Marine sediments contain a high diversity of micro- and macro-organisms which are important in the functioning of biogeochemical cycles. Traditionally, anthropogenic perturbation has been investigated by identifying macro-organism responses along gradients. Environmental DNA (eDNA) analyses have recently been advocated as a rapid and cost-effective approach to measuring ecological impacts and efforts are underway to incorporate eDNA tools into monitoring. Before these methods can replace or complement existing methods, robustness and repeatability of each analytical step has to be demonstrated. One area that requires further investigation is the selection of sediment DNA extraction method. Environmental DNA sediment samples were obtained along a disturbance gradient adjacent to a Chinook (Oncorhynchus tshawytscha) salmon farm in Otanerau Bay, New Zealand. DNA was extracted using four extraction kits (Qiagen DNeasy PowerSoil, Qiagen DNeasy PowerSoil Pro, Qiagen RNeasy PowerSoil Total RNA/DNA extraction/elution and Favorgen FavorPrep Soil DNA Isolation Midi Kit) and three sediment volumes (0.25, 2, and 5 g). Prokaryotic and eukaryotic communities were amplified using primers targeting the 16S and 18S ribosomal RNA genes, respectively, and were sequenced on an Illumina MiSeq. Diversity and community composition estimates were obtained from each extraction kit, as well as their relative performance in established metabarcoding biotic indices. Differences were observed in the quality and quantity of the extracted DNA amongst kits with the two Qiagen DNeasy PowerSoil kits performing best. Significant differences were observed in both prokaryotes and eukaryotes (p < 0.001) richness among kits. A small proportion of amplicon sequence variants (ASVs) were shared amongst the kits (~3%) although these shared ASVs accounted for the majority of sequence reads (prokaryotes: 59.9%, eukaryotes: 67.2%). Differences were observed in the richness and relative abundance of taxonomic classes revealed with each kit. Multivariate analysis showed that there was a significant interaction between “distance” from the farm and “kit” in explaining the composition of the communities, with the distance from the farm being a stronger determinant of community composition. Comparison of the kits against the bacterial and eukaryotic metabarcoding biotic index suggested that all kits showed similar patterns along the environmental gradient. Overall, we advocate for the use of Qiagen DNeasy PowerSoil kits for use when characterizing prokaryotic and eukaryotic eDNA from marine farm sediments. We base this conclusion on the higher DNA quality values and richness achieved with these kits compared to the other kits/amounts investigated in this study. The additional advantage of the PowerSoil Kits is that DNA extractions can be performed using an extractor robot, offering additional standardization and reproducibility of results.

Proliferations of the benthic anatoxin-producing cyanobacterium Phormidium are increasing in prevalence in cobble-bed rivers worldwide. This proliferation is of particular concern when rivers are used as sources of drinking water or for recreation. Little is known about the physicochemical variables promoting proliferations, and our existing knowledge is based on data from only a few rivers. We assessed Phormidium cover, physicochemical variables, and anatoxin concentrations at 10 sites in 7 New Zealand rivers every week for 2 y. Generalized additive mixed models (GAMMs) identified dissolved inorganic N (DIN) over the accrual period <0.8 mg/L, dissolved reactive P accrual 15°C, and conductivity as having positive and statistically significant effects on % Phormidium cover. Flow intensity, expressed relative to the long-term median, had a positive effect up to 0.4× the median flow and a negative effect when >0.5× the median flow. Quantile regression models showed marked variability among sites in relation to the flow intensity required to reduce % Phormidium cover (90^th percentile ranged 0.65–249× the long-term median flow). Anatoxins were detected in variable concentrations in samples from 7 of the 10 sites. GAMMs identified strong relationships between elevated toxin concentrations and low conductivity and increasing % Phormidium cover, and significantly lower toxin concentrations when DIN was <0.2 mg/L. These data demonstrate that multiple physicochemical variables influence Phormidium proliferations and toxin concentrations and indicate that the relative importance of these variables differs among rivers and sites.