Assessment of genetic markers for multilocus sequence typing (MLST) of Fasciola isolates from Iran
Abstract Background Several markers have been described to characterise the population structure and genetic diversity of Fasciola species (Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica). However, sequence analysis of a single genomic locus cannot provide sufficient resolution...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2023-03-01
|
| Series: | Veterinary Medicine and Science |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/vms3.995 |
| _version_ | 1797866287978774528 |
|---|---|
| author | Naser Nazari Mohamad Bagher Rokni Madoka Ichikawa‐Seki Saber Raeghi Homa Hajjaran Shahab Falahi Yazdan Hamzavi Peyman Heydarian Afshin Davari Keyphobad Ghadiri Arezoo Bozorgomid |
| author_facet | Naser Nazari Mohamad Bagher Rokni Madoka Ichikawa‐Seki Saber Raeghi Homa Hajjaran Shahab Falahi Yazdan Hamzavi Peyman Heydarian Afshin Davari Keyphobad Ghadiri Arezoo Bozorgomid |
| author_sort | Naser Nazari |
| collection | DOAJ |
| description | Abstract Background Several markers have been described to characterise the population structure and genetic diversity of Fasciola species (Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica). However, sequence analysis of a single genomic locus cannot provide sufficient resolution for the genetic diversity of the Fasciola parasite whose genomes are ∼1.3 GB in size. Objectives To gain a better understanding of the gene diversity of Fasciola isolates from western Iran and to identify the most informative markers as candidates for epidemiological studies, five housekeeping genes were evaluated using a multilocus sequence typing (MLST) approach. Methods MLST analysis was developed based on five genes (ND1, Pepck, Pold, Cyt b and HSP70) after genomic DNA extraction, amplification and sequencing. Nucleotide diversity and phylogeny analysis were conducted on both concatenated MLST loci and each individual locus. A median joining haplotype network was created to examine the haplotypes relationship among Fasciola isolates. Results Thirty‐three Fasciola isolates (19 F. hepatica and 14 F. gigantica) were included in the study. A total of 2971 bp was analysed for each isolate and 31 sequence types (STs) were identified among the 33 isolates (19 for F. hepatica and 14 for F. gigantica isolates). The STs produced 44 and 42 polymorphic sites and 17 and 14 haplotypes for F. hepatica and F. gigantica, respectively. Haplotype diversity was 0.982 ± 0.026 and 1.000 ± 0.027 and nucleotide diversity was 0.00200 and 0.00353 ± 0.00088 for F. hepatica and F. gigantica, respectively. There was a high degree of genetic diversity with a Simpson's index of diversity of 0.98 and 1 for F. hepatica and F. gigantica, respectively. While HSP70 and Pold haplotypes from Fasciola species were separated by one to three mutational steps, the haplotype networks of ND1 and Cyt b were more complex and numerous mutational steps were found, likely due to recombination. Conclusions Although HSP70 and Pold genes from F. gigantica were invariant over the entire region of sequence coverage, MLST was useful for investigating the phylogenetic relationship of Fasciola species. The present study also provided insight into markers more suitable for phylogenetic studies and the genetic structure of Fasciola parasites. |
| first_indexed | 2024-04-09T23:21:37Z |
| format | Article |
| id | doaj.art-3e72729e5a2141058e862524b190b6cb |
| institution | Directory Open Access Journal |
| issn | 2053-1095 |
| language | English |
| last_indexed | 2024-04-09T23:21:37Z |
| publishDate | 2023-03-01 |
| publisher | Wiley |
| record_format | Article |
| series | Veterinary Medicine and Science |
| spelling | doaj.art-3e72729e5a2141058e862524b190b6cb2023-03-21T15:53:53ZengWileyVeterinary Medicine and Science2053-10952023-03-019292493310.1002/vms3.995Assessment of genetic markers for multilocus sequence typing (MLST) of Fasciola isolates from IranNaser Nazari0Mohamad Bagher Rokni1Madoka Ichikawa‐Seki2Saber Raeghi3Homa Hajjaran4Shahab Falahi5Yazdan Hamzavi6Peyman Heydarian7Afshin Davari8Keyphobad Ghadiri9Arezoo Bozorgomid10Infectious Diseases Research Center Health Institute Kermanshah University of Medical Sciences Kermanshah IranDepartment of Medical Parasitology and Mycology School of Public Health Tehran University of Medical Sciences Tehran IranLaboratory of Veterinary Parasitology Faculty of Agriculture Iwate University Morioka JapanDepartment of Laboratory Sciences Maragheh University of Medical Sciences Maragheh IranDepartment of Medical Parasitology and Mycology School of Public Health Tehran University of Medical Sciences Tehran IranZoonotic Diseases Research Center Ilam University of Medical Sciences Ilam IranInfectious Diseases Research Center Health Institute Kermanshah University of Medical Sciences Kermanshah IranDepartment of Medical Parasitology and Mycology School of Medicine Qazvin University of Medical Sciences Qazvin IranDepartment of Medical Parasitology and Mycology School of Public Health Tehran University of Medical Sciences Tehran IranInfectious Diseases Research Center Health Institute Kermanshah University of Medical Sciences Kermanshah IranInfectious Diseases Research Center Health Institute Kermanshah University of Medical Sciences Kermanshah IranAbstract Background Several markers have been described to characterise the population structure and genetic diversity of Fasciola species (Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica). However, sequence analysis of a single genomic locus cannot provide sufficient resolution for the genetic diversity of the Fasciola parasite whose genomes are ∼1.3 GB in size. Objectives To gain a better understanding of the gene diversity of Fasciola isolates from western Iran and to identify the most informative markers as candidates for epidemiological studies, five housekeeping genes were evaluated using a multilocus sequence typing (MLST) approach. Methods MLST analysis was developed based on five genes (ND1, Pepck, Pold, Cyt b and HSP70) after genomic DNA extraction, amplification and sequencing. Nucleotide diversity and phylogeny analysis were conducted on both concatenated MLST loci and each individual locus. A median joining haplotype network was created to examine the haplotypes relationship among Fasciola isolates. Results Thirty‐three Fasciola isolates (19 F. hepatica and 14 F. gigantica) were included in the study. A total of 2971 bp was analysed for each isolate and 31 sequence types (STs) were identified among the 33 isolates (19 for F. hepatica and 14 for F. gigantica isolates). The STs produced 44 and 42 polymorphic sites and 17 and 14 haplotypes for F. hepatica and F. gigantica, respectively. Haplotype diversity was 0.982 ± 0.026 and 1.000 ± 0.027 and nucleotide diversity was 0.00200 and 0.00353 ± 0.00088 for F. hepatica and F. gigantica, respectively. There was a high degree of genetic diversity with a Simpson's index of diversity of 0.98 and 1 for F. hepatica and F. gigantica, respectively. While HSP70 and Pold haplotypes from Fasciola species were separated by one to three mutational steps, the haplotype networks of ND1 and Cyt b were more complex and numerous mutational steps were found, likely due to recombination. Conclusions Although HSP70 and Pold genes from F. gigantica were invariant over the entire region of sequence coverage, MLST was useful for investigating the phylogenetic relationship of Fasciola species. The present study also provided insight into markers more suitable for phylogenetic studies and the genetic structure of Fasciola parasites.https://doi.org/10.1002/vms3.995Fasciola hepaticaFasciola giganticagenetic diversityIranmultilocus sequence typing |
| spellingShingle | Naser Nazari Mohamad Bagher Rokni Madoka Ichikawa‐Seki Saber Raeghi Homa Hajjaran Shahab Falahi Yazdan Hamzavi Peyman Heydarian Afshin Davari Keyphobad Ghadiri Arezoo Bozorgomid Assessment of genetic markers for multilocus sequence typing (MLST) of Fasciola isolates from Iran Veterinary Medicine and Science Fasciola hepatica Fasciola gigantica genetic diversity Iran multilocus sequence typing |
| title | Assessment of genetic markers for multilocus sequence typing (MLST) of Fasciola isolates from Iran |
| title_full | Assessment of genetic markers for multilocus sequence typing (MLST) of Fasciola isolates from Iran |
| title_fullStr | Assessment of genetic markers for multilocus sequence typing (MLST) of Fasciola isolates from Iran |
| title_full_unstemmed | Assessment of genetic markers for multilocus sequence typing (MLST) of Fasciola isolates from Iran |
| title_short | Assessment of genetic markers for multilocus sequence typing (MLST) of Fasciola isolates from Iran |
| title_sort | assessment of genetic markers for multilocus sequence typing mlst of fasciola isolates from iran |
| topic | Fasciola hepatica Fasciola gigantica genetic diversity Iran multilocus sequence typing |
| url | https://doi.org/10.1002/vms3.995 |
| work_keys_str_mv | AT nasernazari assessmentofgeneticmarkersformultilocussequencetypingmlstoffasciolaisolatesfromiran AT mohamadbagherrokni assessmentofgeneticmarkersformultilocussequencetypingmlstoffasciolaisolatesfromiran AT madokaichikawaseki assessmentofgeneticmarkersformultilocussequencetypingmlstoffasciolaisolatesfromiran AT saberraeghi assessmentofgeneticmarkersformultilocussequencetypingmlstoffasciolaisolatesfromiran AT homahajjaran assessmentofgeneticmarkersformultilocussequencetypingmlstoffasciolaisolatesfromiran AT shahabfalahi assessmentofgeneticmarkersformultilocussequencetypingmlstoffasciolaisolatesfromiran AT yazdanhamzavi assessmentofgeneticmarkersformultilocussequencetypingmlstoffasciolaisolatesfromiran AT peymanheydarian assessmentofgeneticmarkersformultilocussequencetypingmlstoffasciolaisolatesfromiran AT afshindavari assessmentofgeneticmarkersformultilocussequencetypingmlstoffasciolaisolatesfromiran AT keyphobadghadiri assessmentofgeneticmarkersformultilocussequencetypingmlstoffasciolaisolatesfromiran AT arezoobozorgomid assessmentofgeneticmarkersformultilocussequencetypingmlstoffasciolaisolatesfromiran |