Molecular Characterization of Fasciola spp. from Some Parts of Iran

2020 ◽  
Author(s):  
Hamid HASANPOUR ◽  
Reza FALAK ◽  
Saied Reza NADDAF ◽  
Santiago MAS-COMA ◽  
Mohammad Bagher ROKNI ◽  
...  
Keyword(s):  
East Asia ◽  
Fasciola Hepatica ◽  
Genetic Difference ◽  
Rflp Analysis ◽  
Restriction Fragment Polymorphism ◽  
Pcr Rflp ◽  
The World ◽  
Liver Flukes ◽  
Fasciola Spp ◽  
Different Parts

Background: Identification of liver flukes, Fasciola hepatica, and Fasciola gigantica by morphometric parameters is not always reliable due to the overlapping measurements. This study aimed to characterize the liver flukes of animals from different parts of Iran by the genetic markers, ITS1, and COXI. Methods: We collected flukes from infected livestock in six provinces of Iran from Sep to Nov 2016. The flukes were identified by amplification of a 680 bp sequence of ITS1 locus followed by a restriction fragment polymorphism (RFLP) assay. The genetic diversity among isolates was evaluated by amplification and sequencing of a 493 bp fragment of the COXI gene. Results: We obtained 38 specimens from Khuzestan, 22 from Tehran, 10 from Isfahan, 10 from Mazandaran, 4 from Kurdistan, and 3 from Ardabil provinces. PCR-RFLP analysis revealed two patterns, representing F. hepatica, and F. gigantica. Fifty specimens from cattle and sheep exhibited F. hepatica pattern and 37 from the cattle, sheep, buffalo, and goat that of F. gigantica. The phylogeny based on COXI revealed two distinct clades separating F. hepatica from F. gigantica. In our phylogeny, the Iranian F. gigantica isolates showed a distinct separation from the African flukes, while grouped with the East Asia specimens demonstrating a common ancestor. The F. hepatica isolates clustered with the flukes from different parts of the world, including East Asia, Europe, and South America. Conclusion: The present study revealed a substantial genetic difference between F. gigantica populations of Asia and Africa, while F. hepatica isolates from different parts of the world shared high similarities.

scholarly journals Specific Detection of Fasciola hepatica and F. gigantica in Infected Domesticated Animals Using High-Resolution Melting Analysis (HRM)

2020 ◽  
Author(s):  
Zeinab MOGHADAMIZAD ◽  
Ahmad HOSSEINI-SAFA ◽  
Mehdi MOHEBALI ◽  
Peyman HEYDARIAN ◽  
Mojgan ARYAEIPOUR ◽  
...  
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Fasciola Hepatica ◽  
High Resolution Melting Analysis ◽  
Universal Primers ◽  
Specific Primer ◽  
Universal Primer ◽  
Melting Analysis ◽  
Liver Flukes ◽  
Fasciola Spp

Background: It is difficult to make an exact morphological distinction between Fasciola hepatica and Fasciola gigantica. We used High Resolution Melting analysis (HRM) method to differentiate the F. hepatica species from F. gigantica in order to differentiate them. Methods: Overall, 80 adult liver flukes were collected from infected slaughtered animals including cattle, sheep and goats from Lorestan Province, western Iran from Sep 2015 to Aug 2017. Genomic DNA was extracted using commercial DNA extraction kit. The multilocus sequences of mDNA including COX1, COX3 and ND6 were amplified employing real-time PCR & HRM analysis. Specific and universal primer pairs were designed for differentiation Fasciola spp. Results: Universal primers cannot be used to distinguish between these two species, but in the contrary, specific primer pairs of each species could differentiate them properly. Molecular identification using specific primer pairs were consistent. Conclusion: HRM is a simple, fast and reliable method for detecting and differentiating F. hepatica from F. gigantica and can be used for diagnostic and epidemiological purposes.

scholarly journals Occurrence, Morphometric, and Molecular Investigation of Cattle and Buffalo Liver Adult Fluke in Peninsular Malaysia Main Abattoirs

2020 ◽  
Vol 2020 ◽  
pp. 1-6
Author(s):  
J. Nur Alia Diyana ◽  
M. I. Nur Mahiza ◽  
H. Latiffah ◽  
S. H. Nur Fazila ◽  
I. H. Lokman ◽  
...  
Keyword(s):  
Molecular Analysis ◽  
Fasciola Hepatica ◽  
Peninsular Malaysia ◽  
Adult Fluke ◽  
Parasitic Disease ◽  
Body Width ◽  
Body Area ◽  
Pcr Rflp ◽  
Molecular Investigation ◽  
Liver Flukes

Fascioliasis is a parasitic disease of human and animal caused by Fasciola gigantica (F. gigantica) and Fasciola hepatica (F. hepatica). More than 700 million of grazing animals and over 180 million human population are at the risk of fascioliasis. This study was conducted in Banting, Ipoh, Shah Alam, and Taiping abattoirs within Peninsular Malaysia to determine the occurrence and identify the species of liver flukes, causing liver condemnation in cattle and buffaloes. Within a study period from January to December 2018, a total of 25 condemned bovine livers were collected from Banting, Ipoh, Shah Alam, and Taiping abattoirs and analyzed. Taiping abattoir had the highest occurrence of fascioliasis [14/1014 (1.38%)], and Shah Alam abattoir had the least occurrence [1/3377 (0.03%)]. From all the sampled livers, the average number of adult flukes recorded ranged from 1 to 83. A total of 440 adult flukes were studied morphometrically, using parameters such as body length (BL), body width (BW), cone length (CL), cone width (CL), body area (BA), body perimeter (BP), the distance between the ventral sucker and posterior end of body (VS-P), BL/BW ratio, BL/V-SP, and body roundness (BR). Furthermore, molecular analysis was conducted using PCR-RFLP methods to distinguish between F. gigantica and F. hepatica involving ITS1 primer and RsaI restriction enzyme. RFLP pattern with RsaI produced a consistent pattern of 360,100 and 60 bp fragments in F. hepatica, whereas F. gigantica worms had a profile of 360,170, and 60 bp in size. The morphometric and molecular analysis results indicated that cattle and buffaloes slaughtered at Banting, Shah Alam, Taiping, and Ipoh abattoirs were infected with F. gigantica.

Genotyping of Fasciola hepatica Isolated from Domestic Animals in the West of Iran

2019 ◽  
Vol 19 (4) ◽  
pp. 409-413
Author(s):  
Sahar Shokouhi ◽  
Jahangir Abdi ◽  
Reza Valizadeh
Keyword(s):  
Restriction Enzyme ◽  
Fasciola Hepatica ◽  
Animal Husbandry ◽  
Domestic Animals ◽  
The West ◽  
Accurate Identification ◽  
Its1 Sequence ◽  
Pcr Rflp ◽  
Control And Prevention ◽  
Fasciola Spp

Objective: The most common parasites responsible for fascioliasis are F. hepatica and F. gigantica. Fascioliasis is an important problem in terms of health and economics. The western part of Iran is one of the major hubs of animal husbandry and the prevalence of fascioliasis is high. The aim of the current study was to determine the Fasciola spp. present in the western part of Iran. In the present study, 45 samples were collected from slaughterhouses in three provinces in the western part of Iran, including Ilam, Lorestan and Kermanshah. Methods: The flukes were detected using morphological methods and the DNA of all samples was extracted. The ribosomal internal transcribed spacer (ITS1) was identified by PCR and PCR-RFLP techniques using the Rsa1 restriction enzyme. Then, 15% of the samples were sequenced. Results: Based on their ITS1 sequence, all samples showed 700 bp bands. The results of sequencing showed a similarity of 99% to 100% across samples. Using the Rsa1 restriction enzyme, all samples produced three distinct bands (60 bp, 100 bp, and 360 bp). These results demonstrated that the most common fasciola in the western part of Iran in sheep and cattle is F. hepatica. Conclusion: The results of the present study showed that only the species F. hepatica is parasitizing livestock in the western part of Iran. Further studies using new molecular markers for more accurate identification of fasciola-causing species will be useful in the control and prevention of fascioliasis.

scholarly journals Fascioliasis-An Uncommon Cause of Recurrent Cholangitis

2019 ◽  
Vol 19 (1) ◽  
pp. 35-38
Author(s):  
Bidhan C Das ◽  
Arif Salam Khan ◽  
Md Saief Uddin ◽  
Md Mohsen Chowdhury ◽  
Md Zulfiqur Rahman Khan
Keyword(s):  
East Asia ◽  
Pancreatic Surgery ◽  
Fasciola Hepatica ◽  
Far East ◽  
Biliary Tree ◽  
Definite Diagnosis ◽  
Liver Flukes ◽  
Medical University

We report a case of Fascioliasis presented with recurrent cholangitis in a female necessitating multiple consultations and hospitalizations over a period of two years. Investigative profile failed to confirm any definite diagnosis prior to her admission in Hepato-Biliary-Pancreatic Surgery Division of the Department of Surgery of Bangabandhu Sheikh Mujib Medical University where she was provisionally diagnosed as a case suffering from extrahepatic cholagiocarcinoma. With the intent of extrahepatic excision with Rouxen-Y hepaticojejunostomy abdomen was explored and surprisingly 28 mature liver flukes (fasciola hepatica) were found lodged in the extra and intrahepatic biliary tree. Although fasciola hepatica infestation is more common in far East Asia, this case may represent the tip of an iceberg of endemic infestation of this trematode in Bangladesh which warrants further investigations. Journal of Surgical Sciences (2015) Vol. 19 (1) : 35-38

scholarly journals Fascioliasis

2019 ◽  
Vol 2 (1) ◽  
pp. 1-5
Author(s):  
Kittiyod Poovorawan
Keyword(s):  
Aquatic Plants ◽  
Fasciola Hepatica ◽  
Clinical Symptoms ◽  
Intestinal Wall ◽  
Fasciola Gigantica ◽  
Mild Disease ◽  
Drug Of Choice ◽  
Liver Flukes ◽  
Fasciola Spp ◽  
The Right

Fascioliasis is infection by liver flukes, Fasciola spp. The two species that infect humans are Fasciola hepatica and Fasciola gigantica. Other herbivorous animals such as cattle, goats, water buffalo, horses, camels, hogs, rabbits, and deer can be infected with Fasciola spp. Humans acquire Fasciola after ingestion of food and water that are contaminated with Fasciola metacercaria, especially aquatic plants. Fascioliasis is usually mild disease in human. Clinical manifestrations are related with the the phases of infection. In migratory phase, symptoms may occur when metacercariae excyst in the duodenum and young liver flukes migrate through the intestinal wall to the liver capsule and hepatic parenchyma before reaching the bile ducts. Clinical symptoms include abdominal pain in the right hypochondrium, dyspepsia and anorexia, nausea, vomiting, and hepatomegaly. Systemic manifestration such as urticaria and eosinophilia are also common presentation. After Fasciola reside in the biliary tract, most of cases are asymptomatic. Biliary track obstruction can accidentally occur and might lead to cholangitis. Triclabendazole 10 mg/kg as a single dose is the current drug of choice for fascioliasis treatment and triclabendazole two doses of 10 mg/kg for 1 day is effective in severe or persistent infections, with cure rates of 79% and 92%, respectively. Currently, there is no vaccine for prevent Fasciola infection. Proper cooking or cleaning food especially aquatic plants before consumption will prevent accidental Fasciola infection. Adequate mass treatment among infected animals in endemic areas and proper sanitary in the livestock farm are important for disease control.   Figure 1  ภาพถ่ายตัวเต็มวัยพยาธิ Fasciola hepatica (Linnaeus, 1758) (A) และ Fasciola gigantica (Cobbold, 1855) (B)(ได้รับการอนุเคราะห์ภาพจาก รศ.ดร.อุรุษา แทนขำ ภาควิชาปรสิตหนอนพยาธิ คณะเวชศาสตร์เขตร้อน มหาวิทยาลัยมหิดล)                 

scholarly journals Fasciola Hepatica Extract Induces Cell Death of Mammalian Cells

2018 ◽  
Vol 16 (2) ◽  
pp. 144-146 ◽  
Author(s):  
Stephanie Ferreira ◽  
Ruben Fernandes ◽  
Monica C. Botelho
Keyword(s):  
Cell Proliferation ◽  
Mammalian Cells ◽  
Fasciola Hepatica ◽  
Cell Number ◽  
International Agency ◽  
Mitochondrial Activity ◽  
Poor People ◽  
Unexpected Death ◽  
The World ◽  
Liver Flukes

Background: Fascioliasis is a neglected tropical disease that affects poor people from poor and developing countries. In the world, it has been estimated that at least 2.6 million people are affected with this disease. The International agency for Research on Cancer, states that O. viverrini and C. sinensis, also liver flukes, are considered as definitive causes of cholangiocarcinoma. However, fascioliasis caused by F. hepatica has not been associated with cancer to date. There are not any known causative associations between this parasite and liver cancer (cholangiocarcinoma). Methods: Chine Hamster Ovary (CHO) cells were treated with F. haepatica extracts and cell proliferation was assessed by using the indirect method for estimating cell number based on the mitochondrial activity with MTS cell proliferation reagent. We observed unexpected death of these cells when treated with F. haepatica extracts. Results and Conclusion: We now hypothesize that this parasite could be used as a medically-important trematode pathogen in cancer therapy.

scholarly journals Molecular Characterization of Fasciola spp. from a Donkey (Equus asinus) Using Partial Sequencing of cox1 and nad1

2020 ◽  
Author(s):  
Mohammad ASADPOUR ◽  
Hassan SHARIFIYAZDI ◽  
Mohammad MOAZENI ◽  
Seyed Hossein MALEKPOUR
Keyword(s):  
Fasciola Hepatica ◽  
Sequence Data ◽  
Nucleotide Polymorphisms ◽  
Equus Asinus ◽  
The World ◽  
Mitochondrial Sequences ◽  
Southwestern Iran ◽  
Fasciola Spp ◽  
Important Parasite

Background: Fasciola hepatica as an important parasite affects health of humans and animals in some tropical and subtropical areas of the world, including Iran. Little is known about the molecular diversity of Fasciola in Equidae. Therefore, this study aimed to characterize the genetic polymorphisms among parasites. Methods: Eight adult Fasciola spp. isolates were collected from a working donkey after necropsy in Shiraz, southwestern Iran, in 2018. Primarily, various parameters were measured morphologically. Subsequently, DNA was extracted from each fluke and molecular markers of cytochrome C oxidase (cox1) and NADH dehydrogenase 1(nad1) from individual Fasciola isolates were amplified using PCR assay and sequence data were employed for molecular and phylogenetic analysis. Genetic diversity between isolates was evaluated by comparing the sequences of these two mitochondrial regions. Results: Based on the morphological and analyzed mitochondrial sequences, all of eight donkey isolates (100%) were identified as F. hepatica. Moreover, nine and five nucleotide polymorphisms were identified in the cox1and nad1 region sequences, respectively. Conclusion: Accordingly, phylogenetic data revealed five and four haplotypes among donkey isolates based on the cox1and nad1 markers. Similarly, some of these haplotypes have been previously reported from different host species in Iran as well as all around the world.

Variations in cercarial production and the level of in vitro activation of metacercariae of two different isolates of Fasciola hepatica

2015 ◽  
Vol 60 (3) ◽  
Author(s):  
Kamil Januszkiewicz ◽  
Luke J. Norbury ◽  
Przemysław Wilkowski ◽  
Anna Zawistowska-Deniziak ◽  
Agnieszka Wesołowska ◽  
...  
Keyword(s):  
National Park ◽  
Fasciola Hepatica ◽  
Economic Losses ◽  
Cercarial Production ◽  
The World ◽  
In Vitro Activation ◽  
Białowieża National Park ◽  
Liver Flukes ◽  
Veterinary Laboratory

AbstractFasciola hepatica infections cause large economic losses and are a serious veterinary medicine problem in many regions of the world. Recent studies examining fascioliasis in the bison population from Bialowieza National Park have shown that the prevalence of infection with this parasite is up to 100%. Liver flukes isolated from bison from Bialowieza National Park in Poland were compared with a fluke strain originally obtained from the Central Veterinary Laboratory, Weybridge, UK, to determine variations in cercarial production and establish the ability of their metacercariae to activate in vitro. Some small differences in cercarial production between the two isolates are shown, while significant differences in the ability of their metacercariae to activate in vitro were observed.

scholarly journals Analysis of Kinetoplast DNA from Mexican Isolates ofLeishmania (L.) mexicana

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Omar Hernández-Montes ◽  
Saúl González Guzmán ◽  
Federico Martínez Gómez ◽  
Douglas C. Barker ◽  
Amalia Monroy-Ostria
Keyword(s):  
Rflp Analysis ◽  
Restriction Enzymes ◽  
The Other ◽  
Polymorphism Analysis ◽  
Specific Primers ◽  
Restriction Fragment Polymorphism ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Similar Restriction ◽  
Polymerase Chain

This study analyzed DNA minicircles of Mexican isolates ofL. (Leishmania) mexicanato look for genetic differences between strains isolated from patients with diffuse cutaneous (DCL) and localized (LCL) leishmaniasis. The kDNA was analyzed using polymerase chain reaction (PCR), restriction fragment polymorphism analysis of the PCR products (PCR-RFLP) and the PCR products were sequenced. In the PCR with primers specific for the subgenusLeishmania, the Mexican isolates gave higher amplification products than the otherL. mexicanacomplex strains and with specific primers for theL. mexicanacomplex they were poorly amplified. In the PCR-RFLP analysis with theEco RV,Hae III, andMbo Iendonucleases, the Mexican isolates displayed similar restriction patterns, but different from the patterns of the other members of theL. mexicanacomplex. In the phylogenetic tree constructed, the kDNA sequences of the Mexican clones formed two groups including sequences of LCD or LCL clones, apart from the otherL. mexicanacomplex members. These results suggest that the kDNA minicircles of the Mexican isolates are more polymorphic than the kDNA of other members of theL. mexicanacomplex and have different recognition sites for the restriction enzymes used in this study.

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