Abstract
Succession patterns of carrion insects on large mammal’s carrion has been widely studied, notably to estimate the post-mortem interval in forensic investigations as accurately as possible. However, little attention has been paid to the carrion insects living inside these bones once a carcass is skeletonized. One very recent study documented flies emerging from pig carcasses, and only scarce authors reported the presence of other carrion insects taking advantage of the bone marrow. We, thus, aimed to (1) estimate the frequency of inner-bone space colonization by carrion insects, with particular attention to bone-skipper flies; (2) identify the insects living inside the carrion bones; and (3) determine whether or not carrion insects found within the bones can successfully exit the bones and complete their development. We extensively sampled 185 large mammals’ bones collected from twelve vulture feeding stations and four isolated carcasses in southwest France and northern Spain. Sampled bones were opened, and the insects found inside were identified. For two bones, foramen, i.e., the holes providing a natural entrance and exit to the bone’s inner cavity, was monitored with a camera to assess the insect’s putative exit. We describe the entomofauna, i.e., the set of insect species, living within the bones, and illustrate insects’ ability to exit the bones for their subsequent development and maturity. These results are discussed in the framework of carrion insect conservation and forensic entomology perspectives.
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References
Adair TW, Kondratieff BC (1996) The occurrence of Nitidula flavomaculata (Coleoptera: Nitidulidae) on a human corpse. Entomological News 107(4): 233–236 https://www.biodiversitylibrary.org/partpdf/31173
Arrêté du 28 février 2008 relatif aux modalités de délivrance de l’agrément sanitaire et de l’autorisation des établissements visés par le règlement (CE) n° 1774/2002 du Parlement européen et du Conseil du 3 Octobre 2002 établissant des règles sanitaires applicables aux sous-produits animaux non destinés à la consommation humaine https://www.legifrance.gouv.fr/loda/id/LEGIARTI000018230530/2008-03-08/
Azémar F, Cazaban F, Pelozuelo L (2020) Breaking the silence: how shedding light on the bone-skipper fly Thyreophora cynophila (Diptera: Piophilidae) demonstrated it still has a large distribution area in the Pyrenees mountains. France Biodivers Data J 8:e54868. https://doi.org/10.3897/BDJ.8.e54868
Beaumont C, Cabon F, Larquier C, Azémar F, Pelozuelo L (2023) Observation of the elusive bone-skipper fly Centrophlebomyia furcata (Fabricius, 1794) (Diptera: Piophilidae) at least 128 years after its last record in France, and additional data on Thyreophora cynophila (Panzer, 1794) (Diptera: Piophilidae) in France. Ann Soc Entomol Fr 59(5):326–336. https://doi.org/10.1080/00379271.2023.2239213
Bonicelli A, Mickleburgh HL, Chighine A, Locci E, Wescott D, Procopio N (2022) The ‘ForensOMICS’ approach for postmortem interval estimation from human bone by integrating metabolomics, lipidomics, and proteomics. eLife 11:e83658. https://doi.org/10.7554/eLife.83658
Cabanillas D (2020) Revision of the distribution and ecology of Necrobia violacea (Linnaeus, 1758) (Coleoptera, Cleridae) in the Iberian Peninsula and the Balearic Islands. Bol. Asoc Esp Ent 44(1–2):183–195 https://www.researchgate.net/publication/342571944_Revision_de_la_distribucion_y_ecologia_de_Necrobia_violacea_Linnaeus_1758_Coleoptera_Cleridae_en_la_peninsula_iberica_e_Islas_Baleares
Cardoso P, Barton PS, Birkhofer K, Chichorro F, Deacon C, Fartmann T, Fukushima CS, Gaigher R, Habel JC, Hallmann CA, Hillh MJ, Hochkirch A, Kwak ML, Mammola S, Noriega JA, Orfingern AB, Pedraza F, Pryke JS, Roqueq FO, Settele J, Simaika JP, Stork NE, Suhling F, Vorsterd C, Samways MJ (2020) Scientists’ warning to humanity on insect extinctions. Biol Conserv 242:108426. https://doi.org/10.1016/j.biocon.2020.108426
Carles-Tolrá M, Rodríguez PC, Verdú J (2010) Thyreophora cynophila (Panzer, 1794): collected in Spain 160 years after it was thought to be extinct (Diptera: Piophilidae: Thyreophorini). Bol Soc Ent Aragonesa 46: 1–7. http://sea-entomologia.org/PDF/001007BSEA46Thyreophorabr.pdf
Chimeno C, Morinière J, Podhorna J, Hardulak L, Husmann A, Reckel F, Grunwald JE, Penning R, Haszprunar G (2018) DNA barcoding in forensic entomology – establishing a DNA reference library of potentially forensic relevant arthropod species. JFS 64(2):593–601. https://doi.org/10.1111/1556-4029.13869
Cruise A, Watson DW, Schal C (2018) Ecological succession of adult necrophilous insects on neonate Sus scrofa domesticus in central North Carolina. PLoS One 13(4):e0195785. https://doi.org/10.1371/2Fjournal.pone.0195785
Díaz-Aranda LM, Martín-Vegal D, Baz A, Cifrián B (2018) Larval identification key to necrophagous Coleoptera of medico-legal importance in the western Palaearctic. Int J Legal Med 132(6):1795–1804. https://doi.org/10.1007/s00414-018-1909-2
Duff (2012) Checklist of beetles of the British Isles, Wells, by. Lompe (2012) Käfer Europas Nitidulidae. http://www.coleo-net.de/coleo/texte/nitidulidae.htm
Franceschetti L, Amadasi A, Bugelli V, Bolsi G, Tsokos M (2023) Estimation of late postmortem interval: where do we stand? A Literature Review Biology 12(783):2–18. https://doi.org/10.3390/biology12060783
Hallmann CA, Sorg M, Jongejans E, Siepel H, Hofland N, Schwan H, Stenmans W, Müller A, Sumser H, Hörren T, Goulson D, de Kroon H (2017) More than 75 percent decline over 27 years in total flying insect biomass in protected areas. PLoS 1–21. https://doi.org/10.1371/journal.pone.0185809
Hammel A (2018) Understanding and controlling the red legged ham beetle (Necropia Rufipes). Dissertation University of Nebraska-Lincoln Department of Entomology Online Masters Program Final Project. Lincoln, Nebraska. https://digitalcommons.unl.edu/entodistmasters/37/
Harvey M, Gasz NE, Voss S (2016) Entomology-based methods for estimation of postmortem interval. Res Rep Forensic Med Sci 6:1–9. https://doi.org/10.2147/RRFMS.S68867
Hasan MdM, Athanassiou CG, Schilling MW, Phillips TW (2020) Biology and management of the red-legged ham beetle, Necrobia rufipes DeGeer (Coleoptera: Cleridae). J Stored Prod Res 88:101635. https://doi.org/10.1016/j.jspr.2020.101635
Horenstein MB, Rosso B, García MD (2012) Seasonal structure and dynamics of sarcosaprophagous fauna on pig carrion in a rural area of Cordoba (Argentina): Their importance in forensic science. J Forensic Sci 217:146–156. https://doi.org/10.1016/j.forsciint.2011.10.043
Hu G, Wang M, Wang Y, Tang H, Chen R, Zhang Y, Zhao Y, Jin J, Wang Y, Wu M, Wang J (2020) Development of Necrobia rufipes (De Geer, 1775) (Coleoptera: Cleridae) under constant temperatures and its implication in forensic entomology. Forensic Sci Int 311:110275. https://doi.org/10.1016/j.forsciint.2020.110275
Iwasa M, Okura M (2023) Fauna of flies (Diptera) on bones and emergence succession: forensic implications in skeletal remains. Forensic Sci Int 348:111705. https://doi.org/10.1016/j.forsciint.2023.111705
Kelly JA, van der Linde TC, Anderson GS (2009) The influence of clothing and wrapping on carcass decomposition and arthropod succession during the warmer seasons in Central South Africa. J Forensic Sc 54(5):1105–1112. https://doi.org/10.1111/j.1556-4029.2009.01113.x
Keshavarzi D, Moemenbellah-Fard MD, Fereidooni M, Zarenezhad M, Fakoorziba MR (2015) New record of sap beetle, Nitidula flavomaculata Rossi (Coleoptera: Nitidulidae) on an outdoor mummified human corpse, South of Iran. J Entomol Zool Stud 3(3):396–399. https://www.entomoljournal.com/vol3Issue3/pdf/3-4-10.1.pdf
Kirinoki M, Hitotsugi M, Chigusa Y, Kurosu A, Tokudome S (2010) Larvae of the family Piophilidae found in the marrow space of skeletal remains during a forensic autopsy. Med Entomol Zool 61(2):115–119. https://doi.org/10.7601/mez.61.115
Kirinoki M, Hitotsugi M, Kato-Hayashi N., Iwasa M, Chigusa Y (2015) Discovery of Liopiophila varipes and Protopiophila contecta (Diptera: Piophilidae) from human cadavers. Forensic Sci Int e8–e12. https://doi.org/10.1016/j.forsciint.2014.12.026
Kulshrestha P, Satpathy DK (2001) Use of beetles in forensic entomology. Forensic Sci Inter 120(1–2):15–17. https://doi.org/10.1016/S0379-0738(01)00410-8
Larquier C, Azémar F, Braet Y, Weiss L, Compin A, Pelozuelo L (2022) Presence of the European bone-skipper fly Thyreophora cynophila (Panzer, 1798) (Diptera: Piophilidae) larvae in the cerebral cavity of large mammals and report of different insect larvae living inside mammal bones. Ann Soc Entomol Fr 59(1):1–8. https://doi.org/10.1080/00379271.2022.2156385
Léal X, Mourieres P, Lamarque F, Azémar F, Pelozuelo L (2020) Back from the dead II: Thyreophora cynophila (Panzer, 1798) (Diptera: Piophilidae) resurfaces in France after a 183-year-long absence. Ann Soc Ent Fr 56(1):15–18. https://doi.org/10.1080/00379271.2019.1702478
Lee S, Kirejtshuk A, Lee S (2015) Review of the genus Omosita Erichson (Coleoptera: Nitidulidae: Nitidulinae) in Korean fauna, with key to the Palaearctic species. J Asia-Pac Entomol 18(4):837–843. https://doi.org/10.1016/j.aspen.2015.10.009
Maisonhaute JE, Forbes SL (2023) Decomposition and insect succession on human cadavers in a humid, continental (Dfb) climate (Quebec, Canada). Int J Legal Med 137(2):493–509. https://doi.org/10.1007/s00414-022-02903-0
Marshall AS, Kirk-Spriggs HA, Muller SB, Paiero MS, Yau T, Jackson DM (2016) Key to Diptera families-adults 12. Man Afrotropical Diptera 2106(1):1–81
Martín-Vega D, Baz A (2011) Could the ‘vulture restaurants’ be a lifeboat for the recently rediscovered bone-skippers (Diptera: Piophilidae)? J Insect Conserv 15:747–753. https://doi.org/10.1007/s10841-011-9429-0
Martín-Vega D, Baz A, Michelsen V (2010) Back from the dead: Thyreophora cynophila (Panzer, 1798) (Diptera: Piophilidae) ‘globally extinct’ fugitive in Spain. Syst Entomol 35(4):607–613. https://doi.org/10.1111/j.1365-3113.2010.00541.x
Martín-Vega D, Baz A, Cifrián B, Gómez-Gómez A, Díaz-Aranda LM (2019) Long-term insect successional patterns on pig carcasses in central Spain. Int J Legal Med 133:1581–1592. https://doi.org/10.1007/s00414-019-02088-z
Martín-Vega D, Díaz-Aranda LM, Baz A (2014) The immature stages of the necrophagous fly Liopiophila varipes and considerations on the genus Liopiophila (Diptera: Piophilidae). Dtsch Entomol Z 61(1):37–42. https://doi.org/10.3897/dez.61.7534
Matuszewski S (2021) Post-mortem interval estimation based on insect evidence: current challenges. Insects 12(4):314. https://doi.org/10.3390/insects12040314
Menier JJ (2003) Thyreophora cynophila (Diptera Thyreophoridae) has disappeared from France ! What’s the police doing? Proceedings of the first European forensic Entomology seminar. p. 114. https://inpn.mnhn.fr/fichesEspece/OISIN_2002.pdf
Oliva A (2001) Insects of forensic significance in Argentina. Forensic Sci Int 120(1–2):145–154. https://doi.org/10.1016/S0379-0738(01)00423-6
Pastula EC, Merritt RW (2013) Insect arrival pattern and succession on buried carrion in Michigan. J Med Entomol 50(2):432–439. https://doi.org/10.1603/me12138
Pohjoismaki JL, Karhunen PJ, Goebeler S, Saukko P, Saaksjarvi IE (2010) Indoors forensic entomology: colonization of human remains in closed environments by specific species of sarcosaprophagous flies. Forensic Sci Int 199(1–3):38–42. https://doi.org/10.1016/j.forsciint.2010.02.033
Powell GS, Weidner LM (2023) A Key to the forensically important sap beetles (Coleoptera: Nitidulidae) of North America. JFE 1: https://jfe-ojs-tamu.tdl.org/jfe/article/view/7
R Core Team (2021) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. https://www.R-project.org/
Rochefort S, Giroux M, Savage J, Wheeler TA (2015) Key to forensically important Piophilidae (Diptera) in the Nearctic region. Can J Arthropod Identif 27:1–37. https://doi.org/10.3752/cjai.2015.27
Sessa F, Varotto E, Salernod M, Sine S, Bertozzia G, Galassi FM, Maglietta F, Salernoh M, Tuccia F, Pomarah C, Riccia P (2019) First report of Heleomyzidae (Diptera) recovered from the inner cavity of an intact human femur. J Forensic Leg Med 66:4–7. https://doi.org/10.1016/j.jflm.2019.05.021
Sharanowski BJ, Walker EG, Anderson GS (2008) Insect succession and decomposition patterns on shaded and sunlit carrion in Saskatchewan in three different seasons. Forensic Sci Int 179:219–240. https://doi.org/10.1016/j.forsciint.2008.05.019
Sikes DS, Raithel CJ (2002) A review of hypotheses of decline of the endangered American burying beetle (Silphidae: Nicrophorus americanus Olivier). J Insect Conserv 6:103–113. https://doi.org/10.1023/A:1020947610028
Skowronek R, Tomsia M, Szpila K, Droździok K, Kabiesza J (2015) The presence of Diptera larvae in human bones. Forensic Sci Int Genet Suppl Ser 5:235–237. https://doi.org/10.1016/j.fsigss.2015.09.094
Szpila K (2010) Key for the identification of third instars of European blowflies (Diptera: Calliphoridae) of forensic importance. In: Jens Amendt J, Goff ML, Campobasso CP, Grassberger M (eds) Current concepts in forensic entomology. Springer Science and Business Media, BV. https://doi.org/10.1007/978-1-4020-9684-6_3
Vogt GB (1950) Occurrence and records of Nitidulidae. The Coleopterist’s Bulletin 4(6):81–91. https://www.jstor.org/stable/3998495
Wagner DL, Grames EM, Forister ML, Berenbaum MR, Stopak D (2021) Insect decline in the Anthropocene: death by a thousand cuts. PNAS 118(2):e2023989118. https://doi.org/10.1073/pnas.2023989118
Wang JF, Li Z, Chen Y, Chen Q, Yin X (2008) The succession and development of insects on pig carcasses and their significances in estimating PMI in south China. Forensic Sci Int 179(179):11–18. https://doi.org/10.1016/j.forsciint.2008.04.014
Wells JD, La Motte LR (2019) Estimating the postmortem interval. In: Byrd JH, Tomberlin JK (eds) Forensic entomology, the utility of arthropods in legal investigations, 3rd edn. Boca Raton. https://doi.org/10.4324/9781351163767
Acknowledgements
We thank Jean-Pierre Mercier and Jérôme Lafitte from the “Parc National des Pyrénées,” Marine Piana from the “Commission syndicale du pays de Cize,” Lise Dauverne, and Anna Terras from the “Ligue de Protection des Oiseaux de l’Aude,” Jocelyn Fonderflinck from the “Parc National des Cévennes,” and the farmers Bruno et Valérie Molines, Dominique Rives, Sylvie Dejean, and Louis Richard for providing the necessary authorization and information to access the vulture feeding stations. We also warmly thank Yves Roullaud and Matthieu Vaslin for their kindness and their help during the visit to vulture restaurants in Aude. We are grateful to Yvan Bonneau, master’s degree student, Ingrid Forey from the “Parc Régional des Pyrénées Catalanes,” and unknown breeders from Porté-Puymorens for their help in locating isolated carcasses. We also thank Thierry Otto, who was in our laboratory, for his technical assistance with the band saw we used to open the bones and Nicolas Romet for identifying the Heteroptera specimens.
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Clément Beaumont participated in the field work and data acquisition: he opened the 185 bones and counted and identified the insects inside. He also prepared the data tables and figures and ran the statistical analysis. Flavien Cabon participated in the field work and insect identification. Corentin Larquier contributed in the field work and insect identification. Frédéric Azémar contributed in the field work and helped in producing the figures. Yves Braet provided help regarding the forensic aspects of the discussion in the revised version. Laurent Pelozuelo led the study. He obtained the vulture feeding station coordinates and the access authorizations required for bone collection. He led and participated in all field work, produced the first draft of this paper and edited the last version. All co-authors contributed to the revision of the initial draft.
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Beaumont, C., Cabon, F., Larquier, C. et al. Carrion insects living within the bones of large mammals: insect conservation and forensic entomology implications. Sci Nat 111, 30 (2024). https://doi.org/10.1007/s00114-024-01916-6
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DOI: https://doi.org/10.1007/s00114-024-01916-6