Comparative assay of protease enzyme activity in protoscolices (Hydatid cyst) and liver tissues

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 - 20°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 hydatidosi

In Vitro Study of Mebendazole (Anthelmintic drug) Effects on the Aspartate Aminotransferase (AST) Enzyme Activity of Hydatid Cyst Parasite

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 µg 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

A study on cercarial dermatitis in Khuzestan province, south western Iran

BACKGROUND: Cercarial dermatitis' or swimmer's itch' is an itchy inflammatory response to the penetration of the skin by non-human schistosome parasites. In the hot season, (May to September) in Khuzestan province in the south west of Iran, swimming in canals and agriculture activities in swampy areas are common. This survey was made on people from villages north of Ahwaz city in south west Iran, to estimate cercarial dermatitis in this region. METHODS: 2000 people were observed for clinical signs of cercarial dermatitis. Also 2000 Lymnaea gedrosiana snails were collected from agriculture canals and examined for animal schistosome cercariae during 1998–2000. RESULTS: From this survey 1.1% of people had pruritic maculopapular rash on their feet, hands or other parts of body. From the total of examined snails, 2.4% were found to be infected with bird schistosome cercariae including Trichobilharzia species. CONCLUSION: Cercarial dermatitis could be a health problem in this area. This is the first report of cercarial dermatitis from this region of Iran

Experimental Life Cycle of Hypoderaeum conoideum (Block, 1872) Diez, 1909(Trematoda: Echinostomatidae) Parasite from the North of Iran

Background: Human Echinostomiasis is an intestinal disease caused by the members of family Echinostomatidae parasites. The aim of present research was to identify echinos­tomatidae cercariae emitted by Lymnaea palustris snails from Mazandaran province in the north of Iran based on the morphological and morphometrical charac­teristics of the different stages of experimental parasite life cycle. Methods: Echinostomatidae cercariae were collected from L. palustris (Gastropoda: Lymnaeidae) of the north of Iran. To collect metacercaria, 50 healthy snails were infected with cercariae experimentally (50 cercariae for each). To obtain the adult stage, 9 laboratory animals (3 ducks, 2 rats, 2 mice and 2 quails) were fed with 60 metacercaria for each. To identify parasite, the different stages of worm were exam­ined using light microscope and then the figures were draw under camera Lucida microscope and measures were determined. Results: Averagely, 15metacercaria were obtained from each snail that had been previously exposed with cercariae. Ducks presented worm eggs in feces after 10-15 days post-infection. Intestinal worms were collected and identified as Hypoderaeum conoideum on the bases of figures and measures of cephalic collar, the number of collar spine, suckers diameter ratio, testes arrangement, etc. Conclusion: H. conoideum cercariae and adult worm are described. This is the first report of the different stages of the experimental life cycle of this parasite in Iran

Comparative Assay of Glutathione S- Transferase (GSTs) Activity of Excretory-Secretory Materials and Somatic Extract of Fasciola spp Parasites

Fascioliasis is a worldwide parasitic disease in human and domestic animals. The causative agents of fascioliasis are Fasciola hepatica and Fasciola gigantica. In the recent years, fasciola resistance to drugs has been reported in the many of publications. Fasciola spp has detoxification system including GST enzyme which may be responsible for its resistance. Therefore , the aim of the study was to assay of GST enzyme activity in fasciola parasites. Fasciola gigantica and Fasciola hepatica helminths were collected from abattoir as a live and cultured in buffer media for 4 h at 37 °C. Excretory-Secretory products were collected and stored in -80◦C. F. gigantica and Fasciola hepatica were homogenized with homogenizing buffer in a glass homogenizer to prepare of somatic extract. Suspension was then centrifuged and supernatant was stored at -80°C. In order to assay the enzyme activity, excretory-secretory and somatic extracts in the form of cocktails (potassium phosphate buffer, reduced glutathione and 1-chloro-2,4-dinitrobenzene substrates) were prepared and their absorbance recorded for 5 minutes at 340 nm. The total and specific GST activity of F. gigantica somatic and ES products were obtained as 2916.00, 272.01 micromole/minute and 1.33, 1.70 micromole/minute/mg protein, respectively. Fasciola hepatica also showed 2705.00, 276.86 micromole/minute and 1.33, 1.52 micromole/minute/mg protein, respectively. These results are important for analysis of parasite survival / resistance to drugs which use for treatment of fascioliasis

In vitro assay of alkaline phosphatase enzyme activity in liver and fasciola hepatica parasite

  The present work was designed to determine alkaline phosphatase (ALP) activity level in Fasciola hepatica parasite and Fasciola infected livers to evaluate the effects of liver infection on enzyme activity and to compare enzyme activity in liver and parasite. The sheep livers were collected and adult Fasciola hepatica parasites were isolated and washed with PBS buffer. Collected healthy and infected livers and parasite were homogenized and extract solutions were centrifuged and stored at -20°C. ALP enzymes activity was measured in the extract solution of samples. Proteins of the samples were measured and protein bands were detected through using Bradford and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) methods respectively. Independent two samples t-test was performed to determine significant difference between different groups. The mean values of the assayed ALP specific activities of infected and healthy livers and Fasciola hepatica parasite were estimated 0.163,0.133,0.048 U/mg protein/ml correspondingly. Gel electrophoresis (SDS-PAGE) of Fasciola hepatica and liver extract solution samples displayed different protein bands, including ALP enzyme. Statistical analysis did not show significant difference between enzyme’s activity in infected and healthy livers (P>0.05). However, the liver demonstrated more than ALP activity level to parasite (P<0.05). The findings show that ALP enzymes activity in Fasciola hepatica infected livers could not be concerned as a specific pathological biomarker in fascioliasis, but meanwhile this enzyme displays interest activity in parasite

Seasonal and Geographic Distribution of Cercarial Infection in Lymnaea Gedrosiana (Pulmunata: Lymnaeidae) In North West Iran

Background: 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.Methods: 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.Results: 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.Conclusion: 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 trema­todes’ cercariae, and establish control programs in each regio

Effect of Albendazole and Mebendazole on the Viability of Hydatid Cyst Protoscoleces in Vitro

Background and Objectives: Hydatidosis is caused by larval stage of the cestoda Echinococcus granulosus in humans and domestic animals. Many protoscolicidal agents have been used to treat it. The purpose of this study was to investigate the effect of albendazole and mebendazole on the viability of protoscoleces in vitro. Methods: In this experimental study, after collection of hydatid cyst livers, protoscoleces were aseptically removed and their viability was examined; then, certain volume of protoscoleces was poured into culture medium (RPMI-1640/PBS, pH 7.2) and albendazole and mebendazole solution with final concentration of 1 &micro;g/ml, was added to them. After a certain time interval, in order to determine the significant statistical difference between protoscoleces viability, one-way ANOVA (with bootstrap method) and Tukey post-hoc tests (for comparing two culture media RPMI and PBS), were used. Results: In this study, albendazole was more effective in removing protoscoleces. In the group in which albendazole were used, the viability percentage of protoscoleces reached zero after 35 days, while the viability of protoscoleces in the mebendazole group reached zero after more than 42 days (p<0.05). Conclusion: The obtained results of this research showed the favorable effect of albendazole on protoscoleces, therefore, it is likely that albendazole can be used as an appropriate drug for elimination of hydatid cyst protoscoleces and for prevention of the relapse of this disease

Study of Triclabendazole (TCBZ) Effect on Aspartate Transaminase (AST) Activity of Fasciola gigantica Parasite and Liver Enzyme Activity Assay

Background: Aspartate transaminase (AST) is an important enzyme in parasite and liver tissue. The purpose of this investigation is to evaluate triclabendazole (TCBZ) effect on AST activity of Fasciola gigantica parasite. To compare of enzyme level of parasite and its host tissue, enzyme activity of F. gigantica parasite and liver tissues were also determined. Method: The livers were collected from sheep slaughtered in local slaughterhouse and living F. gigantica parasites were isolated. The washed parasites were cultured in buffe rmedia with or without Triclabendazole (Egaten®); 15μg/mL in an incubator at 37° C. Extractions of collected parasites and liver tissues were prepared by homogenizing buffer in a Mortar and pestle. Extraction samples were examined for protein measurement, AST activity assay and protein recognition. Results: The results of AST assay revealed, enzyme activity for treated and untreated is not significant. Healthy liver tissue shows significantly higher enzyme activity than parasite. Enzyme activity for healthy and infected liver tissues was significant. Enzymatic proteins including Cathepsin L & B (Protease) were recognized in parasite samples. Conclusion: Although AST could not be concerned as an indicator for efficiency treatment, however may be involved as a biomarker for biochemical comparison of parasite and host tissue

Abnormality detection and localization schemes using molecular communication systems : a survey

Abnormality detection and localization (ADL) have been studied widely in wireless sensor networks (WSNs) literature, where the sensors use electromagnetic waves for communication. Molecular communication (MC) has been introduced as an alternative approach for ADL in particular areas such as healthcare, being able to tackle the shortcomings of conventional WSNs, such as invasiveness, bio-incompatibility, and high energy consumption. In this paper, we introduce a general framework for MC-based ADL, which consists of multiple tiers for sensing the abnormality and communication between different agents, including the sensors, the fusion center (FC), the gateway (GW), and the external node (e.g., a local cloud), and describe each tier and the agents in this framework. We classify and explain different abnormality recognition methods, the functional units of the sensors, and different sensor features. Further, we describe different types of interfaces required for converting the internal and external signals at the FC and GW. Moreover, we present a unified channel model for the sensing and communication links. We categorize the MC-based abnormality detection schemes based on the sensor mobility, cooperative detection, and cooperative sensing/activation. We also classify the localization approaches based on the sensor mobility and propulsion mechanisms and present a general framework for the externally-controllable localization systems. Finally, we present some challenges and future research directions to realize and develop MC-based systems for ADL. The important challenges in the MC-based systems lie in four main directions as implementation, system design, modeling, and methods, which need considerable attention from multidisciplinary perspectives