Mitogenomic Insights into Temperature Adaptation: A Comparative Study of the Subfamily Corydalinae Davis, 1903 (Megaloptera: Corydalidae)
Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Sequencing and Assembly
2.2. Bioinformatic Analysis of Mitogenomes
2.3. Phylogenetic Analysis
2.4. Detecting Selective Pressure
3. Results
3.1. Protein-Coding Genes
3.2. Non-Coding Regions
3.3. Phylogenetic Analysis of Corydalinae
3.4. Results of Detecting Selective Pressure
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Cameron, S.L. Insect mitochondrial genomics: Implications for evolution and phylogeny. Annu. Rev. Entomol. 2014, 59, 95–117. [Google Scholar] [CrossRef]
- Curole, J.P.; Kocher, T.D. Mitogenomics: Digging deeper with complete mitochondrial genomes. Trends Ecol. Evol. 1999, 14, 394–398. [Google Scholar] [CrossRef]
- Cameron, S.L.; Sullivan, J.; Song, H.; Miller, K.B.; Whiting, M.F. A mitochondrial genome phylogeny of the Neuropterida (lacewings, alderflies and snakeflies) and their relationship to the other holometabolous insect orders. Zool. Scr. 2009, 38, 575–590. [Google Scholar] [CrossRef]
- Jiang, Y.L.; Yang, F.; Yue, L.; Hayashi, F.; Yang, D.; Liu, X.Y. Origin and spatio-temporal diversification of a fishfly lineage endemic to the islands of East Asia (Megaloptera: Corydalidae). Syst. Entomol. 2021, 46, 124–139. [Google Scholar] [CrossRef]
- Jiang, Y.L.; Yue, L.; Yang, F.; Gillung, J.P.; Winterton, S.L.; Price, B.W.; Contreras-Ramos, A.; Hayashi, F.; Aspöck, U.; Aspöck, H.; et al. Similar pattern, different paths: Tracing the biogeographical history of Megaloptera (Insecta: Neuropterida) using mitochondrial phylogenomics. Cladistics 2022, 38, 374–391. [Google Scholar] [CrossRef]
- Ma, C.; Yang, P.C.; Jiang, F.; Chapuis, M.P.; Shali, Y.; Sword, G.A.; Kang, L. Mitochondrial genomes reveal the global phylogeography and dispersal routes of the migratory locust. Mol. Ecol. 2012, 21, 4344–4358. [Google Scholar] [CrossRef] [PubMed]
- Simon, S.; Hadrys, H. A comparative analysis of complete mitochondrial genomes among Hexapoda. Mol. Phylogenet. Evol. 2013, 69, 393–403. [Google Scholar] [CrossRef]
- Kang, N.; Hu, H. Adaptive evidence of mitochondrial genes in Pteromalidae and Eulophidae (Hymenoptera: Chalcidoidea). PLoS ONE 2023, 18, e0294687. [Google Scholar] [CrossRef] [PubMed]
- Li, X.D.; Jiang, G.F.; Yan, L.Y.; Li, R.; Mu, Y.; Deng, W.A. Positive selection drove the adaptation of mitochondrial genes to the demands of flight and high-altitude environments in grasshoppers. Front. Genet. 2018, 9, 605. [Google Scholar] [CrossRef]
- Boore, J.L.; Collins, T.M.; Stanton, D.; Daehler, L.L.; Brown, W.M. Deducing the pattern of arthropod phylogeny from mitochondrial DNA rearrangements. Nature 1995, 376, 163–165. [Google Scholar] [CrossRef]
- Colinet, H.; Nguyen, T.T.A.; Cloutier, C.; Michaud, D.; Hance, T. Proteomic profiling of a parasitic wasp exposed to constant and fluctuating cold exposure. Insect Biochem. Mol. Biol. 2007, 37, 1177–1188. [Google Scholar] [CrossRef]
- Joanisse, D.R.; Storey, K.B. Mitochondrial enzymes during overwintering in two species of cold-hardy gall insects. Insect Biochem. Mol. Biol. 1994, 24, 145–150. [Google Scholar] [CrossRef]
- Ramirez, L.; Luna, F.; Mucci, C.A.; Lamattina, L. Fast weight recovery, metabolic rate adjustment and gene-expression regulation define responses of cold-stressed honey bee brood. J. Insect Physiol. 2021, 128, 104178. [Google Scholar] [CrossRef]
- Rider, M.H.; Hussain, N.; Dilworth, S.M.; Storey, J.M.; Storey, K.B. AMP-activated protein kinase and metabolic regulation in cold-hardy insects. J. Insect Physiol. 2011, 57, 1453–1462. [Google Scholar] [CrossRef]
- Teets, N.M.; Dalrymple, E.G.; Hillis, M.H.; Gantz, J.D.; Spacht, D.E.; Lee, R.E.; Denlinger, D.L. Changes in energy reserves and gene expression elicited by freezing and supercooling in the Antarctic midge. Insects 2020, 11, 18. [Google Scholar] [CrossRef]
- Luo, Y.J.; Gao, W.X.; Gao, Y.Q.; Tang, S.; Huang, Q.Y.; Tan, X.L.; Chen, J.; Huang, T.S. Mitochondrial genome analysis of Ochotona curzoniae and implication of cytochrome c oxidase in hypoxic adaptation. Mitochondrion 2008, 8, 352–357. [Google Scholar] [CrossRef]
- Xu, S.Q.; Yang, Y.Z.; Zhou, J.; Jing, G.E.; Chen, Y.T.; Wang, J.; Yang, H.M.; Wang, J.; Yu, J.; Zheng, X.G.; et al. A mitochondrial genome sequence of the Tibetan antelope (Pantholops hodgsonii). Genom. Proteom. Bioinf. 2005, 3, 5–17. [Google Scholar] [CrossRef]
- Zhang, J.Y.; Luu, B.E.; Yu, D.N.; Zhang, L.P.; Al-Attar, R.; Storey, K.B. The complete mitochondrial genome of Dryophytes versicolor: Phylogenetic relationship among Hylidae and mitochondrial protein-coding gene expression in response to freezing and anoxia. Int. J. Biol. Macromol. 2019, 132, 461–469. [Google Scholar] [CrossRef] [PubMed]
- Cai, L.N.; Zhang, L.H.; Lin, Y.J.; Wang, J.Y.; Storey, K.B.; Zhang, J.Y.; Yu, D.N. Two-fold ND5 genes, three-fold control regions, lncRNA, and the “missing” atp8 found in the mitogenomes of Polypedates megacephalus (Rhacophridae: Polypedates). Animals 2023, 13, 2857. [Google Scholar] [CrossRef] [PubMed]
- Tu, Y.Z.; Lin, A.L.; Jiang, Y.L.; Liu, X.Y. Comparative mitochondrial genomics and phylogenetics among species of the oriental dobsonfly genus Neoneuromus van der Weele, 1909 (Megaloptera: Corydalidae). J. Asia Pac. Entomol. 2021, 24, 1257–1265. [Google Scholar] [CrossRef]
- Balloux, F.; Handley, L.J.L.; Jombart, T.; Liu, H.; Manica, A. Climate shaped the worldwide distribution of human mitochondrial DNA sequence variation. Proc. Biol. Sci. 2009, 276, 3447–3455. [Google Scholar] [CrossRef] [PubMed]
- Mishmar, D.; Ruiz-Pesini, E.; Golik, P.; Macaulay, V.; Clark, A.G.; Hosseini, S.; Brandon, M.; Easley, K.; Chen, E.; Brown, M.D.; et al. Natural selection shaped regional mtDNA variation in humans. Proc. Natl. Acad. Sci. USA 2003, 100, 171–176. [Google Scholar] [CrossRef]
- Sun, J.T.; Jin, P.Y.; Hoffmann, A.A.; Duan, X.Z.; Dai, J.; Hu, G.; Xue, X.F.; Hong, X.Y. Evolutionary divergence of mitochondrial genomes in two Tetranychus species distributed across different climates. Insect Mol. Biol. 2018, 27, 698–709. [Google Scholar] [CrossRef] [PubMed]
- Lamb, A.M.; Gan, H.M.; Greening, C.; Joseph, L.; Lee, Y.P.; Moran-Ordonez, A.; Sunnucks, P.; Pavlova, A. Climate-driven mitochondrial selection: A test in Australian songbirds. Mol. Ecol. 2018, 27, 898–918. [Google Scholar] [CrossRef]
- Martins, C.C.; Ardila-Camacho, A.; Rivera-Gasperín, S.L.; Oswald, J.D.; Liu, X.; Contreras-Ramos, A. A world checklist of extant and extinct species of Megaloptera (Insecta: Neuropterida). Eur. J. Taxon. 2022, 812, 1–93. [Google Scholar] [CrossRef]
- New, T.R.; Theischinger, G. Megaloptera, alderflies and dobsonflies. Handb. Zool. 1993, 1, 516–520. [Google Scholar]
- Beutel, R.G.; Friedrich, F. Comparative study of larval head structures of Megaloptera (Hexapoda). Eur. J. Entomol. 2008, 105, 917–938. [Google Scholar] [CrossRef]
- Glorioso, M.J. Systematics of the dobsonfly subfamily Corydalinae (Megaloptera: Corydalidae). Syst. Entomol. 1981, 6, 253–290. [Google Scholar] [CrossRef]
- Contreras-Ramos, A. Systematics of the Dobsonfly Genus Corydalus Latreille (Megaloptera: Corydalidae); Entomological Society of America: Lanham, MD, USA, 1998. [Google Scholar]
- Penny, N.D. The phylogenetic position of Chloroniella peringueyi (Megaloptera: Corydalidae) and its zoogeographic significance. Entoml. News 1993, 104, 17–30. [Google Scholar]
- Contreras-Ramos, A. Phylogenetic review of dobsonflies of the subfamily Corydalinae and the genus Corydalus Latreille (Megaloptera: Corydalidae). Zootaxa 2011, 2862, 1. [Google Scholar] [CrossRef]
- Liu, X.Y.; Yang, D.; Ge, S.Q.; Yang, X.K. Phylogenetic review of the Chinese species of Acanthacorydalis (Megaloptera, Corydalidae). Zool. Scr. 2005, 34, 373–387. [Google Scholar] [CrossRef]
- Wang, Y.R.; Wang, Y.Y.; Chang, W.C.; Liu, X.Y.; Yang, D. Sequencing and analysis of the complete mitochondrial genome of the giant dobsonfly Acanthacorydalis orientalis (McLachlan) (Insecta: Megaloptera: Corydalidae). Zool. Scr. 2014, 39, 209–223. [Google Scholar]
- Jiang, Y.L.; Yang, F.; Yang, D.; Liu, X.Y. Complete mitochondrial genome of a neotropical dobsonfly Chloronia mirifica Navás, 1925 (Megaloptera: Corydalidae), with phylogenetic implications for the genus Chloronia Banks, 1908. Zootaxa 2016, 4162, 46–60. [Google Scholar] [CrossRef] [PubMed]
- Bolger, A.M.; Lohse, M.; Usadel, B. Trimmomatic: A flexible trimmer for Illumina sequence data. Bioinformatics 2014, 30, 2114–2120. [Google Scholar] [CrossRef]
- Kearse, M.; Moir, R.; Wilson, A.; Stones-Havas, S.; Cheung, M.; Sturrock, S.; Buxton, S.; Cooper, A.; Markowitz, S.; Duran, C.; et al. Geneious basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 2012, 28, 1647–1649. [Google Scholar] [CrossRef]
- Lowe, T.M.; Chan, P.P. tRNAscan-SE on-line: Search and contextual analysis of transfer RNA genes. Nucleic Acids Res. 2016, 44, 54–57. [Google Scholar] [CrossRef]
- Kumar, S.; Stecher, G.; Tamura, K. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol. Biol. Evol. 2016, 33, 1870–1874. [Google Scholar] [CrossRef] [PubMed]
- Perna, N.T.; Kocher, T.D. Patterns of nucleotide composition at fourfold degenerate sites of animal mitochondrial genomes. J. Mol. Evol. 1995, 41, 353–358. [Google Scholar] [CrossRef]
- Sueoka, N. Directional mutation pressure and neutral molecular evolution. Proc. Natl. Acad. Sci. USA 1988, 85, 2653–2657. [Google Scholar] [CrossRef]
- Rozas, J.; Ferrer-Mata, A.; Sánchez-DelBarrio, J.C.; Guirao-Rico, S.; Librado, P.; Ramos-Onsins, S.E.; Sánchez-Gracia, A. DnaSP 6: DNA sequence polymorphism analysis of large datasets. Mol. Biol. Evol. 2017, 34, 3299–3302. [Google Scholar] [CrossRef]
- Xia, X.H. DAMBE7: New and improved tools for data analysis in molecular biology and evolution. Mol. Biol. Evol. 2018, 35, 1550–1552. [Google Scholar] [CrossRef] [PubMed]
- Benson, G. Tandem repeats finder: A program to analyze DNA sequences. Nucleic Acids Res. 1999, 27, 573–580. [Google Scholar] [CrossRef] [PubMed]
- Beier, S.; Thiel, T.; Münch, T.; Scholz, U.; Mascher, M. MISA-web: A web server for microsatellite prediction. Bioinformatics 2017, 33, 2583–2585. [Google Scholar] [CrossRef]
- Katoh, K.; Standley, D.M. MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Mol. Biol. Evol. 2013, 30, 772–780. [Google Scholar] [CrossRef] [PubMed]
- Lanfear, R.; Frandsen, P.B.; Wright, A.M.; Senfeld, T.; Calcott, B. PartitionFinder 2: New methods for selecting partitioned models of evolution for molecular and morphological phylogenetic analyses. Mol. Bio. Evol. 2017, 34, 72–773. [Google Scholar] [CrossRef]
- Ronquist, F.; Teslenko, M.; van der Mark, P.; Ayres, D.L.; Darling, A.; Höhna, S.; Larget, B.; Liu, L.; Suchard, M.A.; Huelsenbeck, J.P. MRBAYES 3.2: Efficient bayesian phylogenetic inference and model selection across a large model space. Syst. Biol. 2012, 61, 539–542. [Google Scholar] [CrossRef]
- Nguyen, L.T.; Schmidt, H.A.; von Haeseler, A.; Minh, B.Q. IQ-TREE: A Fast and Effective Stochastic Algorithm for Estimating Maximum-Likelihood Phylogenies. Mol. Biol. Evol. 2015, 32, 268–274. [Google Scholar] [CrossRef]
- Gao, F.L.; Chen, C.; Arab, D.A.; Du, Z.; He, Y.; Ho, S.Y.W. EasyCodeML: A visual tool for analysis of selection using CodeML. Ecol. Evol. 2019, 9, 3891–3898. [Google Scholar] [CrossRef]
- Yang, Z.H. PAML 4: Phylogenetic analysis by maximum likelihood. Mol. Biol. Evol. 2007, 24, 1586–1591. [Google Scholar] [CrossRef]
- NASA Langley Research Center. NASA POWER Data Access Viewer (Version 2024.07); NASA POWER Project; NASA Langley Research Center: Hampton, VA, USA, 2024. Available online: https://power.larc.nasa.gov/data-access-viewer/ (accessed on 31 October 2025).
- Hua, J.M.; Dong, P.Z.; Xie, Q.; Li, M.; Bu, W.J. The mitochondrial genome of Protohermes concolorus Yang et Yang 1988 (Insecta: Megaloptera: Corydalidae). Mol. Biol. Rep. 2009, 36, 1757–1765. [Google Scholar] [CrossRef]
- Jiang, Y.L.; Zhou, Y.J.; Wang, Y.Y.; Yue, L.; Yan, Y.; Wang, M.Q.; Liu, X.Y. Complete mitochondrial genomes of two oriental dobsonflies, Neoneuromus tonkinensis (van der Weele) and Nevromus exterior (Navás) (Megaloptera: Corydalidae), and phylogenetic implications of Corydalinae. Zootaxa 2015, 3964, 44–62. [Google Scholar] [CrossRef] [PubMed]
- Li, Q.; Wei, S.; Shi, M.; Chen, X.X. Complete mitochondrial genome of Neochauliodes bowringi (MacLachlan) (Megaloptera: Corydalidae). Mitochondrial DNA 2013, 26, 112–113. [Google Scholar] [CrossRef]
- Yang, F.; Jiang, Y.L.; Yang, D.; Liu, X.Y. Mitochondrial genomes of two Australian fishflies with an evolutionary timescale of Chauliodinae. Sci. Rep. 2017, 7, 4481. [Google Scholar] [CrossRef] [PubMed]
- Zhao, Y.Y.; Zhang, H.L.; Zhang, Y.H. Complete mitochondrial genome of Neochauliodes parasparsus (Megaloptera: Corydalidae) with phylogenetic consideration. Biochem. Syst. Ecol. 2017, 70, 192–199. [Google Scholar] [CrossRef]
- Bai, H.J.; Zhang, J.R.; Dietrich, C.H.; Li, Y.P.; Yuan, X.Q. Structural features of mitochondrial genomes of eight treehoppers (Hemiptera: Membracidae: Centrotinae) and their phylogenetic implications. Genes 2023, 14, 1510. [Google Scholar] [CrossRef] [PubMed]
- Ma, L.Y.; Liu, F.F.; Chiba, H.; Yuan, X.Q. The mitochondrial genomes of three skippers: Insights into the evolution of the family Hesperiidae (Lepidoptera). Genomics 2020, 112, 432–441. [Google Scholar] [CrossRef]
- Yan, L.P.; Xue, W.T.; Zhang, D.; Li, J.Q. Comparative analysis of the mitochondrial genomes of flesh flies and their evolutionary implication. Int. J. Biol. Macromol. 2021, 174, 385–391. [Google Scholar] [CrossRef]
- Zou, Y.; Ma, W.Q.; Zhang, L.F.; He, S.L.; Zhang, X.Y.; Tao, Z. The complete mitochondrial genome of the bean pod borer, Maruca testulalis (Lepidoptera: Crambidae: Spilomelinae). Mitochondrial DNA A DNA Mapp. Seq. Anal. 2014, 27, 740–741. [Google Scholar]
- Louis, D.; Vincent, L.; Philippe, V.; Bastien, B. Evaluation of methods to detect shifts in directional selection at the genome scale. Mol. Bio. Evol. 2023, 40, msac247. [Google Scholar]
- Steffen, R.; Ogoniak, L.; Grundmann, N.; Pawluchin, A.; Soehnlein, O.; Schmitz, J. paPAML: An improved computational tool to explore selection pressure on protein-coding sequences. Genes 2022, 13, 1090. [Google Scholar] [CrossRef]
- Álvarez, H.A.; Rivas-Soto, M.A. Potential geographic distribution and ecological niche of new world dobsonflies (Megaloptera: Corydalidae): The case of the Nearctic-Neotropical transition zone. Aquat. Insects 2022, 44, 182–194. [Google Scholar] [CrossRef]
- Zhang, X.; Chen, J.; Luo, H.Y.; Chen, X.; Zhong, J.; Ji, X. Climate-driven mitochondrial selection in lacertid lizards. Ecol. Evol. 2024, 14, e11176. [Google Scholar] [CrossRef]
- Beckenbach, A.T.; Stewart, B. Insect mitochondrial genomics 3: The complete mitochondrial genome sequences of representatives from two neuropteroid orders: A dobsonfly (order Megaloptera) and a giant lacewing and an owlfly (order Neuroptera). Genome 2008, 52, 31–38. [Google Scholar] [CrossRef]
- Hassan, M.A.; Shen, R.R.; Ali, M.; Liu, X.Y. The first complete mitochondrial genomes of three dobsonfly species (Megaloptera: Corydalidae) from Pakistan with phylogenetic implications. Rev. Bras. Entomol. 2022, 66, e20220112. [Google Scholar] [CrossRef]
- Xu, X.; Cheng, C.; Yan, B.; Su, E.; He, N. The complete mitochondrial genome of the Neochauliodes fraternus (Megaloptera: Corydalidae). Mitochondrial DNA Part A 2016, 27, 2163–2164. [Google Scholar] [CrossRef] [PubMed]





| Codon Position | Iss | Iss.cSym | P cSym | Iss.cAsym | P cAsym |
|---|---|---|---|---|---|
| 1st position | 0.530 | 0.849 | 0.000 | 0.837 | 0.000 |
| 2nd position | 0.142 | 0.842 | 0.000 | 0.758 | 0.000 |
| 3rd position | 0.057 | 0.849 | 0.000 | 0.837 | 0.000 |
| All positions | 0.221 | 0.858 | 0.000 | 0.846 | 0.000 |
| Genus | LRT p-Value | Positive Sites | Genes |
|---|---|---|---|
| Acanthacorydalis | 0.000000023 | 2190 E 0.972 * | nad2 |
| Corydalus | 0.000010332 | 260 S 0.981 *, 751 T 0.964 *, 1913 S 0.991 **, 2040 E 0.968 *, 2300 M 0.978 *, 2826 V 0.981 *, 3600 N 0.988 *, 3631 E 0.959 * | atp8, cox1, nad2, nad4, nad6 |
| Neoneuromus | 0.000000364 | 1835 A 0.950 * | nad1 |
| Nevromus | 0.000000017 | 267 N 0.969 *, 2906 N 0.957 * | atp8, nad4l |
| Protohermes | 0.000000048 | 2056 D 0.960 *, 2754 C 0.987 *, 2782 S 0.987 *, 3356 S 0.984 * | nad2, nad4, nad5 |
| Genus | Distribution | Highest Temperature (°C) | Lowest Temperature (°C) |
|---|---|---|---|
| Acanthacorydalis | Oriental Region, Northeastern India to Vietnam and China | 36.51–40.19 (Northeast India) | −31.32–−26.18 (Northeast China) |
| Chloronia | Neotropical Region, Brazil through Mexico, Lesser Antilles | 37.21–42.49 (Amazon Basin) | 0.39–1.93 (Amazon Basin) |
| Chloroniella | South Africa | 41.96–45.46 | 4.3–9.43 |
| Corydalus | North, Central, and South America | 37.21–42.49 (Amazon Basin) | −44.62–−39.48 (South-central Canada) |
| Neoneuromus | Oriental Region, Northeastern India to Malay Peninsula and China | 36.51–40.19 (Northeast India) | −31.32–−26.18 (Northeast China) |
| Nevromus | Oriental Region, Northwest India to Indonesia and China | 44.58–48.42 (Northwest India) | −31.32–−26.18 (Northeast China) |
| Platyneuromus | Central America to Northeastern Mexico | 43.38–46.84 (Northeast Mexico) | 2.71–6.01 (Northeast Mexico) |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Wang, W.; Tian, S.; Wang, Z.; Wang, Y.; Liu, X.; Jiang, Y. Mitogenomic Insights into Temperature Adaptation: A Comparative Study of the Subfamily Corydalinae Davis, 1903 (Megaloptera: Corydalidae). Insects 2025, 16, 1151. https://doi.org/10.3390/insects16111151
Wang W, Tian S, Wang Z, Wang Y, Liu X, Jiang Y. Mitogenomic Insights into Temperature Adaptation: A Comparative Study of the Subfamily Corydalinae Davis, 1903 (Megaloptera: Corydalidae). Insects. 2025; 16(11):1151. https://doi.org/10.3390/insects16111151
Chicago/Turabian StyleWang, Wenye, Shuo Tian, Zifan Wang, Yuyu Wang, Xingyue Liu, and Yunlan Jiang. 2025. "Mitogenomic Insights into Temperature Adaptation: A Comparative Study of the Subfamily Corydalinae Davis, 1903 (Megaloptera: Corydalidae)" Insects 16, no. 11: 1151. https://doi.org/10.3390/insects16111151
APA StyleWang, W., Tian, S., Wang, Z., Wang, Y., Liu, X., & Jiang, Y. (2025). Mitogenomic Insights into Temperature Adaptation: A Comparative Study of the Subfamily Corydalinae Davis, 1903 (Megaloptera: Corydalidae). Insects, 16(11), 1151. https://doi.org/10.3390/insects16111151

