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Ophiotaenia alessandrae sp. n. (Eucestoda: Proteocephalidea), a parasite of Hyla boans (Anura: Hylidae) from Amazonia in Ecuador. Ophiotaenia alessandrae sp. n., is described from the intestine of the hylid frog, Hyla boans (Linnaeus, 1758) (Anura: Hylidae), from Amazonia in Ecuador. The new species differs from the 24 known species of the genus Ophiotaenia La Rue, 1911, parasitic in amphibians by the presence of an additional layer egg envelope forming a three layers embryophore. Furthermore, it differs from these species by one to several other morphological characters such as absence of apical organ, anterior and posterior position of the vagina, number of testes and the cirrus-sac length/proglottis width ratio. The presence of two kinds of secondary canals in proteocephalideans is discussed. This is the first observation of a three-layered embryophore in NewWorld amphibians.