Profiling Analysis of Filter Feeder Polypedilum (Chironomidae) Gut Contents Using eDNA Metabarcoding Following Contrasting Habitat Types-Weir and Stream.

We analyzed the dietary composition of Polypedilum larvae among two contrasting habitats (river and weir). Our approach was (i) to apply eDNA-based sampling to reveal the gut content of the chironomid larvae, (ii) the diversity of gut contents in the two aquatic habitats, and (iii) assessment of habitat sediment condition with the food sources in the gut. The most abundant food was Chlorophyta in the gut of the river (20%) and weir (39%) chironomids. The average ratio of fungi, protozoa, and zooplankton in river chironomids gut was 5.9%, 7.2%, and 3.8%, while it was found decreased to 1.2%, 2.5%, and 0.1% in weir chironomids. Aerobic fungi in river midge guts were 3.6% and 10.34% in SC and IS, while they were in the range of 0.34-2.58% in weir midges. The hierarchical clustering analysis showed a relationship of environmental factors with food contents. Abiotic factors (e.g., pH) in the river and weir habitats correlated the clustered pattern with phytoplankton and minor groups of fungi. This study could help understand the food source diversity in the chironomid and habitat environmental conditions by using eDNA metabarcoding as an effective tool to determine dietary composition.

Application of Combined Analyses of Stable Isotopes and Stomach Contents for Understanding Ontogenetic Niche Shifts in Silver Croaker (Pennahia argentata).

Stable isotope analysis (SIA) and stomach content analysis (SCA) were conducted to understand ontogenetic niche shifts in silver croaker Pennahia argentata inhabiting the southern coastal waters of the Korean peninsula. Sampled P. argentata were classified into three groups based on their total length (TL; 60-80 mm TL, 80-120 mm TL, and 120-210 mm TL). Carbon isotope (δ13C) ratios were distinguishable, whereas nitrogen isotope (δ15N) ratios were not significantly different among size classes, and Standard Ellipse Area (SEA), estimated by δ13C and δ15N, was expanded with increasing TL from 0.2 ‰2 (60-80 mm TL) to 2.0 ‰2 (120-210 mm TL). SCA results showed variable contribution of dietary items to each size class. In particular, higher dietary contribution of Polychaeta to P. argentata of 80-120 mm TL than 120-210 mm TL mirrored variation in δ13C values of P. argentata in those size classes. Based on the combined analyses involving SIA and SCA, we concluded that P. argentata underwent ontogenetic niche shifts, particularly dietary shifts, with growth stages. Ontogenetic niche shifting is a representative survival strategy in fish, and, therefore, represents essential information for managing fisheries. The present study demonstrated applicability of combined SIA and SCA analyses, not only for dietary resource tracing, but also for ecological niche studies.

Analyzing the Response Behavior of Lumbriculus variegatus (Oligochaeta: Lumbriculidae) to Different Concentrations of Copper Sulfate Based on Line Body Shape Detection and a Recurrent Self-Organizing Map.

Point detection (e.g., the centroid of the body) of species has been conducted in numerous studies. However, line detection (i.e., the line body shape) of elongated species has rarely been investigated under stressful conditions. We analyzed the line movements of an Oligochaeta Lumbriculus variegatus in response to treatments with a toxic chemical, copper sulfate, at low concentrations (0.01 mg/L and 0.1 mg/L). The automatic line-tracking system was devised to identify the movement of body segments (body length) and the movements of segments (i.e., the speed and angles between segments) were recorded before and after treatment. Total body length was shortened from 31.22 (±5.18) mm to 20.91 (±4.65) mm after the 0.1 mg/L treatment. The Shannon entropy index decreased from 0.44 (±0.1) to 0.28 (±0.08) after treatment. On the other hand, the body and movement segments did not significantly change after the 0.01 mg/L treatment. Sequential movements of test organisms were further analyzed with a recurrent self-organizing map (RSOM) to determine the pattern of time-series line movements. The RSOM made it feasible to classify sequential behaviors of indicator organisms and identify various continuous body movements under stressful conditions.