Effects of essential oil nanoemulsion supplementation on growth, meat quality, and digestive gland histology of farmedCornuaspersummaximum

Abstract

Aim: This study aimed to evaluate the effects of essential oil nanoemulsion-enriched diets on feed acceptability, growth performance, proximate composition, and digestive gland histology of farmedCornuaspersummaximum. Methods: A total of 2,000 juvenile snails were divided into four dietary treatments for a two-month feeding experiment. The control group received a commercial poultry feed, while the experimental groups were fed the same diet supplemented with an essential oil nanoemulsion at concentrations of 1 mL/kg (T1), 3 mL/kg (T2), and 5 mL/kg (T3). Eight samplings were conducted to measure snail diameter, weight, mortality, and feed residues. At the end of the experiment, fillet proximate composition and histopathological alterations of the digestive gland tissue were evaluated. Results: A tendency toward increased snail weight was observed across all treatments. Diet enrichment led to significantly higher fillet protein content across all treatment groups (T1, T2, and T3) compared to the control, while lipid content was highest in T1. Histopathological examination revealed enlargement of hepatic ducts in T1, apoptosis of digestive cells in T2, and necrosis of digestive and calcium cells along with thinned epithelium lining in T3. Conclusions: Dietary supplementation with essential oil nanoemulsion did not negatively affect snail growth and resulted in heavier snails with increased fillet protein content. However, histopathological evidence of toxicity rendered higher supplementation levels (T2 and T3) unsuitable. Enrichment at 1 mL/kg appears to be suitable for use in commercial snail farming.

Keywords

farmed snails / essential oils / growth / meat quality / histomorphology

References

[1] Barbosa IDS, Brito GB, Dos Santos GL, Santos LN, Teixeira LSG, Araujo RGO, et al. Multivariate data analysis of trace elements in bivalve molluscs: Characterization and food safety evaluation. Food Chem. 2019;273:64-70.

[2] Margret MS, Santhiya M, Mary MT, Jansi M. Comparative study on the biochemical compositions of four gastropods along the Kanyakumari Coast. World J Fish Mar Sci. 2013;5:637—40.

[3] Cumplido M, Marinho C, Bigatti G. Nutritional composition of Patagonian marine gastropods during reproductive seasonality. J Mar Biol Assoc U K. 2020;100:567-76.

[4] Berillis P, Hatziioannou M, Karapanagiotidis IT, Neofitou C. Morphological study of muscular tissue collagen of wild and reared Cornu aspersum (Müller, 1774). Molluscan Res. 2013;33:6-13.

[5] Staikou A, Feidantsis K, Gkanatsiou O, Bibos MN, Hatziioannou M, Storey KB, et al. Seasonal cellular stress phenomena and phenotypic plasticity in land snail Helix lucorum populations from different altitudes. J Exp Biol. 2021;224:jeb243298.

[6] Hatziioannou M, Kokkinos K. Evaluation of Sustainability Determinants of Small Farming Systems via Participatory Modeling and Fuzzy Multi—Criteria Processes: The Case Study of Heliciculture in Greece. Front Sustain. 2021;2:629408.

[7] Hatziioannou M, Theodorou A, Apostolou K, Kougiagka E, Giannouli P, Karapanagiotidis IT, et al. Grape Marc as a Functional Feed Ingredient for Farmed Snails (Cornuaspersummaximum): Effects on Production Performance, Parasitological Status, and Meat Quality . Animals. 2025;15:2680.

[8] Farfar K, Bouraiou C, Youbi A, Fetnaci I, Khebbeb ME, Djebar MR, et al. Histopathology of the digestive gland of the snail Cantareus aspersus exposed chronically to cadmium and copper fungicide. Research Square [Preprint]. 2022 [cited 2024 Dec 10]. Available from: https://doi.org/10.21203/rs.3.rs—2280356/v1

[9] Kougiagka E, Apostologamvrou C, Giannouli P, Hatziioannou M. Quality Factors of Commercial Snail Fillets as Affected by Species. Food Technol Biotechnol. 2022;60:330-7.

[10] Kougiagka E, Apostologamvrou C, Hatziioannou M, Giannouli P. Quality characteristics and microstructure of boiled snail fillet meat. J Food Process Preserv. 2022;46:e17079.

[11] Apostolou K, Staikou A, Sotiraki S, Hatziioannou M. An Assessment of Snail—Farm Systems Based on Land Use and Farm Components. Animals (Basel). 2021;11:272.

[12] Kougiagka E, Gkafas GA, Exadactylos A, Hatziioannou M. Morphology and Genetic Structure Profile of Farmed SnailsCornuaspersumaspersum andCornuaspersummaximum in Greece . Sustainability. 2022;14:15965.

[13] Apostolou K, Klaoudatos D, Staikou A, Sotiraki S, Hatziioannou M. Evaluation of production performance between two heliciculture farming systems. Molluscan Research. 2023;43:211-21.

[14] Hatziioannou M, Eleutheriadis N, Lazaridou—Dimitriadou M. Food preferences and dietary overlap by terrestrial snails in Logos area (Edessa, Macedonia, Northern Greece). J Molluscan Stud. 1994;60:331-41.

[15] Milinsk MC, das Graças Padre R, Hayashi C, de Souza NE, Matsushita M. Influence of diets enriched with different vegetable oils on the fatty acid profiles of snailHelixaspersamaxima . Food Chem. 2003;82:553—8.

[16] Milinsk MC, das Graças Padre R, Hayashi C, de Oliveira CC, Visentainer JV, de Souza NE, et al. Effects of feed protein and lipid contents on fatty acid profile of snail (Helixaspersamaxima) meat . J Food Compos Anal. 2006;19:212—6.

[17] Toader—Williams A, Bentea M. Comparative study on the adaptation and growth dynamics of the Helix pomatia and Helix aspersa Muller terrestrial snails under different feeding regimes. Sci Pap Anim Sci Biotechnol. 2010;43:133.

[18] Luis AIS, Campos EVR, de Oliveira JL, Fraceto LF. Trends in aquaculture sciences: from now to use of nanotechnology for disease control. Rev Aquacult. 2019;11:119-32.

[19] Sharifi—Rad J, Sureda A, Tenore GC, Daglia M, Sharifi—Rad M, Valussi M, et al. Biological Activities of Essential Oils: From Plant Chemoecology to Traditional Healing Systems. Molecules. 2017;22:70.

[20] Sutili FJ, Gatlin III DM, Heinzmann BM, Baldisserotto B. Plant essential oils as fish diet additives: benefits on fish health and stability in feed. Rev Aquacult. 2018;10:716—26.

[21] Laparra JM, Sanz Y. Interactions of gut microbiota with functional food components and nutraceuticals. Pharmacol Res. 2010;61:219-25.

[22] Freccia A, de Negreiros Sousa SM, Meurer F, Butzge AJ, Mewes JK, Bombardelli RA. Essential oils in the initial phase of broodstock diets of Nile tilapia. Rev Bras Zootec. 2014;43:1-7.

[23] Zhai H, Liu H, Wang S, Wu J, Kluenter AM. Potential of essential oils for poultry and pigs. Anim Nutr. 2018;4:179-86.

[24] Peng QY, Li JD, Li Z, Duan ZY, Wu YP. Effects of dietary supplementation with oregano essential oil on growth performance, carcass traits and jejunal morphology in broiler chickens. Anim Feed Sci Technol. 2016;214:148-53.

[25] Dalle Zotte A, Celia C, Szendrő Z. Herbs and spices include as feedstuff or additive in growing rabbit diets and as additive in rabbit meat: A review. Livest Sci. 2016;189:82-90.

[26] Hanczakowska Ε, Swiatkiewicz Μ. Effect of herbal extracts on piglet performance and small intestinal epithelial villi. Czech J Anim Sci. 2012;57:420—9.

[27] Ashraf A, Sabu S, Sasidharan A, Sunooj KV. Natural Feed Supplements From Crustacean Processing Side Streams for Improved Growth of Finfishes and Crustaceans: A Review. J Anim Physiol Anim Nutr (Berl). 2025;109:376-401.

[28] Kirkpinar F, Ünlü HB, Serdaroğlu M, Turp GY. Effects of dietary oregano and garlic essential oils on carcass characteristics, meat composition, colour, pH and sensory quality of broiler meat. Br Poult Sci. 2014;55:157-66.

[29] Ranucci D, Beghelli D, Trabalza—Marinucci M, Branciari R, Forte C, Olivieri O, et al. Dietary effects of a mix derived from oregano (Origanum vulgare L.) essential oil and sweet chestnut (Castanea sativa Mill.) wood extract on pig performance, oxidative status and pork quality traits. Meat Sci. 2015;100:319—26.

[30] Rivaroli DC, Guerrero A, Valero MV, Zawadzki F, Eiras CE, Campo MDM, et al. Effect of essential oils on meat and fat qualities of crossbred young bulls finished in feedlots. Meat Sci. 2016;121:278-84.

[31] Ferreira P, Soares GL, Dávila S, Bessa EC. The influence of caffeine and thymol on the survival, growth and reproduction of Subulina octona (Brugüière, 1789) (Mollusca, Subulinidae). Braz Arch Biol Technol. 2009;52:945—52.

[32] Ferreira P, Soares GL, Dávila S, Bessa EC. The influence of thymol+DMSO on survival, growth and reproduction of Bradybaena similaris (Mollusca: Bradybaenidae). Zoologia (Curitiba). 2011;28:145—50.

[33] Mc Donnell R, Yoo J, Patel K, Rios L, Hollingsworth R, Millar J, et al. Can essential oils be used as novel drench treatments for the eggs and juveniles of the pest snail Cornu aspersum in potted plants? J Pest Sci. 2016;89:549-55.

[34] Klein ML, Chastain TG, Garbacik CJ, Qian YPL, Mc Donnell RJ. Acute toxicity of essential oils to the pest slug Deroceras reticulatum in laboratory and greenhouse bioassays. J Pest Sci. 2020;93:415-25.

[35] Linhart YB, Thompson JD. Terpene—based selective herbivory by Helix aspersa (Mollusca) on Thymus vulgaris (Labiatae). Oecologia. 1995;102:126—32.

[36] Vokou D, Tziolas M, Bailey SER. Essential—oil—mediated interactions between oregano plants and Helicidae grazers. J Chem Ecol. 1998;24:1187-202.

[37] Lemjallad L, Chabir R, Kandri Rodi Y, El Ghadraoui L, Ouazzani Chahdi F, Errachidi F. Improvement of Heliciculture by Three Medicinal Plants Belonging to the Lamiaceae Family. ScientificWorldJournal.2019;2019:2630537.

[38] Association of Official Analytical Chemists International. Official methods of analysis. Maryland, USA: AOAC International; 2020.

[39] Apostologamvrou C, Hatziioannou M, Exadactylos A, Vafidis D. Reproductive biology of the commercial sea cucumberHolothuria (Roweothuria) poli, in the Central Aegean Sea, Greece . Fish Res. 2024;279:107157.

[40] El Khayari A, Rour E, Bouymajane A, Mzoudi N, Chenni Y. Use of medicinal plants to enhance the breeding and microbiological quality of the flesh in the snailCryptomphalusaspersus . Invert Reprod Dev. 2024;68:27-35.

[41] Ali SM, Mohammed TA, Mandour AM, Abd EL—Malek AR. Effect of Acacia nilotica and Cuminum cyminum extracts on Eobania vermiculata and Monacha obstructa (land snails). Egypt J Zool. 2015;64:117-30.

[42] Dias Junior PCG, dos Santos IJ, Gasparina JM, Bandoria NA, Soares LCB, da Silva AL, et al. Essential oil fromArnicamontana alters the protein metabolism in lambs fed with high—concentrate diets . Small Rumin Res. 2023;219:106894.

[43] Cheng C, Xia M, Zhang X, Wang C, Jiang S, Peng J. Supplementing Oregano Essential Oil in a Reduced—Protein Diet Improves Growth Performance and Nutrient Digestibility by Modulating Intestinal Bacteria, Intestinal Morphology, and Antioxidative Capacity of Growing—Finishing Pigs. Animals (Basel). 2018;8:159.

[44] Dennis MM, Molnár K, Kriska G, Lőw P. Mollusca: Gastropoda. Invert Histol. 2021;87-132.

[45] Delakorda SL, Letofsky—Papst I, Novak T, Giovannelli M, Hofer F, Pabst MA. Application of elemental microanalysis to elucidate the role of spherites in the digestive gland of the helicid snail Chilostoma lefeburiana. J Microsc. 2008;231:38-46.

[46] Ziegler M, Eckstein H, Köhler HR, Tisler S, Zwiener C, Triebskorn R. Effects of the antidepressants citalopram and venlafaxine on the big ramshorn snail (Planorbariuscorneus) . Water. 2021;13:1722.

[47] Abdel—Azeem HH, Osman GY, Mohamed AH. Potential toxic effects of titanium oxide (TiO₂) nanoparticles on the biological, biochemical, and histological aspects of the land snail Helix aspersa . Environ Sci Pollut Res Int. 2023;30:78127—38.

[48] Ngo PA, Neurath MF, López—Posadas R. Impact of Epithelial Cell Shedding on Intestinal Homeostasis. Int J Mol Sci. 2022;23:4160.

[49] Hamed SS, Abdelmeguied NE, Essawy AE, Radwan MA, Hegazy AE. Histological and ultrastructural changes induced by two carbamate molluscicides on the digestive gland ofEobaniavermiculata . J Biol Sci. 2007;7:1017-37.

[50] Mohammadein A, El—Shenawy NS, Al—Fahmie ZHH. Bioaccumulation and histopathological changes of the digestive gland of the land snailEobaniavermiculata (Mollusca: Gastropoda), as biomarkers of terrestrial heavy metal pollution in Taif city . It J Zool. 2013;80:345-57.