Animal hydatid cyst genotypes as a potential substitute for human hydatid cyst as a source of antigen for diagnosis of zoonotichydatidosis.

The Diagnosis of hydatidosis is still an unsolved issue due to difficulties in obtaining of patient's hydatid cyst appropriate for antigen extraction. This study evaluated the suitability of HC protoscolices somatic antigens (HCPsS-Ag) fractions from animal origin to substitute that extracted from HC of patients in diagnosis of hydatidosis using enzyme-linked immunoelectro-transfer blot and Enzyme-linked immunosorbent assay (ELISA). Eight fractions in HC-G6 from patients react specifically versus HC-G6 infected patient's sera. Five of them (28, 32, 38, 59 and 89 Kilo Dalton (KDa) and two of them (28 KDa and 45 KDa) reacted versus HC-G1 and HC-G4 infected sheep and equine sera, respectively. Six fractions in HCPsS-Ag-G1 of sheep react versus HC-G1 sheep infected sera, four (28, 32, 52 and 58 KDa) and two of them reacted versus HC-G6 and HC-G4 infected patient and equine sera, respectively. Two fractions only in HCPsS-Ag-G4 of equine reacted versus infected human and sheep sera. This fraction displayed the same degree of ELISA value versus different infected sera with a significantly perfect classification for kappa agreement and non-statistically significant difference (p ≥ 0.05) for ELISA Optical density values of the positive samples without cross-reaction versus other parasites antibodies in sera. HCPsS-Ag from HC genotypes that developed in humans and animals as HC-G6 and HC-G1 can substitute each other for diagnosis of infection than antigens extracted from non-zoonotic HC-G4. The fraction at 28 KDa is the only fraction that can be extracted from any animals HC and used in diagnosis of zoonotic hydatidosis.

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.

Modified dot-ELISA for diagnosis of human trichinellosis.

This study aimed to modify Dot-Enzyme-linked immunosorbent assay (dot-ELISA) for the diagnosis of human trichinellosis and to compare its performance with indirect ELISA and Western-blot assay (EITB). A total of 175 human serum samples were enrolled in the study. Indirect ELISA was used for the primary diagnosis. EITB versus fractionated 1st larval stage excretory-secretory antigens (TL-1 ESA) revealed three specific protein fractions at MW of 45, 50, and 55 kDa (kDa). Dot-ELISA was performed in two ways. In the first one, sera were dotted on the separated three specific protein fractions, while in the second one the three fractions were eluted, concentrated at one pooled antigen that used in classic dot-ELISA. Both types of dot-ELISA proved absolute (100%) sensitivity and specificity in comparison with the gold standard EITB reaction. While sensitivity of ELISA was 100% and its specificity was 79.5%. The fraction at 45 kDa was the most sensitive one. The use of the pooled antigen improved the test results. The described dot-ELISA is an easy applicable diagnostic tool gathering the benefits of both ELISA and EITB.

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.