Fig. 2

Alignment of heterozygous ITS2 sequences of D. immitis from Portuguese canine samples. Haplotypes 7, 9, 10 and 18 include Portuguese sequences with codes LN626262-7 that are from infected mosquitoes. Haplotypes were inferred using the programme PHASE, by comparison with homozygous sequences, with 100 iterations, 100 thinning interval and 100 burn-in settings. The first position on the alignment corresponds to position 162 of the sequence, EU087699 (H18). The nucleotide codes are as standard (M, R, Y and W, correspond, respectively to A/C, A/G, C/T and A/T). Base assignment in position 11 (R) is considered uncertain (probability of 58-9%), as is for position 26 (Y) of sample 723 (50%), position 43 (Y) of sample 7, and position 51 (R) of sample 52 (50%). Haplotypes: H1: JX481279, JX866681, EU182329; H4: U182331, JN084166, JX889634, JX8896351, JX889636, JX889637, JX889638; H6: JN084168; H7: LN626265; H8: FJ263455; H9: LN626264/66; H10: LN626262; H12: FJ263458/66/67; H13: FJ263459/60/63; H14: FJ263457/64; H15: FJ263468, H16. FJ263461; H17: FJ263465; H18: EU087699, FJ263456, LN626263.1, LN626267.1; H19: EU182330. Haplotypes H2, H3, H5 and H11 result from inference