Table 4.
Polymorphic sites in the sequence comparison of the nuclear rDNA ITS-2 between haplotypes of “genetically pure” Fasciola hepatica from livestock in Europe and the Americas, “genetically pure” F. gigantica from livestock in African countries, F. nyanzae from hippopotamuses in the nature reserves in South Africa, and from snails and one hippopotamus in Zimbabwe, Fascioloides jacksoni from elephants in Sri Lanka, and Fs. magna from cervids in USA.
| Fasciolid species | haplotype (H) or isolate (I) | Country | Acc. No. | ITS-2 variable positions | Length (bp) | GC (%) | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| 11111 | 1111112222 | 2222222222 | 2222223333 | 3333333333 | 333 | ||||||
| 257822234 | 4558990001 | 3333455777 | 8888890001 | 2233333334 | 555 | ||||||
| 6779001232 | 6182340341 | 3579803234 | 0168913457 | 5801235897 | 169 | ||||||
| F. hepatica | Fh-ITS2-1 | Spain + a | MG569980 | T A T T G A T G T A | G G A T C T G A G T | C T T A A A T G T C | C A G C A T G A A G | C T T T C A A G C C | TGT | 365 | 48.49 |
| F. hepatica | Fh-ITS2-2 | Spain + b | MG569981 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . . | . . . T . . . . . . | . . . . . . . . . . | . . . | 365 | 48.22 |
| F. hepatica | Fh-ITS2-Htz* | Ecuador | MK212150 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . . | . . . Y . . . . . . | . . . . . . . . . . | . . . | 365 | 48.35 |
| F. gigantica | Fg-ITS2-1 | Burkina Faso + c | AJ853848 | . . . . . . . . . . | . . . . . . . . . . | . C . . . . . . . T | T . . . . . . . . . | . . . - . . . A . . | . . . | 364 | 48.08 |
| F. gigantica | Fg-ITS2-2 | southern Algeria | ON661090 | . . . . . . . . . . | . . . . . . . . . . | . C . . C . . . T | T . . . . . . . . . | . . . - . . . A . . | . . . | 364 | 48.35 |
| F. gigantica | Fg-ITS2-Htz* | southern Algeria | ON661091 | . . . . . . . . . . | . . . . . . . . . . | . C . . M . . . T | T . . . . . . . . . | . . . - . . . A . . | . . . | 364 | 48.21 |
| F. nyanzae | Fn-ITS2-1 | South Africa-K | ON661092 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . T | . . . . . . . . . . | . . . . . . . A . . | . . . | 365 | 47.95 |
| F. nyanzae | Fn-ITS2-1 | South Africa-S | ON661092 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . T | . . . . . . . . . . | . . . . . . . A . . | . . . | 365 | 47.95 |
| F. nyanzae | I S8bLNA1 | Zimbabwe | MT893595 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . T | . . . . . . . . . . | . . . . . . . A . . | . . . | 365 | 47.95 |
| F. nyanzae | I 483 | Zimbabwe | MT893588 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . T | . . . . . . . . . . | . . . . . . . A . . | . . . | 365 | 47.95 |
| F. nyanzae | I 481 | Zimbabwe | MT893587 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . T | . . . . . . . . . . | . . . . . . . A . . | . . . | 365 | 47.95 |
| F. nyanzae | I 480 | Zimbabwe | MT893586 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . T | . . . . . . . . . . | . . . . . . . A . . | . . . | 365 | 47.95 |
| F. nyanzae | I 394 | Zimbabwe | MW046872 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . T | . . . . . . . . . . | . . . . . . . A . . | . . . | 365 | 47.95 |
| F. nyanzae | I 393 | Zimbabwe | MW046870 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . T | . . . . . . . . . . | . . . . . . . A . . | . . . | 365 | 47.95 |
| F. nyanzae | I 1 | Zimbabwe | MT909820 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . T | . . . . . . . . . . | . . . . . . . A . . | . . . | 365 | 47.95 |
| F. nyanzae | I 2 | Zimbawbe | MT909821 | . . . . . . . . . . | . . . . . . . . . . | . . . . . . . . . T | . . . . . . . . . . | . . . . . . . A . . | . . . | 365 | 47.95 |
| Fs. jacksoni | Fsj-ITS2-1 | Sri Lanka | ON661093 | . T. - . GCA. G | TAGA . CA . TC | T . . GG . C . . T | TGA . GC . GG . | T . A . A . GAAT | GA. | 364 | 48.90 |
| Fs. magna | n.d. | Oregon, USA | KU232369 | CTG- A. . . CG | . . GAACAGTG | T . C . . GCACT | . GA . GCAGGA | . CG . TGTAAT | . AA | 364 | 50.00 |
| Fs. magna | n.d. | MS, USA | EF051080 | CTG- A. . . CG | . . GAACAGTG | T . C . . GCACT | . GA . GCAGGA | . CG . TGTAAT | . AA | 364 | 50.00 |
Differences observed in a 365-bp-long alignment including 314 conserved and 51 variable positions plus 3 gapped positions, of which 50 were parsimony informative positions and 1 was a singleton site. Positions, numbers (to be read in vertical) refer to variable positions obtained in the alignment made with MEGA X; . = identical; - = insertion/deletion; n.d., not determined.
*Htz = pure species with heterozygotic position not differentiating between F. hepatica and F. gigantica (represented with corresponding symbol of IUPAC code for incomplete nucleic acid specification).
a, France, Poland, Mexico, Venezuela, Peru, Bolivia, Uruguay, Argentina.
b, Andorra, Mexico, Ecuador, Bolivia, Uruguay.
c, Niger, Nigeria, Senegal, Cameroon.
K, Klaserie Private Nature Reserve; S, Sabi Sand Nature Reserve.