First integrative study of the diversity and specificity of metacercariae of Posthodiplostomum Dubois, 1936 from native and introduced fishes in the Caribbean.

Metacercariae of the genus Posthodiplostomum are often recorded in freshwater fish hosts. While the diversity and taxonomy of this genus are receiving increasing attention in molecular phylogenetic studies, available data remain geographically biased. Most molecular studies of Posthodiplostomum and morphologically similar (neascus) worms originate in North America and Europe and Asia (more than 60% of DNA sequences are from USA and Canada), with few data currently available from the Neotropics, where high host diversity suggests high and under-sampled parasite diversity. In this study, we report molecular and morphological data from metacercariae of Posthodiplostomum in fish in Puerto Rico, where only a single species has been previously reported. Partial sequences of cytochrome c oxidase subunit 1 from metacercariae from Dajaus monticola (native to Puerto Rico) and the introduced fishes Poecilia reticulata, Parachromis managuensis, Lepomis macrochirus and Micropterus salmoides revealed 7 genetically distinct species-level lineages, of which 4 were novel. We report novel molecular life-cycle linkages in Posthodiplostomum macrocotyle (metacercariae in muscle of the cichlid Pa. managuensis), a species previously known only from adults in birds from South America; and in Posthodiplostomum sp. 23 (metacercariae in poeciliids), which has recently been found in Ardea herodias in Georgia, USA. We also report the first molecular data from Posthodiplostomum sp. 8 in M. salmoides in the Caribbean. Metacercariae of most species were morphologically distinguished and all displayed narrow specificity for fish hosts, with no indication of parasite sharing among introduced and native fishes.

Autochthonous transmission of the Indomalayan parasite, Transversotrema patialense, in the Caribbean: Molecular, morphological, and experimental evidence.

The Asian freshwater snail Melanoides tuberculata has been established since the 1960s in the Americas, where it transmits cercariae of a small number of digenetic trematode species from its native range. In 2021-2022, 24 M. tuberculata were discovered shedding transversotrematid cercariae in Puerto Rico, where parasites of this snail have not been previously studied. Adult transversotrematids (in some cases, gravid) were found on field-caught fish and on fish exposed to shedding snails, including on fish species native to Puerto Rico. Adults and cercariae were identified as Transversotrema patialense (Soparkar, 1924), a species native to the Indomalayan region. Morphological identification was supported with 28S rDNA sequences closely matching that from unidentified transversotrematid cercariae in Thailand. The absence of T. patialense in snails collected prior to 2021, increasing prevalence of infection in snails collected thereafter, and lack of variation in parasite DNA sequences (28S, internal transcribed spacer 2, cytochrome c oxidase I) from three isolates are consistent with a recently introduced and possibly expanding parasite population. Transversotrema patialense has been recorded outside its native range before, but most studies (including a prior record in the Americas) reported the parasite from captive hosts from commercial sources such as pet shops. The present results thus provide the first demonstration of natural transmission of T. patialense in the Americas. Phylogenetic analysis of 28S but not of ITS2 show the transversotrematid genus Transversotrema Witenberg, 1944 is paraphyletic, with Crusziella Cribb, Bray and Barker 1992 nested within it.