Identification and Confirmation of Chironomus circumdatus Kieffer, 1916 (Diptera: Chironomidae) Using DNA Barcoding

Present study was performed to identify and confirm collected larval Chironomid up to species level. To assess molecular taxonomy and phylogeny, DNA barcoding was done using Sanger dideoxy sequencing of mitochondrial cytochrome c oxidase subunit I (COI) gene of larval sample. Bioinformatics analysis was done by using NCBI’s BLAST software. By evidence of DNA barcoding, it was confirmed that present larval chironomid species was Chironomus circumdatus Kieffer, 1916.

An analysis on the divergence of Chironomid spp. based on the study of 18S rRNA and polytene chromosome organization in the species revealing the role of environment on speciation

Background Nine species of Chironomus evolved throughout the world were measured for their divergence with regard to their DNA sequences concerning 18S rRNA since it is conserved for a specific species. With the advancement of the field of molecular evolution, cytogenetics requires further correlation between molecular architecture and morphological features of a species to compare amongst others to decipher their role in speciation. Therefore, divergence of DNA sequences of the Chironomus were compared with differences in the polytene chromosome features of most of the species under this investigation to evaluate underlying correlation among them, if any, to finally establish a novel method of molecular classification broadly applicable in cytogenetics studies. Results When Chironomus javanus Kieffer was considered as a reference organism, an in silico pair-wise alignment of sequences for the 18S rRNA gene regions of the other eight different species of the same genus exhibited nucleotide sequence homology ranging from 67 to 98%. This divergence of the species under consideration might be due to environmental impact causing alteration of nitrogenous bases probably due to mismatch pairing in DNA replication. This may be suggested as a cause of evolution of species in nature. A concomitant study on the polytene chromosome band patterns of majority of these species belonging to this series also indicated a divergence ranging from 10% to 30%. Conclusions Sequence analysis based on 18S rRNA of nine species of Chironomus under this investigation shows a similarity in the polytene chromosome organization in most of the Chironomid species of the series. Hence, molecular divergence in the species is consistent with cytological difference among Chironomid species. Therefore, molecular data based on 18S rRNA and cytological characters based on the polytene chromosome features of the Chironomid species may be useful for their taxonomical recognition. Moreover, variations concerning two aspects of this study may be correlated to their environmental distinctions.

DNA barcoding reveals an unknown Chironomidae diversity from the freshwater biodiversity hot-spot: comparison between local and the European datasets

In the present study, we developed and evaluated the first reference barcode library for non-biting midges (Diptera: Chironomidae) as flagship taxa of freshwater ecology from Skadar Lake system (Montenegro and Albania), a well-known hotspot of freshwater biodiversity composed by the young lake Skadar (originated 1200 before present) and by its old system of springs (originated during Pliocene). Using an expanded reference library and records deposited in Barcode of Life Database (BOLD), we estimated DNA barcoding efficiency for the European Chironomidae. Study provides COI barcodes for 770 Chironomidae individuals assigned, based on morphology, to 77 species collected in the Skadar Lake basin. Molecular analyses assigned sequences to 100 BINs and 104 OTUs (all records from this area are new for online repositories) and confirms the usefulness of DNA barcoding for the identification of non-biting midges. Additionally, we explored chironomid species distribution patterns in Europe using universal Barcode Index Number (BIN) with a discussion of problematic species groups, both for traditional taxonomy and DNA barcoding. The results of our study provide the first insight into the factual chironomid species diversity of the Lake Skadar basin, in comparison with chironomid fauna at the European scale. The results fill a significant gap in knowledge of biodiversity in the Balkan region. Based on the results of Chironomidae fauna investigation, we conclude that the Skadar Lake basin is now well sampled and such a high representation of species from various sampling sites provides reliable estimation of the non-biting midges fauna.

Ecological impacts caused by the alternance of wet/dry episodes occurred in the last 2,000 years in southern Tibetan Plateau: A paleoecological record from Lake Nam Co

<p>High altitudinal aquatic ecosystems are subject to environmental change due to global warming and increasing solar radiation. The Nam Co catchment is part of the highest and largest alpine plateau on Earth, where the effects of climate change are expressed stronger than the global average. Thus, this area has experienced rapid changes in biodiversity. Fluctuations between wetter and drier periods during the last 2,000 calibrated (cal.) years were detected. These changes may alter the dynamics in ecosystems and therefore their resilience to climate change.</p><p>A ~65 cm sediment record from Nam Co spanning the late Holocene, was analyzed to evaluate the assemblage composition of three of the most abundant and diverse benthic taxa (Arcellinidae, Ostracoda and Chironomidae) and the diverse family of small bivalves (Sphaeriidae). In general, the presence of the bivalve <em>Pisidium stewarti</em>, together with a high abundance of black-coated ostracod shells, and high Ca/Ti and Zr/Rb ratios correspond to the driest period (~ 1,000 - 1,860 cal. years BP) detected in our sediment record. For the last 256 cal. years, higher lake levels were inferred from aquatic fauna composition and geochemical analysis (XRF and XRD) suggesting a more humid environment. This period was characterized by higher temperatures and a higher input of organic matter. Species not previously reported for Lake Nam Co such as Arcellinida species, the ostracod <em>Ilyocypris angulata</em>, several chironomid species, and the bivalve <em>P. stewarti</em>, were observed. These new records, as well as the detection of varieties in morphological structures (e.g. spines, aggregate material, valve ornamentations, etc.) highlight the probable existence of cryptic species in the ecosystem, which is an important factor to take into account for biodiversity evaluation and paleoenvironmental inferences, due to potential misleading ecological interpretation.</p><p>Therefore, emphasis should be placed on combining ecology, morphology and DNA analysis to corroborate the taxonomy of species already described, and determine the accurate richness and distribution of the species in an environment where endemism is expected. This is essential in order to evaluate possible losses or gains in terms of diversity that climate change may exert on aquatic ecosystems in the future.</p>

Identification of chironomid species as natural reservoirs of toxigenic Vibrio cholerae strains with pandemic potential

Vibrio cholerae causes the fatal cholera diarrhea. Chironomids (Diptera; Chironomidae) are abundant in freshwater aquatic habitats and estuaries and are natural reservoirs of V. cholerae. Until now, only the non-O1/O139 serogroups of V. cholerae were identified in chironomids. Here, we explored whether chironomids are natural reservoirs of V. cholerae O1/O139 serogroups, which are associated with cholera endemics and pandemics. All four life stages of chironomids were sampled from two rivers, and a laboratory culture in Pune, India, and from a pond in Israel. In total, we analyzed 223 chironomid samples. The presence of V. cholerae O1/O139 serogroups was verified using molecular tools. Nine chironomid species were identified; of them, Chironomus circumdatus was the most abundant. The presence of V. cholerae serogroup O1 and the cholera toxin genes were detected in samples from all chironomid species. However, serogroup O139 was detected in only two chironomid species. Besides PCR to detect specific genes, a metagenomic analysis that was performed in three selected C. ramosus larvae, identified a list of virulence genes associated with V. cholerae. The findings provide evidence that chironomids are natural reservoirs of toxigenic V. cholerae O1/O139. Chironomid populations and V. cholerae show biannual peak patterns. A similar pattern is found for cholera epidemics in the Bengal Delta region. Thus, we hypothesize that monitoring chironomids in endemic areas of the disease may provide a novel tool for predicting and preventing cholera epidemics. Moreover, serogroup O139 was detected only in two chironomid species that have a restricted distribution in the Indian subcontinent, possibly explaining why the distribution of the O139 serogroup is limited.

MACROFAUNA OF INVERTEBRATE OF LAKE NERO

The taxonomic composition of macrobenthos of Lake Nero and the mouth area of the river Sarah are shown. The main part of the benthos in the summer in the lake is formed by larvae of amphibiotic insects – chironomids, biting midges and mayflies. The macrobenthos of the lake and the mouth of the Sara River can be characterized as chironomid species complex. Relative to the results of previous studies, changes occurred in the dominant complex, a decrease in the total biomass of macroinvertebrates was noted. Together with a decrease in the species diversity, the only one dominant species (Chironomus f.l. plumosus) indicates a decrease in the homogeneity of the structure of benthic communities and its simplification. A decrease in the species richness, occurrence and density of oligochaetes was observed in comparison with the data of other studies. In contrast benthic communities, aquatic plants biotopes can be characterized as more favorable for both benthic and fitophilic invertebrates. The large number of invertebrate species makes it possible to evaluate the thicket as a biotope with very favorable environmental conditions. Rheophilic, limnophilic and obligate benthic species occurring here can coexist, rising from the bottom into thickets with a favorable oxygen regime, which allows them to survive in conditions of oxygen deficiency.

Identification and expression pattern of chemosensory genes in the transcriptome of Propsilocerus akamusi

Chironomidae is the most ecologically diverse insects in aquatic and semi-aquatic habitats. Propsilocerus akamusi (Tokunaga) is a dominant and ubiquitous chironomid species in Eastern Asia and its morphologically unique larvae are also considered as indicator organisms to detect water contamination, potential toxicity and waterborne pathogens. Since few studies to date have focused on the olfactory system of P. akamusi, our study aims to elucidate the potential functions of chemosensory genes in P. akamusi. In our study, we found that although signals released from male groups might attract female swarmers, it was a completely male-dominated mating process. Sequencing the transcriptome of P. akamusi on an Illumina HiSeq platform generated 4.42, 4.46 and 4.53 Gb of clean reads for heads, legs, and antennae, respectively. 27,609 unigenes, 20,379 coding sequences (CDSs), and 8,073 simple sequence repeats were finally obtained. The gene-level differential expression analysis demonstrated variants among three different tissues, including 2,019 genes specifically expressed in heads, 1,540 genes in legs, and 2,071 genes in antennae. Additionally, we identified an assortment of putative olfactory genes consisting of 34 odorant binding proteins, 17 odorant receptors, 32 gustatory receptors, 22 ionotropic receptors, six chemosensory proteins as well as 3 sensory neuron membrane proteins; their relative abundances in the above three tissues were also determined by RT-qPCR. Our finding could allow a more plausible understanding of certain olfaction-mediated behaviors in groups of this macroinvertebrate.

DNA barcodes and morphology confirm a new species of Rheocricotopus (Psilocricotopus) orientalis group (Diptera: Chironomidae)

Morphology and DNA barcodes confirm a new chironomid species within the Rheocricotopus (Psilocricotopus) orientalis group (Diptera: Chironomidae). Rheocricotopus (Psilocricotopus) kongi Lin et Wang sp. n. is described and illustrated based on adult male from Hainan, China. Key to adult males of the R. orientalis group is given.

A new marine intertidal chironomid from the Brazilian coast (Diptera: Chironomidae: Telmatogetoninae)

The male, female, pupa and larva of a new Neotropical chironomid species, Telmatogeton yamaguchiae sp. n., are described and illustrated. Specimens were collected in southeastern Brazilian coast.