Advances in high-throughput sequencing (HTS) technologies and their increasing affordability have fueled environmental DNA (eDNA) metabarcoding data generation from freshwater, marine and terrestrial ecosystems. Research institutions worldwide progressively employ HTS for biodiversity assessments, new species discovery and ecological trend monitoring. Moreover, even non-scientists can now collect an eDNA sample, send it to a specialized laboratory for analysis and receive in-depth biodiversity record from a sampling site. This offers unprecedented opportunities for biodiversity assessments across wide temporal and spatial scales. The large volume of data produced by metabarcoding also enables incidental detection of species of concern, including non-indigenous and pathogenic organisms. We introduce an online app—Pest Alert Tool—for screening nuclear small subunit 18S ribosomal RNA and mitochondrial cytochrome oxidase subunit I datasets for marine non-indigenous species as well as unwanted and notifiable marine organisms in New Zealand. The output can be filtered by minimum length of the query sequence and identity match. For putative matches, a phylogenetic tree can be generated through the National Center for Biotechnology Information’s BLAST Tree View tool, allowing for additional verification of the species of concern detection. The Pest Alert Tool is publicly available at https://pest-alert-tool-prod.azurewebsites.net/.
Symbiodiniaceae are a diverse group of dinoflagellates, the majority of which are free-living and/or associated with a variety of protists and other invertebrate hosts. Maintenance of isolated cultures is labour-intensive and expensive, and cryopreservation provides an excellent avenue for their long-term storage. We aimed to cryopreserve 15 cultured isolates from six Symbiodiniaceae genera using dimethyl sulfoxide (DMSO) as the cryoprotectant agent (CPA). Under 15% DMSO, 10 isolates were successfully cryopreserved using either rapid freezing or controlled-rate freezing. Cultures that failed or had low survival, were subjected to (1) a reduction of CPA to 10%, or (2) increased salinity treatment before freezing. At 10% DMSO, three further isolates were successfully cryopreserved. At 15% DMSO there were high cell viabilities in Symbiodinium pilosum treated with 44 parts per thousand (ppt) and 54 ppt culture medium. An isolate of Fugacium sp. successfully cryopreserved after salinity treatments of 54 ppt and 64 ppt. Fatty acid (FA) analyses of S. pilosum after 54 ppt salinity treatment showed increased saturated FA levels, whereas Fugacium sp. had low poly-unsaturated FAs compared to normal salinity (34 ppt). Understanding the effects of salinity and roles of FAs in cryopreservation will help in developing protocols for these ecologically important taxa.
The increasing spread of marine non-indigenous species (NIS) due to the
growth in global shipping traffic is causing widespread concern for the
ecological and economic impacts of marine bioinvasions. Risk management
authorities need tools to identify pathways and source regions of
priority concern in order to better target efforts for preventing NIS
introduction. The probability of a successful NIS introduction is
affected by the probability that a marine species entrained in a
transport vector will survive the voyage between origin and destination
locations, and establish an independently reproducing population at the
destination. Three important risk factors are voyage duration, range of
environmental conditions encountered during transit, and environmental
similarity between origin and destination. In this study, we aimed for a
globally comprehensive approach of assembling quantifications of
source-destination risk factors from every potential origin to every
potential destination. To derive estimates of voyage-related marine
biosecurity risk, we used computer-simulated vessel paths between pairs
of ecoprovinces in the Marine Ecoregions Of the World biogeographic
classification system. We used the physical length of each path to
calculate voyage duration risk, and the cross-latitudinal extent of the
path to calculate voyage path risk. Environmental similarity risk was
based on comparing annual average sea surface temperature and salinity
within each ecoprovince to those of other ecoprovinces. We derived three
separate sets of risk quantifications, one each for voyage duration,
voyage path, and environmental similarity. Our quantifications can be
applied to studies that require source-destination risk estimates. They
can be used separately or combined, depending on the importance of the
types of source-destination risks that might be relevant to particular
scientific or risk management questions or applications.
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