ABSTRACT
Hydatid disease is caused by the larva of Echinococcus granulosus parasite. Mebendazole [MBZ] is used as an alternative choice for the treatment of the disease
Aspartate aminotransferase [AST] is an essential enzyme in amino acid metabolism
The aim of the present study is to evaluate the effect of MBZ on AST activity of hydatid cyst parasite in order to detect enzymatic parameter for drug efficiency. In the present study, AST activity was estimated in the extracts of untreated parasite [hydatid cyst protoscolices] and treated samples by MBZ [100 microg final concentration]
Samples' protein quantity and quality were detected by Bradford and sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE] methods respectively
For the purpose of determining the significant difference between the two independent samples, t-test was conducted. The values of the assayed AST specific activities of treated and untreated parasite samples were measured as 0.18 and 1.53U/ml/mg protein respectively
The difference between AST activities mean values of the two groups proved to be significant [P<0.05]. SDS-PAGE demonstrated protein band of 50 kDa for AST enzyme. Considering the effect of the MBZ drug on AST activity in parasite, it can be concluded that this enzyme is useful for improving the drug efficiency?
ABSTRACT
Hydatidosis is a zoonosis disease caused by the larva of Echinococcus granulosus parasite in man and animals. The Proteases enzymes are necessary for nutrition, migration and evasion of immunity of parasite into host. The aim of this study was to determine the protease activity of hydatid cyst protoscolices [PSC], healthy and cystic liver tissues in order to compare of this biomarker for host and parasite in hydatid disease. In this study, PSC were collected from sheep liver tissue infected with hydatid cysts at an abattoir and washed 3 times with PBS buffer. PSC samples were freeze-thawed and sonicated while collected liver tissues were homogenized. Extract solution samples were centrifuged and stored at - 20degree C. Protease enzymes activity was measured in the extract solutions of PSC and sheep liver tissue samples [healthy and cystic livers]. Samples protein concentrations and protein bands were detected using Bradford and sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE] methods respectively. To determine significant difference between two groups, statistical t-test was performed. The values of protease enzyme activities in healthy, cystic and PSC were determined 0.0028, 0.0087 and 0.50U/ml/mg respectively. Elevation of protease enzyme activity in cystic liver as compared with healthy was not significant. Statistical T-test showed higher protease enzyme activity for PSC as compared to healthy [p<0.05]. SDS-PAGE confirmed 24 kDa and 54 kDa bands for protease enzyme in PSC samples and 24 kDa band in liver samples. Protease enzyme activity and molecular weight as compared to healthy liver tissues could be concerned as a comparative metabolic biomarker for host and parasite in hydatidosis
ABSTRACT
Fasciola hepatica is one of the most important helminthes parasites and triclabendazole [TCBZ] is routinely used for treatment of infected people and animals. Secreted protease enzymes by the F hepatica plays a critical role in the invasion, migration, nutrition and the survival of parasite and are key targets for novel drugs and vaccines. The aim of study was to determine the protease activity of excretory- secretory products [ESP] of F. hepatica in the presence of TCBZ anthel-mintic. F hepatica helminthes were collected and cultured within RPMI 1640 [TCBZ treated [test] and untreated [control]] for 6 h at 37 degree C. ESP of treated and control were collected, centrifuged and supernatants were stored at -20 degree C. Protein concentrations were measured according to Bradford method. Protease enzymes activities of ESP samples were estimated by using sigma's non-specific protease activity assay. ESP protein bands were detected by sodium dodecyl sulfate poly-acrylamide gel electrophoresis [SDS-PAGE]. Mean protein concentrations in control and treated of ESP samples were determined 196.1 +/- 14.52 and 376.4 +/- 28.20 microg/ml, respectively. Mean protease enzymes activities in control and treated were 0.37 +/- 0.1 and 0.089 +/- 0.03 U/ml, respectively. Significant difference between proteins concentrations and protease enzymes activities of two groups was observed [P<0.05] SDS-PAGE showed different patterns of protein bands between treated and control samples. The TCBZ reduced secreted protease enzymes activities and possibly effects on invasion, migration, nutrition and particularly survival of the parasite in the host tissues
ABSTRACT
The aim of this study was to detect the Glutathione S-Transferase [GST] enzyme activity of healthy / cystic liver as a diagnostic biomarker for hydatidosis. In order to compare with liver tissue, the level of the GSTs enzyme activityof parasite was also determined. Parasites were collected from sheep liver tissue with hydatid cysts at a local abattoir and washed with PBS buffer. Collected parasites and liver tissues were sonicated or homogenized respectively. Extract solution samples were centrifuged and stored at - 20 degree C. GSTenzyme activities were measured in the extract of parasite and liver tissue samples [healthy and infected livers]. Protein amounts and protein bands were detected using Bradford and sodium do-decyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE] methods respectively. To determine significant difference between two groups,two-sample/-test was performed. GST specific activities of healthy / infected livers and parasites were estimated 304, 1297 and 146 U/ml/mgrespectively. Significant higherGST specific activities in cystic liver than healthy liver was observed [P<0.05]. T-test analysis showed GST activity of parasite was lower than healthy liver tissue. SDS-PAGE showed GST protein bands with 24 kDa in parasite samples and25 kDa in liver tissues. GST activity incystic liver tissue could be concerned as a biomarker for hydatid cyst diagnosis with other hydatid disease parameters
ABSTRACT
On the continuation of sorting the puzzle of the situation of hydatid disease in Iran and considering that so far no survey was conducted in this context in Arak City, Markazi Province central Iran, the present study was conducted to determine the seroprevalence of human hydatidosis using AgB-ELISA test. Totally 578 serum samples randomly were collected from patients referred to hospitals and different health centers in the city and 3 nearby villages of Arak. All sera were examined by ELISA tests using AgB. Before sampling, a questionnaire was filled out for each case. Data were analyzed using Chi-square test and multivariate logistic regression for risk factors analysis. P<0.05 was considered significant. Cut-off value was calculated 0.32. Twenty cases [3.46%] were seropositive for hydatidosis in the region. This rate for females was 3.99% and for males 2.26%, respectively. There was no significant difference as regards all factors studied except location [P<0.001]. As for job, farmers and ranchmen had the highest rate of infection as 6.67%. The seropositivity rate infection was 4.8% in illiterate people, which showed the highest rate. As regards residency, rural life showed significant difference with urban life [1.5% vs. 7%]. Age group of 40-49 yr old [6.25%] had the highest rate of positivity. The rate of prevalence in this region shows more or less the same range with other cities of Iran. Obtained result might assist the policy makers to take sanitary measures to control the disease
ABSTRACT
Trematodes are a diverse group of endoparasites which require molluscan and vertebrate animals as intermediate and definitive hosts in their life cycle. The present study was carried out to determine the diversity and geographic distribution of infection with trematodes'cercariae in the snail Lymnaea gedrosiana from north-west Iran. A total number of 6759 Lymnaeidae snails were collected from 28 snail habitats; of these L. gedrosiana was the prevalent snail [74.37%] which examined for cercarial infection by shedding method. The overall infection rate was 8.03%. The most frequent trematodes cercariae in the snail were xiphidiocercariae [81.98%], furcocercariae [32.26%], echinostome cercariae [5.19%], and monostome cercariae [1.24%]. The highest infection rate in L. gedrosiana [100%] was with echinostome cercariae from Golestaneh in autumn. Due to the important role of pond snails in transmission of cercariae to fish as a source of zoonotic diseases, it is essential to estimate the distribution and abundance of the snails and the rate of their infection with different trema-todes' cercariae, and establish control programs in each region
ABSTRACT
Fascioliasis is an endemic disease in Iran and triclabendazole [TCBZ] is using for treatment of domestic animals and infected people. Excretory-secretory products [ESP] play an important role in the host biochemical defense by means of activities of detoxifying and antioxidant glutathione S-transferase [GST] and superoxide dismutase [SOD] enzymes respectively. Therefore, the aim of this comparative study was to evaluate fasciola protection against TCBZ drug by detection of enzymatic activities, GST and SOD, in TCBZ treated Fasciola hepatica / Fasciola gigantica and control ESP samples. F. gigantic and F. hepatica helminthes were collected and cultured within buffer media [TCBZ treated and untreated or control] for 4 h at 37[degree]C. Three TCBZ treated and 1 control ESP samples for each species were collected, centrifuged and supernatants were stored at-20°C. ESP samples protein concentrations were measured by Bradford method. SOD and GST enzymes activities of ESP samples were estimated photometrically. To determine the statistically significant difference between ESP of treated and control samples, t-test was conducted. ESP protein bands were detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE]. Protein concentrations in treated F. hepatica and F. gigantica ESP samples were estimated 204.88, 428, 130.4 and 288.2, 488.2, 308.2 micro g/ml respectively. Protein concentrations in control samples were estimated 488.18 and 124.8 micro g/ml respectively. SOD enzyme specific activities level in treated F.hepatica and F. gigantica ESP samples were determined 0.14, 0.31, 3.96 and 11.11, 13.54, 19.95 U/mg/protein respectively. SOD activities level in control samples were detected 70.69 and 10.92 U/mg/protein. GST specific activities level in treated F.hepatica and F. gigantica ESP samples were calculated 25.3, 85.5, 37.3 and 1823, 1314.3, 1320.8 U/mg respectively. GST activities levels in control samples were detected 98.6 and 1083.9 U/mg/protein respectively. Statistical analysis reveal the significant different between proteins concentrations, GST and SOD enzyme specific activities of TCBZ treated ESP samples of F. gigantic in comparison to the control samples [P<0.05], however, is not significant for treated F. hepatica and control ESP samples [P>0.05]. There is no difference between SDS-PAGE results of treated and control samples. Based on the results of the present work, significant increase of enzymatic activities of GST and SOD in TCBZ treated F. gigantica ESP, it seems, the protection of this species against drug is higher than F. hepatica