Immunodetection of Fasciola gigantica circulating antigen in sera of infected individuals for laboratory diagnosis of human fascioliasis.

Currently, the laboratory diagnosis of human fascioliasis is based on the parasitological examination of parasite eggs in stool specimens and serological detection of specific antibodies in serum samples, which are often unreliable diagnostic approaches. Ideally, a sensitive and specific diagnostic test for Fasciola infection should be based on the detection of circulating Fasciola antigen, which implies active infection. Here, a 27-kDa-molecular-mass antigen was identified in a Fasciola gigantica adult worm antigen preparation, excretory-secretory products, and sera from F. gigantica-infected individuals, and it was not detected in antigenic extracts of other parasites and sera from noninfected individuals. The target antigen was isolated and partially characterized as a protein. Immunoperoxidase staining located the target epitope within teguments and guts of F. gigantica adult worms. The performance characteristics of a newly developed enzyme-linked immunosorbent assay (ELISA) based on F. gigantica circulating antigen detection in serum (FgCA-27 ELISA) were investigated using sera of 120 parasitologically diagnosed F. gigantica-infected individuals and 80 noninfected individuals. The area under the receiving operating characteristic (ROC) curve (AUC) for ELISA was significantly high (AUC = 0.961, P < 0.0001) for discriminating Fasciola-infected and noninfected individuals. The developed assay showed high degrees of sensitivity, specificity, and efficiency (>93%), and a significant correlation (r = 0.715, P < 0.0001) between antigen level and parasite egg count was shown. In conclusion, a 27-kDa Fasciola antigen was identified in sera of F. gigantica-infected individuals. A highly sensitive and specific Fasciola antigen detection assay, FgCA-27 ELISA, was developed for laboratory diagnosis of human fascioliasis.

Clinical, parasitological and social studies on Wuchereria bancrofti in Egypt.

This study was conducted on patients attending Al-Hussein University Hospital and Mansura University Hospitals. Sheets were filled out on each patient (age, residence, occupation, family similar condition, travelling, disease duration, symptoms suggesting acute dermatolymphangitis attacks, frequency and duration. They were 25 lymphoedema patients and 8 controls (17 males & 16 females) and subjected to clinical and parasitological examinations. According to residence, five patients were from Giza Govemorate (18.18%), four from Dakahlia G. (12.12%), four from Gharbia G. (12.12%), five from Menoufia G. (15.15%), four from Sharkia G. (12.12%), two cases from Kafr Elsheikh G. (6.06%) and one patient from Assiut G. (3.03%). In controls, 6 were from Al-Hussein Cairo G. (15.15%), and two cases (6.06) from Al-Abbassia. They were 48% farmers (10 male & 2 female); 4% grocers (1 male); 8% carpenters (2 male); and 40% house wife (10 female). According to lymphedema site: 3 cases (12%) were bilateral lower limbs lymphedema (2 female & 1 male), 13 cases (52%) had right lower limb lymphedema (6 female & 7 male), and 9 cases (36%) had left lower limb lymphedema (4 female & 5 male). They were 18 (72%) with below knee extension of lymphedema (10 male & 8 female) and 7 (28%) above knee extension of lymphedema (3 male & 4 female). Grade of lymphedema was one case (male) with grade II lymphedema (4%), 13 cases (52%) with grade III (7 male & 6 female) and 11 cases (44%) with grade IV (5 male & 6 female). Duration of lymphedema was one case with grade II lymphedema in a period of 3 years, 13 cases with grade III lymphedema and the durations of lymphedema for these cases were (one case with 5 years duration, 3 cases were > 6-9 years,6 cases were > 9-12 years and 3 cases were > 12 years) and 11 cases with grade IV lymphedema and duration of lymphedema for these cases were (one case with 6 years duration, 2 cases were > 6-9 years,3 cases were > 9-12 years and 5 cases were > 12 years).

Seroprevalence of human toxocariasis (visceral larva migrans).

A total of 455 patients who fulfilled the inclusion criteria were included in the study. The enrolled patients were subjected to a questionnaire (including sociodemographic and other risk factors) and thorough clinical examination was done for the patients. Sera were collected from patients and tested for anti-Toxocara IgG antibodies using ELISA. The overall anti- Toxocara seropositive was (7.7%). It was significantly higher than among the randomly selected 30 healthy controls. There were no significant differences between the seropositive and seronegative patients regarding age, sex, educational level and monthly family income of the patient. However, rural residence, poor house, pet's ownership and frequent contact with soil were found to be significant. Patients who had confirmed bronchial asthma were more than 2 times at higher risk of developing toxocariasis (OR, 2.33; 95% CI, 1.09-4.98) than those with other clinical diagnosis (PUO, hepatomegaly or heptosplenomegaly, lymphadenopathy, neurological disorders, gastrointestinal troubles and dermatitis). Patients with eosinophilia were at 149 times greater risk of being Toxocara seropositive compared to those without eosinophilia (OR, 148.7; 95% CI: 53.5-413.3). Multivariate regression analysis showed eosinophilia and contact with soil were the most important predictors of toxocariasis. OD of anti-Toxocara antibodies (ELISA) was significantly positive with eosinophilia level.

Soil-parasites particularly Toxocara eggs in Egypt.

Soil-transmitted helminths (STHs) remain a major threat to the human health throughout the world, mostly in developing nations. Soil is usually contaminated by different parasitic stages through promiscuous defecation of man particularly children. This is more liable to occur in fields, public gardens and sides of streets or canals. Also, feces of domestic and wild animals & birds play a role in soil contamination. This study was conducted at Mansoura city and Batra village & El-Minia city and Zohra village representing the urban and rural areas of Dakahlia Governorate, Lower Egypt and Minia Governorate, Upper Egypt respectively. The most common helminthic stage presents in the soil of the two Governorates was Toxocara eggs. No ova was detected in-door yards in the urban areas, but only in the sides of streets, crop fields and fruit gardens, around houses and canal banks of both Governorates, and the highest intensity was 13-19 eggs/10 gm soil at canal banks at Zohra village, El-Minia Governorate, Upper Egypt.

Past and present situation of human fascioliasis in Dakahlia Governorate, Egypt.

Zoonotic fascioliasis is a problem not only in Dakahlia Governorate, but also in other Egyptian Governorates. This work was conducted in ten centers at Dakahlia Governorate, Egypt. Three thousand cases were randomly selected annually from each center representing five rural and five urban areas. Cases were examined for Fasciola infection by stool examination using, direct smear, formol ether sedimentation and Kato-Katz technique. Infection was higher in rural areas than urban areas. Female /male ratio was 1.5:1. Children between five and ten years old represent the highest infected group. The prevalence of infection was more among illiterate compared to educated individuals examined. Remarkable reduction in the prevalence rate was recorded in the last year of study compared to previous years, which is a good index of excellent control measures conducted by the Egyptian governorate to overcome parasitic infections.

The kinetics of microfilaraemia and antigenaemia status among asymptomatic bancroftian filariasis before and after treatment.

In this study, a total of 54 male patients with asymptomatic cases of lymphatic filariasis were selected from the known endemic governorates; Damietta, Sharkia, Gharbia and Dakahlia. Medical sheets were filled out for each case. Diagnosis was based on detection of circulating microfilariae by specific ELISA. Some were treated with Ivermectin, and Albendazole. The treated cases were parasitologic and serologic evaluated pre-treatment and post-treatment for two years follow-up.

Kinetics of eosinophilia and IgE production in experimental murine toxocariasis.

Eosinophilia and immunoglobulin E (IgE) antibody are considered the first and important elements in host responses to helminth infection. Toxocara canis, which elicits prominent eosinophilia and IgE production in normal mice, may be useful in understanding the kinetics of circulating peripheral blood eosinophils and IgE antibody during infection. The onset, magnitude and duration of peripheral blood eosinophilia and total IgE production after primary T. canis infection in female outbred albino mice was investigated. Mice were either sham inoculated (controls) or were orally infected with 1000 embryonated eggs of T. canis. Patterns in leucocytic changes include significant increase in total WBC count between weeks 6 and 13 post infection (PI) with a peak on week 8 PI. Mice showed eosinophilia between weeks 2 to 17 PI with a peak on week 7. The development of eosinophilia in T. canis-infected mice was accompanied by the release of prominent level of serum IgE between weeks 2 to 21 with a peak at week 6 PI. These findings showed that eosinophilia in T. canis infected outbred mice can be T-cell dependent.

Mirazid alone or combined with Paromomycin in treating cryptosporidiosis parvum in immunocompetent hospitalized patients.

Sixty cryptosporidiosis patients from Mansoura University Hospitals, 36 males and 24 females, with age from few months to ten years (mean age 6.1) were divided into three cross-matched groups of 20 patients each. All patients received the glutamine-based oral rehydration solution with 111 mmol/l glutamine, 20 mg zinc acetate once a day and vitamin A supplementation (200,000 IU) once a day for 2 weeks. For cryptosporidiosis treatment, G1 received Mirazid (10 mg/kg for 2 weeks), G2 received Paromomycin (500 mg qid for 2 weeks), and G3 received a combination of Mirazid (10 mg/kg) and Paromomycin (500 mg) for two weeks. The result was assessed according to the scales: 0 = no improvement, 1 = symptoms began improvement (reduction of diarrhea frequency and stool volume, less abdominal pain, less nausea & vomiting), 2 = diarrhea eradication, 3 = weight gain, 4 = oocyst counts reduction, 5 = reduction in diarrhea and oocyst counts, 6 = eradication of diarrhea and oocysts. G3 showed significantly higher difference than G1 & G2 in the 1st week (p = .036, 0.025 respectively), no significant difference in 2nd week, a significantly higher difference than in G1 (0.003), & G2 (0.006) in 3rd week, and a significantly higher difference than G1 (0.014), & G2 (0.01) in 4th week, but without significant differences in oocyst shedding in the 3 groups.

Biochemical parameters in chronic fascioliasis.

Serum lipids, liver enzymes and total bile acids were measured in chronic Fasciola (cholestatic and non-cholestatic) patients. Variations in the biochemical parameters between infected and control groups were detected and typified by considerably higher serum triglycerides (TG, P < 0.001), alkaline phosphatase (ALP, P< 0.001) and total bile acids (TBA, P < 0.001) in the infected group. For cholestatic patients, TG, very low density lipoprotein (VLDL), aspartate aminotransferase (AST), alanine aminotransferase (ALT), gammaglutamyl transferase (GGT) and ALP were statistically higher (P < 0.001 for each comparison) and total cholesterol (TC); low density lipoprotein (LDL) and high density lipoprotein (HDL), were statistically lower than non-cholestatic patients (P = 0.02, < 0.001, < 0.001 respectively). No statistically significant differences were found for the TC, HDL, VLDL, ALT, AST and GGT between non-cholestatic patients and healthy controls.

Phylogenetic evidence for recombination in SAG5 locus in Toxoplasma gondii.

The present study evaluated the hypothesis that genetic diversity in SAG5 genes was generated by recombination events. Three lines of evidence suggested that recombination occurred in SAG5 genes in T. gondii. The permutation test revealed strong signature of intragenic recombination, pairwise comparisons of nucleotide sequences of SAG5 genes revealed that SAG5A alleles have chimerical structures composed of segments derived through recombination events between different alleles, and phylogenetic trees reconstructed based on SAG5 sequences using neighbor-joining and maximum parsimony methods, showed statistically well-supported consensus clusters of T. gondii strains specific to each SAG5 gene. Topological discrepancies between trees based on the N-terminal variable domain and C-terminal conserved domain sequences, were observed, suggesting intragenic recombinetion between SAG5A and SAG5B/C genes. The results showed that recombination within SAG5 in T. gondii was a major evolutionary mechanism generating both allelic variation at SAG5 locus and contributing to genotypic diversity and to emergence of new T. gondii variants, allowing them to evade the host immune defence mechanism.

Zoonotic hydatidosis in donkeys: post mortum examination in the Zoo, Giza, Egypt.

A total of 17 out of 160 donkeys slaughtered had hepatic hydatidosis as proved macroscopically and microscopically. The epidemiology of zoonotic echinococcosis/hydatidosis was reviewed and discussed.

Mirazid (Commiphora molmol) in treatment of human heterophyiasis.

A total of 3278 patients attended Mansoura University Hospitals' Clinics with gastro-intestinal troubles suggesting parasitosis were examined by direct smear and by Kato-Katz methods for parasites especially Heterophyes heterophyes. Fifty clinically and parasitologically proved pure heterophyiasis patients were given Mirazid as two capsules for 9 successive days on an empty stomach an hour before breakfast. All the cases were subjected to history taking and clinical examination before treatment and were followed-up for four weeks post-treatment. There was an overt clinical and parasitological improvement. A total of 47 out of 50 (94%) were cured. Another course was given to the three patients who were still positive, but only two of them were cured (66.7%). The overall cure rate was 49/50 (98%) and none had any side effect. The history and treatment of this zoonotic parasite was critically discussed.

Commiphora molmol extracts as plant molluscicide against Lymnaea natalensis.

Two extracts from the herbal plant, Commiphora molmol showed a high molluscicidal effect against Lymnaea natalensis. The oil extract was more potent than the oleo-resin. A concentration of 10 ppm of the oil extract killed 100% of L. natalensis after 5th day, but the oleo-resin extract killed 100% of them at a concentration 20 ppm after 5th day.

Diagnostic criteria for house dust mites sensitized allergic patients.

House dust mites (HDM) contain a large number of components that react with IgE in individuals with allergies and induced sensi-tization and allergic diseases. The work aimed to correlate between clinical manifestations of HDM allergic patients and skin prick test (SPT) grading, and to clarify some markers denoting allergic disorders caused by HDM exposure. Thirty allergic patients with positive SPT reaction to HDM were selected, grading of disease severity according to SPT and clinical presentations. The results revealed bronchial asthma (40%), atopic dermatitis (26.7%), rhinitis (20%) and conjuncttivitis (13.3%). Serum ELISA-IgE showed mean level of 398.41 +/- 84.73, 369.87 +/- 86.76, 332.16 +/- 65.57 and 300.25 +/- 79.39 IU/ml respecttively, with highly statistically significant difference between cases and controls (p < 0.001). Eosinophila showed highly significant difference between atopic dermatitis cases and controls (p < or = 0.001).

Ectoparasites of commensal rodents in Talkha Center, Dakahlia Governorate, Egypt.

The ecto-parasites infesting commensally rodents were collected from the different localities in Talkha Center (Dakahlia Governorate) from April 2006 to March 2007. The seasonal abundance of rodent number and rat index was 52 (0.58) in spring, 27 (0.3) in summer, 39 (0.22) in autumn and 17 (0.05) in winter. From 135 rodent species and rat index was Rattus norvegicus N=33 (0.24), R. r. frugivorous N=39 (0.29); R. r. alexandrinus N=48 (0.36) and Mus musculus N=15 (0.11). From 388 ecto-parasite infested rodent collected number and ecto index was fleas N= 114 (0.84 flea/rat), Lice N=93 (0.69 lice/rat), Mites N = 165 (1.2 mite/rat) and larva of ticks N=16 (0.12 tick/rat). No doubt, rodents and their ectoparasites played a serious role in spreading and transmission of zoonotic diseases to human and animal.

Echinococcosis in dogs in urban and rural areas in Dakahlia Governorate, Egypt.

A sample of one hundred and ninety stray dogs was captured from Mansoura city (urban) and three hundreds and fifty stray dogs were captured from Meet El-Korama, Mansheit El-Badawy villages (rural). The total prevalence of E. granulosus was 5%, with a worm burden ranging from 4 to 1010 (mean = 421). The significant prevalence was 6% in the rural area and 3.2% in the urban one. E. granulosus showed higher prevalence in young than old dogs and in males than females but without significant difference in both variants. The overall Echino-ELISA sensitivity was 61.5% and specificity was 97.5%. The major cross reactivity was with Taenia spp., and Diplydium caninum, but neither with Toxocara canis or Trichurus vulpis nor Ancylostoma caninum. There was a negative correlation between ELISA and Echinococcus granulosus burden in dogs.

Correlation of ELISA copro-antigen and oocysts count to the severity of cryptosporidiosis parvum in children.

A total of one thousand and fifty (1050) young children from Mansoura Pediatric Hospital, July 2005 to July 2006 were examined for cryptosporidiosis. Detailed history was taken from each participant, stool examination by direct smear, Formalin-ether sedimentation, Sheather's floatation, modified Ziehl-Nel-seen (MZN) stain, Cryptospordium oocysts count, coproantigen detection by ELISA. Other infections were excluded. Out of 1050 children examined, cryptosporidial oocysts were detected in 90 cases by modified Zeihl Nelseen (MZN) stain, and copro-antigen ELISA added another 10 cases. There was a significant difference between age group (1-2 years), rural locality, nutrition status, and diarrhea regarding Cryptosporidium infection. There was highly significant difference in malnourished children regarding Cryptosporidium oocysts and ELISA-OD. There is a significant positive correlation between oocysts count, ELISA-OD and malnutrition. Only diarrhea had a highly significant difference in relation to other symptoms in both mean oocysts count and ELISA-OD. There was highly significant positive correlation between symptoms, oocysts count and ELISA-OD.

Risk factors contributing to fascioliasis endemicity in a focus in Dakahlia Governorate 1-human host.

Zoonotic fascioliasis is a problem not only in Dakahlia Governorate, but also in other Egyptian Governorates. Two hundreds & twenty patients randomly selected with suggesting manifestations were examined for fascioliasis. A total of 23 (10.4%) were positive by Kato thick smears, of which 21 were from Kafr El-Hessah and two from Oweish El-Hager. The proven human fascioliasis was examined for anti-Fasciola antibodies by IHAT & ELISA (Fhes), haemoglobin level, eosinophils percent, serum bilirubin & liver function tests. IHAT gave 82.61% positive results (19/23), and ELISA gave 100% positive results (23/23). The clinical signs ranged between splenomegaly & ascitis in 4.34% for each up to hepatomegaly in 73.91% & mild fever in 78.26% but 2 cases were asymptomatic. Mild eosinophilia and moderate anaemia were recorded with means of 11 +/- 5.8 and 10 +/- 1.3 respectively. Mean serum bilirubin was not elevated (0.91 +/- 0.51 gm/dl). Liver function tests (AST within normal range in all cases; <40 unit/ml but one patient had ALT above normal; >45 unit/ml).

New trends in diagnosis and treatment of chronic intestinal strongyloidiasis stercoralis in Egyptian patients.

Strongyloidiasis, caused by Strongyloides stercoralis, is diagnosis considered as a challenge to clinician and laboratory technician. Because the auto-infective larvae are difficult to eradicate, one regimen dose may be in-sufficient and re-treatment of patients on two occasions, at 1 and 2 months after the initial treatment dose was recommended. This re-treatment regimen has yet to be proven in clinical trials. This study was performed on 24 patients who completed the study and having Strongyloides larvae in their stool obtained from Mansoura University Hospitals. Each stool sample was examined by direct saline smear, the formalin-ether sedimentation technique and agar plate culture. Patients were treated with Mirazid double course for a month to be followed up by stool examination by traditional method and agar plate culture for three consecutive months. In this study five cases out of 24 were asymptomatic (20.8%). Symptoms include abdominal manifestations as nausea and vomiting (16.7%), epi-gastric pain and nausea (12.5%), generalized abdominal pain (12.5%), chronic diarrhea (16.7%), irregular bowel habit (8.3%), and urticaria with abdominal pain (4.2%). Agar plate culture gave 100% positivity, even in cases were negative by coprological methods either direct smear and/or sedimenttation technique. All cases were cured by Mirazid given for one month except three resistant cases. Only one case responded to repeated course of Mirazid, while the other two cases still had larvae in their stool by agar culture plate. On combined therapy of both Mirazid and Mebendazole, larvae could be eliminated from their stool as approved by agar plate culture.

The liver profile in patients with hepatitis C virus and/or fascioliasis.

In this study, twenty HCV/PCR-RNA positive patients with neither infection nor infestation were 15 males and five females with ages ranging between 17-78 years old. Liver function tests: S. albumin was decreased in 15 patients (75%), total protein decreased in 3 patients (15%), total bilirubin increased in 7 patients (35%), AST/ALT increased in 3 patients (15%). Globulin value increased in 15 patients (75%). A/G ratio decreased in 12 (60%). Ten normal individuals (five males & five females), gave normal findings. However, a 19 years-old female had non significant elevation (0.26 mg/dl) in direct bilirubin. Liver function tests: one patient had increased AST/ALT, Globulin value decreesed (-0.2) in another one (10%) and A/G ratio increased (+0.3 to +0.6) in three (30%) individuals. Twenty randomly selected patients (15 males and five females) HCV/3rd generation ELISA showed ALT elevation in 17 (85%), AST in all (100%), S. albumin decreased in 9 (45%), and increased in 1 (5%). Total protein decreased in 3 (15%), total bilirubin increased in 7 (35%) and direct bilirubin in 4 (20%), AST/ALT value increased in 3 patients (15%), Globulin value increased in 15 patients (75%). A/G ratio decreased in 12 (60%). Three patients had schistosomiasis, one 30 years old male had increased AST/ALT, normal globulin, increased A/G and positive HBs-Ag. The second one was a 33-year-old male had normal AST/ALT, normal globulin, increased A/G and positive HBs-Ag. The third patient was a 19-year-old female with normal AST/ALT and normal globulin, increased A/G and positive HBs-Ag. Two patients had fascioliasis, one was a 20-year-old male with increased AST/ALT, globulin normal, A/G normal and negative HBs-Ag. The second one was a 26-year-old female with normal AST/ALT and normal globulin, increased A/G and positive HBs-Ag. The other four positive HBs-Ag patients were parasite-free. Also, the other HCV/ELISA positive were negative for HBs-Ag. HCV/ELISA may have cross-reacted with HBs-Ag and/or with elevated ALT and gave false positive HCV. The 3rd generation ELISA in detection of HCV was not as sensitive as PCR/RNA. Out of 41 fascioliasis patients (26 males & 15 females), 14 were positive HCV/ELISA (34.1%), but only six were positive HCV/PCR (14.6%). ALT increased in 18 fascioliasis patients (43.9%), AST in 23 patients (56.1%) but, albumin decreased in 7 patients (17.1%), total protein decreased in 5 (12.2%), but total bilirubin increased in 14 (34.1%) and direct bilirubin increased in 2 (4.9%). Liver function tests of 14 fascioliasis and ELISA positive HCV showed AST/ALT increased in 6 (42.9%), globulin increased in 3 (21.4%) and decreased in 6 (42.8%). A/G decreased in 4 (28.6%) and increased in 8 (57.2%). Liver function tests of pure 27 fascioliasis patients showed that AST /ALT increased in 8 (29.6%), globulin increased in one patient (3.7%) but decreased in 10 (37.0%) and A/G ratio increased in 13 (48.1%).