Activity of superoxide dismutase [SOD] enzyme in the excretory-secretory products of Fasciola hepatica and F. gigantica parasites

The aim of this study was to compare superoxide dismutase [SOD] activity in Fasciola hepatica and fasciola gigantica parasites. F. gigantica and F. hepatica helminths were collected from abattoir and cultured in buffer media for 4 h at 37 °C. Excretory-Secretory [ES] products were collected, centrifuged and stored at 20°C. E-S protein concentration was measured by Bradford method and SOD activity was detected using RANSOD kit [Randox Lab. Crumlin, UK]. Statistical t-test was conducted for analysis of results. Protein concentration for F. hepatica and F. gigantica were obtained 7.293 ug/ml and 19.65 ug/ml respectively and SOD activity as 0.721 U/ml and 1.189 U/ml, in that order. ES protein concentration of two species was significantly different [P<0.05], however the difference of SOD activity of two species was not significant. Two species of Fasciola have comparable SOD biochemical defense enzyme and can help us explain the parasite survival in host tissue

Survey of migratory birds [anatidae: anas platyrhynchos] for schistosome parasites from mazandaran province, northern Iran

The aim of the present study was to survey birds' schistosomes in migratory birds [Anatidae: Anas platyrhynchos] which are the source of the disease in Mazandaran Province, Northern Iran. A number of mallards were bought from the markets of hunted birds. The respiratory tracts [nasal mucosa] and intestinal blood vessels were studied for adult worms. The nasal mucosa was separated and observed by a microscope. In order to separate the visceral schistosomes, after separating intestine, vessel mesenteric was studied under the lamp light and then in saline. The parasite sample was collected for subsequent observation. Fifteen [13.6%] cases out of 110 studied birds had nasal mucosa contaminated with Trichobilharzia sp. egg. Besides that, two birds had adult worms schistosome visceral i.e. Bilharziella sp. The elements that cause cercarial dermatitis in aforementioned region are Trichobilharzia sp. and Bilharziella sp. parasites. Thus, it is necessary for the authorities of health, environmental and agricultural organization of the province to cooperate in order to control this disease.

Superoxide dismutase [SOD] enzyme activity assay in fasciola spp. parasites and liver tissue extract

The purpose of this comparative study was to detect superoxide dismutase [SOD] activities in Fasciola hepatica, F. gigantica parasites, infected and healthy liver tissues in order to determine of species effects and liver infection on SODs activity level. Fasciola spp. parasites and sheep liver tissues [healthy and infected liver tissues], 10 samples for each, were collected, homogenized and investigated for protein measurement, protein detection and SOD enzyme activity assay. Protein concentration was measured by Bradford method and SODs band protein was detected on SDS-PAGE. SODs activity was determined by iodonitrotetrazolium chloride, INT, and xanthine substrates. Independent samples t-test was conducted for analysis of SODs activities difference. Protein concentration means were detected for F. hepatica 1.3 mg/ ml, F. gigantica 2.9 mg/ml, healthy liver tissue 5.5 mg/ml and infected liver tissue 1.6 mg/ml [with similar weight sample mass]. Specific enzyme activities in the samples were obtained 0.58, 0.57, 0.51, 1.43 U/mg for F. hepatica, F. gigantica, healthy liver and infected liver respectively. Gel electrophoresis of Fasciola spp. and sheep liver tissue extracts revealed a band protein with MW of 60 kDa. The statistical analysis revealed significant difference between SOD activities of Fasciola species and also between SOD activity of liver tissues [P<.05]. Fasciola species and liver infection are effective causes on SOD enzyme activity level

Haemolymph components of infected and none infected lymnaea snails with xiphidiocercariae

In this study the haemolymph components of infected and none infected Lymnaea gedrosiana with xiphidiocercaria larvae was compared. Five hundred Fifty Lymnaea snails were collected from Ilam and Mazandaran provinces, Iran, during 2008-2009. The snails were transported to the lab at Tehran University of Medical Sciences and their cercarial sheddings were studied. Haemolmyphs of snails were extracted and cells were counted using haemocytometer and cell-surface carbohydrate were recognized by conjugated lectin [Lentil]. Haemolymph protein concentrations were measured by Bradford protein assay method and soluble protein compositions were determined on sodium dodecyl sulphate polyacrilamide gel electrophoresis [SDS-PAGE]. From the 550 examined Lymnaea snails for cercariae, 27 snails were infected with xiphidiocercariae. Mean of haemolymph cells [haemocyte] number were obtained 93480 +/- 2.43 [cells/ml] for none infected snails [25 snail] and 124560 +/- 2800 [cells/ml] for infected snails [25 snail]. Mannose carbohydrate was recognized on haemocyte of none infected and infected snails. Mean of protein concentration of haemolymph plasma was obtained as 1354 +/- 160 micro g/ml [1.4 mg/ml] for none infected snails [25 snails] and 1802 +/- 138 micro g/ml [1.8 mg/ml] for infected snail [25 snails]. Comparing to none infected snails, the SDS-PAGE results of haemolymph plasma of infected snails, showed an extra protein band [70 kDa]. The results showed a significant difference between the amounts and the kinds of proteins in haemolymph of infected and none infected snails. This information might be useful to understand of parasite detection, adhesion, engulfment and antigen agglutination by snail

Carbohydrate detection and lectin isolation from TEGU-mental tissue of fasciola hepatica

Fascioliasis is a chronic hepatic disease and may be resulted from mechanical/molecular parasite adhesion to host liver tissue. The aim of this study was to detect surface carbohydrate and lectin, carbohydrate-binding protein isolation that might be responsible of this molecular binding. The present experimental work was conducted in the Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. Fasciola hepatica parasites were collected from abattoir [Saman, Tehran, Iran] and surface mannose-carbohydrate was detected by fluorescein isothiocyanate [FITC] conjugated lectin [Lentil]. Lectin of tegumental tissue from F. hepatica was isolated by affinity chromatography and detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE]. Mannose carbohydrate was observed on the surface of tegumental tissue from parasite under fluorescence microscope. Carbohydrate-binding protein or lectin with MW of 50 kDa also was isolated from homogenized tegument of helminth. These results are important for understanding of molecular pathogenesis of F. hepatica at the chronic phase of fascioliasis

Seroepidemiology of human hydatidosis in Golestan province, Iran

Hydatidosis is one of the most prevalent zoonotic diseases worldwide. So far no survey was conducted to determine the rate of human hydatidosis in Golestan Province, so using IFA and ELISA tests the prevalence of this disease was detected in patients referred to health centers in this province. Totally 1024 serum samples were collected from patients referred to different health centers in 4 cities of Gloestan Province including Gorgan, Gonbad kawoos, Aliabad Katool and Kordkoy. All the sera were examined using IFA and ELISA tests. Twenty four cases [2.34%] were positive for hydatidosis in Golestan Province using IFA, whereas 22 cases [2.15%] showed positivity using ELISA. Gorgan, Gonbadkaoos, Aliabad Katool and Kordkoy demonstrated the rate of positivity as 1.41%, 2.40%, 5.36% and 2.30%, respectively, but no significant difference was seen. As to positivity, there was no significant difference between age groups, sex, different cities and rural or urban life, but a significant different was seen according to job and literacy [P< 0.001]. According to Job and literacy, housewives and illiterates had the highest rate of infection as 3.67% and 3.72%, respectively. As regards residency, urban life showed no significant difference with rural life [2.47% vs. 2.45%]. Age group of 40-49 years old had the highest rate of positivity [3.95%]. Females were more infected than males [3.16% vs. 1.93%]. The rate of prevalence in this province shows somehow a resemblance with the other cities in Iran. Considering the lifestyle in this province a complementary study is suggested in all related cities

Study on glutathione S-transferase inhibition assay by triclabendazole. III: nematodirus parasite and sheep liver tissue

The most important and widely prevalent nematodes of sheep are the trichostrongyle group parasites, including nematodirus parasite. Accidental infection of man by nematodirus has been reported in Iran. Glutathione S-Transferase enzymes [GSTs] are detoxification enzymes in parasites such as nematodirus. Therefore, GST enzymes of these parasites could be a target for evaluation of drugs effect as triclabendazole [C[14]H[9]CL[3]N[[2]OS]. For this reason, GST enzymes were purified from nematodirus parasite and sheep liver tissue by glutathione affinity chromatography and prepared their SDS-PAGE banding pattern for GST fraction separation. GST enzymes specific activity levels are also assayed in the whole extract and purified solutions with reduced glutathione [GSH] and l-chloro-2, 4-dinitrobenzen [CDNB] secondary substrate. Finally, GST inhibition assay was investigated in the solutions by powder and bolus forms of triclabendazole. The level of GST specific activity in purified solutions was detected 9.86 micromol / min/ mg protein for nematodirus parasite and 37.84 micromol/ min/ mg protein for liver tissue. Comparison of the effect of powder and bolus of triclabendazole on solutions revealed inhibition concentration [IC[50]] 5.54 and 6.01 microg/ml for nematodirus GST and 8.65 and 9.70 microg/ml for liver tissue GST, respectively. These findings revealed the possibility of isolation and inhibition of nematodirus GST by triclabendazole, and more tolerance of liver tissue than parasite against this drug in vitro situation

faunistic survey of cercariae from fresh water snails: melanopsis spp. and their role in disease transmission

Snail transmitted diseases are one of the major group of helminth parasitic diseases which have been established by trematode parasites. The larvae of trematodes [cercariae] use the snails as host. The purpose of the present study was to identify of cercariae released from Melanopsis spp. [M. doriae, M. costata, M. praemorsa, and M. nodosa] and evaluate their medical importance. Accordingly, 2, 266 Melanopsis spp. [fresh water snails] were collected from various agriculture canals in the central area of Khuzestan Province in the south west of Iran. 72 [3.1%] infected Melanopsis spp. snails were isolated and the cercariae were obtained by emerging or crushing methods. Subsequently, measurement and drawing were made on cercariae specimens and recognized. In some cases experimental infections were established in the animals for further iden-tification. A total of 4 cercarial families and 1 cercarial group were identified as follows: Heterophyidae: Haplorchis pumilio, H. taithui, Stellantchasmus falcatus and Centrocestus formosanus; Echinostomatidae: Echinochasmus milvi; Cyathocotyli-dae, Philophthalmidae and Monostome group cercariae [probably Notocotylidae]. These results have been recorded for the first time and these cercariae are of medical and veterinary importance

Characterization of purified glutathione s-transferase [GSTs] from fasciola hepatica and liver tissue by two-dimensional electrophoresis [2-DE]

Two-dimensional electrophoresis [2-D electrophoresis] is a powerful and extensively used method for analysis of complex protein mixtures extracted from cells, tissue, or other biological samples such as helminth parasites including, F. hepatica. Each spot on the resulting two-dimensional collection corresponds to a single protein species in the sample. This study was carried out to detect of GSTs isoenzyme spots map for collection of highly specific proteins. For this purpose, GSTs were purified from adult parasite of F. hepatica and sheep liver tissue as an enzyme pool by a glutathione affinity matrix using a wash-bath method and investigated for sodium dodecyl sulphate poly aery lam ide gel electrophoresis [SDS-PAGE] pattern. For 2-DE, purified GSTs from F. hepatica and sheep liver tissue were resuspended in sample buffer and then run on a IPG strip in the first dimension and then on an Excel Gel SDS in the second dimension before protein spots staining with Coomassie blue. The obtaining spots in the gels were compared and GSTs protein spots were detected with similar molecular weight, 26 kDa. The protein spots which are recorded in this paper could be GSTs isoenzymes and are highly specific peptids. These findings may be considered for vaccination or chemotherapeutic targets in sheep and human fascioliasis

Study on glutathione S-tranferase [GST] inhibition assay by triclabendazole.1: Protoscoleces [Hydatid cyst; Echinococcus granulosus] and sheep liver tissue

Hydatid disease is a term used to refer infection with the methacestode of Echinococcus granulosus parasite in humans, and echinococcusis is restricted to infection with the adult stage in carnivores.Glutathione S-Transferase [GST] represents the major class of detoxification enzymes from helminth parasites such as Echinococcus protoscoleces [PSC] and it is candidate for chemotherapeutic and vaccine design. Therefore, GST of protoscoleces could be a target for evaluation of drug effect as triclabendazole in hydatid cyst. For this purpose, GST enzymes were purified from protoscoleces of hydatid cyst and sheep liver tissue by glutathione affinity chromatography using a wash-batch method and subsequently detected their SDS-PAGE pattern. Afterward, GST specific activity levels were assayed in the whole extract and purified solutions spectrophotometrically at 30 C with reduced glutathione [GSH] and 1-chloro-2, 4-dinitrobenzen [CDNB] substrate. Finally, GST inhibition assay was investigated in the solutions by powder and bolus of triclabendazole. GST fraction as a 26 kDa [MW] band was obtained on SDS-PAGE. The level of GST specific activity in purified solutions was detected 10.24 mmol/min/mg proteins for protoscoleces and 37.84 mmol/min/mg protein for liver tissue. Comparison of the effect of powder and bolus of triclabendazole in solutions revealed inhibition concentration [IC50] 8.71 and 11.16 mg/ml for protoscoleces GST and 8.65 and 9.70 mg/ml for liver tissue GSTs, respectively. These findings suggest the possibility of selective inhibition of protoscoleces. GSTs by triclabendazol in vitro and use of these results for understanding of its molecular effect in vivo

faunistic survey on the cercariae of bellamya [viviparus] bengalensis snails and their zoonotic importance

Limited studies have been made on the cercariae from freshwater snails in Iran. This study was conducted to determine the cercariae fauna of Bellamya [Viviparus] bengalensis and evaluation of their zoonotic importance in Khuzestan province, south-west of Iran. For this purpose, 1143 Bellamya snails were collected from Ahoudasht region in central areas of the province during 2002-2003. Bellamya snails were examined for cercariae with shedding or crushing methods, and identified by systematic key references. Findings showed that 5 [0.4%] Bellamya snails were infected with Xiphidiocercariae belonging to Plagiorchiidae helminth parasites. The zoonotic importance of these cercariae and life cycle of Plagiorchiidae helminth [Trematode] parasites are discussed

faunistic survey on the Bird Helminth Parasites and their medically importance

Khuzestan province in the south west of Iran having several seasonal and permanent lagoons which are shelter for domestic and migratory birds including, fish-eating birds. This research study was carried out to find the intestinal helminth parasites of birds in this ecosystem and evaluation of their medically importance with emphasis on heterophyid trematodes. For these reasons, the total of 37 birds including; Himantopus himantopus, Fulica atra, Egretta grazetta, Bubulcus ibis, Ceryle rudis, Vanellus indicus, Vanellus vanellus, Charadrius sp. Calidris sp. and Saher [Local name] were hunted and transported to Ahwaz Health Research Center as alive or freshly dead after having been shot. Helminthes collected as alive or dead and fixed in ethanol or formaldehyde. Parasites were identified using morphometric measurements and morphological descriptions. 24 species of intestinal helminth parasites were found as follow: trematodes [Haplorchis taichui, Haplorchis pumilio, Stellantchasmus falcatus, Centrocestus formosanus. Psiloterma marki, Echinostoma revolutum, Parechinostomum cinctum, Echinochasmus coaxatus, Paramonostomum alveatum, Uvitellina pseudocotylea, Cyclocoelum mutabile, Apharyngostrigea cornu, Cardiocephallus br and esi, Cotylurus cornutus, Pseudostrigea buteonis] and nematodes [Amidostomum fuligulae, Cosmocephalus diesingii, Microtetrameres accipiter, Strongyloides minimus] and cestodes [Gyrocoelia perversa, Infula burhini, Dirorchis tringae, Echinocotyle nitida, Spiniglans microsoma]. These results have suggested that, the birds are reservoir for helminth parasitic diseases such as heterophyiasis for man and animals in the areas. These helminthes are reported for the first time in the region