Development of a Rainbow Trout () Intestinal Platform for Profiling Amino Acid Digestion and Absorption of a Complete Diet

Overview

Journal Animals (Basel)

Date 2023 Jul 29

PMID 37508055

Authors

Rolando Pasquariello

Radmila Pavlovic

Marcelo A Chacon

Federica Camin

Nicole Verdile

Guro Lokka

Sara Panseri

Massimo Faustini

Amos Tandler

David Peggs

Trond M Kortner

Amir Bitan

Tiziana A L Brevini

Fulvio Gandolfi

Affiliations

Soon will be listed here.

Abstract

The ever-increasing number and variation of raw materials utilized to provide alternative feed formulations continues to allow for a more sustainable and flexible approach. Testing all these options is still the most robust and reliable manner to pick the best raw material candidates, but it requires the use of large numbers of animals and is time-consuming and expensive. Therefore, we are developing an platform that can provide a reliable evaluation of new ingredients. The main aim of this work was to combine an digestion protocol of extruded, commercially relevant aquafeeds with the exposure of intestinal epithelial cells to the extracted bio-available fraction (BAF). The results show that 250,000 cells/cm represents the optimal seeding density and that up to 50% BAF concentration for up to 24 h had no negative effects on the epithelial barrier morphology and function. It is possible to determine amino acid digestibility and bioavailability in all the experimental conditions (with and without BSA, at 25% and 50% dilution) and at all time points (0, 6, and 24 h). However, BAF concentration, the medium used for its dilution, and the length of exposure to the different epithelial cell lines can all influence the results and, therefore, must be selected according to the final aim of the experiment.

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References

Borenfreund E, Puerner J . Toxicity determined in vitro by morphological alterations and neutral red absorption. Toxicol Lett. 1985; 24(2-3):119-24. DOI: 10.1016/0378-4274(85)90046-3. View

Fedi A, Vitale C, Ponschin G, Ayehunie S, Fato M, Scaglione S . In vitro models replicating the human intestinal epithelium for absorption and metabolism studies: A systematic review. J Control Release. 2021; 335:247-268. DOI: 10.1016/j.jconrel.2021.05.028. View

Brodkorb A, Egger L, Alminger M, Alvito P, Assuncao R, Ballance S . INFOGEST static in vitro simulation of gastrointestinal food digestion. Nat Protoc. 2019; 14(4):991-1014. DOI: 10.1038/s41596-018-0119-1. View

Minghetti M, Drieschner C, Bramaz N, Schug H, Schirmer K . A fish intestinal epithelial barrier model established from the rainbow trout (Oncorhynchus mykiss) cell line, RTgutGC. Cell Biol Toxicol. 2017; 33(6):539-555. PMC: 5658468. DOI: 10.1007/s10565-017-9385-x. View

Xu Y, Shrestha N, Preat V, Beloqui A . An overview of in vitro, ex vivo and in vivo models for studying the transport of drugs across intestinal barriers. Adv Drug Deliv Rev. 2021; 175:113795. DOI: 10.1016/j.addr.2021.05.005. View

Bjorgen H, Li Y, Kortner T, Krogdahl A, Koppang E . Anatomy, immunology, digestive physiology and microbiota of the salmonid intestine: Knowns and unknowns under the impact of an expanding industrialized production. Fish Shellfish Immunol. 2020; 107(Pt A):172-186. DOI: 10.1016/j.fsi.2020.09.032. View

Reshkin S, Vilella S, Cassano G, Ahearn G, Storelli C . Basolateral amino acid and glucose transport by the intestine of the teleost, Anguilla anguilla. Comp Biochem Physiol A Comp Physiol. 1988; 91(4):779-88. DOI: 10.1016/0300-9629(88)90965-6. View

Lokka G, Dhanasiri A, Krogdahl A, Kortner T . Bile components affect the functions and transcriptome of the rainbow trout intestinal epithelial cell line RTgutGC. Fish Shellfish Immunol. 2022; 131:1144-1156. DOI: 10.1016/j.fsi.2022.10.049. View

Lesmes U . In vitro digestion models for the design of safe and nutritious foods. Adv Food Nutr Res. 2023; 104:179-203. DOI: 10.1016/bs.afnr.2022.10.006. View

10.

Song C, Chai Z, Chen S, Zhang H, Zhang X, Zhou Y . Intestinal mucus components and secretion mechanisms: what we do and do not know. Exp Mol Med. 2023; 55(4):681-691. PMC: 10167328. DOI: 10.1038/s12276-023-00960-y. View

11.

Menard O, Lesmes U, Shani-Levi C, Araiza Calahorra A, Lavoisier A, Morzel M . Static digestion model adapted to the general older adult population: an INFOGEST international consensus. Food Funct. 2023; 14(10):4569-4582. DOI: 10.1039/d3fo00535f. View

12.

Francis G . Albumin and mammalian cell culture: implications for biotechnology applications. Cytotechnology. 2010; 62(1):1-16. PMC: 2860567. DOI: 10.1007/s10616-010-9263-3. View

13.

OBrien J, Wilson I, Orton T, Pognan F . Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem. 2000; 267(17):5421-6. DOI: 10.1046/j.1432-1327.2000.01606.x. View

14.

Haddad M, Sztupecki W, Delayre-Orthez C, Rhazi L, Barbezier N, Depeint F . Complexification of In Vitro Models of Intestinal Barriers, A True Challenge for a More Accurate Alternative Approach. Int J Mol Sci. 2023; 24(4). PMC: 9958734. DOI: 10.3390/ijms24043595. View

15.

Schirmer K, Chan A, Greenberg B, Dixon D, Bols N . Methodology for demonstrating and measuring the photocytotoxicity of fluoranthene to fish cells in culture. Toxicol In Vitro. 2010; 11(1-2):107-19. DOI: 10.1016/s0887-2333(97)00002-7. View

16.

Pasquariello R, Verdile N, Pavlovic R, Panseri S, Schirmer K, Brevini T . New Stable Cell Lines Derived from the Proximal and Distal Intestine of Rainbow Trout () Retain Several Properties Observed In Vivo. Cells. 2021; 10(6). PMC: 8235179. DOI: 10.3390/cells10061555. View

17.

Drieschner C, Minghetti M, Wu S, Renaud P, Schirmer K . Ultrathin Alumina Membranes as Scaffold for Epithelial Cell Culture from the Intestine of Rainbow Trout. ACS Appl Mater Interfaces. 2017; 9(11):9496-9505. DOI: 10.1021/acsami.7b00705. View

18.

Terova G, Robaina L, Izquierdo M, Cattaneo A, Molinari S, Bernardini G . PepT1 mRNA expression levels in sea bream (Sparus aurata) fed different plant protein sources. Springerplus. 2013; 2(1):17. PMC: 3579422. DOI: 10.1186/2193-1801-2-17. View

19.

Daniel H . Molecular and integrative physiology of intestinal peptide transport. Annu Rev Physiol. 2004; 66:361-84. DOI: 10.1146/annurev.physiol.66.032102.144149. View

20.

Bohn T, Carriere F, Day L, Deglaire A, Egger L, Freitas D . Correlation between in vitro and in vivo data on food digestion. What can we predict with static in vitro digestion models?. Crit Rev Food Sci Nutr. 2017; 58(13):2239-2261. DOI: 10.1080/10408398.2017.1315362. View