New mitogenomes from the genus Cricotopus (Diptera: Chironomidae, Orthocladiinae): Characterization and phylogenetic implications.

Cricotopus is a large and diverse genus of non-biting midges composed of several subgenera. Complete mitogenome sequences are available for very few Cricotopus species. The subgenus Pseudocricotopus unites species with unusual morphological structures in adult male and pupal stages, however, molecular methods are needed to verify the placement of this subgenus within Cricotopus. We obtained mitogenomes of C. (Pseudocricotopus) cf. montanus and nine other Cricotopus species for phylogenetic analysis, coupled with two Rheocricotopus species and one Synorthocladius species as outgroups. The structure of the mitogenome was similar among these Cricotopus species, exhibiting A+T bias and retaining ancestral gene order. Mutation rate, estimated as Ka/Ks, varied among genes, and was highest for ATP8 and lowest for COI. The phylogenetic relationships among species of Cricotopus, Rheocricotopus and Synorthocladius was reconstructed using Bayesian inference and maximum likelihood estimation. The phylogenetic trees confirmed placement of subgenus Pseudocricotopus, represented by Cricotopus cf. montanus, within Cricotopus. Our study increases the library of chironomid mitogenomes and provides insight into the properties of their constituent genes.

Stream ecosystem puzzle: understanding how water column and sediment variables shape macroinvertebrate patterns in some Afrotropical streams.

Although the interaction of water column and sediment variables in streams is intricate, minimal studies have been conducted on how they influence macroinvertebrate community patterns. This study, therefore, aimed to evaluate the influence of water column and sediment variables on macroinvertebrate community patterns in selected Afrotropical streams. Spatiotemporal scales of water column and sediment variables were analysed following standard methods while macroinvertebrates were sampled using the kick sampling technique. Redundancy analysis (RDA) and variation partitioning were used to assess the relationship of macroinvertebrates with water column and sediment variables. Significant differences were observed between seasons amongst water column variables such as total dissolved solids (p=0.046), turbidity (p=0.027), dissolved oxygen (p=0.011), chemical oxygen demand (p=0.002), bank vegetation (p=0.013), velocity (p=0.04), phosphates (p=0.031), and sediment variables such as total organic matter (p=0.01), pH (p=0.024), electrical conductivity (p=0.014). This accounted for the shift in biotic communities across the two seasons. In the studied area and seasons, Baetidae, Chironomidae, and Thiaridae were the most abundant families of macroinvertebrates representing 21.5%, 17.8%, and 6.9% of the 5266 recorded individuals belonging to 68 families. The water column was the most important predictor of macroinvertebrate community patterns (57%) compared to sediments (35%). Therefore, the use of both water column and sediment variables in ecological studies and biomonitoring should be emphasised because the two compartments provide complementary information. This enables researchers to gain a more complete understanding of the ecological health of aquatic habitats, useful in the development of effective management strategies.

New data on mitogenomes of Thienemanniella Kieffer, 1911 (Diptera: Chironomidae, Orthocladiinae).

The mitochondrial genome (mitogenome) has been widely used as a powerful marker in phylogenetic and evolutionary studies of various Dipteran groups. However, only a few mitogenomes from the Thienemanniella genus have been reported till now. Furthermore, there is still indeterminacy in the phylogenetic relationships of the genus Thienemanniella. In this study, mitogenomes of five Thienemanniella species were sequenced and analyzed newly. Combined with the published mitogenome of Thienemanniella nipponica, the obtained results showed that mitogenomes of Thienemanniella were conserved in structure, and all genes were observed to be arranged in the same gene order as the ancestral mitogenome. Nucleotide composition varied significantly among different genes, and the control region displayed the highest A + T content. All protein coding genes are subjected to purification selection, and the fastest evolving gene is ATP8. Maximum likelihood and Bayesian inference analyses showed the phylogeny of Thienemanniella which was supported in five topologies. Our present study provides valuable insight into the phylogenetic relationships of Thienemanniella species.

Energy status of chironomid larvae (Diptera: Chironomidae) from high alpine rivers (Tyrol, Austria).

Chironomids (non-biting midges) inhabit almost every wet or semi-wet continental environment on Earth with probably 10,000 different species. Species occurrence and composition are undoubtedly limited by environmental harshness and food availability being reflected in their energy stores. Most animals store energy as glycogen and lipid. They enable the animals to survive adverse situations and to continue growth, development, and reproduction. This general statement is also true for insects and also particularly true for chironomid larvae. The rationale behind this research was, that probably any stress, any environmental burden, and any harmful influence increases the energy requirement of individual larvae depleting energy stores. We developed new methods to measure the glycogen and lipid content in small tissue samples. Here we show how to apply these methods to single chironomid larvae to demonstrate their energy stores. We compared different locations of the high Alpine rivers along harshness gradient densely populated and dominated by chironomid larvae. All samples show very low energy stores without any major differences. We found glycogen concentrations below 0.01% of dry weight (DW) and lipid concentrations below 5% of DW irrespective of the specific sampling point. These values are among the lowest ever observed in chironomid larvae. We demonstrate that individuals living in extreme environment are stressed leading to reduced energy stores in their bodies. This appears to be a general feature of high altitude regions. Our results provide new insights and a better understanding of population and ecological dynamics in harsh mountainous areas, also in view of a changing climate.

Species-specific traits predict whole-assemblage detritus processing by pond invertebrates.

Functional trait diversity determines if ecosystem processes are sensitive to shifts in species abundances or composition. For example, trait variation suggests detritivores process detritus at different rates and make different contributions to whole-assemblage processing, which could be sensitive to compositional shifts. Here, we used a series of microcosm experiments to quantify species-specific coarse and fine particulate organic matter (CPOM and FPOM) processing for ten larval caddisfly species and three non-caddisfly species in high-elevation wetlands. We then compared trait-based models including life history, dietary, and extrinsic traits to determine which traits explained interspecific variation in detritus processing. Finally, we compared processing by mixed caddisfly assemblages in microcosms and natural ponds to additive predictions based on species-specific processing to determine if single-species effects are additive in multi-species assemblages. We found considerable interspecific variation in biomass-specific CPOM (13-fold differences) and FPOM (8-fold differences) processing. Furthermore, on a mass-specific basis, amphipods, chironomids, and caddisflies processed similar amounts of detritus, suggesting non-shredder taxa could process more than previously recognized. Trait models including dietary percent detritus, development rate, body size, and wetland hydroperiod explained 81 and 57% of interspecific variation in CPOM and FPOM processing, respectively. Finally, species-specific additive predictions were strikingly similar to mixed-assemblage processing in microcosms and natural ponds, with the largest difference being a 15% overestimate. Thus, additivity of species-specific processing suggests single-species rates may be useful for understanding functional consequences of shifting assemblages, and a trait-based approach to predicting species-specific processing could support generating additive predictions of whole-assemblage processing.

Multi-Level Gene Expression in Response to Environmental Stress in Aquatic Invertebrate Chironomids: Potential Applications in Water Quality Monitoring.

In freshwater ecosystems, aquatic invertebrates are influenced continuously by both physical stress and xenobiotics. Chironomids (Diptera; Chironomidae), or non-biting midges, are the most diverse and abundant invertebrates in freshwater habitats. They are a fundamental link in food chains of aquatic ecosystems. Chironomid larvae tolerate stress factors in their environments via various physiological processes. At the molecular level, environmental pollutants induce multi-level gene responses in Chironomus that regulate cellular protection through the activation of defense processes. This paper reviews literature on the transcriptional responses of biomarker genes to environmental stress in chironomids at the molecular level, in studies conducted from 1991 to 2020 (120 selected literatures of 374 results with the keywords "Chironomus and gene expression" by PubMed search tool). According to these studies, transcriptional responses in chironomids vary depending on the type of stress factor and defensive responses associated with antioxidant activity, the endocrine system, detoxification, homeostasis and stress response, energy metabolism, ribosomal machinery, apoptosis, DNA repair, and epigenetics. These data could provide a comprehensive overview of how Chironomus species respond to pollutants in aquatic environments. Furthermore, the transcriptomic data could facilitate the development of genetic tools for water quality and environmental monitoring based on resident chironomid species.

Generational sensitivity alteration in Chironomus yoshimatsui to carbamate and pharmaceutical chemicals and the effect on Catalase, CYP450, and hemoglobin gene transcription.

To ascertain the tolerance mechanisms of aquatic organisms to artificial chemicals, intergenerational sensitivity changes of Chironomus yoshimatsui to a carbamate pesticide (pirimicarb) and pharmaceutical chemical (diazepam) were investigated. The larvae (<48-h-old) in each generation were exposed to both chemicals for 48 h and then the surviving chironomids were cultured until the fifth generation (F0-F4) without chemical addition. The 48-h 50% effective concentration (EC50) value of chironomids was determined for each generation. In the pirimicarb treatment group, the EC50 values significantly increased in F3 and F4, and those in the diazepam treatment group slightly increased. Catalase, Cytochrome P450 and hemoglobin (Hb) mRNA levels were monitored to see whether these were related to the trans-generational sensitivity. Although the generalized linear model results showed that the sensitivity to diazepam was slightly increased in the diazepam treatment, we could not find any mRNA levels related to sensitivity alteration. In contrast, the model approach showed that the chironomids exposed to pirimicarb trans-generationally became tolerant with increasing Hb mRNA levels. Therefore, they might decrease their chemical stress by modifying Hb gene transcription.

Rapid adaptation of Chironomus yoshimatsui to acetylcholinesterase inhibitors (pyraclofos and pirimicarb) in a multi-generation study.

We evaluated the real effects of pollutants through a multi-generation study. We tested whether short-term exposure (48 h) of successive (first and second) generations of Chironomus yoshimatsui neonates (<24-h-old) to two acetylcholinesterase inhibitor insecticides, pyraclofos, and pirimicarb, would change insecticide sensitivity and life-cycle parameters over four generations. Additionally, we tested whether acetylcholinesterase (AChE) activity levels would be associated with this sensitivity change. Sensitivities (48 h EC50 value, using immobility as the endpoint) in chironomids (<24-h-old) and insect life-cycle parameters (the number of larvae per egg mass and adult size) were investigated. Parental chironomids produced larvae that were less sensitive than those in the control group following the two 48 h pirimicarb exposure events, whereas exposure to pyraclofos did not affect sensitivity. The AChE activity in larvae with low sensitivity to pirimicarb was significantly higher than that in the control. Thus, increased AChE activity might be associated with low sensitivity. The life-cycle parameters in chironomids recovered from the effects of pyraclofos and pirimicarb suggested they could adapt to the insecticides by changing biomass allocation. Our study suggested potential chemical risks of insecticide stress and how aquatic organisms adapt to it.

Chironomus sancticaroli (Diptera, Chironomidae) as a Sensitive Test Species: Can We Rely on Its Use After Repeated Generations, Under Laboratory Conditions?

In ecotoxicological assays, previously selected and standardized organism tests are exposed to an environmental sample. Some species of the Chironomus genus have been extensively used in ecotoxicological assays. Among these, Chironomus tentans is usually utilized in the USA and Chironomus sancticaroli in Brazil. We conducted ecotoxicological bioassays to compare a population of C. sancticaroli, kept for 6 years under laboratory conditions, with a sylvatic population of the same species, collected in the field. The aim was to test the hypothesis that populations of C. sancticaroli, maintained in the laboratory for long periods, could have a different response to stressors/substances. We analyzed the responses of C. sancticaroli for potassium chloride, zinc chloride, potassium dichromate, linear alkylbenzene sulphonate (LAS) and caffeine. The results showed no significant differences between the two populations in the analyses and seems to indicate the possible use of C. sancticaroli from populations kept in the laboratory for long periods.

NGS barcoding reveals high resistance of a hyperdiverse chironomid (Diptera) swamp fauna against invasion from adjacent freshwater reservoirs.

BACKGROUND: Macroinvertebrates such as non-biting midges (Chironomidae: Diptera) are important components of freshwater ecosystems. However, they are often neglected in biodiversity and conservation research because invertebrate species richness is difficult and expensive to quantify with traditional methods. We here demonstrate that Next Generation Sequencing barcodes ("NGS barcodes") can provide relief because they allow for fast and large-scale species-level sorting of large samples at low cost. RESULTS: We used NGS barcoding to investigate the midge fauna of Singapore's swamp forest remnant (Nee Soon Swamp Forest). Based on > 14.000 barcoded specimens, we find that the swamp forest maintains an exceptionally rich fauna composed of an observed number of 289 species (estimated 336 species) in a very small area (90 ha). We furthermore barcoded the chironomids from three surrounding reservoirs that are located in close proximity. Although the swamp forest remnant is much smaller than the combined size of the freshwater reservoirs in the study (90 ha vs. > 450 ha), the latter only contains 33 (estimated 61) species. We show that the resistance of the swamp forest species assemblage is high because only 8 of the 314 species are shared despite the close proximity. Moreover, shared species are not very abundant (3% of all specimens). A redundancy analysis revealed that ~ 21% of the compositional variance of midge communities within the swamp forest was explained by a range of variables with conductivity, stream order, stream width, temperature, latitude (flow direction), and year being significant factors influencing community structure. An LME analysis demonstrates that the total species richness decreased with increasing conductivity. CONCLUSION: Our study demonstrates that midge diversity of a swamp forest can be so high that it questions global species diversity estimates for Chironomidae, which are an important component of many freshwater ecosystems. We furthermore demonstrate that small and natural habitat remnants can have high species turnover and can be very resistant to the invasion of species from neighboring reservoirs. Lastly, the study shows how NGS barcodes can be used to integrate specimen- and species-rich invertebrate taxa in biodiversity and conservation research.

Rhabdostylid Ciliates (Ciliophora, Peritrichia, Epistylididae) as Epibionts on Chironomid Larvae: Evidence of High Specificity and Association with Organic Pollution.

Although epibiont ciliates make up a significant part of the biomass in aquatic ecosystems and may cause perceptible alterations in the population dynamics of their hosts, studies on the extrinsic and intrinsic factors that control the abundance of these microorganisms are scarce in literature. In the present study, we investigated the colonization site and intensity of rhabdostylid epibiont upon chironomid larvae and assessed the influence of organic pollution and chironomid communities on the prevalence and abundance of epibiont ciliates at five sampling stations (two in rural areas and three in urban areas) along a neotropical urban stream over a period of 12 months. Among the 24 genera of chironomids found and the 31,976 larvae analyzed, 96.12% belonged to the Chironomus genus, of which 16.95% (5212) were colonized by Rhabdostyla aff. chironomi. The infestation intensity varied from one to 67 individuals per host with an average intensity of 4.86 (± 33.45). Ciliates were only found colonizing the chironomids' ventral tubules. The high number of chironomid larvae, high host- and site-specificity, low infestation intensity, and absence of apparent structural damage to hosts evidence an intimate relationship between epibiont and basibiont as well as a possibly long coevolutionary history. Both prevalence and abundance of epibiont ciliates were correlated to the pollution rate of the studied stream. There was an alteration in the composition and structure of the chironomid community along the sampling stations. Numeric dominance of Chironomus-tolerant chironomids and its direct correlation to infestation prevalence and to ciliates abundance highlights the predilection of both peritrich ciliates and Chironomus larvae for organically-enriched environments.

A multi-year study following BACI design reveals no short-term impact of Bti on chironomids (Diptera) in a floodplain in Eastern Austria.

Short-term impacts of aerial application of Bacillus thuringiensis israelensis (Bti) on Culicidae and Chironomidae were investigated over several years in temporary waters of the Dyje and Morava floodplains in Eastern Austria. The sampling followed a Before-After-Control-Impact (BACI) approach with sampling dates immediately before and shortly after the application and was repeated for 3 years. To test for effects of the Bti treatment on the two Diptera families, linear mixed-effects models were used. Data analysis included the factors Before-After and Control-Impact as fixed effects, while general temporal and spatial variables were random effects. One hundred sixteen taxa of chironomids were identified. Abundance varied between 2 and 1125 larvae per m2, while culicid densities reached values of several 100 ind. per liter. Total culicid abundance significantly decreased after the Bti treatment, whereas no significant effects were found on the abundance of total chironomids and dominant chironomid subfamilies, tribes, and genera, on relative proportions of chironomid feeding guilds, diversity, and species composition. Further studies from this area are needed to extend the investigation over a period of several weeks in order to reveal possible delayed effects of the larvicide application.

Trophic state changes can affect the importance of methane-derived carbon in aquatic food webs.

Methane-derived carbon, incorporated by methane-oxidizing bacteria, has been identified as a significant source of carbon in food webs of many lakes. By measuring the stable carbon isotopic composition (δ13C values) of particulate organic matter, Chironomidae and Daphnia spp. and their resting eggs (ephippia), we show that methane-derived carbon presently plays a relevant role in the food web of hypertrophic Lake De Waay, The Netherlands. Sediment geochemistry, diatom analyses and δ13C measurements of chironomid and Daphnia remains in the lake sediments indicate that oligotrophication and re-eutrophication of the lake during the twentieth century had a strong impact on in-lake oxygen availability. This, in turn, influenced the relevance of methane-derived carbon in the diet of aquatic invertebrates. Our results show that, contrary to expectations, methane-derived relative to photosynthetically produced organic carbon became more relevant for at least some invertebrates during periods with higher nutrient availability for algal growth, indicating a proportionally higher use of methane-derived carbon in the lake's food web during peak eutrophication phases. Contributions of methane-derived carbon to the diet of the investigated invertebrates are estimated to have ranged from 0-11% during the phase with the lowest nutrient availability to 13-20% during the peak eutrophication phase.

Pollution-tolerant invertebrates enhance greenhouse gas flux in urban wetlands.

One of the goals of urban ecology is to link community structure to ecosystem function in urban habitats. Pollution-tolerant wetland invertebrates have been shown to enhance greenhouse gas (GHG) flux in controlled laboratory experiments, suggesting that they may influence urban wetland roles as sources or sinks of GHG. However, it is unclear if their effects can be detected in highly variable conditions in a field setting. Here we use an extensive data set on carbon dioxide (CO2 ), methane (CH4 ), and nitrous oxide (N2 O) flux in sediment cores (n = 103) collected from 10 urban wetlands in Melbourne, Australia during summer and winter in order to test for invertebrate enhancement of GHG flux. We detected significant multiplicative enhancement effects of temperature, sediment carbon content, and invertebrate density on CH4 and CO2 flux. Each doubling in density of oligochaete worms or large benthic invertebrates (oligochaete worms and midge larvae) corresponded to ~42% and ~15% increases in average CH4 and CO2 flux, respectively. However, despite exceptionally high densities, invertebrates did not appear to enhance N2 O flux. This was likely due to fairly high organic carbon content in sediments (range 2.1-12.6%), and relatively low nitrate availability (median 1.96 µmol/L NO3- -N), which highlights the context-dependent nature of community structural effects on ecosystem function. The invertebrates enhancing GHG flux in this study are ubiquitous, and frequently dominate faunal communities in impaired aquatic ecosystems. Therefore, invertebrate effects on CO2 and CH4 flux may be common in wetlands impacted by urbanization, and urban wetlands may make greater contributions to the total GHG budgets of cities if the negative impacts of urbanization on wetlands are left unchecked.

Impact of routine Bacillus thuringiensis israelensis (Bti) treatment on the availability of flying insects as prey for aerial feeding predators.

Since 1980, mosquito breeding habitats in the Upper Rhine Valley were routinely treated with Bacillus thuringiensis var. israelensis (Bti). Bti is considered to significantly reduce the number of mosquitoes, and - especially when used in higher dosages - to be toxic to other Nematocera species, e.g. Chironomidae, which could be food sources for aerial feeding predators. To investigate direct and indirect effects of routine Bti treatment on food sources for aerial feeding predators, the availability of flying insects in treated and untreated areas was compared. A car trap was used for insect collection, which allowed their exact spatiotemporal assignment. The statistical analysis revealed that insect taxa abundance was influenced differently by the factors season, temperature and time of day. Nematocera (Diptera) were the most frequently collected insects in all areas. Chironomidae were the predominant aquatic Nematocera. The comparison of treated and untreated sites did not show significant differences that would indicate any direct or indirect effect of routine Bti treatment on the availability of flying insects. Additional to food availability, food selection must be considered when investigating food resources for aerial feeding predators. In this study, food selection of Delichon urbicum (House Martin) as an example was investigated with the help of neck ring samples. The preferred prey of the investigated D. urbicum colony consisted of diurnal insects with terrestrial larvae (Aphidina, Brachycera, Coleoptera). Chironomidae were consumed, but not preferred.

A Drosophila heat shock response represents an exception rather than a rule amongst Diptera species.

Heat shock protein 70 (Hsp70) is the major player that underlies adaptive response to hyperthermia in all organisms studied to date. We investigated patterns of Hsp70 expression in larvae of dipteran species collected from natural populations of species belonging to four families from different evolutionary lineages of the order Diptera: Stratiomyidae, Tabanidae, Chironomidae and Ceratopogonidae. All investigated species showed a Hsp70 expression pattern that was different from the pattern in Drosophila. In contrast to Drosophila, all of the species in the families studied were characterized by high constitutive levels of Hsp70, which was more stable than that in Drosophila. When Stratiomyidae Hsp70 proteins were expressed in Drosophila cells, they became as short-lived as the endogenous Hsp70. Interestingly, three species of Ceratopogonidae and a cold-water species of Chironomidae exhibited high constitutive levels of Hsp70 mRNA and high basal levels of Hsp70. Furthermore, two species of Tabanidae were characterized by significant constitutive levels of Hsp70 and highly stable Hsp70 mRNA. In most cases, heat-resistant species were characterized by a higher basal level of Hsp70 than more thermosensitive species. These data suggest that different trends were realized during the evolution of the molecular mechanisms underlying the regulation of the responses of Hsp70 genes to temperature fluctuations in the studied families.

Bioturbation enhances the aerobic respiration of lake sediments in warming lakes.

While lakes occupy less than 2% of the total surface of the Earth, they play a substantial role in global biogeochemical cycles. For instance, shallow lakes are important sites of carbon metabolism. Aerobic respiration is one of the important drivers of the carbon metabolism in lakes. In this context, bioturbation impacts of benthic animals (biological reworking of sediment matrix and ventilation of the sediment) on sediment aerobic respiration have previously been underestimated. Biological activity is likely to change over the course of a year due to seasonal changes of water temperatures. This study uses microcosm experiments to investigate how the impact of bioturbation (by Diptera, Chironomidae larvae) on lake sediment respiration changes when temperatures increase. While at 5°C, respiration in sediments with and without chironomids did not differ, at 30°C sediment respiration in microcosms with 2000 chironomids per m(2) was 4.9 times higher than in uninhabited sediments. Our results indicate that lake water temperature increases could significantly enhance lake sediment respiration, which allows us to better understand seasonal changes in lake respiration and carbon metabolism as well as the potential impacts of global warming.

Prevalence and dynamics of the zoosporic pathogen Catenaria uncinata in a natural population of the midge Glyptotendipes lobiferus.

A qPCR assay specific for zoospores of Catenaria uncinata, a fungal parasite in eggs of the midge Glyptotendipes lobiferus, was developed and used in parallel with traditional microscopic methods in a season-long study of a C. uncinata/G. lobiferus association in a local pond. Twenty-six consecutive weekly collections of egg masses were screened with a microscope to obtain percentages of infection and mortality in organogenetic egg masses and weekly water samples were processed by absolute quantification using qPCR to obtain estimates of zoospore density. Overall, 36.0% of G. lobiferus egg masses were infected to varying degrees and 11.2% of eggs were killed by C. uncinata. Continuous infection of egg masses occurred during a 6-wk period in May-June and a 7-wk period in September-October. Infection by C. uncinata was absent during a 10-week interval between periods of infection. Abrupt declines in zoospore density occurred during both infection periods and occurred only when water temperatures met or exceeded the viability threshold for zoospores (⩾31.0°C). The episodic death of zoospores during weeks in which egg infection and mortality levels were continuous likely resulted from distribution of zoospores throughout the water column and a temperature gradient in which zoospores sampled near the surface were subjected to lethal temperatures while non-sampled zoospores at lower depths were provided low temperature sanctuary. The hiatus of infection during the 10-week interval was likely due to lethal temperatures throughout the water column as average water temperatures exceeded 31.0°C over the period. A positive correlation between weekly zoospore densities obtained from qPCR and levels of infection/mortality in egg masses obtained from counts with a microscope supports the use of the qPCR assay alone in future studies that can rapidly and accurately determine parasite presence, prevalence and geographical range.

Morphological identification and COI barcodes of adult flies help determine species identities of chironomid larvae (Diptera, Chironomidae).

Establishing reliable methods for the identification of benthic chironomid communities is important due to their significant contribution to biomass, ecology and the aquatic food web. Immature larval specimens are more difficult to identify to species level by traditional morphological methods than their fully developed adult counterparts, and few keys are available to identify the larval species. In order to develop molecular criteria to identify species of chironomid larvae, larval and adult chironomids from Western Lake Erie were subjected to both molecular and morphological taxonomic analysis. Mitochondrial cytochrome c oxidase I (COI) barcode sequences of 33 adults that were identified to species level by morphological methods were grouped with COI sequences of 189 larvae in a neighbor-joining taxon-ID tree. Most of these larvae could be identified only to genus level by morphological taxonomy (only 22 of the 189 sequenced larvae could be identified to species level). The taxon-ID tree of larval sequences had 45 operational taxonomic units (OTUs, defined as clusters with >97% identity or individual sequences differing from nearest neighbors by >3%; supported by analysis of all larval pairwise differences), of which seven could be identified to species or 'species group' level by larval morphology. Reference sequences from the GenBank and BOLD databases assigned six larval OTUs with presumptive species level identifications and confirmed one previously assigned species level identification. Sequences from morphologically identified adults in the present study grouped with and further classified the identity of 13 larval OTUs. The use of morphological identification and subsequent DNA barcoding of adult chironomids proved to be beneficial in revealing possible species level identifications of larval specimens. Sequence data from this study also contribute to currently inadequate public databases relevant to the Great Lakes region, while the neighbor-joining analysis reported here describes the application and confirmation of a useful tool that can accelerate identification and bioassessment of chironomid communities.

Comparing the impacts of sediment-bound bifenthrin on aquatic macroinvertebrates in laboratory bioassays and field microcosms.

We conducted two laboratory bioassays and two field microcosm exposures with bifenthrin (a synthetic pyrethroid) in order to evaluate the capacity of single-species laboratory bioassays to predict lethal and sublethal impacts on aquatic invertebrates in microcosms. For the laboratory species, Chironomus tepperi, larval survival was reduced by 24% at 53.66µg/g OC, while adult emergence was reduced at concentrations of 33.33µg/g OC and higher, with a 61% decrease at 77.78µg/g OC and no emergence at 126.67µg/g OC. The abundance of several other microcosm taxa was reduced in the microcosms at a similar concentration range (33.33µg/g OC and above), however there was no impact on the abundance of the congeneric species, Chironomus oppositus. The differences in impacts between test systems were potentially due to both differing species sensitivity and the interaction of ambient temperature with bifenthrin toxicity. Bifenthrin also was associated with early emergence of Chironomus sp. in both test systems, at concentrations of 10µg/g OC and higher (laboratory) and 43.90µg/g OC (microcosm), and with a significant decrease in the proportion of C. oppositus males in a microcosm. These findings indicate that while laboratory bioassays accurately predict many impacts in the field, there are some limitations to the predictive capacity of these tests.