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1.

Background

The aim of the present study was to determine the molecular characteristics of Echinococcus granulosus from paraffin-embedded tissues of hydatid cysts isolated from human and protoscoleces of hydatid cysts from sheep, cattle and camel isolates using PCR- RFLP of ITS1- rDNA analysis in Golestan Province, northern Iran.

Methods

E. granulosus isolates from human patients infected with hydatid cyst and protoscoleces from hydatid cysts of sheep, cattle and camel isolates were collected from different hospitals and the abattoir throughout the Golestan Province. In all, 60 E. granulosus genomic DNA were extracted and examined by PCR - ITS1 of rDNA and amplified using BD1 / 4S and EGF1 / EGR2 primers, followed by RFLP using Alu1, Msp1 and TaqI restriction enzymes.

Results

The PCR-ITS1 products obtained from sheep, cattle and human isolates were similar to sheep strain (1000 bp and 391 bp). Majority of the camel samples yielded 295 bp DNA bands. RFLP -ITS1 of E. granulosus with Taq1 in human, sheep and cattle isolates showed similar patterns in the number and size of DNA. RFLP methods in camel isolates showed a different genotype, using Taq1, whereas no DNA bands were observed using Alu1 in camel and human isolates. Therefore, two clearly distinguishable banding patterns of E. granulosus were obtained with the three enzymes, which separating human, sheep and cattle isolates from the camel origin.

Conclusion

The results indicate the possible of transmission of the G1 and G6 genotypes of E. granulosus between livestock animals and human in Golestan Province.  相似文献   

2.

Background

The aim of this study was to assess the performance of Antigen B (AgB) isolated from different Echinococcus granulosus intermediate hosts and from different cyst locations for immunodiagnosis of human cystic echinococcosis (CE).

Methods

Hydatid cyst fluids were collected from lung and liver cysts of sheep, liver cysts of goats, lung cysts of camels and cattle, and liver cysts of human. AgB was purified from each of these hydatid cysts fluids. Serum samples obtained from 47 pathologically confirmed cases of CE along with 30 sera samples from non-CE patients and 40 sera from healthy individuals were tested by ELISA using AgB prepared from different hosts or cyst locations.

Results

The highest sensitivity (97.8%) for diagnosis of CE was seen with AgB prepared from human liver cysts. This maximal sensitivity was followed by AgB isolated from those of sheep liver and lung cysts. The least sensitivity was found with AgB prepared from bovine lung cysts. The highest specificities (97.1%) were observed with AgB isolated from human liver cysts followed by those of sheep and goat liver cysts while the lowest specificity was seen with AgB isolated from bovine lung cysts. In view of the specificities and sensitivities of the different AgB, the best validity was found for AgB prepared from human liver cysts while the least validity was found with AgB prepared from bovine lung cysts.

Conclusion

For any AgB-based tests, obtaining of the antigen from one of these sources will significantly increase the diagnostic sensitivity and specificity of the assay.  相似文献   

3.

Background

Cystic echinococcosis (CE), caused by hydatid cysts, is a widespread and hazardous disease in humans and animals worldwide. The disease is very common in Turkey, causing serious economic losses and public health problem. In this study, the seroprevalence of equine CE was determined by enzyme-linked immunosorbent assay (ELISA).

Methods

Partially purified cyst fluid antigen from sheep hydatid cyst fluid was used as antigen in ELISA. A total of 250 equids consisting of 206 donkeys and 44 horses from various regions of Erzurum province of Eastern Turkey.

Results

Anti- Echinococcus granulosus antibodies were detected in 78 (31.2%) of 250 equids. The prevalence rate was 20.4% for horses and 33.5% for donkeys. There was no statistically difference between sex and ages groups for both horses and donkeys (P>0.05).

Conclusion

Equine CE is quite endemic in Eastern Turkey. The high prevalence of CE suggests that equids in the transmission cycles is possible as a source of infection for definitive hosts.  相似文献   

4.

Background

Antigen B (AgB) is frequently used for immuno-diagnosis of human cystic echinococcosis (CE). Echinococcus granulosus AgBs show a high degree of genetic variability in different hosts or in different CE endemic areas. The present study aimed to evaluate the genetic polymorphisms of encoding antigen B2 gene (AgB2) among different Iranian isolates of E. granulosus.

Methods

A total of 50 CE isolates were collected from human, sheep, cattle, goat and camels, 10 isolates from each intermediate host of E. granulosus. Total genomic DNA from either protoscolices or germinal layer was extracted from each cyst and PCR-RFLP followed by DNA sequencing was used to evaluate sequence variation and polymorphism of AgB2 in the isolates.

Results

After the PCR amplification, using AgB2 primers, an almost 400 bp band was amplified in all of the isolates. The PCR products were digested with Alu1 restriction endonuclease. After restriction enzyme digestion with Alu1‚ sheep and human isolates gave a similar pattern of RFLP with the gene size of approximately 140 and 240bp and camel and goat isolates gave a similar pattern, but different from sheep and human, with the gene size of approximately 150 and 250bp. Sequence analysis showed the most genetic similarity of AgB2 between human and sheep isolates.

Conclusion

Findings of this study revealed the differences in the sequences of AgB2 within and between the Iranian isolates of E. granulosus. These differences may affect the performance of any diagnostic test which uses AgB.  相似文献   

5.

Background

Cystic echinococcosis (CE), a zoonotic parasitic infection caused by the metacestode (larvae) stage of dog tapeworm Echinococcus granulosus and recognized as a major economic and public health concern in the world. This study aimed to investigate the in vitro scolicidal effect of methanolic extract of Berberis vulgaris L. roots and its main compound, berberine against protoscoleces of hydatid cysts.

Methods

For this purpose, protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the methanolic extract (0.25-2 mg/ml) and berberine (0.062- 0.5 mg/ml) were used for 5 to 30 min. Viability of protoscoleces was confirmed by eosin exclusive test.

Results

In the present study, all of the various concentrations of the B. vulgaris methanolic extract (0.25, 0.5, 1 and 2 mg/ml) and berberine (0.062, 0.125, 0.25 and 0.5 mg/ml) revealed significant (P<0.05) scolicidal effects against protoscoleces of E. granulosus in a dose-dependent manner. Both berberine and methanolic extract exhibited 100% inhibition against protoscoleces of E. granulosus at the concentration of 2.0 and 0.5 mg/ml after 10 min incubation, respectively.

Conclusion

According to the results, both B. vulgaris methanolic extract and berberine alone demonstrated high scolicidal activities against protoscoleces of hydatid cysts in low concentration and short exposure time on in vitro model. However, in vivo efficacy of B. vulgaris and berberine also requires to be evaluated using an animal model with hydatid infection.  相似文献   

6.

Background

Echinococcosis or hydatidosis is a chronic, zoonotic worldwide infection that occurs by the larval stages of taeniid cestodes of the genus Echinococcus. Iran is known as endemic region for this infection in the world. Vaccination has been considered as a good prevention method for this disease. Recombinant vaccines containing EG95 protein, against E. granulosus, has shown a high degree of protection against E. granulosus infection. In this study EG95 gene was extracted from Iranian isolates of E. granulosus and then cloned and expressed in expression vector.

Methods

Protoscoleces were collected from sheep hydatid cysts. Then DNA and RNA were extracted from protoscoleces, and amplified by PCR and RT-PCR with specific primer. Afterward the purified RT-PCR products were successfully ligated into pTZ57R/T plasmid vector. The pcDNA3 plasmid was used as expression vector and Eg95 fragment sub cloned into this plasmid. The pcEG95 plasmid was digested by restriction enzymes to confirm cloning of this gene in pcDNA3 plasmid. In last step, the subcloned gene was expressed in CHO as eukaryotic cell.

Results

EG95 fragment successfully was subcloned in pcDNA3 and EG95 protein was expressed by eukaryotic cell. The recombinant EG95 protein was confirmed by SDS-PAGE and Western blot.

Conclusion

Recombinant plasmid of pcEG95 was constructed successfully and express of recombinant EG95 protein was confirmed.  相似文献   

7.

Background

Regarding that accurate diagnosis of human hydatidosis still needs more investigations, the present study was conducted to clone, express, and evaluate the gene encoding AgB subunits (EgAgB16 kDa) from Echinococcus granulosus (Iranian G1 strain) and its evaluation by ELISA test.

Methods

DNA was extracted from protoscoleces and was utilized by PCR for strain identification. Total RNA was prepared with RNeasy protect mini kit from E. granulosus (Iranian G1 strain) protoscoleces collected from naturally infected sheep with hydatid cyst. Recombinant AgB16 kDa was produced using pETDuet as vector and evaluated by ELISA method. A panel of sera including hydatid cyst-infected individuals (n=72), healthy individual (n=48), toxoplasmosis (n=4), strongyloidosis (n=4), kala-azar (n=5) and tuberculosis (n=5) were examined using this recombinant antigen.

Results

Recombinant protein was purified by affinity chromatography using His-Tag column. After purification, recombinant protein was confirmed by western blot analysis using His Tag monoclonal antibody or hydatid positive human serum. The sensitivity, specificity; positive and negative predictive values were calculated as 93.5%, 95.6%, 96% and 92.9%, in that order. The cut-off point was detected 0.3 for rAgB16.

Conclusion

While the produced recombinant AgB16 kDa showed promising results in diagnosing human hydatidosis, but more investigations should be implemented to reach an accurate gold standard.  相似文献   

8.

Background

Hydatidosis is the larval stage of the Echinococcus granulosus. This disease is endemic in Iran. There are many studies about hydatidosis in different regions of the country, but there is not any information about the disease in Kermanshah Province. This article will review all available data about hydatidosis in this province.

Methods

Using web based search engines and a survey on medical student''s theses, all the information about hydatid cysts in the province from 1986 -2008 was collected.

Results

During these twenty years, at least 482 proven cases of hydatid cyst have been identified in the province. Accordingly, the trend of hydatid cyst operation in the province has been growing and the average annual number of cases has reached 1.41/100,000. Frequency of disease in urban areas was slightly higher than rural areas and the rate of infection in housewives was more than others.

Conclusion

Because of the growing trend of hydatid cyst operation in Kermanshah Province, which may be due to many different reasons, this province should be considered as one of the important endemic regions of hydatidosis in Iran.  相似文献   

9.

Background

The aim of this study was DNA extraction from protoscolecses of Echinococcus granulosus and identification of these strains in West-Azerbaijan Province, north western Iran.

Methods

Thirty one livestock isolates from sheep and cattle were collected from abattoirs of the province. To investigate the genetic variation of the isolates, after DNA extraction by Glass beads-phenol chloroform method; PCR-RLFP analysis of rDNA-ITS1 was performed using three different restriction enzymes of Taq 1, Rsa 1 and Alu 1.

Result

Amplified PCR products for all isolates were 1000bp band which is expected band in sheep strains (G1-G3 complex). The results of RFLP analysis also were the same for all isolates. PCR-RFLP patterns restriction enzymes were identical as follows, Rsa1 bands under UV showed two bands approximately 655bp and 345bp. Alu1 bands were as follows: two approximately 800bp and 200bp and Taq1 did not cut any region and bands were approximately 1000 bp in all samples.

Conclusions

Based on PCR-RFLP patterns of ITS1 fragment produced with endonucleases enzyme digestion in animal isolates, it can be concluded that a single strain of E. granulosus (sheep strain or G1-G3 complex) is dominant genotype in this province.  相似文献   

10.

Background

Modulation of the immune response is an important strategy by which establishment and growth of hydatid cyst in the internal organs of human is warranted. Induction of apoptosis in the lymphocytes might be a considerable component. This study was designed to evaluate apoptotic impact of hydatid fluid (HF) on human lymphocytes.

Methods

Human lymphocytes were treated with hydatid fluid. After 6 hours of exposure, caspase-3 activity, the central enzyme of apoptosis cascade, was measured by fluorometric assay in the HF-treated lymphocytes and control cells. In addition, the expression of Bax (a pro-apoptotic protein) and Bcl-2 (an anti-apoptotic protein) mRNA was assessed by RT-PCR after 12 hours of exposure.

Results

Both the ratio of Bax/Bcl-2 mRNA expression and Caspase-3 activity were higher in the HF-treated lymphocytes relative to the control group.

Conclusion

Apoptosis could be as a possible mechanism by which Echinococcus granulosus overwhelms host defenses.  相似文献   

11.

Background

Echinococcus granulosus, a zoonotic cestode parasite, causative agent of hydatid cyst is endemic in many parts of the world including the Middle East. Study on different aspects of this parasite is very important and valuable. However, working with adult worms which their habitat situated in the small intestine of canids, is dangerous and risky. Achieving such risky situation needs a controlled condition which is cultivation of the organisms in the laboratory. In this regard, cultivation of E. granulosus protoscoleces leading to adult worms was established in the laboratory for the first time in Iran.

Methods

Under aseptic conditions a number of protoscoleces were cultivated in diphasic S.10E.H medium using CO2 incubator to produce adult worms.

Results

Different forms of parasites including pre-segmentation stages (PS1 - PS4) and segmentation stages (S5-S8) and developing stages in segmented worms (S10-S11) were observed and evaluated in these medium. Finally adult worms contained four proglottids with a large and distinct genital pore were observed 50-55 days post cultivation. These parasites do not produce fertile eggs and conclusively do not have risk of hydatid disease transmission to the researchers.

Conclusion

The mentioned method for producing E. granulosus adult worms can open a new window for researches and facilitate working on different aspects of hydatidosis especially for diagnosis, protection and treatment studies.  相似文献   

12.

Background

Toxoplasma gondii is a zoonotic protozoan parasite found worldwide and responsible for major economic losses in most classes of livestock. This study was aimed to determine the prevalence of T. gondii infection in sheep, cattle and horses in Urmia, north-west of Iran, using MAT.

Methods

Blood samples of 276 livestock and 26 horses were collected from July 2009 till April 2010. The data were analyzed by the Chi-square, Fisher''s Exact and Cochran''s and Mantel-Haenszel Tests. The level of significance was set at P<0.05.

Results

Thirty-three (21.1%) sheep, 2 (1.6%) cattle and 3 (11.5%) horses were seropositive to T. gondii. Analysis showed that sheep were 15 times more likely to be seropositive comparing to cattle also 2 times more likely to be seropositive than horses.

Conclusion

This study showed seroprevalence of equine T. gondii infection with a considerable rate in sheep in Urmia, northwest of Iran. More comprehensive studies on livestock toxoplasmosis are required for further analysis of the parasite reservoir for human infection.  相似文献   

13.

Introduction

Rotavirus vaccine was introduced in Mauritania in December 2014. We investigated hospitalizations with diarrhea during pre and post-vaccination periods among children aged 0–5?years in Nouakchott, the capital of Mauritania.

Methods

We conducted a retrospective review of hospital admission registries from November 1st 2012 through October 31th 2017 at all referral hospitals in Nouakchott. We described admissions of children aged 0–5?years by diagnosis, data of admission, age and sex, and compared the proportion of all childhood hospitalizations with diarrhea before and after rotavirus vaccine introduction.

Results

In total, 6552 (19%) of all 34,329 hospitalizations in 0–5?year-olds had diarrhea. Of these, 3523/16,952 (20.7%) were recorded during the pre-vaccine period, 1373/6897 (19.9%) during the transition period (November 2014-October 2015), and 1656/10,480 (15.8%) during the post-vaccination period. The proportion of all childhood hospitalizations with diarrhea during the pre-vaccine period was 22.6% among males and 18.7% among females. Approximately one third (32.3%) of hospitalizations with diarrhea occurred in children aged 6–11?months. During the post-vaccination period, the proportion of hospitalizations with diarrhea declined by 24%, and the highest reduction (74%) was observed in children aged 2 to 5?years (P?<?0.001).

Conclusions

The proportion of childhood hospitalizations with diarrhea in Nouakchott was reduced by about one fourth after introduction of rotavirus vaccination in Mauritania, indicating a major impact for public health for children in the capital city.  相似文献   

14.

Background

Although infection of dromedary camels with Cryptosporidium spp. is rare in Iran, it is considered a zoonotic threat to the keepers and herders of camels. Thus we investigated the prevalence of Cryptosporidium in these two hosts in Yazd Province, a semi-arid region in center of Iran.

Methods

This study was conducted during 4 seasons (winter 2008, summer 2009, winter 2009 and summer 2010). Fecal samples (n=200) were collected from live camels. Also, 100 abomasal mucosa and related fecal samples of the slaughtered camels were investigated. Stool samples from 100 individuals who were in persistent contact with camels were also obtained. After staining by modified Ziehl-Neelsen method, the prepared specimens were studied microscopically. Results were analyzed using SPSS 16.

Results

The rate of infection in feces and abomasal mucosa of camels were 20.33% and 12%, respectively. In addition, simultaneous fecal and mucosal infection was detected in 3 cases in winter. Statistical analyses showed no significant relation between infection and age of camels, as well as their sex and the season. Cryptosporidiosis in people who were in long-term contact with camels was also investigated microscopically by obtaining stool samples of 100 individuals (50 in summers, 50 in winters), 24 of them being infected with Cryptosporidium spp. The rate of infection was higher in winter than summer (16/50 compared with 8/50).

Conclusion

The prevalence of Cryptosporidium spp. in camels and involved humans in Yazd Province is relatively considerable and of public health importance.  相似文献   

15.

Background

No data is available on morphology and genetic characteristics of Echinococcus granulosus derived from donkeys of Iran, despite of its existence in donkeys. In the present study morphometric variations of the rostellar hooks of protoscoleces and genotype characteristics of hydatid cyst of donkey from Iran were determined.

Methods

Protoscoleces prepared from hydatid cyst of donkey of Iran were morphometric and genetic analyzed. The genetic analysis was done using Cox 1 gene by comparative sequence analysis.

Results

Our morphometric results showed that donkey of Iran shares 6 out of 7 determined parameters with donkeys of Jordan and 4 out of 7 with 4 available data with Switzerland donkeys. Morphological similarities and dissimilarities were observed with sheep-dog (G1) and camel-dog strains (G6) of Iran. The nucleotide sequence alignment showed that the partial sequence of Cox 1 from donkey had 91% homology with query coverage of 99% to the corresponding sequence of E. equinus, 90% homology to the E. felidis, 90% homology to E. ortleppi, 89% homology to the E. shiquinus, 89% homology to the E. vogeli, 89% homology to the E. oligarthrus, 88% homology to the E. canadensis and 83% homology to the Taenia solium. Additionally, the amino acid sequence of this gene has also some differences between this strain and all known strains of E. granulosus even with E. equinus (G4).

Conclusion

Despite of common morphological characteristics of Iranian donkey hydatid cyst with those of donkeys of other parts of the world, genetically it has its own entity.  相似文献   

16.

Background

Hydatidosis is a zoonotic disease of global prevalence. It causes considerable health problems and economic losses throughout the world, including Iran. The objective of this study was to assess the current status of echinococcosis/hydatidosis in the province of Ilam (western Iran).

Methods

From April to September 2011, 65 stray dogs were collected from urban and rural areas of Ilam City. Parasites were isolated from the dogs and stained with carmine. A taxonomic study was carried out by measuring different parts of helminths. Meat inspection documents from slaughterhouses in Ilam were used to assess the prevalence of hydatidosis during a 3-year period in sheep, cattle, and goats. ELISA test was used to detect the presence of antibodies to hydatidosis in human sera. Clinical records from 2000 to 2010 of either treated or diagnosed patients from public hospitals of this province were reviewed.

Results

The prevalence of Echinococcus granulosus infection in stray dogs was 9%. A total of 81,726 animals were assessed for hydatidosis; 2.94% (2403 cases) had liver hydatidosis and 2.34% (1918 cases) had lung hydatidosis. Within a 10-year period, 140 patients (91 females and 49 males) were treated for hydatidosis. Of 1200 human sera, 2.25% (27 patients) were seropositive for hydatidosis.

Conclusion

Hydatidosis is endemic in Ilam Province especially in rural area. The health and economic losses caused by the disease are significant; thus, our efforts need to be focused on the control of this disease.  相似文献   

17.

Background

Fascioliasis is considered as the most important helminthic infection of cattle and sheep. Traditional approaches using morphological and biologic characters cannot cause a certainty in the accurate and precise identification and intra-specific differences of Fasciola spp. In this study, we identified Fasciola species using ITS-1 marker and described genetic variation of each species of the parasite in isolates from Tabriz slaughterhouse in West Azerbaijan Province, north- western Iran.

Methods

Overall, 100 samples (50 from sheep and 50 from cattle) morphologically detected as Fasciola worms were studied for identification of Fasciola species by PCR-RFLP method and intra-species variation of the parasite using RAPD-PCR technique.

Results

A region of approximately 460bp in all samples was successfully amplified. There were no identifiable variations among the size of PCR products. Two and three fragments in samples correspond to F. hepatica and F. gigantica was seen, respectively, through PCR-RFLP method. No difference was seen in digestion pattern according to host (sheep or cattle). Different types of each species of the parasite was observed using RAPD-PCR technique.

Conclusion

We could have an estimate of frequency of F. hepatica and F. gigantic and different genotypes of the parasite in isolates from one locality in north- western of Iran. By extension of such studies in future to other animal hosts (buffalo and goat) and including more regions to sampling, the reliability of the results and their application for control programs in zoonotic diseases will be increased.  相似文献   

18.

Background

Cryptosporidium spp. is a coccidian parasite infected humans and animals. Prevalence rate of Cryptosporidium spp. infection associated with is some parameters such as sampling, age, season, country and contact to domestic animals. This study aimed to determine Cryptosporidium spp. Infection in humans and some animals in rural areas of Shushtar district from Khuzestan Province, south- west of Iran.

Methods

In this study, Stool specimens were randomly collected from 45 cattle, 8 buffalos, 35 calves, 22 turkeys, 3 sheep, 2 geese as well as 62 humans in different seasons selected from rural areas of Shushtar district located in Khuzestan in the south- west of Iran from August 2009 to April 2011. The collected stool samples were examined by modified Ziehl-Neelsen staining method.

Results

Altogether, 68/115 (59.1%) domestic animals and 9/62 (14.5%) of humans were showed Cryptosporidium spp. infection in the study areas.

Conclusion

In this study we found the high frequency of Cryptosporidium spp. infection in the studied areas.  相似文献   

19.

Background

The present study was aimed to investigate molecular diversity of Echinococcus granulosus isolates collected from human clinical samples using two mitochondrial genes cox1 and nad1 in Iran.

Methods

Forty seven human hydatid cysts were collected through surgery from two hospitals in Tehran during 2010-2012. To determine the fertility of protoscoleces, the cyst fluids were subjected to morphological microscopic examinations. Protoscoleces were removed from each cyst and their total genomic DNAs were extracted. PCR was performed to amplify fragments of 450 and 400 base pair (bp) for cox1 and nad1 genes, respectively. Genotype diversity and sequence variation of the strains were studied by bioinformatics software and in comparison with those mtDNA sequences already deposited in GenBank.

Results

Sixteen, (53.3%), 13 (43.3%), and 1 (3.3%) samples were related to lung, liver, and spleen, respectively. The remained 17 unfertile samples were excluded from the study. From the 29 isolates, 86.7% (n=26) and 10% (n=3) were related to G1, and G3 genotypes, respectively. The sole isolate with G6 genotype was obtained from lung sample. Analysis of concatenated sequences of cox1+nad1 indicated the presence of 11 haplotypes among our strains that were related to genotypes G1 (n=9), G3 (n=1) and G6 (n=1).

Conclusion

In consistent to other reports from Iran, genotypes G1, G3, and G6 were observed in our human isolates. The rate of G3 genotype was however higher than other studies implying that human can be considered as a new appropriate host for G3 genotype. Further studies with more sample size from different geographic areas of Iran are needed for E. granulosus mapping.  相似文献   

20.

Background

The aim of this study was to determine the seroprevalence of antibody to Neospora caninum in healthy and aborted dairy cattle in Tabriz, capital of East-Azarbaijan in northwest of Iran.

Methods

In this cross-sectional study serum samples were collected from 266 healthy and aborted Holestein-Feriesisnc cows from September 2008 to August 2009. The sera were analyzed to detect of antibody against N. caninum using the commercially ELISA kit.

Results

Seroprevalence of antibody to N. caninum was 10.5% in Tabriz dairy cattle. Also the abortion rate in all cattle sampled was 33.6% but percentage of seropositive aborted cattle was 18.4%.

Conclusion

Neosporosis could be one of the possible causes of abortion in dairy cattle in Tabriz and regarding the distribution in dogs as definitive host for the parasite, further studies in dog and cattle are recommended.  相似文献   

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