Efficient Extraction of Small Microplastic Particles from Rat Feed and Feces for Quantification

Overview

Journal Heliyon

Specialty Social Sciences

Date 2023 Jan 30

PMID 36711289

Authors

Benuarda Toto

Alice Refosco

Jutta Dierkes

Tanja Kogel

Affiliations

Soon will be listed here.

Abstract

To date, microplastic is ubiquitously encountered in the environment. Studies analyzing microplastic in terrestrial ecosystems, including animal feces and feed, are few. Microplastic quantification method validation and harmonization are not yet far developed. For the analysis of small microplastic, approximately <0.5 mm, extraction from organic and inorganic materials is fundamental prior to quantitative and qualitative analysis. Method validation, including recovery studies, are necessary throughout the analytical chain. In this study, we developed an optimized, efficient protocol with a duration of 72 h for the digestion of laboratory rat feed and feces. A combination of a mild acidic (HO 15%; HNO 5%) and an alkaline treatment (10% KOH) dissolving the previous filter, followed by enzymatic digestion (Viscozyme®L) proved to be efficient for the extraction and identification of spiked polyamide (15-20 μm) and polyethylene (40-48 μm) from feed and feces samples from rats, showing high recovery rates. Extracted rat feces samples from an study in which Wistar rats were fed with feed containing microplastic were analyzed with pyrolysis-gas chromatography-Orbitrap™ mass spectrometry, quantifying recovered microplastic in rat feces in environmentally relevant concentrations. The presented three-step protocol provides a suitable, time and cost-effective method to extract microplastic from rat feed and feces.

Citing Articles

Influence of selected dosages of plastic microparticles on the porcine fecal microbiome.

Galecka I, Rychlik A, Calka J Sci Rep. 2025; 15(1):1269.

PMID: 39779716 PMC: 11711237. DOI: 10.1038/s41598-024-80337-x.

Using feces to indicate plastic pollution in terrestrial vertebrate species in western Thailand.

Teampanpong J, Duengkae P PeerJ. 2024; 12:e17596.

PMID: 38948236 PMC: 11212639. DOI: 10.7717/peerj.17596.

References

Lusher A, Munno K, Hermabessiere L, Carr S . Isolation and Extraction of Microplastics from Environmental Samples: An Evaluation of Practical Approaches and Recommendations for Further Harmonization. Appl Spectrosc. 2020; 74(9):1049-1065. DOI: 10.1177/0003702820938993. View

Perez-Guevara F, Kutralam-Muniasamy G, Shruti V . Critical review on microplastics in fecal matter: Research progress, analytical methods and future outlook. Sci Total Environ. 2021; 778:146395. DOI: 10.1016/j.scitotenv.2021.146395. View

Donohue M, Masura J, Gelatt T, Ream R, Baker J, Faulhaber K . Evaluating exposure of northern fur seals, Callorhinus ursinus, to microplastic pollution through fecal analysis. Mar Pollut Bull. 2019; 138:213-221. DOI: 10.1016/j.marpolbul.2018.11.036. View

von Friesen L, Granberg M, Hassellov M, Gabrielsen G, Magnusson K . An efficient and gentle enzymatic digestion protocol for the extraction of microplastics from bivalve tissue. Mar Pollut Bull. 2019; 142:129-134. DOI: 10.1016/j.marpolbul.2019.03.016. View

Liu S, Guo J, Liu X, Yang R, Wang H, Sun Y . Detection of various microplastics in placentas, meconium, infant feces, breastmilk and infant formula: A pilot prospective study. Sci Total Environ. 2022; 854:158699. DOI: 10.1016/j.scitotenv.2022.158699. View

Leslie H, van Velzen M, Brandsma S, Vethaak A, Garcia-Vallejo J, Lamoree M . Discovery and quantification of plastic particle pollution in human blood. Environ Int. 2022; 163:107199. DOI: 10.1016/j.envint.2022.107199. View

Jin Y, Lu L, Tu W, Luo T, Fu Z . Impacts of polystyrene microplastic on the gut barrier, microbiota and metabolism of mice. Sci Total Environ. 2018; 649:308-317. DOI: 10.1016/j.scitotenv.2018.08.353. View

Toto B, Refosco A, OKeeffe M, Barkhald O, Bronstad A, Lied G . Intestinal permeability and gene expression after polyethylene and polyamide microplastic ingestion in Wistar rats. Toxicol Lett. 2022; 370:35-41. DOI: 10.1016/j.toxlet.2022.09.002. View

Provencher J, Vermaire J, Avery-Gomm S, Braune B, Mallory M . Garbage in guano? Microplastic debris found in faecal precursors of seabirds known to ingest plastics. Sci Total Environ. 2019; 644:1477-1484. DOI: 10.1016/j.scitotenv.2018.07.101. View

10.

Dehaut A, Cassone A, Frere L, Hermabessiere L, Himber C, Rinnert E . Microplastics in seafood: Benchmark protocol for their extraction and characterization. Environ Pollut. 2016; 215:223-233. DOI: 10.1016/j.envpol.2016.05.018. View

11.

Carr K, Smyth S, McCullough M, Morris J, Moyes S . Morphological aspects of interactions between microparticles and mammalian cells: intestinal uptake and onward movement. Prog Histochem Cytochem. 2012; 46(4):185-252. DOI: 10.1016/j.proghi.2011.11.001. View

12.

Ragusa A, Svelato A, Santacroce C, Catalano P, Notarstefano V, Carnevali O . Plasticenta: First evidence of microplastics in human placenta. Environ Int. 2021; 146:106274. DOI: 10.1016/j.envint.2020.106274. View

13.

Bai C, Liu L, Hu Y, Zeng E, Guo Y . Microplastics: A review of analytical methods, occurrence and characteristics in food, and potential toxicities to biota. Sci Total Environ. 2021; 806(Pt 1):150263. DOI: 10.1016/j.scitotenv.2021.150263. View

14.

Deng Y, Zhang Y, Lemos B, Ren H . Tissue accumulation of microplastics in mice and biomarker responses suggest widespread health risks of exposure. Sci Rep. 2017; 7:46687. PMC: 5402289. DOI: 10.1038/srep46687. View

15.

Catarino A, Thompson R, Sanderson W, Henry T . Development and optimization of a standard method for extraction of microplastics in mussels by enzyme digestion of soft tissues. Environ Toxicol Chem. 2016; 36(4):947-951. DOI: 10.1002/etc.3608. View

16.

Zuccarello P, Ferrante M, Cristaldi A, Copat C, Grasso A, Sangregorio D . Exposure to microplastics (<10 μm) associated to plastic bottles mineral water consumption: The first quantitative study. Water Res. 2019; 157:365-371. DOI: 10.1016/j.watres.2019.03.091. View

17.

Avio C, Gorbi S, Regoli F . Experimental development of a new protocol for extraction and characterization of microplastics in fish tissues: First observations in commercial species from Adriatic Sea. Mar Environ Res. 2015; 111:18-26. DOI: 10.1016/j.marenvres.2015.06.014. View

18.

Rose C, Parker A, Jefferson B, Cartmell E . The Characterization of Feces and Urine: A Review of the Literature to Inform Advanced Treatment Technology. Crit Rev Environ Sci Technol. 2015; 45(17):1827-1879. PMC: 4500995. DOI: 10.1080/10643389.2014.1000761. View

19.

Cole M, Webb H, Lindeque P, Fileman E, Halsband C, Galloway T . Isolation of microplastics in biota-rich seawater samples and marine organisms. Sci Rep. 2014; 4:4528. PMC: 3970126. DOI: 10.1038/srep04528. View

20.

Lwanga E, Mendoza Vega J, Ku Quej V, Chi J, Sanchez Del Cid L, Chi C . Field evidence for transfer of plastic debris along a terrestrial food chain. Sci Rep. 2017; 7(1):14071. PMC: 5658418. DOI: 10.1038/s41598-017-14588-2. View