Potential zoonotic role of the tick Amblyomma cf. oblongoguttatum (Ixodida: Ixodidae) in the bacterial transmission of Ehrlichia chaffeensis (Rickettsiales: Anaplasmataceae) in a deciduous tropical forest in Mexico.

Ehrlichia chaffeensis is a bacterium belonging to the Anaplasmataceae family. In Mexico, only 2 species have been recorded in association with tick species and humans. The objective of the present study was to detect the presence of bacteria of the genus Ehrlichia in ticks collected from the Chamela-Cuixmala Biosphere Reserve, Jalisco, Mexico. The collected ticks were identified and analyzed individually by polymerase chain reaction to amplify a fragment of the Anaplasmataceae 16S rRNA gene and the Ehrlichia-specific dsb gene. A total of 204 ticks, corresponding to 5 species of Ixodidae and 1 of Argasidae, were collected from 147 mammals of 6 species and 4 orders; 57 ticks collected from vegetation were also included. Among the total ticks collected, 1.47% (3/204) was positive for Ehrlichia sp. DNA was obtained using the primers EHR 16SD and EHR 16SR for 16S rRNA and DSB-330 and DSB-728 for dsb. The positive samples corresponded to a larva (Amblyomma sp.) associated with Didelphis virginiana and 2 nymphs (Amblyomma cf. oblongoguttatum) infesting Nasua narica. None of the ticks collected from the vegetation tested positive for Ehrlichia sp. DNA on the basis of the 16S rRNA and dsb genes. The sequences from the larvae of Amblyomma sp. and the nymphs of A. cf. oblongoguttatum were similar to those of E. chaffeensis. The phylogenetic analysis inferred with maximum likelihood corroborated the identity as E. chaffeensis. Although the role of these tick species as vectors of E. chaffeensis is still undetermined, the presence of infected ticks in the area indicates a potential zoonotic risk.

Evolution of an Alternative Genetic Code in the Providencia Symbiont of the Hematophagous Leech Haementeria acuecueyetzin.

Strict blood-feeding animals are confronted with a strong B-vitamin deficiency. Blood-feeding leeches from the Glossiphoniidae family, similarly to hematophagous insects, have evolved specialized organs called bacteriomes to harbor symbiotic bacteria. Leeches of the Haementeria genus have two pairs of globular bacteriomes attached to the esophagus which house intracellular "Candidatus Providencia siddallii" bacteria. Previous work analyzing a draft genome of the Providencia symbiont of the Mexican leech Haementeria officinalis showed that, in this species, the bacteria hold a reduced genome capable of synthesizing B vitamins. In this work, we aimed to expand our knowledge on the diversity and evolution of Providencia symbionts of Haementeria. For this purpose, we sequenced the symbiont genomes of three selected leech species. We found that all genomes are highly syntenic and have kept a stable genetic repertoire, mirroring ancient insect endosymbionts. Additionally, we found B-vitamin pathways to be conserved among these symbionts, pointing to a conserved symbiotic role. Lastly and most notably, we found that the symbiont of H. acuecueyetzin has evolved an alternative genetic code, affecting a portion of its proteome and showing evidence of a lineage-specific and likely intermediate stage of genetic code reassignment.

New species of Foleyellides (Nematoda: Onchocercidae: Waltonellinae), parasite of Lithobates brownorum (Amphibia: Ranidae) from South-eastern Mexico and genetic barcodes of the Mexican species of the genus.

Specimens of Foleyellides were collected from the body cavity of frogs in different regions of Mexico; Lithobates brownorum from Yucatán, Quintana Roo and Campeche; L. megapoda from Jalisco and Rhinella marina, from Guerrero. Foleyellides calakmulesis n. sp. is described based on specimens found parasitizing L. brownorum. The new species is distinguished from the other members of the genus by the combination of the following male characters: four pairs of caudal papillae different in size and the presence of a preanal plaque. Partial DNA sequences of the mitochondrial Cytochrome Oxidase C, subunit I of the four known Mexican species of Foleyellides and two potentially new species collected in this study were generated and compared, validating the erection of the new species.

Diversity of myxozoans (Cnidaria) infecting Neotropical fishes in southern Mexico.

Myxozoans are a unique group of microscopic parasites that infect mainly fishes. These extremely reduced cnidarians are highly diverse and globally distributed in freshwater and marine habitats. Myxozoan diversity dimension is unknown in Mexico, a territory of an extraordinary biological diversity. This study aimed to explore, for the first time, myxozoan parasite diversity from fishes of the Neotropical region of Mexico. We performed a large morphological and molecular screening using host tissues of 22 ornamental and food fish species captured from different localities of Veracruz, Oaxaca and Chiapas. Myxozoan infections were detected in 90% of the fish species, 65% of them had 1 or 2 and 35% had 3 and up to 8 myxozoan species. Forty-one putative new species were identified using SSU rDNA phylogenetic analyses, belonging to two main lineages: polychaete-infecting (5 species) and oligochaete-infecting (36 species) myxozoans; from those we describe 4 new species: Myxidium zapotecus sp. n., Zschokkella guelaguetza sp. n., Ellipsomyxa papantla sp. n. and Myxobolus zoqueus sp. n. Myxozoan detection increased up to 6 × using molecular screening, which represents 3.7 × more species detected than by microscopy. This study demonstrated that Neotropical fishes from Mexico are hosts of a multitude of myxozoans, representing a source of emerging diseases with large implications for economic and conservation reasons.

Temporal variation in helminth infracommunities of the Gafftopsail pompano, Trachinotus rhodopus (Pisces: Carangidae) off the Pacific coast of Mexico.

Temporal variation of the helminth infracommunity structure in the Gafftopsail pompano Trachinotus rhodopus was studied during bi-monthly revisions of samples collected offshore from Puerto Ángel, Oaxaca (Mexican Pacific) in 2018. In total, 110 specimens of T. rhodopus were subjected to a parasitic review. Helminths found were identified to the lowest possible taxonomic level (six species and three genera) by means of morphological and molecular data. Attributes of the helminth infracommunities are described through statistical analyses, showing stability in terms of their richness throughout the year. However, variations were found in helminth abundance related to the seasonality of samplings, which may be associated with the life cycles of the parasites, the host species' gregarious behavior, the availability of intermediate hosts, and/or the diet of T. rhodopus.

Phylogenetic relationships of the Geoplaninae land planarians (Platyhelminthes, Tricladida) assessed with a total evidence approach, with the description of a new species of Gigantea.

Several studies have focused on the phylogenetic relationships within the Geoplaninae land planarians (Tricladida). In those studies, ancient phylogenetic relationships remained obscure. In this work, the phylogeny of Geoplaninae is assessed through three different datasets, namely morphological, molecular, and both datasets combined, i.e, a total evidence approach (TE). The data matrix consisted of six DNA regions, including a newly developed marker (DOM5), and a morphological matrix with 37 characters. The study produced the best-resolved hypothesis so far for the phylogeny of Geoplaninae, although ancient clades still remain elusive. The effect of the morphological data on the TE tree topology and clade support is seemingly negligible. The phylogenetic tree also suggests that most of the diagnostic morphological characters of the genera are homoplastic, while unambiguous unique synapomorphies can characterize some supra-generic informal groupings.

Phylogenetic relationships and species delimitation in Haemopis (Annelida: Hirudinea: Haemopidae).

The Holarctic leech genus Haemopis currently includes 11 species, all of which are macrophagous, as opposed to their more infamous bloodfeeding counterparts among hirudiniform leeches. In spite of their ecological importance as fish food and predators of freshwater invertebrates, there is a paucity of data regarding morphology and genetic variation that might guide future identification efforts for members of the genus. The lack of detailed descriptions of distinguishing morphological features, coupled with the absence of a robust phylogenetic hypothesis for the genus, have conspired to prevent meaningful inferences on the natural history of the group. In an attempt to remedy this, we present new genetic (using COI, 12S rDNA, 28S rDNA and 18S rDNA) data for the majority of the known species diversity within the genus in order to both infer a phylogenetic hypothesis and to introduce authoritative DNA barcodes for the newly collected species. The potential of these barcodes is increased through rigorous morphological investigations of the specimens, with comparisons to the original literature. Our resulting phylogenetic hypothesis is agnostic as to the geographic origin of the genus, with equal probability afforded to both a Nearctic and Palearctic origin. Beyond this, we show that there is a strong tendency towards a barcoding gap within the genus, but that a distinct gap is lacking due to the relatively high genetic variation found within H. marmorata. Taken together, our results shed light on species delimitation within, and evolutionary history of, this often-neglected group of leeches.

A new species of Hystrignathus (Nematoda: Thelastomatoidea: Hystrignathidae) associated with the Bess Beetle Passalus Interruptus Linnaeus (Coleoptera: Passalidae) from The Peruvian Amazonia.

Bess beetles (Passalidae) are known to maintain biological interactions with a diverse gut symbiotic community, which is far from being well described since these communities from most of the species of Passalidae have not been studied. In order to improve our knowledge of these communities, we describe a new species of Nematoda of the genus Hystrignathus (Hystrignathidae) associated with Passalus interruptus, a bess beetle species of the tribe Passalini collected in Hunuco, Peru, in the Peruvian Amazonia. Hystrignathus nunashae n. sp. is characterized by the presence of cuticular spines beginning just after the end of the first cephalic annule, a posterior cephalic ring dilated and longer than the anterior ring, as well as ornamented eggs. The new species has a didelphic-amphidelphic reproductive system, lateral alae extending from the posterior end of procorpus almost reaching the anus, and a short subulate tail. The external and internal morphological traits of the new species are described, and additionally, based on nuclear ribosomal DNA, we investigated the phylogenetic relationships of the new species.

Finding a needle in a haystack: larval stages of Didymozoidae (Trematoda: Digenea) parasitizing marine zooplankton.

Larval didymozoids (Trematoda: Digenea) were discovered parasitizing the hemocoel of the heteropod Firoloida desmarestia (redia mean intensity = 13) and the chaetognaths Flaccisagitta enflata and Flaccisagitta hexaptera (metacercaria mean intensity = 1) during a 2014-2016 systematic study of parasites of zooplankton collected in the central and southern regions of the Gulf of California, Mexico. Didymozoid infection route during the early life cycle was inferred combining morphological (light microscopy) and molecular (mitochondrial cytochrome c oxidase subunit I gene, cox1) evidence. Didymozoid rediae parasitizing F. desmarestia were observed, just after field collection of the host, containing hundredths of completely developed cystophorous cercariae, releasing them though the birth pore at approximately one cercaria every 12 s. Cercariae lost their tails developing into a 'young metacercaria' in 1 d at 22 °C without need of an intermediate host. Molecular analysis of cox1 showed that rediae found in F. desmarestia belong to two distinct didymozoid species (Didymozoidae sp. 1 and sp. 2). Metacercariae parasitizing chaetognaths were morphologically identified as Didymozoidae type Monilicaecum and cox1 sequences showed that metacercariae of chaetognaths matched with these two Didymozoidae sp. 1, and sp. 2 species found parasitizing F. desmarestia, plus a third distinct Didymozoidae sp. 3. These are the first DNA sequences of cox1 gene from didymozoid larvae for any zooplankton taxonomic group in the world. We concluded that F. desmarestia is the first intermediate host of rediae and cercariae, and the chaetognaths are the second intermediate hosts where non-encysted metacercariae were found. The definitive host is still unknown because cox1 sequences of present study did not genetically match with any available cox1 sequence of adult didymozoid. Our results demonstrate a potential overlap in the distribution of two carnivorous zooplankton taxonomic groups that are intermediate hosts of didymozoids in the pelagic habitat. The didymozoid specimens were not identified to species level because any of the cox1 sequences generated here matched with the sequences of adult didymozoids currently available in GenBank and Bold System databases. This study provides baseline information for the future morphological and molecular understanding of the Didymozoidae larvae that has been previously based on the recognition of the 12 known morphotypes.

PERUANOCOTYLE PELAGICA N. SP. (MONOGENEA: MONOCOTYLIDAE), PARASITE OF THE PACIFIC COWNOSE RAY RHINOPTERA STEINDACHNERI EVERMANN AND JENKINS, 1891 (BATOIDEA: RHINOPTERIDAE) FROM THE SOUTHERN MEXICAN PACIFIC OCEAN.

Peruanocotyle pelagica n. sp. is described based on specimens collected from the wall of the pharyngeal cavity of the Pacific cownose ray Rhinoptera steindachneri offshore Oaxaca and Guerrero, Mexico. The new species is distinguished from Peruanocotyle chisholmae by its anchors, which include a slender guard that curves towards the tip of the blade and which lack an accessory piece, morphological differences of the seminal vesicle, the lack of a male copulatory organ accessory piece and a greater number of spines, and an unsclerotized vagina. Molecular data of Peruanocotyle pelagica were generated to place the phylogenetic position of the genus within Monocotylidae.

Rickettsia spp. in ticks from a tropical dry forest reserve on Mexico's Pacific Coast.

The present study analyzes the diversity of bacteria of the genus Rickettsia in ticks collected from wild mammals and vegetation at the Chamela Biological Station (EBCh), Jalisco, Mexico, a core area in the Chamela-Cuixmala Biosphere Reserve, which is characterized by the presence of tropical dry forest, one of the most threatened ecosystems in the world. Ticks were collected during April and November 2018 and 2019, determined to species, and subsequently processed for rickettsiae. Genomic DNA was extracted from each of 349 ticks (198 from mammals, 151 from vegetation) and screened for Rickettsia by targeting the gltA and ompA genes. Overall, 75 ticks (21.5%) were positive for rickettsiae (46 from mammals, 29 from vegetation). Through BLASTn and maximum likelihood analysis, four Rickettsia taxa were identified. Only Rickettsia amblyommatis had been previously recorded in the vicinity of the EBCh, and we confirm its presence in this reserve, associated with Amblyomma mixtum, Amblyomma cf. oblongoguttatum and Amblyomma parvum. Additionally, we record for the first time at EBCh the taxa Rickettsia parkeri strain Atlantic rainforest, which is known to cause spotted fever illness in humans, detected in Amblyomma ovale; Rickettsia bellii in A. parvum and Ornithodoros talaje; and an undescribed Rickettsia sp. in Ixodes sinaloa.

The potential of aquatic bloodfeeding and nonbloodfeeding leeches as a tool for iDNA characterisation.

Leeches play important roles in food webs due to their abundance, diversity and feeding habits. Studies using invertebrate-derived DNA (iDNA) extracted from leech gut contents to target vertebrate DNA have focused on the Indo-Pacific region and mainly leveraged the leech family Haemadipsidae, composed of bloodfeeding terrestrial leeches, while predatory, fluid/tissue-feeding and aquatic bloodfeeding species have been largely disregarded. While there is some general knowledge regarding the taxonomic groups that leeches prefer to feed on, detailed taxonomic resolution is missing and, therefore, their potential use for monitoring animals is unknown. In this study, 116 leeches from 12 species (six families) and spanning the three feeding habits were collected in Mexico and Canada. We used DNA metabarcoding to investigate their diet and assess their potential use for biodiversity monitoring. We detected vertebrates from five orders including fish, turtles and birds in the diet of aquatic bloodfeeding leeches; eight invertebrate orders of annelids, arthropods and molluscs in leeches that feed on body fluids and tissues; and 10 orders of invertebrates belonging to Arthropoda and Annelida, as well as one vertebrate and one parasitic nematode, in predatory leeches. These results show the potential use of iDNA from aquatic bloodfeeding leeches for retrieving vertebrate taxa, and from predatory and fluid-feeding leeches for invertebrates. Our study provides information about the dietary range of freshwater leeches and one terrestrial leech and contributes proof-of-concept for the use of these leeches for animal monitoring, expanding our knowledge of the use of iDNA from leech gut contents to North America.

Phylogenetic position of Acanthobothrium cleofanus (Cestoda: Onchoproteocephalidea) using molecular evidence.

Despite the large number of species described to date for the onchoprotepcephalid genus Acanthobothrium (207), only 16 named species have a genetic sequence. With this background, specimens of adult cestodes of the stingray Hypanus longus were collected off San Blas, Nayarit, and onchoproteocephalid larvae in the carangid fish Trachinotus rhodopus from Puerto Ángel, Oaxaca, both located on the Pacific coast of Mexico. The objective of this work is to investigate the phylogenetic position of these adults and larvae using nuclear ribosomal markers (18S rDNA and 28S rDNA). Morphologically, adult specimens were identified as Acanthobothrium cleofanus; larvae were identified only to family level. The phylogenetic position of both taxa was investigated based on the information of two nuclear molecular markers analyzed under Parsimony (PA) and Bayesian Inference (BI) methods. The newly generated sequences of A. cleofanus from Nayarit are identical to the sequences of several samples of Acanthobothrium sp. collected in the Mexican Pacific, which sequence are available in GenBank; DNA sequences obtained from onchoproteocephalid larva clearly place this taxon within Acanthobothrium but representing an independent lineage. In the resulting phylogenetic trees, Uncibilocularis okei was found nested within Acanthobothrium with an unstable position depending on the optimality criteria, indicating the need for more molecular analyzes with a greater number of species of both genera prior to define its phylogenetic relationships.

Segmented worms (Phylum Annelida): a celebration of twenty years of progress through Zootaxa and call for action on the taxonomic work that remains.

Zootaxa has been the leading journal on invertebrate systematics especially within Annelida. Our current estimates indicate annelids include approximately 20,200 valid species of polychaetes, oligochaetes, leeches, sipunculans and echiurans. We include herein the impact of Zootaxa on the description of new annelid species in the last two decades. Since 2001, there have been over 1,300 new annelid taxa published in about 630 papers. The majority of these are polychaetes (921 new species and 40 new genera) followed by oligochaetes (308 new species and 10 new genera) and leeches (21 new species). The numerous papers dealing with new polychaete species have provided us a clear picture on which polychaete families have had the most taxonomic effort and which authors and countries have been the most prolific of descriptions of new taxa. An estimated additional 10,000+ species remain to be described in the phylum, thus we urge annelid workers to continue their efforts and aid in training a new generation of taxonomists focused on this ecologically important group.

Molecular Identification of Plerocercoids of Clistobothrium montaukensis (Cestoda: Phyllobothriidea) Parasitizing the King of Herrings Regalecus glesne.

PURPOSE: Endo-parasites of the bathypelagic king of herrings Regalecus glesne and oarfish Regalecus russelii are only known from few specimens opportunistically examined. As a consequence, there are few records of parasites from either Regalecus species. We report plerocercoid larvae of phyllobothriidean cestodes parasitizing an adult R. glesne stranded in Bahía de La Paz, Baja California Sur, Mexico. METHODS: Sixty-three plerocercoids were obtained from the intestine of R. glesne and characterized using morphological and molecular methods (nuclear 28S rDNA and mitochondrial cytochrome c oxidase I gene sequences). RESULTS: Following the morphological diagnostic criteria of scolex and muscle bands in the strobila, plerocercoids specimens were preliminary assigned to the genus Clistobothrium. Mitochondrial and nuclear DNA sequences indicate these plerocercoids correspond to Clistobothrium montaukensis Ruhnke, 1993. CONCLUSION: Regalecus glesne is a new host known for C. montaukensis and this report is a new geographical record of C. montaukensis parasitizing species of the genus Regalecus previously known only from California and Florida, USA.

Molecular Phylogeny of Thelastomatoidea (Nematoda) with the Description of a New Genus and Two New Species of Hystrignathidae Associated with Bess Beetles (Coleoptera: Passalidae) from Oaxaca, Mexico.

Bess beetles (Passalidae) display important roles in forestall ecosystems, particularly in energy extraction from dead wood. These organisms maintain complex biological interactions with their gut symbiotic communities, including bacteria, protists, and metazoans. Very little is known about symbionts since most of the species of Passalidae haven't been studied from a parasitological point of view. Here we describe a new genus and 2 new species of nematodes of the family Hystrignathidae associated with 2 beetle species of the tribe Proculini collected in the State of Oaxaca, Mexico. Tuhmai garciaprietoi n. gen., n. sp., found in Vindex agnoscendus is characterized by the presence of an unarmed cervical cuticle, a subcylindrical procorpus and a conspicuous isthmus, a monodelphic-prodelphic reproductive system, and a short subulate tail. Urbanonema osorioi n. sp., found in Verres hageni mainly differs from other species of Urbanonema by the number and disposition of cervical spines, as well as by a subulate tail. For each new taxon, we describe the external and internal morphology, and we generated molecular data (nuclear ribosomal DNA) to place the new taxa in a phylogenetic context.

Morphological and molecular data reveal a new species of Lueheia (Acanthocephala: Plagiorhynchidae) from Turdus migratorius (Turdidae) in central Mexico and its phylogenetic implications within the family.

Members of the genus Lueheia Travassos, 1919, are endoparasites of birds, particularly passerines, throughout the Americas. Adults of Lueheia sp., (Plagiorhynchidae Golvan, 1960; Porrorchinae Golvan, 1956) were recovered from the intestine of the American robin (Turdus migratorius phillipsi Bangs) in Mexico City, and two other species of acanthocephalans identified as Porrorchis nickoli, (Plagiorhynchidae: Porrorchinae) Salgado-Maldonado and Cruz-Reyes, 2002 and Centrorhynchus microcephalus (Bravo-Hollis, 1947) Golvan, 1956 (Centrorhynchidae Van Cleave, 1916), were recovered from the Virginia opossum (Didelphis virginiana Allen) and groove-billed ani (Crotophaga sulcirostris Swainson), respectively in southeastern Mexico. Specimens of three species were sequenced at two molecular markers, the small subunit (SSU) and large subunit (LSU) of the nuclear rDNA and compared with other sequences available in GenBank. Maximum likelihood and Bayesian inference analyses of the combined (LSU + SSU) dataset and each individual dataset revealed that the specimens of Lueheia sp. formed an independent lineage, which is recognized herein as a new species, Lueheia aztecae n. sp., representing the fifth species of the genus in the Americas, and the second in the Nearctic region. The new species can be morphologically distinguished from the other five species in the genus by having a cylindrical proboscis, armed with 24-26 longitudinal rows with 9-10 hooks each. Phylogenetic inference performed with the combined dataset consisting of two genes (LSU + SSU) revealed that Lueheia aztecae n. sp. and P. nickoli belonging to subfamily Porrorchinae, formed two independent lineages, indicating that the subfamily is paraphyletic. Porrorchis nickoli and C. microcephalus formed a clade with other species of the genus Centrorhynchus, suggesting that P. nickoli should be transferred to genus Centrorhynchus, to form C. nickoli n. comb. In addition, we briefly discuss the ecological associations between the members of the families Plagiorhynchidae and Centrorhynchidae.

New records of Ichthyostraca Zrzavý, Hypsa amp; Vlásková, 1997 (Pancrustacea) from Mexico with an annotated checklist of North America.

In this work, we document for the first time four species of Ichthyostraca in Mexico, one Branchiura: Argulus foliaceus (fish parasite), and three Pentastomida: Porocephalus clavatus (reptile parasite), Raillietiella hebitihamata (reptile parasite) and Raillietiella orientalis (frog parasite). In addition, a total of 11 new host records and 16 new localities are presented for Ichthyostraca in Mexico. Based on the compilation of published information and on specimens deposited in scientific collections, the first checklist of Ichthyostraca from North America (Canada, United States of America and Mexico) is compiled. To March 2019, records of Ichthyostraca from North America include 64 species and 8 undetermined taxa, parasitizing 293 host species (64 of them identified only to a supra-specific level) in 77 provinces and states (8 from Canada, 47 from the USA, and 22 from Mexico). The subclass Branchiura is represented by 1 order, 1 family, 1 genus, 35 species and 1 undetermined taxa; the subclass Pentastomida includes records of 4 orders, 9 families, 14 genera, 29 species and 7 undetermined taxa. Ten species reported in this work are thought to be introduced into North America with only a few records of them parasitizing native hosts. This checklist summarizes the available records of this class of crustaceans in North America and represents a base line for future studies.

Narcinecotyle longifilamentus n. gen., n. sp. (Monogenea: Hexabothriidae), gill parasite of the numbfish Narcine entemedor (Torpediniformes: Narcinidae) from the Mexican Pacific coast.

A new genus and species of Hexabothriidae Price, 1942 is described from specimens infecting the gill of the numbfish Narcine entemedor Jordan and Starks from the Pacific coast of Mexico. In addition to the new taxon described here, species of 8 genera also display symmetrical haptors and have eggs with two polar filaments. However, they differ in the combination of the following features: distal portion of the male copulatory organ tubular, unarmed and proximally dilated confining an internal coiled duct, as well as dorsal origin of haptoral appendix. Based on molecular data derived from 3 loci, the mitochondrial Cytochrome C oxidase subunit 1 as well as the nuclear ribosomal 18S and 28S, the new species was found nested within Hexabothriidae together with the other 4 genera with representatives in GenBank. This is the first species of Hexabothriidae reported from a species of Narcinidae.

Phylogeny and biogeography of the Cavernicola (Platyhelminthes: Tricladida): Relicts of an epigean group sheltering in caves?

The planarian suborder Cavernicola Sluys, 1990 was originally created to house five species of triclad flatworms with special morphological features and a surprisingly discontinuous and broad geographic distribution. These five species could not be accommodated with any degree of certainty in any of the three taxonomic groups existing at that moment, viz., Paludicola Hallez, 1892, Terricola Hallez, 1892, and Maricola Hallez, 1892. The scarce representation of the group and the peculiarities of the morphological features of the species, including several described more recently, have complicated new tests of the monophyly of the Cavernicola, the assessment of its taxonomic status, as well as the resolution of its internal relationships. Here we present the first molecular study including all genera currently known for the group, excepting one. We analysed newly generated 18S and 28S rDNA data for these species, together with a broad representation of other triclad flatworms. The resulting phylogenetic trees supported the monophyly of the Cavernicola, as well as its sister-group relationship to the Maricola. The sister-group relationship to the Maricola and affinities within the Cavernicola falsify the morphology-based phylogeny of the latter that was proposed previously. The relatively high diversity of some cavernicolan genera suggests that the presumed rarity of the group actually may in part be due to a collecting artefact. Ancestral state reconstruction analyses suggest that the ancestral habitat of the group concerned epigean freshwater conditions. Our results point to an evolutionary scenario in which the Cavernicola (a) originated in a freshwater habitat, (b) as the sister clade of the marine triclads, and (c) subsequently radiated and colonized both epigean and hypogean environments. Competition with other planarians, notably members of the Continenticola, or changes in epigean habitat conditions are two possible explanations -still to be tested- for the loss of most epigean diversity of the Cavernicola, which is currently reflected in their highly disjunct distributions.