Fasciola gigantica of ruminants: the phylogenetic analysis based on COX1 sequence

Background: fasciola species are parasitic trematode with worldwide distribution that infects wild and domesticated herbivores, particularly ruminants

Objectives: the aim of the present study was to investigate the intra species variations of F. gigantica, from goats and buffalo isolates in two common geographic climates of Iran

Methods: fasciola species were collected from goat, buffalo, sheep, and cattle in different regions. Cytochrome c oxidase I [COX1] of mitochondrial DNA [mt-DNA] was amplified from individual trematodes by polymerase chain reaction [PCR], using universal primers, and the amplicons were consequently sequenced and sequencing data were analyzed, using Clutal W software against the GenBank database

Results: a monomorphic DNA segment of approximately 499bp was seen in Fasciola isolates. The results of the amino acid sequence alignment defined strictly conserved amino acid residues in buffalo isolates of F. gigantica and partially conserved residues for goat isolates of F. gigantica. There are four tandem amino-acid replacements in the goat isolates at the position of 135-138, where Leucine [L], F [Phenylalanine], T [Threonine], and D [Aspartate] sequences changed into S [Serine], L [Leucine], H [Histidine], and L [Leucine], respectively. Furthermore, a replacement in the sequence of amino acid was found in isolates from buffalo at the position of 154, where Serine [S] was transformed into Leucine [L]

Conclusions: the findings of our study indicate that the variants of goat and buffalo can be responsible for persistence of Fasciola infection in the endemic areas of Iran. It seems that biological differences could occur by considering a variety of F. gigantica-hosts in Iran. Thus, suitable approaches are required for effective treatments and useful control strategies

[Evaluation of the anthelminitic effects of Quercus robur extract against ovine gastrointestinal nematodes]

Background: recent investigations have identified anthelminitic effects of many medicinal plants particularly from condensed tannin sources. In addition, gastrointestinal nematodes of ruminants have a negative effect on the farming industry worldwide

Objectives: the aim of the present study was to determine the potential anthelmintic effects of Quercus robur extract on alimentary canal nematodes in naturally infected sheep by faecal egg count reduction test [EPGRT]

Methods: the crude aqueous extract was prepared from Quercus robur as tannin extract. The nature and intensity of helminth infection was determined by coprological examination. The faecal samples of 600 sheep were collected from different regions of Kurdistan province. The samples were examined by flotation method [Clyton-Lane technique]. Fifteen sheep with the most count in egg per gram [include Marshallagia, Nematodirus and Trichostrongylids] were divided into three groups of five animals: First group [test group] were drenched with Quercus robur extract at 3.75g/kg, second group [positive control group] received Albendazole 2.5%, orally at 5mg/kg and third group [negative control] without treatment

Results: the results of faecal examination 3 days after administration indicated significant reduction of EPG in both group's treatment and positive control groups, 90.76% and 90.83% respectively, whereas there was no effect in the third group. Results were evaluated by Chi-square analysis and showed significant differences between treatment and negative control groups [p/=0.05]

Conclusions: results reveal that aquatic extract of Quercus robur has anthelminitic activity and further large scale studies are suggested to confirm pharmacologic effects of this herbal extract

[Designing a rapid test to diagnose the ovine hydatid cyst]

Using a rapid and cost benefote test for diagnosing of hydatidosis, a zoonotic, can be beneficial as a diagnostic. The aim of the present study was to design and assess the performance of a dipstick method for diagnosting of cystic echinococcosis. Hydatid cyst fluid antigens and homogenized antigens of protoscolex were prepared and its electrophoretic pattern was determined by SDS-PAGE. Afterthen, for providing the hyperimmune serum, rabbits were injected by hydatid cyst fluid and protoscolex antigens along with complete freund adjuant and then incomplete freund adjuant. The immune sera were evaluated by dot ELISA. Dipstick was prepared based on nitrocellulose paper as solid phase and coated with antigens which were dotted in the upper surface of the nitrocellulose strip. PBS was used as negative control and rabbit sera noninfected were used as positive control. As a negative control the lower part of the strip was coated by PBS. Each strip was floated in the serum [1:100 dilution] for 7 min, washed for 7 min and floated in the second antibody for 7 min. Afterthen, they were washed and incubated in chromogen/substrate diaminobenzidin for 2 min to show the coloured band at the site of coated antigen. Our findings revealed 15 protein fractions in fluid antigens and 9 protein band in protoscolex antigens at the range of 29-130 kDa and 25-90 kDa, respectively. Meanwhile, the protoscolex antigens at in 1:10 dilution and fluid antigens at in 1:20 dilution showed the best results to diagnose hyper immune serum of 1:10 dilution. This rapid Dipstick assay test can be considered as a suitable immunodiagnostic test for hydatid cyst disease; however further investigations should be done to improve its specificity through preparing highly purified antigens

Variations to somatic protein profiles of dicrocoelium dendriticum in Iranian ruminants

Dicrocoelium dendriticum, a liver trematode of herbivores, and man is very prevalent among ruminants of Iran, resulting in causing significant economic losses. Recently its phenotype polymorphism has been found in Iran, a phenomenon that may affect the protein profile of the parasite. The aim of this study was to determine the somatic protein profiles of D.dendriticum in different ruminants in Iran. Adult worms were collected from infected livers of sheep, goat, cattle, buffalo and camel. Somatic antigens were prepared separately by centrifugation of the homogenized adult worms and their patterns were determined using SDS-PAGE. Our findings revealed 18-20 protein fractions in somatic proteins of D. dendriticum of sheep and goat and 12, 11 and 10 in that of buffalo, cattle and camel respectively showing similar molecular weights of 14.4 to 116 kDa. Similar protein bands were seen in sheep and goats samples; but not for in other samples. The results showed polymorphism in the somatic protein patterns of D.dendriticum isolated from different hosts. A different number of protein bands with similar molecular weights of somatic antigens of D.dendriticum are described for the first time in the present study

Animal fascioliasis in coastal regions of the Caspian Sea, Iran [2006-2007]

To determine the prevalence of animal fascioliasis in coastal regions of the Caspian Sea, during 2006-2007, a total of 2368 faecal samples were collected from sheep [n=1250], cattle [n=975] and horse [n=143]. The samples were obtained directly from the rectum of animals. Floatation method was performed for determination of egg per gram of faeces [EPG]. Our findings revealed the presence of Fasciola spp. eggs in 9.53, 7.8 and 2.5% of sheep and 32.5, 12.1 and 3.1% of cattle in Gilan, Mazandaran and Golestan, respectively. Among the horse faecal samples collected from Golestan and Gilan provinces, no eggs were found in Golestan, while 50% of those of Gilan were infected with Fasciola spp. Cattle was one of the most infected animals in the studied areas. A positive correlation was found between climatic conditions and animal fascioliasis. Among different meteorological factors, rainfall seems to be the strongest factor

[Determination of antigenic protein of Dicrocoelium dendriticumin naturally infected sheep]

Dicrocoeliasis is an important livestock disease caused by digenean trematode namely Dicrocoelium dendriticum. The aim of the present study was to identify somatic and metabolic antigens of adult D.dendriticum in naturally infected sheep. Adult parasites collected from the liver of naturally infected sheep, were washed in cold phosphate buffer saline [PBS, pH=7.4] and stored at 20°C until analysis. Antigen used for the detection of antibody included somatic and metabolic of mature trematode. Somatic and excretory-secretory, antigens prepared with haemogenization and incubation of adult helminths, respectively. Electrophoretic patterns of excretory secretory and somatic antigens of D.dendriticum were revealed by sodium dodecyl sulphate polyacrylamid gel electrophoresis [SDS-PAGE] and wester blotting using sera from naturally harbored sheep. In western blot analysis of antigens D. dendriticum demonstrate 1 major antigenic polypeptide 130 kDa in both somatic and metabolic antigens and 6 protein bands ranging from 25 to 60 kDa in excretory-secretory antigens which were recognized by serum of sheep naturally infected. Our findings showed that the 130 kDa molecular weight polypeptide could be used as specific antigen for the immunodiagnosis of sheep dicrocoeliasis

Morphological and morphometrical description of trichostrongylus species isolated from domestic ruminants in Khuzestan province, southwest Iran

Genus Trichostrongylus [Nematoda: Trichostrongylidae] is one of the most important zoonotic nematodes with wide geographic distribution in the world. The purpose of the present study was to describe morphological and morphometrical characteristics of male Trichostrongylus species, currently prevalent in domestic ruminants of Khuzestan Province, southwest Iran. Gastro-inetstinal organs of 1600 sheep, goats, cattle, and buffalos, slaughtered in Khuzestan Province, southwest Iran, were examined for infectivity with Trichostrongylus species. For examination and measurements of helminthes, Azo-carmine staining was performed, followed by camera lucida drawings of morphological characters and measurements of morphometrical criteria with a calibrated microscope. Using valid nematodes systematic keys, almost all the parasites were identified at the level of species. Overall, 114 animals were found infected with at least one species of Trichostrongylus. Considering morphlogifcal characteristics of male Trichostrongylus, six species were identified including T. colubriformis, T. vitrinus, T. probolorus, T. capricola, T. longispicularis and Trichostrongylus sp.. Although, compared to the previous decades, currently Trichostrongylus is much less prevalent in the domestic ruminants of the study area, but still different species occur in these animals

Helminth infections of stray dogs from Garmsar, Semnan province, central Iran

The aim was to study the gastro-intestinal helminths of stray dogs of Garmsar, Semnan Province, Central Iran, and its impacts on human health and animal production. During 2006, the alimentary tracts of 50 stray dogs at necropsy, selected from villages around Garmsar, were removed, and examined for helminth infections. Subsequently helminths were collected from the contents of each part and scraped sample of small intestines of washed materials in a 100-mesh sieve. To identify the species of helminths, the nematodes were cleared in lactophenol and cestodes were stained using carmine acid. Mixed infection was the rule and 40 dogs [80%] harbored more than one species of helminth. Taenia hydatigena was the most prevalent species [80%] followed by Echinococcus granulosus [64%], Toxocara canis [22%], Mesocestoides lineatus [12%], Taenia multiceps [10%] and Dipylidium canium [4%]. The mean intensity of worm infection was low [1-3] except for that of E.granulosus [645]. No significant difference was noticed between sex, age and most helminth infections except for that of sex and T. hydatigena [P=0.001] as well as age and T.canis [P=0.001]. Although human infection with T. hydatigena is unlikely, but other helminths reported in this study are of zoonotic importance, and may pose a threat to community health, and reduce the productions of ruminants harboring taeniid metacestodes

Determination of diagnostic antigens in cattle amphistomiasis using western blotting

Mixed infection with amphistomes seems common in native cattle of Iran. The aim of this study was to determine diagnostic antigens in cattle mixed amphistomiasis. Specific antigens of Cotylophoron cotylophorum, Gastrothylax crumenifer and Paramphistomum cervi [mixed infection], the most common species, were collected from cattle was determined. Adult trematodes were collected from the rumen of naturally infected cattle at meat inspection. After their homogenization and centrifugation, somatic antigens were prepared and analyzed by SDS-PAGE. Specific antigens were determinated by western blot with homologous and heterologous sera. SDS-PAGE of whole worms extract was performed at different concentrations and subsequent gels staining. Immunoblotting analysis using sera from cattle naturally infected with amphistomes, Dicrocoelium dendriticum, Fasciola spp. and hydatid cyst was performed. Electrophorese analysis of somatic antigens revealed the presence of 10 and 21 protein bands at 4 micro gr/ml and 8 micro gr/ml with molecular weights ranging from 25-120 and 25-150 kDa, respectively. The best result was taken at 8 mg/ml concentration. Although western blot of these proteins demonstrate 5 major antigenic polypeptides ranging from 50 to 100 kDa which were recognized by serum of cattle naturally infected with mixed amphistomes. Ninety-kDa protein can be specific diagnostic antigen for mixed amphistomiasis in Iran

Prevalence of parasitic infections in the red fox [Vulpes vulpes] and golden Jackal [Canis aureus] in Iran

Red foxes and golden jackals are the two most abundant wild carnivores of Iran which have the ability to adopt a variety of habitats and human proximity. Despite this, very few investigations on their helminth and none on their external parasites infections have been carried out in Iran. Between 2003 and 2004, a total of 79 jackals and 37 foxes were collected from 3 different climatic zones of Iran and examined for helminth and ectoparasite infections. A number of parasites including: Mesocestoides lineatus, Taenia hydatigena, Dipylidium caninum, Diplopylidium nolleri, Ancylostoma caninum, Uncinaria stenocephala, Toxocara canis, Onicola canis, Dirofilaria immitis, Rhipicephalus sp., Ctenocephalides canis and Pulex irritans were common parasites between jackals and foxes, whereas Echinococcus granulosus, Spirocerca lupi, Rictularia affinis, Macracanthorhynchus hirudinaceus, Dermacentor sp. parasitized jackals and Jeuyoxiella pasquali, Ixodes ricinus, Haemaphysalis sp. were collected only from foxes. All ectoparasites, S. lupi and O. canis reported from jackals and foxes in this study represent new host and distribution records

Animal fasciolosis in north of Iran

Fasciolosis is a well known parasitic disease of animals with public health importance. In Rasht and Bandar- Anzali, in Gilan Province, where experienced two large human fasciolosis outbreaks, no update information is available on animal fasciolosis. Paucity of information on animal fasciolosis in these regions and its possible impacts on human fasciolosis called us for carrying out this study. During 2005, coprologic surveys using flotation method were applied to fecal samples of 156 stray cattle, 171 calves, 178 sheep, 85 buffaloes, 79 horses and 10 samples from 10 different preserved animal manure collections to detect Fasciola egg. Fecal samples of 32% of sheep, 32.1% of cattle, 0% of calves, 17% of buffaloes, 50% of horses and 100% of animal manure samples harbored Fasciola egg. The mean intensity of Fasciola egg per gram of feces [EPG] was low [0-13]. Fasciolosis was very prevalent among animals in studied regions. Because sheep breeding is not a common practice in Rasht and Bandar -Anzali and horse population is low, cattle and to a lesser extent buffalo were the predominant reservoir hosts of infection. Regular treatment of all animals with an effective flukicide and sanitation of animal manure through its preservation for two month should be applied in order to reduce the level of infection in animals, water, wild and cultivated vegetables and consequently human beings

Evaluation of different antigens in western blotting technique for the diagnosis of sheep haemonchosis

To use different methods for serodiagnosis of ruminants' haemonchosis is important because detection of egg in the faeces is not so reliable. Peptide bands of 5 different crude antigens of intestine, uterus, cuticle, whole male and whole female of Hae-monchus contortus were determined using SDS-PAGE and western blotting. Five lambs were infected with 10,000 third stage larvae of H.contortus and 2 parasites free were kept as control. Positive and negative sera collected from infected and no infected animal were tested using western blotting for immunodiagnostic antigens. In electrophoresis the major peptide bands of crud antigens of uterus, intestine, cuticle, whole male and whole fe'male of H. contortus were 7, 2, 6, 8 and 5, respectively at molecular weights of 15 to 110 kDa. In immunoblotting positive and negative sera were compared and the molecular weight of specific protein bands for Haemonchus in sheep was deter'mined. Two major peptide bands belong to intestine and uterus with 35 and 40 kDa molecular weight, respectively, were specific for diagnosis of the parasite infection. Our findings suggest the use of these antigens with different molecular weights for immunodiagnosis of hae'monchosis in sheep as a primary screening test is promising

Biodiversity and prevalence of parasites of Rook [Corvus frugilegus] in Iran

Rooks are distributed all over Iran and no information is available in the literature on their parasitic infections. One hundred twenty five rooks were examined at post-mortem for parasitic infections. Two species of cestodes, 5 species of nematodes and 4 species of protozoa were found of which all were new host and distribution record. Rooks have several parasites of which some are common with other domestic birds and some have zoonotic importance

Gastrointestinal parasites of ostrich [struthio camelus domesticus] raised in Iran

Introduction of ostrich [Struthio camelus domesticus], a new exotic host species to a new ecosystem like Iran, necessitates the study of its infectious diseases including parasitic infestations. For a period of one year, from 2002-03, faecal samples of 254 ostriches on 5 farms located at different parts of Iran, including Garmsar, Semnan province [n = 85]; Nashtaroud, Maz and aran province [n = 29]; Saveh, Central province [n = 47]; Eshtehard, Ghazvin province [n = 8] and around Tehran, Tehran province [n = 85] were examined for gastrointestinal parasites. Our findings revealed the presence of Libostrongylus douglasii in 55% of the examined birds in Nashtaroud and 25% of birds of Eshtehard farms. In the latter farm, 12.5% in the ostriches harboured Heterakis dispar in their alimentary canals. Oocyst of Eimeria sp. was found in ostrich on farm of Garmsar. All these parasites are reported for the first time from ostriches in Iran

[Evaluation the effects of extracts ofzataria, carum and geranium on different stages haemonchus contortus [nematode] invitro and invivo condition]

In Iran different species from Zataria, Carom and Geranium are present and in some areas are endemic. There is no report about effect of this herbal extracts on helminth parasites. This study aims at investigating the effect of three herbal extracts of Zataria, Carom and Geranium on different stages including egg, larvae and mature of Haemonchus contortus, parasitic nematode of sheep. After experimental infection of the lambs was determinated the number of egg per gram [EPG] of feces and its cultured. Evaluating the effect of the extracts breeding, a plant synthesis along with the control group were added to the feces samples separately and after placing the sample in the incubator for its culture and then examined light microscope to evaluate of hatching status. Furthermore, in order to evaluate the effect of extracts on larvae stage, three different volume of herbal extracts were added to the larvae and the percentage of lost larvae was counted through in different period [0, 5, 10, 15, 20, 30, 40 and 60 min]. Infected lambs were fed by hand during their maintenance and a month after contamination the volume of 20 cm3 of each extract was fed to each lamb. The results showed that three extracts of, Zataria, Carom and Geranium have the effect of 89.9%, 21.6% and 6.6% on egg stage of Haemonchus contortus, respectively. Besides, with direct contact of the larvae, the Zataria extract caused 95.7% 97.7% and 100% loss in volume of 0.01, 0.02 and 0.03 cm3 after 10 min and the extracts of Carom and Geranium caused 83.2%, 87.5%, 86% and 65.8%, 77.6% and 78.5% respectively. The field condition and necropsy results showed the lack of extracts effects on the adult stage of Haemonchus contortus

[ prevalence of Giardia spp. among the pet dogs of Isfahan]

Study on Giardia spp, among the pet dogs of Isfahan. Cross-sectional study. One hundered and twenty dogs. Faecal specimens of 120 pet dogs which were referred to the small animal clinic of Isfahan were tested for Giardia spp. The diagnosis of Giardia spp was based on identification of cycts or trophozoietes in faece. Direct and formalin -ether methods as well as trichorme staining were appllied for identification of Giardia spp. Fisher exact test. The results indicated that 4 dogs [3.33%] were infected with Giardia spp. Because of probable infection of human by this species, our findings recommend killing of stray dogs and treatment of infected ones

Comparative study on the accuracy of different methods or diagnosis of cysticercus bovis in slaughtered cattle

To present a more accurate method for diagnosis of Cysticercus bovis in slaughtered cattle. ProjectrSlaughterhouse study. Animal:3436 cattle.To present a more accurate method for diagnosis of C.bovis in slaughtered cattle 1812 cattle were examined at meat inspection using traditional method. Meanwhile by using standard method for detection of c.bovis at meat nspection in South Africa,Germany and England 145 cattle for each region were examined.In each method the percentage of infection with C.bovis was determined. In raditional method heart,external and internal master and ;riceps muscles were inspected.Whereas for the method used in Germany and England,heart,external and internal master,oesophagus,diaphragm, tongue and for that of South Africa in addition to these,triceps was also inspected. The best, among the method used, was selected and according to the latter 1189 carcasses were examined. If the accuracy of selected method was statistically superior or similar to the best method,thus it was considered as an alternative or replacement method to the traditional method.By examining of 1182 cattle at meat inspection using traditional method 47 carcasses [2.6%] harboured .bovis in different organs,whereas using standard method,used in Germany,England and South Africa 7.6%, 6.9% and 6.9% of examined cattle were found to be infected with' C.bovis.There was a significant differences between traditional method and standard methods of three different countries us in X2 test. Standard method used in Germany was more accurate than the others, thus it was chosen as our selected method by which 1189 cattle were examined of which ninty cattle [7.6%] were found to be infected with C.bovis, an infection rate comparable to German method, Thus it could be suggested as an alternative or replacement to the traditional method

[Study on blood filariosis of dog in tonekabon]

To determine the prevalence and species ofblood filaria of dog in Tonekabon.Cross sectional study.80 dogs.After clinical examinations and recording ofnecessary information [age, sex and breed], blood sampleswere collected from cephalic or saphen vein and examinedby modified knott method.Analysis of variance [ANOVA].Out of 80, 14 dogs [17.5%] were infected withmicrofilaria. In this regard, about 15%, 1.25% and 1.25% ofinfectionswere Dirifilariaimmitis, microfilaria ofdipetalonemareconditumandmixedinfection, respectively. Infection showed no correlation with sex, age, breed and rates of breathing.Because of the possibility ofinfection in human by this species, our findings recommendkilling of stray dogs and treatment of infected ones

[Study on stray cats infected with parasitic helminthes in Tehran]

Objectives: a study on the prevalence of stray cats infected with parasitic helminthes in Tehran

Design: cross sectional study

Animals: one hundred stray cats captured from different urban areas of Tehran

Methods: one hundred stray cats were trapped from different geographic regions of Tehran and were necropsied. Different organs including: kidney, heart, liver, lungs, gastrointestinal tract and abdominal cavity were inspected for helminthic infection

Statistical analysis: X[2] test was used to show the relationship between different factors and parasitic infection and E ta coeffeciency was used to presenting effects of these factors on the infection

Results: four species of helminthes including 3 nematodes, Toxocara cat; [23%], Physaloptra praeputialis [7%] and Toxascaris leonine [2%] and one cestode, Diplopylidium nolleri [5%] were found in gastrointestinal tract and only this organ was found infected in examined cats

Conclusion: although for some species [e.g. Tcati 23%], the percentage of infection was relatively high, but with regard to the mild intensity of the collected worms, they could not cause clinical or subclinical symptoms. On the other hand, considering the contact between cat and man especially children, through courtyard environment or direct contact and visceral or ocular larva migration produced by ascarids of dog and cat, the report of two species of Ascaris in the present study could be a health hazard for human especially children