Blue crabs Callinectes sapidus have expanded their geographic range northward in the NW Atlantic with possible trophodynamic effects on benthic communities. In this study, we examined the blue crab’s diet in 2 southern New England tidal rivers (USA) and expounded on their predator-prey interaction with juvenile winter flounder Pseudopleuronectes americanus. Blue crabs (8-185 mm carapace width [CW]; n = 1835) were collected from the Seekonk River, Rhode Island, and Taunton River, Massachusetts, between May and August 2012 to 2016, and their feeding habits were assessed via stomach content, stable isotope, and molecular genetic analyses. Blue crabs were found to be generalist carnivores-omnivores with diets varying throughout ontogeny, yet shifts in prey composition had no effect on size-based nitrogen isotope signatures and trophic position (3.50 ± 0.35, mean ± SD). Carbon isotope values indicated that detritus-macroalgae were the dominant carbon source to the food web, with additional contributions from terrestrially derived organic matter and phytoplankton in oligohaline and polyhaline waters, respectively. The main prey of blue crabs ≤49 mm CW were amphipods, shrimp, and unidentified crustaceans, and larger conspecifics fed on bivalves, crabs, and fish. Winter flounder remains, e.g. sagittal otoliths, were identified in the diet of 2.5% of field-collected blue crabs, whereas PCR-based assays detected winter flounder DNA in 17.7% of crab stomachs. Blue crabs 23 to 160 mm CW preyed on winter flounder ranging from 26 to 66 mm total length, with occurrences of predation most closely associated with increases in crab size. Blue crab predation on winter flounder also varied spatially in the rivers, reflecting site-specific differences in flounder densities, abundances of other preferred prey, and dissolved oxygen concentrations that altered predator-prey dynamics. Lastly, the current predatory impact of blue crabs on juvenile winter flounder is nearly equivalent to other portunid crab species. Anticipated temperature-mediated increases in blue crab densities at northern latitudes, however, will intensify the predator-induced mortality of winter flounder and likely hinder their recovery in southern New England.
A multiplex quantitative PCR (qPCR) assay for the simultaneous detection of 3 eastern oyster Crassostrea virginica parasites, Perkinsus marinus, Haplosporidium nelsoni, and H. costale, was developed using 3 different fluorescently labeled hydrolysis probes. The primers and probe from a previously validated singleplex qPCR for P. marinus detection were combined with newly designed primers and probes specific for H. nelsoni and H. costale. The functionality of the multiplex assay was demonstrated on 2 different platforms by the linear relationship of the standard curves and similar cycle threshold (CT) values between parasites. Efficiency of the multiplex qPCR assay on the Roche and BioRad platforms ranged between 93 and 101%. The sensitivity of detection ranged between 10 and 100 copies of plasmid DNA for P. marinus and Haplosporidium spp., respectively. The concordance between the Roche and BioRad platforms in the identification of the parasites P. marinus, H. nelsoni, and H. costale was 91, 97, and 97%, respectively, with a 10-fold increase in the sensitivity of detection of Haplosporidium spp. on the BioRad thermocycler. The concordance between multiplex qPCR and histology for P. marinus, H. nelsoni, and H. costale was 54, 57, and 87%, respectively. Discordances between detection methods were largely related to localized or low levels of infections in oyster tissues, and qPCR was the more sensitive diagnostic. The multiplex qPCR developed here is a sensitive diagnostic tool for the quantification and surveillance of single and mixed infections in the eastern oyster.
Investigating animal gut microbiomes can lead to a better understanding of their foraging preferences and their overall health. In this study, the fecal and cloacal microbiomes of 4 cold-stunned, frozen loggerhead Caretta caretta, 9 Kemp’s ridley Lepidochelys kempi, and 5 green sea turtles Chelonia mydas that stranded on beaches in Massachusetts, USA, were surveyed. Cloacal swabs and in situ fecal samples were collected from each turtle. From the extracted DNA, the hypervariable V1-V3 regions of the 16S rRNA gene were amplified with PCR, then sequenced using next generation Illumina MiSeq technology. Fecal and cloacal microbiomes were primarily composed of the phyla Proteobacteria, Bacteroidetes, and Firmicutes. Microbial communities varied significantly based on location of the gut sampled. Cloacal samples were largely dominated by Proteobacteria, while fecal samples appeared to have a greater distribution of taxa and higher alpha diversity. Green turtles had a higher abundance of Firmicutes and Bacteroidetes than Kemp’s ridley and loggerhead turtles, but a lower abundance of Proteobacteria. The information gained from this study contributes to knowledge of cold-stunned sea turtle gut microbiomes and may eventually be applied to rehabilitation efforts.
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