Assessing microplastic exposure of large marine filter-feeders.

Large filter-feeding animals are potential sentinels for understanding the extent of microplastic pollution, as their mode of foraging and prey mean they are continuously sampling the environment. However, there is considerable uncertainty about the total and mode of exposure (environmental vs trophic). Here, we explore microplastic exposure and ingestion by baleen whales feeding year-round in coastal Auckland waters, New Zealand. Plastic and DNA were extracted concurrently from whale scat, with 32 ± 24 (mean ± SD, n = 21) microplastics per 6 g scat sample detected. Using a novel stochastic simulation modeling incorporating new and previously published DNA diet information, we extrapolate this to total microplastic exposure levels of 24,028 (95% CI: 2119, 69,270) microplastics per mouthful of prey, or 3,408,002 microplastics (95% CI: 295,810, 10,031,370) per day, substantially higher than previous estimates for large filter-feeding animals. Critically, we find that the total exposure is four orders of magnitude more than expected from microplastic measurements of local coastal surface waters. This suggests that trophic transfer, rather than environmental exposure, is the predominant mode of exposure of large filter feeders for microplastic pollution. Measuring plastic concentration from the environment alone significantly underestimates exposure levels, an important consideration for future risk assessment studies.

Multi-locus DNA metabarcoding of zooplankton communities and scat reveal trophic interactions of a generalist predator.

To understand the ecosystem dynamics that underpin the year-round presence of a large generalist consumer, the Bryde's whale (Balaenoptera edeni brydei), we use a DNA metabarcoding approach and systematic zooplankton surveys to investigate seasonal and regional changes in zooplankton communities and if whale diet reflects such changes. Twenty-four zooplankton community samples were collected from three regions throughout the Hauraki Gulf, New Zealand, over two temperature regimes (warm and cool seasons), as well as 20 samples of opportunistically collected Bryde's whale scat. Multi-locus DNA barcode libraries were constructed from 18S and COI gene fragments, representing a trade-off between identification and resolution of metazoan taxa. Zooplankton community OTU occurrence and relative read abundance showed regional and seasonal differences based on permutational analyses of variance in both DNA barcodes, with significant changes in biodiversity indices linked to season in COI only. In contrast, we did not find evidence that Bryde's whale diet shows seasonal or regional trends, but instead indicated clear prey preferences for krill-like crustaceans, copepods, salps and ray-finned fishes independent of prey availability. The year-round presence of Bryde's whales in the Hauraki Gulf is likely associated with the patterns of distribution and abundance of these key prey items.

PCR enrichment techniques to identify the diet of predators.

The increasing sensitivity of PCR has meant that in the last two decades PCR has emerged as a major tool in diet studies, enabling us to refine our understanding of trophic links and to elucidate the diets of predators whose prey is as yet uncharacterized. The achievements and methods of PCR-based diet studies have been reviewed several times, but here we review an important development in the field: the use of PCR enrichment techniques to promote the amplification of prey DNA over that of the predator. We first discuss the success of using group-specific primers either in parallel single reactions or in multiplex reactions. We then concentrate on the more recent use of PCR enrichment techniques such as restriction enzyme digests, peptide nucleic acid clamping, DNA blocking and laser capture microdissection. We also survey the vast literature on enrichment techniques in clinical biology, to ascertain the pitfalls of enrichment techniques and what refinements have yielded some highly sensitive methods. We find that while there are several new approaches to enrichment, peptide nucleic acid clamping and DNA blocking are generally sufficient techniques for the characterization of diets of predators and highlight the most important considerations of the approach.