Regarding the findings of Vanstreels et al. (2014), later analyses revealed that the blood sample of a tropical screech owl (Megascops choliba) that had been analyzed 4 weeks earlier at the same laboratory yielded a Haemoproteus sp. cyt-b sequence that was 100% identical to that obtained for penguin 586. This owl had been sampled in 2009 at Carapicuíba, SP, Brazil (23°34’S 46°52’W), and blood smears confirmed the infection by Haemoproteus syrnii (3.83 parasites per 10000 erythrocytes).

We reason that (a) H. syrnii is thought to be host-specific to the order level and had never been reported in non-Strigiformes hosts [2], (b) no parasites were identified in the blood smear of penguin 586 whereas they were abundant in the blood of the owl, and (c) the nested PCR method employed uses a total 55 amplification cycles [3], which may theoretically amplify each fragment up to 255 times and thus has a significant probability of contaminating the laboratory equipment and facilities.

We therefore conclude that the positive result obtained for penguin 586 was a false positive due to contamination from the owl sample, and thus the Haemoproteus lineage identified in our study (“lineage D”; GenBank KJ575554) was not identified in penguins.