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Title: EFFECT OF ALCOHOL-BASED PRESERVATIVES ON QUALITY OF BEETLE LARVAL SPECIMENS
EFFECT OF ALCOHOL-BASED PRESERVATIVES ON QUALITY OF BEETLE LARVAL SPECIMENS

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Martin Novak; Pavel Jakubec; Karolina Mahlerova; Santiago Montoya-Molina; Jarin Qubaiova
10.5593/sgem2022/5.1/s20.019
10.5593/sgem2022/5.1
1314-2704
978-619-7603-46-0
English
22
5.1
•    Prof. DSc. Oleksandr Trofymchuk, UKRAINE 
•    Prof. Dr. hab. oec. Baiba Rivza, LATVIA
Ecology and Environmental Protection
The standard procedure for fluid preservation of insects consists of two steps. First, the specimen is killed and prepared in a fixative fluid and then transferred to a storage fluid. Recommendations for using specific chemicals or mixtures differ depending on the size and anatomy of the animal and vary among the authors. However, controlled comparative studies are often missing. The ideal method should preserve the specimen’s morphology as well as its genetic information. We have investigated the effect of 12 combinations of commonly used methods based on ethanol, using four types of fixatives (Hot Water Kill (HWK) for 5 minutes/6 hours; absolute ethanol cold/boiling) and three types of storage fluids (70 %; 80 %; and absolute ethanol) after eight months of storage. The effects were tested on mature larvae of a forensically relevant beetle species. We have evaluated the effectivity in terms of color preservation, shape of the specimen, and quality of genetic material. The color was best preserved by HWK for 5 min with subsequent storage in absolute ethanol, and the worst results were achieved by cold absolute ethanol with subsequent storage in 70 % ethanol. The fixative type has a major effect on the shape compared to the storage fluid. The effect of treatments on shape appears to be inverse to the effect on color preservation. The genetic material was preserved equally well in all treatments. The advantages and disadvantages of specific treatments and the recommendation of the best method are discussed.
[1] Mourek J. & Liskova E. Biologicke sbirky – Metody sberu, preparace a uchovavani, prirucka k projektu Ama Mater Studiorum. Univerzita Karlova v Praze – Pedagogicka fakulta, 55 pp, 2010. [in Czech]
[2] Amendt J., Campobasso C.P., Gaudry E., Reiter C., LeBlanc H.N. & Hall M.J. R. Best Practice in Forensic Entomology — Standards and Guidelines. International Journal of Legal Medicine, vol. 121, pp 90-104, 2007.
[3] Sanford M.R., Byrd J.H., Tomberlin J.K. & Wallace J.R. Entomological Evidence Collections Methods: American Board of Forensic Entomology Approved Protocols. In Byrd J.H. & Tomberlin J.K. (Eds). Forensic Entomology, The Utility of Arthropods in Legal Investigations, Third Edition. CRC Press, Taylor & Francis Group, pp 63-86, 2020.
[4] Simmons J.E. Fluid Preservation, A Comprehensive Reference. Rowman & Littlefield Publishers, 364 pp, 2014.
[5] Hammond P.M., Marshall J.E., Cox M.L., Jessop L., Garner B.H. & Barclay M.V.L. British Coleoptera larvae. A guide to the families and major subfamilies. Royal Entomological Society, 280 pp, 2019.
[6] Riley C.V. Directions for Collecting and Preserving Insects (Classic Reprint). Forgotten Books, 162 pp, 1892.
[7] Krogmann L. & Holstein J. Preserving and Specimen Handling: Insects and other Invertebrates, pp 463-481. In Eymann J., Degreef J., Hauser C., Monje J., Samyn Y. & VandenSpiegel D. (Eds). Manual on field recording techniques and protocols for all Taxa Biodiversity Inventories and Monitoring, AbcTaxa, Brussel, 653 pp, 2010.
[8] Gennard D. Forensic entomology an introduction, Second edition. Wiley-Blackwell, 272 pp, 2014.
[9] Cox M.L., Schray C.L., Luster C.N., Stewart Z.S., Korytko P.J., Khan K.N.M., Paulauskis J.D. & Dunstan R.W. Assessment of fixatives, fixation, and tissue processing on morphology and RNA integrity, Experimental and Molecular Pathology, vol. 80, pp 183-191, 2006.
[10] Fukatsu T. Acetone preservation: a practical technique for molecular analysis, Molecular Ecology, vol. 8, 1935-1945, 1999.
[11] Day D.M. & Wallman J.F. Effect of preservative solutions on preservation of Calliphora augur and Lucilia cuprina larvae (Diptera: Calliphoridae) with implications for post-mortem interval estimates, Forensic Science International, vol. 179, pp 1-10, 2008
[12] Mandrioli M. Insect collections and DNA analyses: how to manage collections? Museum Management and Curatorship, vol. 23, 193-199, 2008.
[13] Novak M., Fratczak-Lagiewska K., Madra-Bielewicz A. & Matuszewski S. Eyebackground contrast as a quantitative marker for pupal age in a forensically important carrion beetle Necrodes littoralis L. (Silphidae), Scientific Reports, vol. 10, 14494, 2010.
[14] Simon C., Frati F., Beckenbach A., Crespi B., Liu H. & Flook P. Evolution, Weighting, and Phylogenetic Utility of Mitochondrial Gene Sequences and a Compilation of Conserved Polymerase Chain Reaction Primers, Annals of the Entomological Society of America, vol. 87, pp 651–701, 1994.
[15] Kumar S., Stecher G., Li M., Knyaz, C. & Tamura, K. MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms, Molecular Biology and Evolution, vol. 35, pp 1547–1549, 2018.
The project was supported by the Ministry of the Interior of the Czech Republic (grant no. VI20192022130).
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https://doi.org/10.5593/sgem2022/5.1/s20.019
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Proceedings of 22nd International Multidisciplinary Scientific GeoConference SGEM 2022
22nd International Multidisciplinary Scientific GeoConference SGEM 2022, 04 - 10 July, 2022
Proceedings Paper
STEF92 Technology
International Multidisciplinary Scientific GeoConference SGEM
SWS Scholarly Society; Acad Sci Czech Republ; Latvian Acad Sci; Polish Acad Sci; Serbian Acad Sci and Arts; Natl Acad Sci Ukraine; Natl Acad Sci Armenia; Sci Council Japan; European Acad Sci, Arts and Letters; Acad Fine Arts Zagreb Croatia; Croatian Acad Sci and Arts; Acad Sci Moldova; Montenegrin Acad Sci and Arts; Georgian Acad Sci; Acad Fine Arts and Design Bratislava; Turkish Acad Sci.
143-150
04 - 10 July, 2022
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fluid preservation, fixative, ethanol, storage, entomology