{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T16:49:07Z","timestamp":1786121347162,"version":"build-2736575974"},"reference-count":80,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Natural Resources","award":["12110600000018003910"],"award-info":[{"award-number":["12110600000018003910"]}]},{"award":["12110600000018003910"],"award-info":[{"award-number":["12110600000018003910"]}],"id":[{"id":"https:\/\/ror.org\/02kxqx159","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["41502021"],"award-info":[{"award-number":["41502021"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["41502021"],"award-info":[{"award-number":["41502021"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Geological Museum of China","award":["12111300000018001-14"],"award-info":[{"award-number":["12111300000018001-14"]}]},{"award":["12111300000018001-14"],"award-info":[{"award-number":["12111300000018001-14"]}],"id":[{"id":"https:\/\/ror.org\/04ax2jp69","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Insects"],"abstract":"<jats:p>Species delimitation and biodiversity assessment in Chironomidae are frequently hindered by morphological conservatism and widespread cryptic diversity. Neozavrelia (Diptera: Chironomidae), a genus inhabiting specialized freshwater microhabitats such as springs and hygropetric seepages, remains poorly understood with respect to its genetic diversity and the environmental processes underlying lineage differentiation. Clarifying lineage diversity is a prerequisite for evaluating the ecological factors associated with genetic differentiation. In this study, we compiled a global COI DNA barcode dataset comprising 137 sequences from newly generated specimens and publicly available records in the Barcode of Life Data Systems (BOLD). Genetic diversity was evaluated using neighbor-joining (NJ) analysis and Automatic Barcode Gap Discovery (ABGD), whereas environmental and geographic drivers of genetic differentiation were assessed using Mantel tests, distance-based redundancy analysis (db-RDA), variation partitioning, and hierarchical partitioning. A total of 26 molecular operational taxonomic units (MOTUs) were identified, with most lineages forming well-supported clades in the NJ tree. Genetic differentiation was significantly associated with both environmental and geographic distances, although environmental variables explained a substantially greater proportion of the observed variation. Hierarchical partitioning identified slope, annual cloud cover, mean precipitation amount of the coldest quarter, and frost frequency as the principal environmental predictors of lineage differentiation. Our findings reveal substantial COI genetic diversity within Neozavrelia and suggest that environmental heterogeneity, particularly variation in water availability and thermal conditions, is significantly associated with lineage differentiation. This study expands the global COI DNA barcode reference library for Neozavrelia and provides new insights into ecological and evolutionary factors associated with diversification in freshwater chironomids.<\/jats:p>","DOI":"10.3390\/insects17080814","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T10:41:42Z","timestamp":1785926502000},"page":"814","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["DNA Barcoding Reveals Hidden Genetic Diversity and Environmental Correlates of Lineage Differentiation in Neozavrelia Goetghebuer &amp; Thienemann, 1941 (Diptera: Chironomidae)"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7059-9141","authenticated-orcid":false,"given":"Tong-Yin","family":"Xie","sequence":"first","affiliation":[{"name":"College of Plant Protection, Northeast Agricultural University, Harbin 150030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-0372-1062","authenticated-orcid":false,"given":"Yan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shanghai Universities Key Laboratory of Marine Animal Taxonomy and Evolution, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi-Zhu","family":"Chen","sequence":"additional","affiliation":[{"name":"Shanghai Universities Key Laboratory of Marine Animal Taxonomy and Evolution, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zheng","family":"Liu","sequence":"additional","affiliation":[{"name":"Laboratory of Geo-Specimens Study and Testing, Geological Museum of China, Beijing 100034, China"},{"name":"Department of Stratigraphy and Paleontology, Geological Museum of China, Beijing 100034, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6544-6204","authenticated-orcid":false,"given":"Xiao-Long","family":"Lin","sequence":"additional","affiliation":[{"name":"Shanghai Universities Key Laboratory of Marine Animal Taxonomy and Evolution, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"216","DOI":"10.4319\/lo.1986.31.1.0216","article-title":"A method for obtaining in situ growth rates of larval Chironomidae (Diptera) and its application to studies of secondary production","volume":"31","author":"Huryn","year":"1986","journal-title":"Limnol. Oceanogr."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Hirabayashi, K. (2025). Annual and seasonal trends in density, biomass, and secondary production of Chironomid midges (Diptera: Chironomidae) in the middle reaches of the Shinano River, Japan, 2015\u20132019. Limnol. Rev., 25.","DOI":"10.3390\/limnolrev25020027"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"403","DOI":"10.21608\/ejabf.2025.415515.6435","article-title":"Diversity and ecological role of aquatic insects as bioindicators of water quality in tropical streams","volume":"29","author":"Suhri","year":"2025","journal-title":"Egypt. J. Aquat. Biol. Fish."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"575","DOI":"10.23818\/limn.38.33","article-title":"Aquatic insects as bioindicators of heavy metals in sediments in Cerrado streams","volume":"38","author":"Souto","year":"2019","journal-title":"Limnetica"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Popovi\u0107, N., \u0110ukni\u0107, J., Marinkovi\u0107, N., Tubi\u0107, B., Atanackovi\u0107, A., Milo\u0161evi\u0107, D., and Rakovi\u0107, M. (2025). Environmental factors determining the distribution pattern of Chironomidae in different types of freshwater habitats. Insects, 16.","DOI":"10.3390\/insects16050501"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Chowdhury, S., Dubey, V.K., Choudhury, S., Das, A., Jeengar, D., Sujatha, B., Kumar, A., Kumar, N., Semwal, A., and Kumar, V. (2023). Insects as bioindicator: A hidden gem for environmental monitoring. Front. Environ. Sci., 11.","DOI":"10.3389\/fenvs.2023.1146052"},{"key":"ref_7","first-page":"125","article-title":"Some aquatic insects and invertebrates as bioindicators for the evaluation of bacteriological pollution in El-Zomor and El-Mariotya canals, Giza, Egypt","volume":"2","author":"Adham","year":"2009","journal-title":"Egypt. Acad. J. Biol. Sci. A Entomol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1007\/BF02255231","article-title":"Freshwater biomonitoring and Chironomidae","volume":"26","author":"Rosenberg","year":"1992","journal-title":"Neth. J. Aquat. Ecol."},{"key":"ref_9","first-page":"74","article-title":"Morphology, cytogenetics and DNA barcode of the Chironomidae (Diptera) in Thailand","volume":"34","author":"Simwisat","year":"2015","journal-title":"J. Sci. Technol. Mahasarakham Univ."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Kang, H.J., Baek, M.J., Kang, J.H., and Bae, Y.J. (2022). DNA barcoding of chironomid larvae (diptera: Chironomidae) from large rivers in south korea to facilitate freshwater biomonitoring and public health surveillance. Int. J. Environ. Res. Public Health, 19.","DOI":"10.3390\/ijerph191912035"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e21060","DOI":"10.3897\/mbmg.2.21060","article-title":"Using DNA metabarcoding for assessing chironomid diversity and community change in mosquito controlled temporary wetlands","volume":"2","author":"Theissinger","year":"2018","journal-title":"Metabarcoding Metagenomics"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Carew, M.E., Pettigrove, V.J., Metzeling, L., and Hoffmann, A.A. (2013). Environmental monitoring using next generation sequencing: Rapid identification of macroinvertebrate bioindicator species. Front. Zool., 10.","DOI":"10.1186\/1742-9994-10-45"},{"key":"ref_13","first-page":"294","article-title":"DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates","volume":"3","author":"Folmer","year":"1994","journal-title":"Mol. Mar. Biol. Biotechnol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"93","DOI":"10.3897\/zookeys.1104.81403","article-title":"Taxonomic study on the genus Stenochironomus Kieffer from the Baishanzu Nature Reserve, China (Diptera, Chironomidae)","volume":"1104","author":"Song","year":"2022","journal-title":"ZooKeys"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"852","DOI":"10.1080\/10635150500354886","article-title":"The promise of DNA barcoding for taxonomy","volume":"54","author":"Hebert","year":"2005","journal-title":"Syst. Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"12161","DOI":"10.1002\/ece3.7979","article-title":"Toward a global DNA barcode reference library of the intolerant nonbiting midge genus Rheocricotopus Brundin, 1956","volume":"11","author":"Lin","year":"2021","journal-title":"Ecol. Evol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Xu, H.F., Lv, M.Y., Zhao, Y., Zhang, Z.C., Liu, Z., and Lin, X.L. (2025). Cryptic diversity and climatic niche divergence of Brillia Kieffer (Diptera: Chironomidae): Insights from a global DNA barcode dataset. Insects, 16.","DOI":"10.3390\/insects16070675"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1002\/mmnd.19540010305","article-title":"Die Gattung Neozavrelia Goetghebuer (Dipt. Chironomidae). Chironomidenstudien II","volume":"1","author":"Fittkau","year":"1954","journal-title":"Dtsch. Entomol. Z."},{"key":"ref_19","first-page":"189","article-title":"New Records of Chironomids (Diptera, Chironomidae) from Czech Republic and Slovak Republic","volume":"48","author":"Bitusik","year":"1993","journal-title":"Biologia"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1","DOI":"10.11646\/zootaxa.1153.1.1","article-title":"A redescription of Neozavrelia cuneipennis (Edwards) comb. nov., with a checklist of Neozavrelia species of the world (Diptera: Chironomidae)","volume":"1153","author":"Ekrem","year":"2006","journal-title":"Zootaxa"},{"key":"ref_21","first-page":"413","article-title":"A systematic review of European Stempellina Thienemann et Bause, 1913 (Diptera: Chironomidae) with description of a new species from Fennoscandia","volume":"55","year":"2005","journal-title":"Ann. Zool."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1111\/j.1440-6055.1998.tb01555.x","article-title":"The Australian species of Neozavrelia Goetghebuer (Diptera: Chironomidae: Tanytarsini)","volume":"37","author":"Cranston","year":"1998","journal-title":"Aust. J. Entomol."},{"key":"ref_23","first-page":"35","article-title":"Notes on the systematics of East Asian Neozavrelia Goetghebuer (Diptera, Chironomidae, Tanytarsini)","volume":"11","year":"2012","journal-title":"Euroasian Entomol. J."},{"key":"ref_24","first-page":"814","article-title":"First Record of the Genus Neozavrelia Goetghebuer with Redescription of Neozavrelia lindbergi Reiss (Diptera: Chironomidae) from India","volume":"88","author":"Mondal","year":"2025","journal-title":"Indian J. Entomol."},{"key":"ref_25","first-page":"121","article-title":"Two new species of Chironomidae from Liaoning Province, China (Diptera: Chironomidae)","volume":"17","author":"Wang","year":"1996","journal-title":"Zool. Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"251","DOI":"10.11646\/zootaxa.4938.3.1","article-title":"Revision of the genus Neozavrelia Goetghebuer, Thienemann, 1941 (Diptera: Chironomidae) from Eastern Siberia and the Russian Far East, with the description of new species","volume":"4938","author":"Orel","year":"2021","journal-title":"Zootaxa"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"349","DOI":"10.3897\/zookeys.1211.125436","article-title":"New faunistic records of chironomids and phantom midges (Diptera, Chironomidae and Chaoboridae) from Ukraine indicate recent climatic refugia in the Eastern Carpathians","volume":"1211","author":"Novikmec","year":"2024","journal-title":"ZooKeys"},{"key":"ref_28","first-page":"11","article-title":"Analysis of faunistic diversity in chironomids of the tribe Tanytarsini in Europe (Diptera: Chironomidae)","volume":"27","year":"2011","journal-title":"Dipteron"},{"key":"ref_29","first-page":"11","article-title":"Ecological patterns of Chironomidae assemblages in Dynaric karst springs","volume":"417","author":"Gadawski","year":"2016","journal-title":"Knowl. Manag. Aquat. Ecosyst."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1080\/00305316.2005.10417432","article-title":"A review of the genus Neozavrelia Goetghebuer from China (Diptera: Chironomidae: Chironominae: Tanytarsini)","volume":"39","author":"Guo","year":"2005","journal-title":"Orient. Insects"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"530","DOI":"10.1016\/j.ympev.2006.11.021","article-title":"A comprehensive DNA sequence library is essential for identification with DNA barcodes","volume":"43","author":"Ekrem","year":"2007","journal-title":"Mol. Phylogenet. Evol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1163\/187631203788964845","article-title":"Towards a phylogeny of Tanytarsus van der Wulp (Diptera: Chironomidae). Is morphology alone sufficient to reconstruct the genealogical relationship?","volume":"34","author":"Ekrem","year":"2003","journal-title":"Insect Syst. Evol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"27","DOI":"10.11646\/zootaxa.2743.1.2","article-title":"Exploring unknown life stages of Arctic Tanytarsini (Diptera: Chironomidae) with DNA barcoding","volume":"2743","author":"Stur","year":"2011","journal-title":"Zootaxa"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Stasiukynas, L., Havelka, J., da Silva, F.L., Torres Jimenez, M.F., Pod\u0117nas, S., and Lekoveckait\u0117, A. (2025). COI insights into diversity and species delimitation of immature stages of Non-Biting midges (Diptera: Chironomidae). Insects, 16.","DOI":"10.3390\/insects16020174"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1673\/031.013.15601","article-title":"Spatial and temporal distribution of non-biting midge larvae assemblages in streams in a mountainous region in southern Brazil","volume":"13","author":"Floss","year":"2013","journal-title":"J. Insect Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1146\/annurev-ento-011613-161958","article-title":"Freshwater biodiversity and aquatic insect diversification","volume":"59","author":"Dijkstra","year":"2014","journal-title":"Annu. Rev. Entomol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1105","DOI":"10.1899\/12-044.1","article-title":"Comparison of intraspecific genetic structure among related chironomids (Diptera) from New Zealand and Patagonia: Disparity between potential and realized dispersal","volume":"31","author":"Krosch","year":"2012","journal-title":"Freshw. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"4957","DOI":"10.1002\/ece3.7419","article-title":"Phylogeography of the northernmost distributed Anisocentropus caddisflies and their comparative genetic structures based on habitat preferences","volume":"11","author":"Takenaka","year":"2021","journal-title":"Ecol. Evol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"19","DOI":"10.3897\/CompCytogen.v16.i1.79182","article-title":"Karyotype characteristics and gene COI sequences of Chironomusbonus Shilova et Dzhvarsheishvili, 1974 (Diptera, Chironomidae) from the South Caucasus (Republic of Georgia, Paravani river)","volume":"16","author":"Karmokov","year":"2022","journal-title":"Comp. Cytogenet."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1328","DOI":"10.1111\/j.1365-2427.2010.02571.x","article-title":"Spatial population genetic structure reveals strong natal site fidelity in Echinocladius martini (Diptera: Chironomidae) in northeast Queensland, Australia","volume":"56","author":"Krosch","year":"2011","journal-title":"Freshw. Biol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1163\/1876312X-00002172","article-title":"DNA barcodes and morphology reveal unrecognized species in Chironomidae (Diptera)","volume":"49","author":"Lin","year":"2018","journal-title":"Insect Syst. Evol."},{"key":"ref_42","first-page":"149","article-title":"Some Nearctic Podonominae, Diamesinae, and Orthocladiinae (Diptera: Chironomidae)","volume":"170","year":"1969","journal-title":"Bull. Fish. Res. Board. Can."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1111\/j.1471-8286.2007.01678.x","article-title":"BOLD: The Barcode of Life Data System (http:\/\/www.barcodinglife.org)","volume":"7","author":"Ratnasingham","year":"2007","journal-title":"Mol. Ecol. Notes"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1647","DOI":"10.1093\/bioinformatics\/bts199","article-title":"Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data","volume":"28","author":"Kearse","year":"2012","journal-title":"Bioinformatics"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"msae263","DOI":"10.1093\/molbev\/msae263","article-title":"MEGA12: Molecular evolutionary genetic analysis version 12 for adaptive and green computing","volume":"41","author":"Kumar","year":"2024","journal-title":"Mol. Biol. Evol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1792","DOI":"10.1093\/nar\/gkh340","article-title":"MUSCLE: Multiple sequence alignment with high accuracy and high throughput","volume":"32","author":"Edgar","year":"2004","journal-title":"Nucleic Acids Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1007\/BF01731581","article-title":"A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences","volume":"16","author":"Kimura","year":"1980","journal-title":"J. Mol. Evol."},{"key":"ref_48","first-page":"406","article-title":"The neighbor-joining method: A new method for reconstructing phylogenetic trees","volume":"4","author":"Saitou","year":"1987","journal-title":"Mol. Biol. Evol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1864","DOI":"10.1111\/j.1365-294X.2011.05239.x","article-title":"ABGD, Automatic Barcode Gap Discovery for primary species delimitation","volume":"21","author":"Puillandre","year":"2012","journal-title":"Mol. Ecol."},{"key":"ref_50","unstructured":"Wickham, H., Bryan, J., Chang, W., McGowan, L., Fran\u00e7ois, R., Grolemund, G., Lemon, J., Kassambara, A., Mundt, F., and Revelle, W. (2020). R: A Language and Environment for Statistical Computing, R Foundation for Statistical Computing."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"526","DOI":"10.1093\/bioinformatics\/bty633","article-title":"Ape 5.0: An environment for modern phylogenetics and evolutionary analyses in R","volume":"35","author":"Paradis","year":"2019","journal-title":"Bioinformatics"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"439","DOI":"10.32614\/RJ-2018-009","article-title":"Simple features for R: Standardized support for spatial vector data","volume":"10","author":"Pebesma","year":"2018","journal-title":"R J."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"e16505","DOI":"10.7717\/peerj.16505","article-title":"phytools 2.0: An updated R ecosystem for phylogenetic comparative methods (and other things)","volume":"12","author":"Revell","year":"2024","journal-title":"PeerJ"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"170122","DOI":"10.1038\/sdata.2017.122","article-title":"Climatologies at high resolution for the earth\u2019s land surface areas","volume":"4","author":"Karger","year":"2017","journal-title":"Sci. Data"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Wilson, A.M., and Jetz, W. (2016). Remotely sensed high-resolution global cloud dynamics for predicting ecosystem and biodiversity distributions. PLoS Biol., 14.","DOI":"10.1371\/journal.pbio.1002415"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/j.isprsjprs.2013.11.002","article-title":"EarthEnv-DEM90: A nearly-global, void-free, multi-scale smoothed, 90m digital elevation model from fused ASTER and SRTM data","volume":"87","author":"Robinson","year":"2014","journal-title":"ISPRS J. Photogramm. Remote Sens."},{"key":"ref_57","first-page":"209","article-title":"The detection of disease clustering and a generalized regression approach","volume":"27","author":"Mantel","year":"1967","journal-title":"Cancer Res."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1179\/sre.1975.23.176.88","article-title":"Direct and inverse solutions of geodesics on the ellipsoid with application of nested equations","volume":"23","author":"Vincenty","year":"1975","journal-title":"Surv. Rev."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1890\/0012-9615(1999)069[0001:DBRATM]2.0.CO;2","article-title":"Distance-based redundancy analysis: Testing multispecies responses in multifactorial ecological experiments","volume":"69","author":"Legendre","year":"1999","journal-title":"Ecol. Monogr."},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Borcard, D., Gillet, F., and Legendre, P. (2018). Numerical Ecology with R, Springer. [2nd ed.].","DOI":"10.1007\/978-3-319-71404-2"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"1045","DOI":"10.2307\/1940179","article-title":"Partialling out the spatial component of ecological variation","volume":"73","author":"Borcard","year":"1992","journal-title":"Ecology"},{"key":"ref_62","doi-asserted-by":"crossref","unstructured":"Lai, J., Zou, Y., Zhang, J., and Peres-Neto, P. (2021). Rdacca. hp: An R package for generalizing hierarchical and variation partitioning in multiple regression and canonical analysis. BioRxiv.","DOI":"10.1101\/2021.03.09.434308"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1098\/rspb.2002.2218","article-title":"Biological identifications through DNA barcodes","volume":"270","author":"Hebert","year":"2003","journal-title":"Ser. B Biol. Sci."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"14812","DOI":"10.1073\/pnas.0406166101","article-title":"Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator","volume":"101","author":"Hebert","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"534","DOI":"10.11646\/zootaxa.4079.5.2","article-title":"Exploring the utility of DNA barcoding in species delimitation of Polypedilum (Tripodura) non-biting midges (Diptera: Chironomidae)","volume":"4079","author":"Song","year":"2016","journal-title":"Zootaxa"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"10723","DOI":"10.1002\/ece3.5590","article-title":"The impact of genetic diversity on the accuracy of DNA barcoding to identify species: A study on the genus Phellodendron","volume":"9","author":"Zhang","year":"2019","journal-title":"Ecol. Evol."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1146\/annurev.ecolsys.34.011802.132421","article-title":"Species-level paraphyly and polyphyly: Frequency, causes, and consequences, with insights from animal mitochondrial DNA","volume":"34","author":"Funk","year":"2003","journal-title":"Annu. Rev. Ecol. Evol. Syst."},{"key":"ref_68","doi-asserted-by":"crossref","unstructured":"Bocek, M., Motyka, M., Kusy, D., and Bocak, L. (2019). Genomic and mitochondrial data identify different species boundaries in aposematically polymorphic Eniclases net-winged beetles (Coleoptera: Lycidae). Insects, 10.","DOI":"10.3390\/insects10090295"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"6617","DOI":"10.1038\/s41598-024-56875-9","article-title":"Species delimitation, discovery and conservation in a tiger beetle species complex despite discordant genetic data","volume":"14","author":"Duran","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1590\/S1415-47572013000400002","article-title":"Mantel test in population genetics","volume":"36","author":"Soares","year":"2013","journal-title":"Genet. Mol. Biol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1080\/00949650008812035","article-title":"Comparison of permutation methods for the partial correlation and partial Mantel tests","volume":"67","author":"Legendre","year":"2000","journal-title":"J. Stat. Comput. Simul."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1899\/07-172.1","article-title":"Environmental factors, spatial variation, and specific requirements of Chironomidae in Mediterranean reference streams","volume":"28","author":"Rieradevall","year":"2009","journal-title":"J. N. Am. Benthol. Soc."},{"key":"ref_73","doi-asserted-by":"crossref","unstructured":"Popovi\u0107, N., Marinkovi\u0107, N., \u010cerba, D., Rakovi\u0107, M., \u0110ukni\u0107, J., and Paunovi\u0107, M. (2022). Diversity patterns and assemblage structure of non-biting midges (Diptera: Chironomidae) in urban waterbodies. Diversity, 14.","DOI":"10.3390\/d14030187"},{"key":"ref_74","doi-asserted-by":"crossref","unstructured":"Vasquez, A.A., Bonnici, B.L., Yusuf, S.H., Cruz, J.I., Hudson, P.L., and Ram, J.L. (2022). Improved chironomid barcode database enhances identification of water mite dietary content. Diversity, 14.","DOI":"10.3390\/d14020065"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"71","DOI":"10.5324\/fn.v31i0.1361","article-title":"Long-term patterns of chironomid assemblages in a high elevation stream\/lake network (Switzerland): Implications to global change","volume":"31","author":"Oertli","year":"2012","journal-title":"Fauna Nor."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"8087","DOI":"10.1007\/s11356-019-04267-2","article-title":"Chironomid genera distribution related to environmental characteristics of a highly impacted basin (Argentina, South America)","volume":"26","author":"Cortese","year":"2019","journal-title":"Environ. Sci. Pollut. Res."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"S444","DOI":"10.1002\/lno.12007","article-title":"Stability of chironomid community structure during historic climatic and environmental change in subarctic Alaska","volume":"67","author":"Mayfield","year":"2022","journal-title":"Limnol. Oceanogr."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1093\/jhered\/esae016","article-title":"Phylogeography of the freshwater crab Potamon persicum (Decapoda: Potamidae): An ancestral ring species?","volume":"115","author":"Naderloo","year":"2024","journal-title":"J. Hered."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"73","DOI":"10.4081\/jear.2013.e14","article-title":"Response of chironomid species (Diptera, Chironomidae) to water temperature: Effects on species distribution in specific habitat","volume":"45","author":"Marziali","year":"2013","journal-title":"J. Entomol. Acarol. Res."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"151","DOI":"10.11646\/zootaxa.5214.2.1","article-title":"Tanytarsini (Diptera: Chironomidae) collected during Polish expeditions to North Korea","volume":"5214","author":"Gadawski","year":"2022","journal-title":"Zootaxa"}],"container-title":["Insects"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4450\/17\/8\/814\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T04:27:31Z","timestamp":1786076851000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4450\/17\/8\/814"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,5]]},"references-count":80,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["insects17080814"],"URL":"https:\/\/doi.org\/10.3390\/insects17080814","relation":{},"ISSN":["2075-4450"],"issn-type":[{"value":"2075-4450","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,5]]}}}