The application of high‐throughput sequencing to retrieve multi‐taxon DNA from different substrates such as water, soil, and stomach contents has enabled species identification without prior knowledge of taxon compositions. Here we used three minibarcodes designed to target mitochondrial COI in plankton, 16S in fish, and 16S in crustaceans, to compare ethanol‐ and tissue‐derived DNA extraction methodologies for metabarcoding. The stomach contents of pygmy devilrays ( Mobula kuhlii cf. eregoodootenkee ) were used to test whether ethanol‐derived DNA would provide a suitable substrate for metabarcoding. The DNA barcoding assays indicated that tissue‐derived operational taxonomic units (OTUs) were greater compared to those from extractions performed directly on the ethanol preservative. Tissue‐derived DNA extraction is therefore recommended for broader taxonomic coverage. Metabarcoding applications should consider including the following: (i) multiple barcodes, both taxon specific (e.g., 12S or 16S) and more universal (e.g., COI or 18S) to overcome bias and taxon misidentification and (ii) PCR inhibitor removal steps that will likely enhance amplification yields. However, where tissue is limited or no longer available, but the ethanol‐preservative medium is still available, metabarcoding directly from ethanol does recover the majority of common OTUs, suggesting the ethanol‐retrieval method could be applicable for dietary studies. Metabarcoding directly from preservative ethanol may also be useful where tissue samples are limited or highly valued; bulk samples are collected, such as for rapid species inventories; or mixed‐voucher sampling is conducted (e.g., for plankton, insects, and crustaceans).
The Mediterranean Sea has a long-lasting history of fishery exploitation that, together with other anthropogenic impacts, has led to declines in several marine organisms. In particular, elasmobranch populations have been severely impacted, with drastic decreases in abundance and species diversity. Based on their experience, fishers can provide information on marine species occurrence, abundance and behavioural traits on a long-term scale, therefore contributing to research on the poorly studied biological aspects of elusive or rare elasmobranch species. In this study, for the first time, the use of the Local Ecological Knowledge (LEK) of fishers was applied to study the behavioural traits of sharks, rays and skates in 12 FAO-GFCM geographical sub-areas (GSAs) of the Mediterranean Sea. This study found both new insight and proved the reliability of LEK-based catch seasonality, reflecting seasonal movements, by comparing LEK-based findings and available literature on five elasmobranch taxa (Mustelus spp., Squalus acanthias, Raja spp., Myliobatis aquila and Scyliorhinus stellaris) in the Adriatic Sea and 7 taxa (Mustelus spp., Raja spp., Prionace glauca, Scyliorhinus canicula, Torpedo spp., Pteroplatytrygon violacea and Isurus oxyrinchus) in the remaining Mediterranean GSAs. In addition, LEK provided new insights into a novel comprehensive representation of species aggregations (Mustelus spp., S. acanthias, M. aquila and S. canicula) in the sampled GSAs and supplied the first descriptions of the size, number of individuals and sex composition of the aggregations. When the limits and shortcomings of LEK-based research are considered, this methodology can be a complementary and cost-effective tool used to study elasmobranchs in either a data-poor scenario or a scenario in which a baseline is missing. LEK can also be useful for the evaluation and inclusion of fishers’ perceptions on bottom-up management and to provide important evidence for conservation plans.
Increasing fishing effort, including bycatch and discard practices, are impacting marine biodiversity, particularly among slow-to-reproduce taxa such as elasmobranchs, and specifically sharks. While some fisheries involving sharks are sustainably managed, collateral mortalities continue, contributing towards > 35% of species being threatened with extinction. To effectively manage shark stocks, life-history information, including resource use and feeding ecologies is pivotal, especially among those species with wide-ranging distributions. Two cosmopolitan sharks bycaught off eastern Australia are the common blacktip shark (Carcharhinus limbatus; globally classified as Near Threatened) and great hammerhead (Sphyrna mokarran; Critically Endangered). We opportunistically sampled the digestive tracts of these two species (and also any whole prey; termed the ‘Russian-doll’ approach), caught in bather-protection gillnets off northern New South Wales, to investigate the capacity for DNA metabarcoding to simultaneously determine predator and prey regional feeding ecologies. While sample sizes were small, S. mokkaran fed predominantly on stingrays and skates (Myliobatiformes and Rajiformes), but also teleosts, while C. limbatus mostly consumed teleosts. Metabarcoding assays showed extensive intermixing of taxa from the digestive tracts of predators and their whole prey, likely via the predator’s stomach chyme, negating the opportunity to distinguish between primary and secondary predation. This Russian-doll effect requires further investigation in DNA metabarcoding studies focussing on dietary preferences and implies that any outcomes will need to be interpreted concomitant with traditional visual approaches.
No fishing gear is entirely selective for the targeted catches, which means that in addition to a global harvest from marine fisheries of ~80 million tons per annum, an additional ~10 million tons of nontarget organisms (termed bycatch) is discarded, mostly dead (Zeller et al., 2018). These collateral mortalities have raised concerns over deleterious cascading effects on ecosystems, especially among priority stocks which include endangered, threatened, and protected (ETP) species (Gray & Kennelly, 2018;Hall, 1996;Pacoureau et al., 2021;Zeller et al., 2018). Recognition of these issues has supported ongoing global bycatch resolution efforts, typically via technical modifications to fishing gears, but also spatial and temporal fish-
Increasing fishing effort, including bycatch and discard practices, are impacting marine biodiversity, particularly among slow-to-reproduce taxa such as elasmobranchs, and specifically sharks. While some fisheries involving sharks are sustainably managed, collateral mortalities continue, contributing towards >35% of species being threatened with extinction. To effectively manage shark stocks, life-history information, including resource use/feeding ecologies is pivotal, especially among those species with wide-ranging distributions and habitats. Two cosmopolitan sharks bycaught off eastern Australia are the common blacktip shark (Carcharhinus limbatus; globally classified as Near Threatened) and great hammerhead (Sphyrna mokarran; Critically Endangered). We opportunistically sampled the digestive tracts of these two species and also any whole prey; (termed the ‘Russian-doll’ approach) caught in bather-protection gillnets off northern New South Wales to investigate their regional feeding ecologies and the capacity for DNA metabarcoding to delineate trophic interactions. Sphyrna mokkaran fed predominantly on Myliobatiformes and Rajiformes, but also teleosts, while C. limbatus mostly consumed teleosts, with some inter-specific dietary overlap of prey items. Extensive cross-contamination of predator and prey digestive tracts, likely via the predator’s stomach chyme, was evident from the metabarcoding assays limiting the opportunity to delineate trophic interactions from these data. This Russian-doll effect requires further investigation in DNA metabarcoding studies focused on dietary preferences, but implies any outcomes will need to be interpreted concomitant with traditional visual approaches.
Gillnets are the world’s most common net-based fishing gear, comprising walls of light mesh designed to entangle fish. Like all fishing gears, gillnets are not 100% effective for the targeted catches, and usually catch similar-sized, unwanted animals that are discarded, often dead. Gillnets are often retrieved with holes in the netting, which means some animals escape or are depredated unseen, but with some mortality. To effectively manage fisheries around the world, information is required on not only the harvested and discarded mortalities, but also problematic interactions and mortalities caused by the fishing gear and especially those involving protected species. This study sought to assess a novel method for determining such interactions by sampling pieces of netting around holes in polyethylene gillnets for environmental deoxyribonucleic acid or ‘eDNA’. Here we show that eDNA correctly identified all previously entangled-and-landed species. Also, eDNA from three uncaptured taxa were recorded: bull shark, Carcharhinus leucas, white shark, Carcharodon carcharias and dolphins (Delphindae), illustrating the potential to reveal previously cryptic gillnet interactions. We propose that as scientific methods evolve and autonomous real-time DNA surveillance becomes routine, eDNA testing of fishing gears and vessels could provide a novel, complementary fishery-monitoring tool.
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