histochemical and molecular investigations were performed on sixty male laboratory bred Swiss Albino mice. A schedule of vaccination and challenge infection was followed and performed on 6 mice groups (each of ten); control normal (G1), control infected (G2), adjuvant received then infected (G3), SEA + adj. received then infected (G4), SWAP + adj. received then infected (G5) and SEA + SWAP + adj. received then infected (G6).Animals were euthanized 10 weeks post infection.Vaccination efficacy was assessed by histopathological, histochemical and molecular studies on murine hepatic tissues.Results showed that:The combined (SEA + SWAP) antigens were better in reducing the number and diameter of the hepatic granulomas, with more protection of the hepatocytes DNA, in addition to more decrease of hepatocytes induced apoptosis and fragmentation as demonstrated by molecular assay.

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.

THE EFFECT OF STEM CELLS AS AN ADJUVANT ON THE IMMUNOGENICITY OF A POTENTIAL ANTI-SCHISTOSOMAL VACCINE IN MICE.

Vaccination against schistosomes can be targeted towards the prevention of infection and/or to the reduction of parasite fecundity and pathology. However, as eggs are responsible mainly for schistosomiasis pathology, so crude soluble egg antigen (SEA) seems suitable to be used as a potential vaccine. Many studies have provided new insights establishing a role for mesenchymal stem cells (MSCs) in liver regeneration and improvement of schistosomiasis hepatic fibrosis, in addition to the need for standardized and effective adjuvant-vaccine formulations. So, the aim of this work is to evaluate the effect of stem cells when used as an adjuvant of a potential anti-schistosomal vaccine (crude SEA) in murine models. The current work was carried out on 100 mice (30 males for harvesting MSCs + 70 females for seven study groups, each of 10). A schedule of vaccination and challenge infection was followed so, GI (control healthy), G2 (control infected only) infected subcutaneously with S. mansoni cercaria (80-90 Schistosoma mansoni cercariae suspended in 0.2 ml distilled Water), G 3 (FCA then infected) received Freund's complete adjuvant (FCA) then infected, G4 (MSCs then infected) received MSCs then infected, G5 (SEA then infected) received SEA vaccine then infected, G6 (SEA+FCA then infected) received SEA vaccine and FCA then infected, G7 (SEA+MSCs then infected) received SEA vaccine and MSCs then infected. The current work was assessed by histopathological study and morphometric analysis (using H&E and Masson's Trichrome stains) to highlight number, size and type of liver granulomas and percentage of liver fibrosis, immunological and molecular studies (RNA extraction, Re- verse Transcriptase and PCR technique) for detection of interleukin-10 mRNA gene expression in liver tissue by reverse transcriptase & polymerase chain reaction (RT & PCR). The results showed that a- SEA alone as a potential anti-schistosomal vaccine was more or less moderately protective, b- MSCs alone before the infection had mild prophylactic effects, c- MSCs as an adjuvant of the crude SEA increased its capabilities with highly significant results regarding the decrease in granuloma number, size, percentage and density of hepatic fi- brosis, and d-There was significant increase in IL-10 mRNA gene expression on using (SEA+MSCs) (G7) if compared to other tested groups.

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.

Hepatitis C virus infection at Sharkia Governorate, Egypt: seroprevalence and associated risk factors.

Because many persons with chronic hepatitis C virus (HCV) infection are asymptomatic, population based serologic studies are needed to estimate the prevalence of infection and to develop and evaluate prevention efforts. A sample of 1422 individuals was included in the study by using multistage sampling technique. Their age ranged from 4-78 years with a mean age (34.7 +/- 18.5), 782 were males (55%) and 640 were females (45%). Exposures and demographic characteristics were obtained through a predesigned questionnaire. Antibody to HCV was assessed using micro-particle enzyme immunoassay (MEIA) enzyme assay by IMX, and the HCV RNA was tested by Real-time PCR technique using ABI Prism 7700 system. The seroprevalence of antibodies to HCV were 23.4% and 27.4% in urban and rural areas respectively, with an overall prevalence (25.8%). This reflects prior HCV infection but not necessarily a current liver disease. Prevalence was higher among males than females and increased sharply with age, from 4.8% in those < 20 years old to (41.9%) in older ages (> or = 40 years). Those who were not educated and farmers had a significantly high prevalence. The significant predictors of HCV infection were previous parenteral therapy for schistosomiasis (OR = 4.3, 95% CI = 3.6-7.9), among those over 20 years of age (3.5, 2.18-5.8), blood transfusion (4.1, 2.4-6.9), invasive procedures (surgery and endoscopy), and use of contaminated syringes and needles. Also, shaving at community barbers added significance to the model. Exposures not significantly related to HCV seropositivity were gender, active infection with Schistosoma mansoni, sutures or intravenous and urinary catheterization, water pipe "goza" smoking in group.

New concept in histopathological grading and staging of chronic hepatitis C infection at Sharkia Governorate, Egypt.

Hepatitis C virus (HCV) has been estimated by the WHO to infect 170 million patients worldwide, with a high prevalence rate (about 24.5%) among Egyptians. The disease could be presented with variable hepatic lesions ranging from mild inflammation, fibrosis, cirrhosis to even end stage liver disease and hepatocellular carcinoma. The Knodell histology activity index, published in 1981, was the first system of its type and is widely regarded as the benchmark for objective, semi-quantitative reproducible description of the various morphological lesions of chronic hepatitis. Other proposals for semi-quantitative evaluation have followed. In this study, when applying these systems on the present cases (109 liver biopsies taken from Egyptian patients infected with HCV), the authors found that the presented histopathological features may be unusual for any of the known scoring systems. Therefore, they suggested a new system for grading and staging of liver diseases in Egyptian patients infected with HCV. Accordingly, the degrees of necroinflammations are classified into 3 grades (1-3) and the progression of fibrosis is classified into 3 stages (1-3). The reduced numbers of grades and stages proposed in this study may be attributed to the rapid course among Egyptians who differ in environmental circumstances from abroad.

HCV and associated concomitant infections at Sharkia Governorate, Egypt.

The natural history of hepatitis C virus (HCV) infection has a highly variable course. Many patients develop chronic infection, with its consequent risk of cirrhosis, liver failure and hepatocellular carcinoma. A key question is whether patients at high risk of disease progression can be distinguished from those with relatively benign disease course. The disease progression is influenced by other factors such as duration of infection, age at infection, sex, co-infection with hepatitis B virus (HBV), Epstein Bar virus (EBV), cytomegalovirus (CMV), the level of HCV viraemia and its type. Other endemic infections in the community as bilharziasis may have a role in progression of the condition to serious complications. These factors are correlated with newly proposed grades and stages of the disease. The studied (109) cases were divided into 6 groups according to the concomitant infection with HCV. The result proved that groups 1, 3 & 5 had a higher level of viraemia than other groups, and to be the high-risk groups as 56.4% and 34.6% were in G2S2 and G3S3, respectively. All cases of liver cell dysplasia and hepatocellular carcinoma in this study were seen in these groups. The conclusion showed that these factors play an important role in the progression of HCV infection. Death of the patients of this progressive condition occurs in younger age and is more due to liver failure than to HCC.

Non-invasive markers and predictors of severity of hepatic fibrosis in HCV patients at Sharkia Governorate, Egypt.

Liver biopsy is thought mandatory for management in patients with hepatitis C virus infection (HCV) especially for histopathological grading and staging of the disease to assess suitability for treatment and monitoring disease progression. However, tracking of liver disease progression can't rely on repeated biopsies. The study aimed to evaluate two significant items, we try to develop and validate a non-invasive predictive tool to assess hepatic necro-inflammation and fibrosis. Also, to determine factors that associate severity of hepatic pathology in HCV infected Egyptian patients particularly at Sharkia G. The study included 109 patients with detectable HCV by Real Time-PCR. The patients were classified into three different pathological stages and grades according to the new concept of histopathoglical staging and grading. The different clinical, biochemical, virological and ultra-sonographic parameters were assessed and analyzed and the variables that showed significant association with histopathological staging and grading were included in multivariate logistic regression analysis. The regression model revealed that, platelet count, matrix metalloproteinase-9 (MMP-9), portal vein diameter, splenic longitudinal axis, alanine transaminase, aspartate transaminase and viral load were the factors that add significance to the model in decreasing order of significance. From these findings we generate a new score ranged from 0-9. The score model was applied to our patients to assess its validity where it proved to be accurate in discriminating patients with mild inflammation and fibrosis (sensitivity 81.8%, specificity 80.5% and accuracy 80.7%) and more accurate in detecting patients with cirrhosis (specificity 96.6%, sensitivity 80% & accuracy 93.6%) but less accurate in detecting patients with moderate to severe fibrosis (specificity 66.7%, sensitivity 68.7% & accuracy 67.9%). Also the results revealed that, co-infection with schistosomiasis, old age > or = 45 years and positive history of blood transfusion as a source of infection was significantly associated with severe hepatic pathology. It is concluded that, the score model can't completely replace liver biopsy but at least it could be used to substantially reduce the number of liver biopsies done in patients with HCV infection in assessing disease progression during follow up. Also, it can be used to make decisions about treatment in patients who have contraindications to or who refused liver biopsy. Co-infection with schistosomiasis, age > or = 45 and positive history of blood transfusion in patients with HCV warrant special attention with more intensive follow up. These factors may play a major role in forecasting the course of HCV as well as in determining the therapeutic approach in each case.

Experimental demonstration of hepatitis C virus (HCV) in an Egyptian strain of Culex pipiens complex.

Reverse transcriptase (RT) polymerase chain reaction (PCR) was used to detect hepatitis C Virus (HCV) RNA among heads, guts, larvae and eggs of Culex pipiens complex. The mosquitoes were trapped from homes of hepatitis C patients or among the same organs of symbiotic (normal gut bacteria) and aposymbiotic (without gut bacteria) mosquitoes fed HCV positive blood by an artificial membrane feeder. The eggs and larvae resulted from symbiotic females fed HCV positive blood was tested for HCV-RNA. Hepatitis C virus RNA was detected only in heads of symbiotic mosquitoes collected from homes of HCV positive patients at 3h and 6h after feeding. The virus was detected at 3d and 8d after being fed on HCV-RNA positive blood in guts of the same group. The virus was not detected in the eggs or larvae resulted from female mosquitoes fed on HCV-RNA positive blood. The results raise the possibility of the mechanical and/or biological transmission of HCV by Cx. pipiens, and pave the way to the ongoing study on the effect of gut bacteria of Cx. pipiens in a trial to identify an anti-HCV agent.