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.

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.

Changes in parasite-chaetognath species assemblages in the Mexican Central Pacific before and during El Niño 1997-1998.

We investigated the seasonal and interannual changes in diversity, abundance, and prevalence of chaetognaths and their parasites collected monthly during 1996-1998 in the Mexican Central Pacific. We tested the hypothesis of a positive relationship between abundance and species richness of chaetognaths and their parasites, and investigated the influence of the 1997-1998 El Niño event on this host-parasite interaction. Of the 9 chaetognath species collected in the present study, only 7 were found to be parasitized. Of 78154 chaetognath specimens collected, 790 were parasitized (1% prevalence) with at least 1 type of epibiont (cysts, perhaps protists) and 6 types of endoparasites: protists (apicomplexans, dinoflagellates, and ciliates), digeneans, cestodes, acanthocephalans, nematodes, and other unidentified endoparasites. Cysts, digeneans, and cestodes were the most abundant parasites. Mean intensity ranged from 1-4 endoparasites and from 1-21 epibionts host-1. Zonosagitta bedoti and Flaccisagitta enflata were the most abundant chaetognath species and had the highest parasite diversity. Mesosagitta minima and Parasagitta euneritica had the highest parasite prevalence (>2%). A 2-way cluster analysis defined sampling month groups as before, during, and after the 1997-1998 El Niño. The highest abundances of chaetognaths and parasites were associated with a high thermal stratification index, salinity, and mixed layer depth. We conclude that there is a positive, non-linear correlation between the abundance of chaetognaths and their parasites. Although El Niño decreased the abundance and diversity of chaetognaths throughout the time series, the abundance and diversity of their parasites were not significantly different among hydro-climatic periods, suggesting that host abundance must decrease orders of magnitude to influence host availability for parasites.

The discovery of acanthocephalans parasitizing chaetognaths.

A comprehensive literature review shows that 12 types of pathogens, micropredators and parasites are reported to interact with chaetognaths, mostly digenean trematodes, cestodes and nematodes larval stages. Through analysis of 78,152 chaetognaths from a monthly zooplankton time series (Jan 1996-Dec 1998) collected in the Mexican Central Pacific twelve acanthocephalan larvae parasitizing chaetognaths were discovered. This is the first record of an acanthocephalan parasitizing chaetognaths, raising to 13 the types of symbionts known to interact with them (excluding predators). Cystacanth larval specimens of Corynosoma sp. (Polymorphidae) were observed parasitizing the head, trunk and caudal cavities of three of the eight chaetognath species inhabiting this tropical coastal region (Flaccisagitta enflata, Flaccisagitta hexaptera and Zonosagitta bedoti). Because Corynosoma sp. parasitized chaetognaths during different months and years (Jan-Feb 1996, Mar and Jul 1997, Jan, Jun, Aug-Sep 1998) and because the total length of these cystacanths varied between 165-480 µm, suggesting growth inside the hosts, we conclude that chaetognaths are intermediate hosts of Corynosoma sp. The twelve parasitized chaetognaths were juveniles (without gonads) or immature adults (none in reproductively mature stage IV); therefore Corynosoma may have a negative influence on host reproduction. Marine crustaceans (amphipods, decapods, copepods, mysids and euphausiids) and fishes are common intermediate or paratenic hosts of acanthocephalans. Fish, sea birds and marine mammals are definitive hosts for marine Corynosoma species. The present discovery implies that acanthocephalans are transmitted trophically through different intermediate hosts (crustaceans, chaetognaths and/or fish); thus chaetognaths can also be part of the marine acanthocephalan life cycle.

Seasonal variation in chaetognath and parasite species assemblages along the northeastern coast of the Yucatan Peninsula.

Chaetognaths are abundant carnivores with broad distributions that are intermediate hosts of trophically transmitted parasites. Monthly variations in chaetognath and parasite species distributions, abundance, prevalence, and intensity related to seasonal environmental changes were recorded in 2004 and 2005 in Laguna Nichupté, a coral reef, and the adjoining continental shelf of Quintana Roo, Mexico. Of 12 chaetognath species plus Sagitta spp., only 5 (Ferosagitta hispida, Flaccisagitta enflata, Sagitta spp., Serratosagitta serratodentata, and Pterosagitta draco) were parasitized. These species were parasitized with 33 types of flatworms and unidentified cysts (likely protozoan ciliates), having an overall mean prevalence of 6%. Digenean metacercaria larvae numerically dominated the parasite assemblages. Cluster analysis defined 2 chaetognath species assemblages. One included 7 species inside Laguna Nichupté, where F. hispida was numerically dominant (98.9%); the other contained 13 chaetognath species in the continental shelf-coral reef region, where F. enflata was abundant (53%). Canonical correspondence analysis showed that Laguna Nichupté had highly variable and hostile conditions (relatively low salinity and high temperature) for chaetognath species except for F. hispida. The higher density of F. hispida promoted greater parasite diversity (23 types), dominated by Brachyphallus sp. metacercariae. F. enflata prevailed in the continental shelf-coral reef area, which had stable high salinity and relatively low temperature. Monilicaecum and unidentified digenean 'type g' infected 5 chaetognath species on the continental shelf. Distinct primary hosts (mollusks and copepods) and contrasting environmental conditions (salinity, dissolved oxygen concentration, and temperature) between Laguna Nichupté and the continental shelf promoted distinct chaetognath species assemblages, resulting in distinct parasite diversity and prevalence patterns.