A molecular and ecological study of Macracanthorhynchus ingens (von Linstow, 1879) (Acanthocephala: Archiacanthocephala), in its paratenic and definitive hosts in southeastern Mexico and the Eastern USA.

The acanthocephalan Macracanthorhynchus ingens (von Linstow 1879) (Acanthocephala: Archiacanthocephala) is a parasite that infects the gut of carnivores (racoons, coyotes, wolves, foxes, badgers, skunks, opossum, mink and bears) as an adult and the body cavity of lizards, snakes, and frogs as a cystacanth in the Americas. In this study, adults and cystacanths of M. ingens from southeastern Mexico and southern Florida, USA, were identified morphologically by having a cylindrical proboscis armed with 6 rows of hooks each with 6 hooks. Hologenophores were used to sequence the small (SSU) and large (LSU) subunits of ribosomal DNA and cytochrome c oxidase subunit 1 (cox 1) from mitochondrial DNA. Phylogenetic analysis of the new SSU and LSU sequences of M. ingens placed them in a clade with other sequences available in GenBank identified as M. ingens. The cox 1 tree showed that the nine new sequences and six previously published sequences of M. ingens from the USA form a clade with other sequences previously identified as M. ingens from GenBank. The intraspecific genetic divergence among isolates from the Americas ranged from 0 to 2%, and in combination with the phylogenetic trees confirmed that the isolates belonged to the same species. The cox 1 haplotype network inferred with 15 sequences revealed 10 haplotypes separated from each other by a few substitutions. Rio Grande Leopard Frogs and Vaillant´s Frogs harbored cystacanths with low prevalence, 28% and 37% respectively, in Mexico. Brown Basilisks, an invasive lizard in Florida, USA, had high values of prevalence, 92% and 93% in males and females, respectively. Females harbored more cystacanths than males (0-39 vs 0-21) for unknown reasons that may, however, be related to ecological differences.

Contrasting the population genetic structure of a specialist (Hexaglandula corynosoma: Acanthocephala: Polymorphidae) and a generalist parasite (Southwellina hispida) distributed sympatrically in Mexico.

Polymorphidae is a monophyletic group of acanthocephalans distributed worldwide. Within this family, Hexaglandula corynosoma is a specialist species that uses a single bird species as a definitive host. Southwellina hispida is a generalist species that uses a broad spectrum of definitive hosts to complete its life cycle. In the current research, sequences of cytochrome c oxidase subunit 1 (cox1) from mitochondrial DNA were generated from 44 specimens of H. corynosoma and 76 of S. hispida distributed sympatrically in 6 biogeographic provinces of Mexico with the objective of characterizing and comparing the population genetic structure of 2 acanthocephalan species with opposing life strategies. The phylogeographic studies indicated that the populations of both species lacked a phylogeographic structure and exhibited high haplotype diversity, low nucleotide diversity and low Fst values among the biogeographic provinces; in combination with negative values on the neutrality test, this suggests that the populations of acanthocephalans are expanding. Paratenic hosts are key for the transmission from intermediate to definitive hosts in the generalist species. However, the inclusion of paratenic hosts does not play a principal role in the population genetic structure of S. hispida within its distribution along the coasts of Mexico.

First record of Patagifer bilobus (Rudolphi, 1819) Dietz, 1909 (Digenea: Echinostomatidae), with a morphological and molecular characterization from two threskiornithid species in Mexico.

Patagifer Dietz, 1909 is a small genus of echinostomatids, with 12 recognized species, mostly parasitising threskiornithid birds, distributed worldwide. In the current research, adult specimens of the type species, Patagifer bilobus (Rudolphi, 1819) Dietz, 1909 from the white faced ibis (Plegadis chihi) and white ibis (Eudocimus albus) were re-described, providing new metrical data for the number of head collar spines. Those specimens were recorded from eight localities in Mexico and compared morphologically with specimens previously identified as Patagifer lamothei. A total of 19 specimens identified as P. bilobus including two hologenophores were sequenced with three molecular markers: domains D1-D3 of the large subunit (LSU), the internal transcribed spacer (ITS1, ITS2) plus 5.8S from the nuclear rDNA, and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (nad1) from mitochondrial DNA. The new sequences were aligned with other sequences of Patagifer spp., downloaded from GenBank. Phylogenetic trees inferred from each data set, placed all the specimens in a clade, confirming that the isolates belonged to the same species. The morphological examination of specimens previously identified as P. lamothei by Ortega-Olivares MP, Hernández-Mena DI, Pérez-Ponce de León G, García-Varela M (2011) Helminths of the white ibis, Eudocimus albus (Aves Therskiornithidae) in Mexico. (Zootaxa 3088, 15-26. 10.11646/zootaxa.3088.1.2) and in combination with molecular data confirms that those specimens should be reassigned to P. bilobus. In addition, this is the first study in P. bilobus using an integrative taxonomy approach.

Host-induced phenotypic plasticity in Saccocoelioides lamothei Aguirre-Macedo and Violante-González, 2008 (Digenea: Haploporidae) a parasite of freshwater, brackish and marine fishes from Middle America.

Saccocoelioides is a genus of trematodes associated with fishes from the Americas. In the current research, morphologically distinct specimens of Saccocoelioides spp. were collected from six countries in Middle America. Specimens were sequenced using three molecular markers, the domains D1-D3 of the large subunit (LSU) from the nuclear rDNA, the cytochrome c oxidase subunit 1 (cox1) and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (nad1) from mitochondrial DNA. A total of 74 new sequences were compared and aligned with other sequences available in GenBank. Maximum likelihood and Bayesian inference analyses were inferred from the LSU and cox1 datasets, revealing unequivocally that all the specimens correspond to S. lamothei. A haplotype network was built with 119 sequences of the nad1 gene. The network detected 57 distinct haplotypes divided into three haplogroups. To explore morphological differences among samples of S. lamothei, 17 morphological features were measured from 53 specimens from three fish families: Eleotridae, Mugilidae and Gobiidae. Principal component analysis yielded three main polygons that corresponded with each family analysed, suggesting host-induced phenotypic plasticity. The current evidence suggests that S. lamothei infects at least five fish families along the Pacific coasts of Mexico, Guatemala, El Salvador, Honduras, Nicaragua and Costa Rica.

Description of a new species and understanding the genetic diversity of Saccocoelioides Szidat, 1954 (Haploporidae) in Middle America using mitochondrial and nuclear DNA sequences.

Members of the genus Saccocoelioides Szidat, 1954, include endoparasites from freshwater and brackish fishes from the Americas. Adult specimens were collected from the intestines of Poecilia catemaconis Miller, 1975, a poeciliid fish endemic to Catemaco Lake, and the white mullet Mugil curema Valenciennes, 1836, from Alvarado Lagoon, Veracruz, Mexico. The specimens were sequenced for three molecular markers, internal transcribed spacer 2 (ITS2) and domains D1-D3 from the large subunit (LSU) of nuclear ribosomal DNA and cytochrome c oxidase subunit 1 (cox 1) from mitochondrial DNA. The newly sequenced specimens were aligned with other sequences downloaded from GenBank. Maximum likelihood and Bayesian inference analyses were inferred with three data sets (a combination of nuclear DNA ITS2 + LSU, cox 1 alone and the concatenated cox 1+ ITS2 + LSU). The phylogenetic analyses inferred with the combined data set of the two nuclear molecular markers (ITS2 + LSU) revealed that Saccocoelioides is monophyletic and formed 11 independent lineages representing 11 valid species previously recognized plus the new lineage that is herein described as a new species named Saccocoelioides macrospinosus n. sp., however, the new species was placed in a basal polytomy in the tree. Therefore, the addition of a mitochondrial gene with a fast rate of substitution was fundamental to clarify the phylogenetic relationships of the new species. The genetic divergences estimated with the cox 1 gene were high, ranging from 8.3 to 15.5% among Saccocoelioides macrospinosus n. sp. and sister taxa. The new species has a slightly elongated body measuring 440-850 µm long and was classified in the diminutive morphotype. In addition, seven adult specimens recovered from the intestines of the banded tetra fish Astyanax aeneus Günther, 1860 from Nicaragua and Costa Rica formed a monophyletic clade with other specimens identified previously as Saccocoelioides tkachi, expanding its distribution range in other areas of Middle America.

Assessing the Taxonomic Validity of Austrodiplostomum SPP. (Digenea: Diplostomidae) Through Nuclear and Mitochondrial Data.

Adults of the genus Austrodiplostomum are parasites in cormorants of the New World, whereas metacercariae are parasites from eye globe and brain of freshwater and brackish water fishes. In this study, specimens of Austrodiplostomum mordax from South America (type-species) were analyzed together with other specimens of Austrodiplostomum spp. collected from several locations across Middle America and North America. Partial DNA sequences of the mitochondrial gene cytochrome c oxidase subunit I ( COI), the internal transcribed spacers ( ITS1, ITS2, and 5.8S gene), and the D2-D3, domains of the large subunit ( LSU) of nuclear ribosomal DNA, were generated for both developmental stages and compared with available sequences of Austrodiplostomum spp. Phylogenetic analyses inferred with each molecular marker using maximum likelihood and Bayesian inference revealed the existence of 4 lineages representing 2 described species, A. mordax and Austrodiplostomum compactum (syn. Austrodiplostomum ostrowskiae) and 2 undescribed species of Austrodiplostomum recognized in previous studies. The COI haplotype network inferred with 172 sequences detected 28 haplotypes divided into 4 clusters, separating each other by 33 and 40 substitutions and with a genetic divergence ranging from 9 to 12%. The largest group included specimens identified as A. compactum plus those identified as A. ostrowskiae, supporting the synonymy of both species. As a result, we conclude that A. compactum is widely distributed across the Americas, in locations of the United States, Mexico, El Salvador, Honduras, Costa Rica, Venezuela, Peru, and Brazil. The other 2 undescribed species of the genus Austrodiplostomum were previously recorded in the United States and now are reported in Mexico. These 2 species cannot be described because adult forms have not been found in their definitive hosts. Finally, the species A. mordax has been found only in some lakes from Argentina, and it was validated in this study through molecular analyses.