Detection of Microplastics in Commercially Important Shrimp via Chemical Digestion and Optical Microscopy

Detection of Microplastics in Commercially Important Shrimp via Chemical Digestion and Optical Microscopy | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Detection of Microplastics in Commercially Important Shrimp via Chemical Digestion and Optical Microscopy Albert Hanchuck-Pereira, Graziele Cristine Silva, Davi Araújo Fernandes, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8745214/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract In aquatic ecosystems, plastics degrade, transforming into smaller particles called microplastics (MPs), which can easily be mistaken for food and ingested by aquatic fauna, contaminating it. Many of these contaminated animals are of great commercial interest and are part of the human food chain. Thus, in an attempt to verify the presence of MPs in the tissues of commercially relevant shrimp, two species of marine shrimp, Xiphopenaeus kroyeri and Farfantepenaeus brasiliensis , collected in the South Atlantic (Ubatuba, São Paulo – Brazil), were used as a biological model. The frozen animals were underwent alkaline digestion and the residual material was analyzed using light microscopy. Particles of varying sizes and colors were found in 80% of the specimens analyzed; in both species, the particles were predominantly blue. This high rate of MPs in commercial species is quite concerning, as these particles can bioaccumulate throughout the food chain, posing a potential risk to the environment and human health. The frequent presence of high levels of contaminants in food-source organisms highlights the need for environmental monitoring and further research into the toxicological risks of indirect MPs ingestion. Plastic Alkaline digestion Crustaceans Marine pollution Shrimp Ecotoxicology. Figures Figure 1 Figure 2 Introduction Over the past few years, the complexity of the modern society has reached new levels, and, as a result, we have become increasingly dependent on technologies that facilitate and promote practicality in daily life, with plastic being one of them due to its versatility, durability, and low production cost. By the year 2060, the production of these polymers may triple, reaching approximately 1,231 million tons per year (OECD, 2022 ). According to Meijer and collaborators (2021), between 0.8 and 2.7 million metric tons of plastics are discharged annually into the oceans through the rivers, due to the improper disposal of these materials. Once in aquatic environments, these materials fragment due to heat, ultraviolet (UV) radiation, humidity, mechanical abrasion, and microbial activity (Maddison et al., 2023 ), generating particles known as microplastics (MPs), commonly defined as fragments less than 5mm in diameter. These tiny particles are present across diverse environments, ranging from aquatic systems to Mount Everest (Napper et al., 2020 ). Typically, MPs recovered from the marine environment range in size from 53µm to3 mm, while in sediments they range from 0.5mm to 2mm (Rathour et al., 2025 ). As plastic production expands and improper disposal persists globally, understanding the ecological and toxicological impacts of these particles on ecosystems and human health has become essential. One of the main pathways of MP bioaccumulation is the trophic transfer, a process in which predators ingest particles present in previously contaminated prey (Palmer & Herat, 2021 ; Li et al., 2023 ). This bioaccumulation is observed in both marine species and freshwater organisms. Even in environments considered natural nurseries for multiple species, including shrimp, the presence of MPs has been reported. In Brazil, Lima and collaborators (2016) reported micro- and macroplastics in all 12 mangrove creeks surveyed in the Goiana estuary (Pernambuco, Brazil). These mangrove creeks act as zones of plastic retention and fragmentation, increasing exposure risk for organisms at lower trophic levels and, consequently, for higher trophic levels. Considering that shrimp play a central role in the trophic chain and are consumed by humans, the detection of MPs in these organisms raises concerns not only for the environment but also for public health. Shrimp constitute one of the most important fishing resources in the southeastern region of Brazil, where fishing is mainly directed towards the seabob shrimp Xiphopenaeus kroyeri and pink shrimp Farfantepenaeus brasiliensis , these being the most consumed species, along with the white shrimp Litopenaeus schmitti (Costa et al., 2016 ). In this regard, documenting these contaminants in species within their natural environments helps clarify the spatial distribution of plastics and identify critical areas of exposure and retention. Despite the growing number of studies on MPs in aquatic organisms, reports assessing their presence in free-living decapods across different natural habitats remain scarce, particularly in tropical regions. Thus, studies like this are essential for elucidating the ecotoxicological consequences of microplastic ingestion and dispersal, not only in decapods but also across the entire trophic chain. Materials and methods Sample collection Adults of Xiphopenaeus kroyeri (n = 181; 07–10cm length and 05–12g weight) and Farfantepenaeus brasiliensis (n = 145; 15-20cm length and 09–16g weight) were caught by local fishermen, between October and December 2019 at different locations (Cedro Beach, Promirim Island and Tenorio Beach) in Ubatuba city (23°26’02’’S, 45°5’09’’W), São Paulo State, Brazil using active capture with dip nets next to the rocky shores. A shrimp-fishing equipped with 2double-rigs nets (mesh size 20mm) was used for trawling. After capture, the specimens were euthanized immediately by immersion in ice water and fixed in 10% formaldehyde in filtered seawater (pH 8.0) for 24h and stored in glass containers. Specimen collection was authorized by the Biodiversity Authorization and Information System (SISBio - nº 7019201) and it was also approved by the Ethics Committee on Animal Experimentation of the Institute of Biosciences of Botucatu –nº 805 – CEEA-IBB/UNESP as bycatch (accompanying fauna). The animals were frozen at a temperature of -20ºC, also in glass tubes, to avoid cross-contamination. From the total number of animals collected, 50 specimens of each species were randomly selected for alkaline digestion. Extraction of microplastics from the tissues The samples were macerated in a lab crucible with a porcelain mortar and washed with a saturated solution of NaCl (0.36 g/mL) and distilled water. The resulting solution was filtered through 0.6µm Whatman glass fiber filters and the filters were placed in Petri dishes containing 15% hydrogen peroxide (H 2 O 2 ) for the digestion of remaining organic matter. The Petri dishes were kept covered in an oven at 60ºC for a period of at least 24h (adapted from Avio et al., 2015 ) until the material was completely dry. After drying, the material obtained on the filter was analyzed using photonic microscopy and the images were documented by a digital image capture system (AMScope MU-1000). Polyester fibers and filaments were excluded from the analysis in order to avoid false positives from the use of animal collection nets. Results and discussion The alkaline digestion procedure allowed the observation of particles of the most varied shapes, colors and sizes (Fig. 1 ) in both X. kroyeri (Fig. 1 A-C) and F. brasiliensis (Fig. 1 D-F). Although not quantified, the detected MPs were predominantly blue (Fig. 1 C-F), consistent with reports that show a higher intake of particles with this pigmentation in relation to other colors (Ogunola et al., 2022 ). Recent studies suggest that this occurs because marine organisms, such as shrimp (Ogunola et al., 2022 ) and fish may confuse blue-MPs with their prey due to visual similarity (Ory et al., 2017 ). In the present study, the MPs were detected in 80% of the tissues of the evaluated samples (Fig. 2 ). Among the species, the rates were 92% for X. kroyeri and 68% for F. brasiliensis (Fig. 2 ). The presence of these particles in the animals analyzed indicates the ability of the MPs to disperse, including through the water column where the specimens were captured, corroborating ongoing studies that describe the presence of these contaminants in aquatic and terrestrial organisms (Rochman et al., 2019 ). An aggravating factor is that the species analyzed here are consumed in large quantities by humans, as are other seafood products. Ogunola and collaborators (2022) found MPs in five species of crustaceans sold in markets, including three species of shrimp. This consumption of contaminated food is one of the most important routes of exposure to MPs – the oral route (Schwabl et al., 2019 ), causing systemic bioaccumulation in the species. In humans, MPs have been detected in a wide variety of tissues of the cardiovascular, digestive, endocrine, lymphatic, respiratory, reproductive and urinary systems, as well as in samples of breast milk, meconium, semen, feces and urine (Roslan et al., 2024 ). Furthermore, studies conducted with economically important species have shown that seafood contains significant amounts of MPs in edible tissues, such as muscles. Shrimp, crabs and fish from the Persian Gulf region showed concentrations ranging from 0.158 to 0.360 items/g of muscle, indicating MPs’s bioaccumulation (Akhbarizadeh et al., 2019 ). Additionally, studies using probabilistic modeling have shown that MPs smaller than 10µm can be partially absorbed in the gastrointestinal tract, accumulating in tissues throughout life, which reinforces the potential for bioaccumulation in the human body (Mohamed Nor et al., 2021 ). Given this evidence, the results of this study reveal important characteristics for the use of shrimp as environmental bioindicators of the presence of MPs. Due to the behavior of these animals, which includes direct contact with contaminated sediment, wide coastal distribution, and economic importance, shrimp meet the criteria recognized in the literature for sentinel species in biomonitoring studies (Redón-Morte et al., 2025 ). Therefore, the applicability of these organisms in assessing marine pollution by MPs should be considered, given the increasing number of studies that detect such contaminants in crustaceans. Thus, future investigations combining morphological screening, spectroscopy and toxicological analyses could consolidate the use of shrimp as a bioindicator model. In summary, the results of this study confirm the presence of MPs in tropical shrimp for human consumption collected from the wild. The potential ecotoxicological and public health risk becomes evident, considering the recurring occurrence of these contaminants at multiple levels of the food chain. Although the criteria applied for the identification of MPs in this study are accepted for screening in environmental studies, it is recognized that the absence of spectroscopic analyses limits the definitive confirmation of the composition of the observed particles. However, even in the absence of spectroscopy, for screening particles > 500µm, the criteria described are recognized as reliable and are widely used in environmental monitoring studies (Hidalgo-Ruz et al., 2012 ). Given the above, the data presented in this study reinforce the need for systematic monitoring and complementary analyses, such as spectroscopy and toxicological tests, in order to elucidate the effects resulting from exposure to MPs. Conclusions This study identified the presence of MPs in most of the collected free-living shrimp specimens, highlighting a relevant warning from both an environmental and public health perspective. Considering that species such X. kroyeri and the F. brasiliensis are widely consumed by the population, the results suggest that these organisms may represent a potential route of human exposure to MPs. The recurrence of these contaminants in high concentrations in organisms of food interest reinforces the need for systematic monitoring strategies and in-depth investigation into the toxicological risks associated with the indirect ingestion of MPs. Thus, this report contributes to the understanding of the ecotoxicological impacts of MPs and highlights the urgency of integrated measures to control marine pollution. Declarations Conflict of interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Funding The authors received no specific funding for this research. Author Contribution A.H.P. implemented the methodology, validated the results and wrote the text; G.C.S. collected the animals, performed chemical digestion, and documented the results; D.A.F. standardized chemical digestion; T.S.M. wrote the final review and editing, managed the project, supervised the team, and secured the resources. All authors reviewed the manuscript. References Akhbarizadeh, R., Moore, F., & Keshavarzi, B. (2019). Investigating microplastics bioaccumulation and biomagnification in seafood from the Persian Gulf: a threat to human health?. Food Additives & Contaminants: Part A , 36 (11), 1696–1708. https://doi.org/10.1080/19440049.2019.1649473 Avio, C. G., Gorbi, S., & Regoli, F. (2015). Experimental development of a new protocol for extraction and characterization of microplastics in fish tissues: first observations in commercial species from Adriatic Sea. Marine environmental research , 111 , 18–26. https://doi.org/10.1016/j.marenvres.2015.06.014 Costa, R. C. D., Carvalho-Batista, A., Herrera, D. R., Pantaleão, J. A. F., Teodoro, S. D. S. A., & Davanso, T. M. (2016). Carcino-bycatch of the seabob shrimp fishery (Xiphopenaeus kroyeri) in Macaé, Rio de Janeiro, Brazilian Southeast. Bol. Inst. Pesca (Impr. ), 611–624. https://doi.org/10.20950/1678-2305.2016v42n3p611 Hidalgo-Ruz, V., Gutow, L., Thompson, R. C., & Thiel, M. (2012). Microplastics in the marine environment: a review of the methods used for identification and quantification. Environmental science & technology , 46 (6), 3060–3075. https://pubs.acs.org/doi/10.1021/es2031505 Li, X., Bao, L., Wei, Y., Zhao, W., Wang, F., Liu, X., … Zhang, R. (2023). Occurrence, bioaccumulation, and risk assessment of microplastics in the aquatic environment: a review. Water , 15 (9), 1768. https://doi.org/10.3390/w15091768 Lima, A. R. A., Barletta, M., Costa, M. F., Ramos, J. A. A., Dantas, D. V., Melo, P. A. M. C., … Ferreira, G. V. B. (2016). Changes in the composition of ichthyoplankton assemblage and plastic debris in mangrove creeks relative to moon phases. Journal of Fish biology , 89 (1), 619–640. https://doi.org/10.1111/jfb.12838 Maddison, C., Sathish, C. I., Lakshmi, D., Wayne, O. C., & Palanisami, T. (2023). An advanced analytical approach to assess the long-term degradation of microplastics in the marine environment. NPJ Materials Degradation , 7 (1), 59. https://doi.org/10.1038/s41529-023-00377-y Meijer, L. J., Van Emmerik, T., Van Der Ent, R., Schmidt, C., & Lebreton, L. (2021). More than 1000 rivers account for 80% of global riverine plastic emissions into the ocean. Science advances , 7 (18), eaaz5803. https://doi.org/10.1126/sciadv.aaz5803 Mohamed Nor, N. H., Kooi, M., Diepens, N. J., & Koelmans, A. A. (2021). Lifetime accumulation of microplastic in children and adults. Environmental science & technology , 55 (8), 5084–5096. https://dx.doi.org/10.1021/acs.est.0c07384 Napper, I. E., Davies, B. F., Clifford, H., Elvin, S., Koldewey, H. J., Mayewski, P. A., … Thompson, R. C. (2020). Reaching new heights in plastic pollution—preliminary findings of microplastics on Mount Everest. One Earth , 3 (5), 621–630. https://doi.org/10.1016/j.oneear.2020.10.020 OECD (2022), Global Plastics Outlook: Policy Scenarios to 2060 , OECD Publishing, Paris, https://doi.org/10.1787/aa1edf33-en . Ogunola, S. O., Reis-Santos, P., Wootton, N., & Gillanders, B. M. (2022). Microplastics in decapod crustaceans sourced from Australian seafood markets. Marine Pollution Bulletin , 179 , 113706. https://doi.org/10.1016/j.marpolbul.2022.113706 Ory, N. C., Sobral, P., Ferreira, J. L., & Thiel, M. (2017). Amberstripe scad Decapterus muroadsi (Carangidae) fish ingest blue microplastics resembling their copepod prey along the coast of Rapa Nui (Easter Island) in the South Pacific subtropical gyre. Science of the Total Environment , 586 , 430–437. https://doi.org/10.1016/j.scitotenv.2017.01.175 Palmer, J., & Herat, S. (2021). Ecotoxicity of microplastic pollutants to marine organisms: A systematic review. Water, Air, & Soil Pollution , 232 (5), 195. https://doi.org/10.1007/s11270-021-05155-7 Rathour, A., Dhar, A., & Pathania, S. (2025). Impact of microplastics pollution on human health and aquatic life: a review. Polymer International . https://doi.org/10.1002/pi.6782 Redón-Morte, M. A., Carreras-Colom, E., Chiacchio, L., Cau, A., Rodríguez-Romeu, O., & Soler-Membrives, A. (2025). Fiber entanglements as a proxy for anthropogenic pollution uptake: Monitoring deep-water rose shrimp (Parapenaeus longirostris) throughout the year. Environmental Pollution , 127337. https://doi.org/10.1016/j.envpol.2025.127337 Roslan, N. S., Lee, Y. Y., Ibrahim, Y. S., Anuar, S. T., Yusof, K. M. K. K., Lai, L. A., & Brentnall, T. (2024). Detection of microplastics in human tissues and organs: A scoping review. Journal of Global Health , 14 , 04179. https://doi.org/10.7189/jogh.14.04179 Rochman, C. M., Brookson, C., Bikker, J., Djuric, N., Earn, A., Bucci, K., … Hung, C. (2019). Rethinking microplastics as a diverse contaminant suite. Environmental toxicology and chemistry , 38 (4), 703–711. https://doi.org/10.1002/etc.4371 Schwabl, P., Köppel, S., Königshofer, P., Bucsics, T., Trauner, M., Reiberger, T., & Liebmann, B. (2019). Detection of various microplastics in human stool: a prospective case series. Annals of internal medicine , 171 (7), 453–457. https://doi.org/10.7326/M19-0618 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8745214","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":600283953,"identity":"6d0443bb-0ed0-42d5-b79a-d214a5908988","order_by":0,"name":"Albert Hanchuck-Pereira","email":"","orcid":"","institution":"Federal University of Alfenas (UNIFAL-MG)","correspondingAuthor":false,"prefix":"","firstName":"Albert","middleName":"","lastName":"Hanchuck-Pereira","suffix":""},{"id":600283954,"identity":"eb0be626-3164-46c4-b1b3-ddaae9d01f9f","order_by":1,"name":"Graziele Cristine Silva","email":"","orcid":"","institution":"São Paulo State University","correspondingAuthor":false,"prefix":"","firstName":"Graziele","middleName":"Cristine","lastName":"Silva","suffix":""},{"id":600283955,"identity":"22233057-86c0-4293-96ee-585366e43c88","order_by":2,"name":"Davi Araújo Fernandes","email":"","orcid":"","institution":"Federal University of Alfenas","correspondingAuthor":false,"prefix":"","firstName":"Davi","middleName":"Araújo","lastName":"Fernandes","suffix":""},{"id":600283956,"identity":"e2ba6419-2265-42be-9074-6f1c6d6ea7a1","order_by":3,"name":"Talita Sarah Mazzoni","email":"data:image/png;base64,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","orcid":"","institution":"Federal University of Alfenas","correspondingAuthor":true,"prefix":"","firstName":"Talita","middleName":"Sarah","lastName":"Mazzoni","suffix":""}],"badges":[],"createdAt":"2026-01-30 22:53:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8745214/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8745214/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":105394324,"identity":"e3200e49-bb1e-44e2-a686-09f69cde459e","added_by":"auto","created_at":"2026-03-25 14:05:29","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":561261,"visible":true,"origin":"","legend":"\u003cp\u003eMicroplastics found in the tissues of \u003cem\u003eXiphopenaeus kroyeri\u003c/em\u003e (A-C) and \u003cem\u003eFarfantepenaeus brasiliensis\u003c/em\u003e (D-F) after alkaline digestion. A) Microparticles (arrowhead) found in different colors from \u003cem\u003eX. kroyeri\u003c/em\u003e. B-C) Detail of A, showing microparticles of different sizes and shapes, predominantly blue. D) A) Microparticles (arrowhead) found in different colors from \u003cem\u003eF. brasiliensis.\u003c/em\u003e E-F) Detail of D, showing microparticles of different sizes and shapes, with a blue hue. Larger microplastics (arrow) Smaller microplastics (arrowhead). Bar: 80μm (A,D), 20μm (B,C,E,F).\u003c/p\u003e","description":"","filename":"OnlineFigure1.png","url":"https://assets-eu.researchsquare.com/files/rs-8745214/v1/9489daacba1def2fc6cd02aa.png"},{"id":105565835,"identity":"1c211d6f-354d-4a72-aaa7-734918f4db49","added_by":"auto","created_at":"2026-03-27 12:54:31","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":116634,"visible":true,"origin":"","legend":"\u003cp\u003ePercentage of microplastics found in the shrimp analyzed, considering both species and the species separately. ADM: Detected microparticles. UDM: Undetected microparticles.\u003c/p\u003e","description":"","filename":"OnlineFigure2.png","url":"https://assets-eu.researchsquare.com/files/rs-8745214/v1/b30cc24796b09cb0ac1d3004.png"},{"id":107220463,"identity":"08f17673-0518-4cd4-9b57-133d28754ef1","added_by":"auto","created_at":"2026-04-18 13:40:16","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":914797,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8745214/v1/bad43844-e9d5-4055-a53a-32e05d85a277.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eDetection of Microplastics in Commercially Important Shrimp via Chemical Digestion and Optical Microscopy\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eOver the past few years, the complexity of the modern society has reached new levels, and, as a result, we have become increasingly dependent on technologies that facilitate and promote practicality in daily life, with plastic being one of them due to its versatility, durability, and low production cost. By the year 2060, the production of these polymers may triple, reaching approximately 1,231\u0026nbsp;million tons per year (OECD, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). According to Meijer and collaborators (2021), between 0.8 and 2.7\u0026nbsp;million metric tons of plastics are discharged annually into the oceans through the rivers, due to the improper disposal of these materials. Once in aquatic environments, these materials fragment due to heat, ultraviolet (UV) radiation, humidity, mechanical abrasion, and microbial activity (Maddison et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), generating particles known as microplastics (MPs), commonly defined as fragments less than 5mm in diameter. These tiny particles are present across diverse environments, ranging from aquatic systems to Mount Everest (Napper et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Typically, MPs recovered from the marine environment range in size from 53\u0026micro;m to3 mm, while in sediments they range from 0.5mm to 2mm (Rathour et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). As plastic production expands and improper disposal persists globally, understanding the ecological and toxicological impacts of these particles on ecosystems and human health has become essential.\u003c/p\u003e \u003cp\u003eOne of the main pathways of MP bioaccumulation is the trophic transfer, a process in which predators ingest particles present in previously contaminated prey (Palmer \u0026amp; Herat, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Li et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). This bioaccumulation is observed in both marine species and freshwater organisms. Even in environments considered natural nurseries for multiple species, including shrimp, the presence of MPs has been reported. In Brazil, Lima and collaborators (2016) reported micro- and macroplastics in all 12 mangrove creeks surveyed in the Goiana estuary (Pernambuco, Brazil). These mangrove creeks act as zones of plastic retention and fragmentation, increasing exposure risk for organisms at lower trophic levels and, consequently, for higher trophic levels.\u003c/p\u003e \u003cp\u003eConsidering that shrimp play a central role in the trophic chain and are consumed by humans, the detection of MPs in these organisms raises concerns not only for the environment but also for public health. Shrimp constitute one of the most important fishing resources in the southeastern region of Brazil, where fishing is mainly directed towards the seabob shrimp \u003cem\u003eXiphopenaeus kroyeri\u003c/em\u003e and pink shrimp \u003cem\u003eFarfantepenaeus brasiliensis\u003c/em\u003e, these being the most consumed species, along with the white shrimp \u003cem\u003eLitopenaeus schmitti\u003c/em\u003e (Costa et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). In this regard, documenting these contaminants in species within their natural environments helps clarify the spatial distribution of plastics and identify critical areas of exposure and retention. Despite the growing number of studies on MPs in aquatic organisms, reports assessing their presence in free-living decapods across different natural habitats remain scarce, particularly in tropical regions. Thus, studies like this are essential for elucidating the ecotoxicological consequences of microplastic ingestion and dispersal, not only in decapods but also across the entire trophic chain.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cp\u003eSample collection\u003c/p\u003e \u003cp\u003eAdults of \u003cem\u003eXiphopenaeus kroyeri\u003c/em\u003e (n\u0026thinsp;=\u0026thinsp;181; 07\u0026ndash;10cm length and 05\u0026ndash;12g weight) and \u003cem\u003eFarfantepenaeus brasiliensis\u003c/em\u003e (n\u0026thinsp;=\u0026thinsp;145; 15-20cm length and 09\u0026ndash;16g weight) were caught by local fishermen, between October and December 2019 at different locations (Cedro Beach, Promirim Island and Tenorio Beach) in Ubatuba city (23\u0026deg;26\u0026rsquo;02\u0026rsquo;\u0026rsquo;S, 45\u0026deg;5\u0026rsquo;09\u0026rsquo;\u0026rsquo;W), S\u0026atilde;o Paulo State, Brazil using active capture with dip nets next to the rocky shores. A shrimp-fishing equipped with 2double-rigs nets (mesh size 20mm) was used for trawling. After capture, the specimens were euthanized immediately by immersion in ice water and fixed in 10% formaldehyde in filtered seawater (pH 8.0) for 24h and stored in glass containers. Specimen collection was authorized by the Biodiversity Authorization and Information System (SISBio - n\u0026ordm; 7019201) and it was also approved by the Ethics Committee on Animal Experimentation of the Institute of Biosciences of Botucatu \u0026ndash;n\u0026ordm; 805 \u0026ndash; CEEA-IBB/UNESP as bycatch (accompanying fauna). The animals were frozen at a temperature of -20\u0026ordm;C, also in glass tubes, to avoid cross-contamination. From the total number of animals collected, 50 specimens of each species were randomly selected for alkaline digestion.\u003c/p\u003e \u003cp\u003eExtraction of microplastics from the tissues\u003c/p\u003e \u003cp\u003eThe samples were macerated in a lab crucible with a porcelain mortar and washed with a saturated solution of NaCl (0.36 g/mL) and distilled water. The resulting solution was filtered through 0.6\u0026micro;m Whatman glass fiber filters and the filters were placed in Petri dishes containing 15% hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) for the digestion of remaining organic matter. The Petri dishes were kept covered in an oven at 60\u0026ordm;C for a period of at least 24h (adapted from Avio et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) until the material was completely dry. After drying, the material obtained on the filter was analyzed using photonic microscopy and the images were documented by a digital image capture system (AMScope MU-1000). Polyester fibers and filaments were excluded from the analysis in order to avoid false positives from the use of animal collection nets.\u003c/p\u003e"},{"header":"Results and discussion","content":"\u003cp\u003eThe alkaline digestion procedure allowed the observation of particles of the most varied shapes, colors and sizes (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) in both \u003cem\u003eX. kroyeri\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA-C) and \u003cem\u003eF. brasiliensis\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eD-F). Although not quantified, the detected MPs were predominantly blue (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC-F), consistent with reports that show a higher intake of particles with this pigmentation in relation to other colors (Ogunola et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Recent studies suggest that this occurs because marine organisms, such as shrimp (Ogunola et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) and fish may confuse blue-MPs with their prey due to visual similarity (Ory et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn the present study, the MPs were detected in 80% of the tissues of the evaluated samples (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Among the species, the rates were 92% for \u003cem\u003eX. kroyeri\u003c/em\u003e and 68% for \u003cem\u003eF. brasiliensis\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The presence of these particles in the animals analyzed indicates the ability of the MPs to disperse, including through the water column where the specimens were captured, corroborating ongoing studies that describe the presence of these contaminants in aquatic and terrestrial organisms (Rochman et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAn aggravating factor is that the species analyzed here are consumed in large quantities by humans, as are other seafood products. Ogunola and collaborators (2022) found MPs in five species of crustaceans sold in markets, including three species of shrimp. This consumption of contaminated food is one of the most important routes of exposure to MPs \u0026ndash; the oral route (Schwabl et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), causing systemic bioaccumulation in the species. In humans, MPs have been detected in a wide variety of tissues of the cardiovascular, digestive, endocrine, lymphatic, respiratory, reproductive and urinary systems, as well as in samples of breast milk, meconium, semen, feces and urine (Roslan et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eFurthermore, studies conducted with economically important species have shown that seafood contains significant amounts of MPs in edible tissues, such as muscles. Shrimp, crabs and fish from the Persian Gulf region showed concentrations ranging from 0.158 to 0.360 items/g of muscle, indicating MPs\u0026rsquo;s bioaccumulation (Akhbarizadeh et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAdditionally, studies using probabilistic modeling have shown that MPs smaller than 10\u0026micro;m can be partially absorbed in the gastrointestinal tract, accumulating in tissues throughout life, which reinforces the potential for bioaccumulation in the human body (Mohamed Nor et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eGiven this evidence, the results of this study reveal important characteristics for the use of shrimp as environmental bioindicators of the presence of MPs. Due to the behavior of these animals, which includes direct contact with contaminated sediment, wide coastal distribution, and economic importance, shrimp meet the criteria recognized in the literature for sentinel species in biomonitoring studies (Red\u0026oacute;n-Morte et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Therefore, the applicability of these organisms in assessing marine pollution by MPs should be considered, given the increasing number of studies that detect such contaminants in crustaceans. Thus, future investigations combining morphological screening, spectroscopy and toxicological analyses could consolidate the use of shrimp as a bioindicator model.\u003c/p\u003e \u003cp\u003eIn summary, the results of this study confirm the presence of MPs in tropical shrimp for human consumption collected from the wild. The potential ecotoxicological and public health risk becomes evident, considering the recurring occurrence of these contaminants at multiple levels of the food chain.\u003c/p\u003e \u003cp\u003eAlthough the criteria applied for the identification of MPs in this study are accepted for screening in environmental studies, it is recognized that the absence of spectroscopic analyses limits the definitive confirmation of the composition of the observed particles. However, even in the absence of spectroscopy, for screening particles \u0026gt;\u0026thinsp;500\u0026micro;m, the criteria described are recognized as reliable and are widely used in environmental monitoring studies (Hidalgo-Ruz et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Given the above, the data presented in this study reinforce the need for systematic monitoring and complementary analyses, such as spectroscopy and toxicological tests, in order to elucidate the effects resulting from exposure to MPs.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThis study identified the presence of MPs in most of the collected free-living shrimp specimens, highlighting a relevant warning from both an environmental and public health perspective. Considering that species such \u003cem\u003eX. kroyeri\u003c/em\u003e and the \u003cem\u003eF. brasiliensis\u003c/em\u003e are widely consumed by the population, the results suggest that these organisms may represent a potential route of human exposure to MPs.\u003c/p\u003e \u003cp\u003eThe recurrence of these contaminants in high concentrations in organisms of food interest reinforces the need for systematic monitoring strategies and in-depth investigation into the toxicological risks associated with the indirect ingestion of MPs. Thus, this report contributes to the understanding of the ecotoxicological impacts of MPs and highlights the urgency of integrated measures to control marine pollution.\u003c/p\u003e"},{"header":"Declarations","content":" \u003ch2\u003eConflict of interest\u003c/h2\u003e \u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThe authors received no specific funding for this research.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eA.H.P. implemented the methodology, validated the results and wrote the text; G.C.S. collected the animals, performed chemical digestion, and documented the results; D.A.F. standardized chemical digestion; T.S.M. wrote the final review and editing, managed the project, supervised the team, and secured the resources. 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Detection of various microplastics in human stool: a prospective case series. \u003cem\u003eAnnals of internal medicine\u003c/em\u003e, \u003cem\u003e171\u003c/em\u003e(7), 453\u0026ndash;457. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.7326/M19-0618\u003c/span\u003e\u003cspan address=\"10.7326/M19-0618\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Plastic, Alkaline digestion, Crustaceans, Marine pollution, Shrimp, Ecotoxicology.","lastPublishedDoi":"10.21203/rs.3.rs-8745214/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8745214/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn aquatic ecosystems, plastics degrade, transforming into smaller particles called microplastics (MPs), which can easily be mistaken for food and ingested by aquatic fauna, contaminating it. Many of these contaminated animals are of great commercial interest and are part of the human food chain. Thus, in an attempt to verify the presence of MPs in the tissues of commercially relevant shrimp, two species of marine shrimp, \u003cem\u003eXiphopenaeus kroyeri\u003c/em\u003e and \u003cem\u003eFarfantepenaeus brasiliensis\u003c/em\u003e, collected in the South Atlantic (Ubatuba, S\u0026atilde;o Paulo \u0026ndash; Brazil), were used as a biological model. The frozen animals were underwent alkaline digestion and the residual material was analyzed using light microscopy. Particles of varying sizes and colors were found in 80% of the specimens analyzed; in both species, the particles were predominantly blue. This high rate of MPs in commercial species is quite concerning, as these particles can bioaccumulate throughout the food chain, posing a potential risk to the environment and human health. The frequent presence of high levels of contaminants in food-source organisms highlights the need for environmental monitoring and further research into the toxicological risks of indirect MPs ingestion.\u003c/p\u003e","manuscriptTitle":"Detection of Microplastics in Commercially Important Shrimp via Chemical Digestion and Optical Microscopy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-25 14:05:24","doi":"10.21203/rs.3.rs-8745214/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"7e272a7b-4b61-4d40-a668-6bc46828b7cb","owner":[],"postedDate":"March 25th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-04-18T13:40:01+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-25 14:05:24","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8745214","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8745214","identity":"rs-8745214","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}