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Community, diversity, and biological index metrics for chironomid surface-floating pupal exuviae (SFPE) were assessed at different subsample sizes and sampling frequencies from wadeable streams in Minnesota (USA). Timed collections of SFPE were made using a biweekly sampling interval in groundwater-dominated (GWD) and surface-water-dominated (SWD) streams. These two types of stream were sampled because they support different Chironomidae communities with different phenologies which could necessitate sampling methodologies specific to each stream type. A subsample size of 300 individuals was sufficient to collect on average 85% of total taxa richness and to estimate most metrics with an error of about 1% relative to 1,000 count samples. SWD streams required larger subsample sizes to achieve similar estimates of taxa richness and metric error compared to GWD streams, but these differences were not large enough to recommend different subsampling methods for these stream types. Analysis of sample timing determined that 97% of emergence occurred from April through September. We recommend in studies where estimation of winter emergence is not important that sampling be limited to this period. Sampling frequency also affected the proportion of the community collected. To maximize the portion of the community, collected samples should be taken across seasons although no specific sampling interval is recommended. Subsampling and sampling frequency was also assessed simultaneously. When using a 300-count subsample, a 4-week sampling interval from April through September was required to collect on average 71% of the community. Due to differences in elements of the chironomid community evaluated by different studies (e.g., biological condition, phenology, and taxonomic composition), richness estimates are documented for five sampling intervals (2, 4, 6, 8, 10, and 12 weeks) and five subsample sizes (100, 200, 300, 500, and 1,000 counts). This research will enhance future studies by providing guidelines for tailoring SFPE methods to study specific goals and resources.

The winter-active Diamesa mendotae Muttkowski (Diptera: Chironomidae) is freeze intolerant in the adult stage with a low mean supercooling point (SCP) of ~−20 °C. However, cold-hardiness strategies for immatures of this species are unknown. In this study, we measured SCP values for D. mendotae larvae, pupae and adults using surface-contact thermometry. In addition, the lower lethal temperature (LLT) was determined for the larval stage. The mean SCPs for larvae (−7.4 °C) and pupae (−9.1 °C) were relatively high compared to adults (−19.7 °C). Our results indicate that the larvae of D. mendotae are freeze tolerant with a LLT99 (−25.4 °C), ~−10 °C lower than their minimum SCP (−15.6 °C). Freeze tolerance in these larvae may be a strategy to provide protection from short-term exposures to ice crystals or to permit diapause within frozen substrates. The change in cold-hardiness strategy from freeze tolerant to freeze intolerant between the larval and adult stages of this species is likely a result of the different habitats occupied by these two life stages.