{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T01:21:03Z","timestamp":1789089663585,"version":"build-2803163510"},"reference-count":99,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T00:00:00Z","timestamp":1788480000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Curr. Issues Mol. Biol.","CIMB"],"abstract":"<jats:p>The increasing demand and limited availability of fishmeal (FM) have intensified the need for sustainable alternative protein sources in shrimp aquafeeds. This study evaluated high FM replacement in juvenile Penaeus vannamei reared under biofloc technology (BFT), integrating in vitro protein digestibility, growth performance, nutrient utilization and microbiome analyses. Six isoproteic and isolipidic diets combined two FM levels (0% and 7.5%) with animal (A), plant (P), or mixed (AP) protein sources, plus a 15% FM Control diet were assayed. Plant-based diets showed higher in vitro protein digestibility and amino acid release than animal-based diets. However, these differences were not reflected in shrimp performance, as final body weight, SGR and survival were similar among dietary treatments. Apparent feed conversion ratio, protein efficiency, and economic performance were improved in shrimp fed the A7.5 and AP7.5 diets, whereas complete FM replacement resulted in poorer performance. Biofloc and shrimp intestinal microbial community composition was primarily driven by temporal succession, whereas dietary protein source modulated the abundance of specific bacterial families. Biofloc and intestinal communities shared limited taxonomic overlap at the family level, suggesting restricted microbial exchange together with strong habitat and host mediated selection. Overall, these findings indicate that a diet containing 7.5% fishmeal (FM), supplemented with either animal-derived proteins or a combination of animal- and plant-derived protein sources, represents an optimal strategy for juvenile Penaeus vannamei cultured under BFT conditions, maintaining performance while preserving intestinal microbial stability and health.<\/jats:p>","DOI":"10.3390\/cimb48090904","type":"journal-article","created":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T15:09:29Z","timestamp":1788534569000},"page":"904","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Combined Animal and Plant Protein Enable High Fishmeal Replacement in Penaeus vannamei Under Biofloc Technology"],"prefix":"10.3390","volume":"48","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0543-3791","authenticated-orcid":false,"given":"S.","family":"Ferrando-Juan","sequence":"first","affiliation":[{"name":"Aquaculture and Biodiversity Research Group, Institute of Science and Animal Technology (ICTA), Universitat Polit\u00e8cnica de Val\u00e8ncia, Cam\u00ed de Vera, s\/n, 46022 Val\u00e8ncia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"software","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6029-3480","authenticated-orcid":false,"given":"J.","family":"G\u00f3mez-Aguilera","sequence":"additional","affiliation":[{"name":"Aquaculture and Biodiversity Research Group, Institute of Science and Animal Technology (ICTA), Universitat Polit\u00e8cnica de Val\u00e8ncia, Cam\u00ed de Vera, s\/n, 46022 Val\u00e8ncia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"A.","family":"Tom\u00e1s-Vidal","sequence":"additional","affiliation":[{"name":"Aquaculture and Biodiversity Research Group, Institute of Science and Animal Technology (ICTA), Universitat Polit\u00e8cnica de Val\u00e8ncia, Cam\u00ed de Vera, s\/n, 46022 Val\u00e8ncia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6767-5676","authenticated-orcid":false,"given":"M.","family":"Rodilla","sequence":"additional","affiliation":[{"name":"Research Institute for Integrated Management of Coastal Areas, Universitat Polit\u00e8cnica de Val\u00e8ncia, Carretera de Nazaret-Oliva, s\/n, Grao de Gand\u00eda, 46730 Val\u00e8ncia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]},{"given":"M.","family":"Jover-Cerd\u00e1","sequence":"additional","affiliation":[{"name":"Aquaculture and Biodiversity Research Group, Institute of Science and Animal Technology (ICTA), Universitat Polit\u00e8cnica de Val\u00e8ncia, Cam\u00ed de Vera, s\/n, 46022 Val\u00e8ncia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0754-1809","authenticated-orcid":false,"given":"F. J.","family":"Moyano","sequence":"additional","affiliation":[{"name":"Department of Biology and Geology, University of Almeria, Agrifood Campus of International Excellence CeiA3, CIAIMBITAL, Carretera Sacramento, s\/n, La Ca\u00f1ada, 04120 Almeria, Spain"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0861-976X","authenticated-orcid":false,"given":"D. S.","family":"Pe\u00f1aranda","sequence":"additional","affiliation":[{"name":"Aquaculture and Biodiversity Research Group, Institute of Science and Animal Technology (ICTA), Universitat Polit\u00e8cnica de Val\u00e8ncia, Cam\u00ed de Vera, s\/n, 46022 Val\u00e8ncia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9824-3267","authenticated-orcid":false,"given":"S.","family":"Mart\u00ednez-Llorens","sequence":"additional","affiliation":[{"name":"Aquaculture and Biodiversity Research Group, Institute of Science and Animal Technology (ICTA), Universitat Polit\u00e8cnica de Val\u00e8ncia, Cam\u00ed de Vera, s\/n, 46022 Val\u00e8ncia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.aquaculture.2014.04.003","article-title":"Practical Supplementation of Shrimp and Fish Feeds with Crystalline Amino Acids","volume":"431","author":"Nunes","year":"2014","journal-title":"Aquaculture"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1038\/s41586-021-03308-6","article-title":"A 20-Year Retrospective Review of Global Aquaculture","volume":"591","author":"Naylor","year":"2021","journal-title":"Nature"},{"key":"ref_3","first-page":"821","article-title":"The Protein Requirement of Penaid Shrimp at Various Life-Cycle Stages in Controlled Environment Systems","volume":"8","author":"Colvin","year":"1977","journal-title":"J. World Aquac. Soc."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1016\/j.aquaculture.2014.05.005","article-title":"Protein Levels for Litopenaeus Vannamei in Semi-Intensive and Biofloc Systems","volume":"432","author":"Vieira","year":"2014","journal-title":"Aquaculture"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"101140","DOI":"10.1016\/j.aqrep.2022.101140","article-title":"Larval Nutritional Stress Affects Trophic Compensation of Juvenile Caridean Shrimp Palaemon Varians","volume":"24","author":"Calado","year":"2022","journal-title":"Aquac. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/j.aquaculture.2016.11.036","article-title":"Assessing the Feasibility of Dietary Soybean Meal Replacement for Fishmeal to the Swimming Crab, Portunus Pelagicus, Juveniles","volume":"469","author":"Taher","year":"2017","journal-title":"Aquaculture"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e26573","DOI":"10.1016\/j.heliyon.2024.e26573","article-title":"Substitution of Fishmeal: Highlights of Potential Plant Protein Sources for Aquaculture Sustainability","volume":"10","author":"Hussain","year":"2024","journal-title":"Heliyon"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Malcorps, W., Kok, B., van\u2019t Land, M., Fritz, M., van Doren, D., Servin, K., van der Heijden, P., Palmer, R., Auchterlonie, N.A., and Rietkerk, M. (2019). The Sustainability Conundrum of Fishmeal Substitution by Plant Ingredients in Shrimp Feeds. Sustainability, 11.","DOI":"10.3390\/su11041212"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1016\/j.aquaculture.2007.04.084","article-title":"Replacement of Fish Meal with Poultry By-Product Meal in Practical Diets for Litopenaeus Vannamei, and Digestibility of the Tested Ingredients and Diets","volume":"272","author":"Scholz","year":"2007","journal-title":"Aquaculture"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1111\/j.1365-2109.2005.01223.x","article-title":"Replacement of Fish Meal by Meat and Bone Meal in Practical Diets for the White Shrimp Litopenaeus Vannamai (Boone)","volume":"36","author":"Tan","year":"2005","journal-title":"Aquac. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1111\/raq.12065","article-title":"Application of in Vitro Digestibility Methods in Aquaculture: Constraints and Perspectives","volume":"7","author":"Moyano","year":"2015","journal-title":"Rev. Aquac."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1387","DOI":"10.2478\/aoas-2025-0090","article-title":"Harnessing Biofloc Technology: A Sustainable Paradigm for Modern Aquaculture\u2014A Review","volume":"25","author":"Dheeran","year":"2025","journal-title":"Ann. Anim. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"101300","DOI":"10.1016\/j.aqrep.2022.101300","article-title":"Microorganisms in Biofloc Aquaculture System","volume":"26","author":"Khanjani","year":"2022","journal-title":"Aquac. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1007\/s10499-025-02414-w","article-title":"Optimization of Protein Requirements and Feed Restriction in Penaeus Vannamei (Boone, 1931) Biofloc Culture","volume":"34","author":"Rodilla","year":"2026","journal-title":"Aquac. Int."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Li, C., Ge, Z., Dai, L., and Chen, Y. (2025). Integrated Application of Biofloc Technology in Aquaculture: A Review. Water, 17.","DOI":"10.3390\/w17142107"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"735159","DOI":"10.1016\/j.aquaculture.2020.735159","article-title":"The Bacteria from Large-Sized Bioflocs Are More Associated with the Shrimp Gut Microbiota in Culture System","volume":"523","author":"Huang","year":"2020","journal-title":"Aquaculture"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Landsman, A., St-Pierre, B., Rosales-Leija, M., Brown, M., and Gibbons, W. (2019). Impact of Aquaculture Practices on Intestinal Bacterial Profiles of Pacific Whiteleg Shrimp Litopenaeus Vannamei. Microorganisms, 7.","DOI":"10.3390\/microorganisms7040093"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1080\/23308249.2018.1440530","article-title":"Gut Microbiota and Its Modulation for Healthy Farming of Pacific White Shrimp Litopenaeus Vannamei","volume":"26","author":"Li","year":"2018","journal-title":"Rev. Fish. Sci. Aquac."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2847","DOI":"10.1007\/s10499-022-00936-1","article-title":"A Review of Shrimp Aquaculture and Factors Affecting the Gut Microbiome","volume":"30","author":"Shehata","year":"2022","journal-title":"Aquac. Int."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1007\/s00248-017-1066-z","article-title":"Intestinal Microbiota of White Shrimp Penaeus Vannamei under Intensive Cultivation Conditions in Ecuador","volume":"75","author":"Gainza","year":"2018","journal-title":"Microb. Ecol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1737","DOI":"10.1111\/are.12628","article-title":"Changes in the Intestinal Bacterial Community during the Growth of White Shrimp, Litopenaeus Vannamei","volume":"47","author":"Huang","year":"2016","journal-title":"Aquac. Res."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Ferrando-Juan, S., Honrado, A., Tom\u00e1s-Vidal, A., Mart\u00ednez-Llorens, S., Rodilla, M., Jover-Cerd\u00e1, M., Gracia, J.A.B., Pe\u00f1aranda, D.S., and Calanche, J. (2026). Effects of Slaughter Methods on the Quality and Refrigerated Shelf Life of Biofloc-Cultured White Shrimp (Penaeus Vannamei). Foods, 15.","DOI":"10.3390\/foods15101695"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1007\/s10499-024-01694-y","article-title":"Potential of Scenedesmus Sp. and Aurantiochytrium Spp. as Protein and n-3 Fatty Acid Sources in Practical Diets for Juvenile Penaeus Vannamei","volume":"33","author":"Nunes","year":"2024","journal-title":"Aquac. Int."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.cbpb.2018.11.002","article-title":"Dietary Replacement of Fish-Meal Impaired Protein Synthesis and Immune Response of Juvenile Pacific White Shrimp, Litopenaeus Vannamei at Low Salinity","volume":"228","author":"Xie","year":"2019","journal-title":"Comp. Biochem. Physiol. B Biochem. Mol. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"5556293","DOI":"10.1155\/are\/5556293","article-title":"Effects of Replacing Fishmeal with Soybean Meal and Peanut Meal in Low-Protein Diets on Production Traits and Nutrient Composition of Penaeus Vannamei","volume":"2026","author":"Dai","year":"2026","journal-title":"Aquac. Res."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Leite, A., Vasconcelos, L., Teixeira, A., and Rodrigues, S.S.Q. (2026). Valorization of By-Products for Functional Ingredients in Meat and Meat Replacers: A Circular Bioeconomy Approach. Foods, 15.","DOI":"10.20944\/preprints202603.2163.v1"},{"key":"ref_27","unstructured":"Waters Corporation (2018). Amino Acid Analysis. Application Notebook, Waters Corporation."},{"key":"ref_28","unstructured":"GLOBEFISH (FAO) (2025). Quartely Fishmeal and Fish Oil Analysis\u2014August, FAO."},{"key":"ref_29","unstructured":"Cereales Montoya, S.L. (2026, January 09). Piensos Ecol\u00f3gicos EcoLUCAT. Available online: https:\/\/www.piensosecolucat.es\/fabrica-piensos-ecologicos\/."},{"key":"ref_30","unstructured":"Vicens i Batllori (2026, February 25). Available online: https:\/\/vicensbatllori.com\/es\/."},{"key":"ref_31","unstructured":"Valgra, S.A. (2026, January 09). Available online: https:\/\/www.valgrasa.com\/."},{"key":"ref_32","unstructured":"(2026, February 25). IndexMundi Soybean Oil\u2014Monthly Price (Euro per Metric Ton). Available online: https:\/\/www.indexmundi.com\/commodities\/?commodity=soybean-oil&currency=eur&months=60&utm."},{"key":"ref_33","unstructured":"(2025, November 22). Pangoo Biotech Import Dicalcium Phosphate China\u2014Understanding Pricing and Costs. Available online: https:\/\/pangoo.biz\/import-dicalcium-phosphate-china\/#understanding-pricing-and-costs."},{"key":"ref_34","unstructured":"Food com, S.A. (2026, February 25). Taurine\u2014Aminosulfonic Acid. Available online: https:\/\/foodcom.pl\/es\/products\/taurina\/."},{"key":"ref_35","unstructured":"(2025, November 22). Florida Aqua Farms Vitamin Premix. Available online: https:\/\/floridaaquafarms.com\/shop\/vitamin-premix\/."},{"key":"ref_36","unstructured":"(2025, November 22). ChemAnalyst Soya Lecithin Prices, Trends, Chart, News Index and Market Demand. Available online: https:\/\/www.chemanalyst.com\/Pricing-data\/soya-lecithin-1609."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3771","DOI":"10.1111\/are.15222","article-title":"Taurine Supplementation Enhances the Replacement Level of Fishmeal by Soybean Concentrate in Diets of Juvenile Pacific White Shrimp (Litopenaeus Vannamei Boone, 1931)","volume":"52","author":"To","year":"2021","journal-title":"Aquac. Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/0003-9861(61)90145-X","article-title":"The Preparation and Properties of Two New Chromogenic Substrates of Trypsin","volume":"95","author":"Erlanger","year":"1961","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1111\/jfbc.12147","article-title":"Purification and Characterization of Trypsin from Hepatopancreas of Pacific White Shrimp","volume":"39","author":"Senphan","year":"2015","journal-title":"J. Food Biochem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1016\/j.aquaculture.2010.05.014","article-title":"Application of an in Vitro Gastrointestinal Model to Evaluate Nitrogen and Phosphorus Bioaccessibility and Bioavailability in Fish Feed Ingredients","volume":"306","author":"Morales","year":"2010","journal-title":"Aquaculture"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1219","DOI":"10.3168\/jds.S0022-0302(83)81926-2","article-title":"Spectrophotometric Assay Using O-Phthaldialdehyde for Determination of Proteolysis in Milk and Isolated Milk Proteins","volume":"66","author":"Church","year":"1983","journal-title":"J. Dairy Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/S0044-8486(01)00649-4","article-title":"Protein Requirement for Maintenance and Maximum Weight Gain for the Pacific White Shrimp, Litopenaeus Vannamei","volume":"204","author":"Kureshy","year":"2002","journal-title":"Aquaculture"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1111\/j.1365-2109.2006.01637.x","article-title":"Soybean Meal as a Protein Source in Gilthead Sea Bream (Sparus Aurata L.) Diets: Effects on Growth and Nutrient Utilization","volume":"38","author":"Vidal","year":"2007","journal-title":"Aquac. Res."},{"key":"ref_44","unstructured":"AOAC (1990). Official Methods of Analysis, Association of Official Analytical Chemists International. [15th ed.]."},{"key":"ref_45","first-page":"441","article-title":"Report of Sub-Committee on Constants and Factors","volume":"Volume 11","author":"Blaxter","year":"1965","journal-title":"Energy Metabolism, Proceedings of the 3rd EAAP Symposium on Energy Metabolism"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"852","DOI":"10.1038\/s41587-019-0209-9","article-title":"Reproducible, Interactive, Scalable and Extensible Microbiome Data Science Using QIIME 2","volume":"37","author":"Bolyen","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1038\/nmeth.3869","article-title":"DADA2: High-Resolution Sample Inference from Illumina Amplicon Data","volume":"13","author":"Callahan","year":"2016","journal-title":"Nat. Methods"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"3059","DOI":"10.1093\/nar\/gkf436","article-title":"MAFFT: A Novel Method for Rapid Multiple Sequence Alignment Based on Fast Fourier Transform","volume":"30","author":"Katoh","year":"2002","journal-title":"Nucleic Acids Res."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Price, M.N., Dehal, P.S., and Arkin, A.P. (2010). FastTree 2\u2014Approximately Maximum-Likelihood Trees for Large Alignments. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0009490"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1186\/s40168-018-0470-z","article-title":"Optimizing Taxonomic Classification of Marker-Gene Amplicon Sequences with QIIME 2\u2019s Q2-Feature-Classifier Plugin","volume":"6","author":"Bokulich","year":"2018","journal-title":"Microbiome"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"D590","DOI":"10.1093\/nar\/gks1219","article-title":"The SILVA Ribosomal RNA Gene Database Project: Improved Data Processing and Web-Based Tools","volume":"41","author":"Quast","year":"2012","journal-title":"Nucleic Acids Res."},{"key":"ref_52","unstructured":"Ernst, F.G.M., Shetty, S.A., Borman, T., and Lathi, L. (2023). Mia: Microbiome Analysis, GitHub."},{"key":"ref_53","unstructured":"Strickland, J.D.H., and Parsons, T.R. (1972). A Practical Handbook of Seawater Analysis, Fisheries Research Board of Canada. [2nd ed.]."},{"key":"ref_54","unstructured":"Koroleff, F. (1969). Determination of Ammonia as Indophenol Blue, International Council for the Exploration of the Sea (ICES)."},{"key":"ref_55","unstructured":"Grasshof, K. (1976). Methods of Seawater Analysis, Verlag Chemie."},{"key":"ref_56","unstructured":"Bendschneider, K., and Robinson, R.J. (1952). A New Spectrophotometric Method for the Determination of Nitrite in Sea Water, University of Washington."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.marchem.2014.01.010","article-title":"Determination of Nitrate plus Nitrite in Small Volume Marine Water Samples Using Vanadium(III)Chloride as a Reduction Agent","volume":"160","author":"Schnetger","year":"2014","journal-title":"Mar. Chem."},{"key":"ref_58","unstructured":"Baumgarten, M.G.Z., Nascimento, V.M., Bastos, G.C.C., Costa, K.M.P., Oliveira, A.E.F., Silva, M.H.L., Dias, J.D., and Lacerda, L.A. (1996). Manual de An\u00e1lises Em Oceanografia: Coletas, Preserva\u00e7\u00e3o e An\u00e1lises F\u00edsico-Qu\u00edmicas e Planct\u00f4nicas, FURG\u2014Funda\u00e7\u00e3o Universidade Federal do Rio Grande."},{"key":"ref_59","unstructured":"Van Wyk, P., Davis-Hodgkins, M., Laramore, R., Main, K.L., Mountain, J., and Scarpa, J. (1999). Farming Marine Shrimp in Recirculating Freshwater Systems, Harbor Branch Oceanographic Institution."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/j.tifs.2005.03.010","article-title":"Integrating the Issues of Folate Bioavailability, Intake and Metabolism in the Era of Fortification","volume":"16","author":"Gregory","year":"2005","journal-title":"Trends Food Sci. Technol."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1111\/anu.12629","article-title":"Apparent Digestibility of Animal, Plant and Microbial Ingredients for Pacific White Shrimp Litopenaeus Vannamei","volume":"24","author":"Qiu","year":"2018","journal-title":"Aquac. Nutr."},{"key":"ref_62","first-page":"e20210177","article-title":"Apparent Digestibility of Protein and Essential Aminoacids from Commonly Used Feed Ingredients in Brazil for Juvenile Shrimp Litopenaeus Vannamei","volume":"51","author":"Pinto","year":"2022","journal-title":"Braz. J. Anim. Sci."},{"key":"ref_63","first-page":"TRJFAS21999","article-title":"Use of Enzyme Mixtures in Diets Based on Animal and Plant Ingredients for Litopenaeus Vannamei: Effect on Digestibility, Growth, and Enzyme Activity","volume":"23","year":"2023","journal-title":"Turk. J. Fish. Aquat. Sci."},{"key":"ref_64","first-page":"119","article-title":"Effect of Hemoglobin Powder Substituted for Fishmeal on Growth Performance, Protein Digestibility, and Trypsin Gene Expression in Litopenaeus Vannamei","volume":"32","author":"Chookird","year":"2010","journal-title":"Songklanakarin J. Sci. Technol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1080\/15222055.2012.716019","article-title":"Apparent Digestibility of 12 Protein-Origin Ingredients for Pacific White Shrimp Litopenaeus Vannamei","volume":"75","author":"Liu","year":"2013","journal-title":"N. Am. J. Aquac."},{"key":"ref_66","first-page":"101675","article-title":"Application of Biofloc Technology in Shrimp Aquaculture: A Review on Current Practices, Challenges, and Future Perspectives","volume":"19","author":"Iber","year":"2025","journal-title":"J. Agric. Food Res."},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Sun, Y., Zhang, S., Feng, W., Zhang, Y., Han, T., and Wang, J. (2025). Impact of Dietary Carbohydrate Levels on Growth Performance, Feed Efficiency, and Immune Response in Litopenaeus Vannamei Cultured in Biofloc Systems. Metabolites, 15.","DOI":"10.3390\/metabo15060409"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.aquaculture.2013.09.051","article-title":"Effect of Dietary Supplementation of Biofloc on Growth Performance and Digestive Enzyme Activities in Penaeus Monodon","volume":"418\u2013419","author":"Anand","year":"2014","journal-title":"Aquaculture"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"740278","DOI":"10.1016\/j.aquaculture.2023.740278","article-title":"Broodstock and Seed Production in Biofloc Technology (BFT): An Updated Review Focused on Fish and Penaeid Shrimp","volume":"579","author":"Khanjani","year":"2024","journal-title":"Aquaculture"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"3107","DOI":"10.1242\/jeb.205.20.3107","article-title":"Protein Turnover, Amino Acid Profile and Amino Acid Flux in Juvenile Shrimp Litopenaeus Vannamei: Effects of Dietary Protein Source","volume":"205","author":"Mente","year":"2002","journal-title":"J. Exp. Biol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"115378","DOI":"10.1016\/j.anifeedsci.2022.115378","article-title":"Dietary Lysine Requirement of Juvenile Pacific White Shrimp, Litopenaeus Vannamei (Boone, 1931) Reared in Inland Saline Water of 10 Ppt Salinity","volume":"291","author":"Maiti","year":"2022","journal-title":"Anim. Feed Sci. Technol."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"928","DOI":"10.1590\/S0100-204X2011000800020","article-title":"Substitution of Fish Meal in Diets for Marine Shrimp Grown in a Biofloc System","volume":"46","author":"Scopel","year":"2011","journal-title":"Pesqui. Agropecu. Bras."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/S0044-8486(99)00354-3","article-title":"Replacement of Fish Meal in Practical Diets for the Pacific White Shrimp, Litopenaeus Vannamei","volume":"185","author":"Arnold","year":"2000","journal-title":"Aquaculture"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1111\/raq.12843","article-title":"Retrospect of Fish Meal Substitution in Pacific White Shrimp (Litopenaeus Vannamei) Feed: Alternatives, Limitations and Future Prospects","volume":"16","author":"Chen","year":"2024","journal-title":"Rev. Aquac."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1111\/raq.12312","article-title":"Innovative Protein Sources in Shrimp (Litopenaeus Vannamei) Feeding","volume":"12","author":"Renteria","year":"2020","journal-title":"Rev. Aquac."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"100914","DOI":"10.1016\/j.aqrep.2021.100914","article-title":"Evaluation of Composite Mixture of Protein Sources in Replacing Fishmeal for Pacific White Shrimp (Litopenaeus Vannamei): Based on the Changing Pattern of Growth Performance, Nutrient Metabolism and Health Status","volume":"21","author":"Yang","year":"2021","journal-title":"Aquac. Rep."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"1087","DOI":"10.2478\/aoas-2025-0006","article-title":"Physiological, Immunological and Nutritional Assessment of Penaeus Vannamei Fed with Different Combinations of Novel Feed Ingredients","volume":"25","author":"Rajalakshmi","year":"2025","journal-title":"Ann. Anim. Sci."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1111\/anu.12838","article-title":"Partial Replacement of Fishmeal by Fermented Soybean Meal in Diets for Juvenile White Shrimp (Litopenaeus Vannamei)","volume":"25","author":"Shao","year":"2019","journal-title":"Aquac. Nutr."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"734630","DOI":"10.1016\/j.aquaculture.2019.734630","article-title":"Effects of Replacing Fishmeal with Dietary Soybean Protein Concentrate (SPC) on Growth, Serum Biochemical Indices, and Antioxidative Functions for Juvenile Shrimp Litopenaeus Vannamei","volume":"516","author":"Ray","year":"2020","journal-title":"Aquaculture"},{"key":"ref_80","doi-asserted-by":"crossref","unstructured":"Lin, H., He, S., Tan, B., Zhang, X., Lin, Y., and Yang, Q. (2022). Effect of Rice Protein Meal Replacement of Fish Meal on Growth, Anti-Oxidation Capacity, and Non-Specific Immunity for Juvenile Shrimp Litopenaeus Vannamei. Animals, 12.","DOI":"10.3390\/ani12243579"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1016\/j.foodres.2017.09.017","article-title":"Effects of Dietary Replacement of Fishmeal by Mealworm Meal on Muscle Quality of Farmed Shrimp Litopenaeus Vannamei","volume":"102","author":"Panini","year":"2017","journal-title":"Food Res. Int."},{"key":"ref_82","doi-asserted-by":"crossref","unstructured":"Mente, E. (2006). Protein Nutrition in Crustaceans. CABI Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources, CABI.","DOI":"10.1079\/PAVSNNR20061043"},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"100294","DOI":"10.1016\/j.aqrep.2020.100294","article-title":"Comparing the Assimilation of Dietary Nitrogen Supplied by Animal-, Plant- and Microbial-Derived Ingredients in Pacific White Shrimp Litopenaeus Vannamei: A Stable Isotope Study","volume":"17","year":"2020","journal-title":"Aquac. Rep."},{"key":"ref_84","doi-asserted-by":"crossref","unstructured":"Kim, S.K., Song, J., Rajeev, M., Kim, S.K., Kang, I., Jang, I.K., and Cho, J.C. (2022). Exploring Bacterioplankton Communities and Their Temporal Dynamics in the Rearing Water of a Biofloc-Based Shrimp (Litopenaeus Vannamei) Aquaculture System. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.995699"},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"985","DOI":"10.1007\/s12275-024-00187-0","article-title":"Rhodobacteraceae Are Prevalent and Ecologically Crucial Bacterial Members in Marine Biofloc Aquaculture","volume":"62","author":"Rajeev","year":"2024","journal-title":"J. Microbiol."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1016\/j.aquaculture.2019.01.026","article-title":"Strain-Specific Changes in the Gut Microbiota Profiles of the White Shrimp Litopenaeus Vannamei in Response to Cold Stress","volume":"503","author":"Liu","year":"2019","journal-title":"Aquaculture"},{"key":"ref_87","first-page":"e02971","article-title":"Profiling the Gut Microbiome of Pacific Whiteleg Shrimp, Litopeneaus Vannamei Raised in an Indoor Biofloc Aquaculture System","volume":"30","author":"Akange","year":"2025","journal-title":"Sci. Afr."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"131584","DOI":"10.1016\/j.biortech.2024.131584","article-title":"Microbiota Succession, Species Interactions, and Metabolic Functions during Autotrophic Biofloc Formation in Zero-Water-Exchange Shrimp Farming without Organic Carbon Supplements","volume":"414","author":"Li","year":"2024","journal-title":"Bioresour. Technol."},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"e0078224","DOI":"10.1128\/msystems.00782-24","article-title":"Genome-Centric Metagenomics Provides Insights into the Core Microbial Community and Functional Profiles of Biofloc Aquaculture","volume":"9","author":"Rajeev","year":"2024","journal-title":"mSystems"},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"126050","DOI":"10.1016\/j.syapm.2019.126050","article-title":"Lacipirellula Parvula Gen. Nov., Sp. Nov., Representing a Lineage of Planctomycetes Widespread in Low-Oxygen Habitats, Description of the Family Lacipirellulaceae Fam. Nov. and Proposal of the Orders Pirellulales Ord. Nov., Gemmatales ord. nov. and Isosphaerales ord. nov","volume":"43","author":"Dedysh","year":"2020","journal-title":"Syst. Appl. Microbiol."},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"362","DOI":"10.1007\/s00253-024-13213-3","article-title":"The Intestine Microbiota of Shrimp and Its Impact on Cultivation","volume":"108","author":"Zeng","year":"2024","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1186\/s40168-020-00879-w","article-title":"Fine-Scale Succession Patterns and Assembly Mechanisms of Bacterial Community of Litopenaeus Vannamei Larvae across the Developmental Cycle","volume":"8","author":"Wang","year":"2020","journal-title":"Microbiome"},{"key":"ref_93","doi-asserted-by":"crossref","unstructured":"Galaviz-Silva, L., Rodr\u00edguez de la Fuente, A.O., Gomez-Flores, R., Ibarra-G\u00e1mez, J.C., Luna-Cruz, I.E., Elizondo-Luevano, J.H., S\u00e1nchez-D\u00edaz, R., and Molina Garza, Z.J. (2025). Characterization of Microbiome Diversity in the Digestive Tract of Penaeus Vannamei Fed with Probiotics and Challenged with Vibrio Parahaemolyticus Acute Hepatopancreatic Necrosis Disease. Pathogens, 14.","DOI":"10.3390\/pathogens14040320"},{"key":"ref_94","first-page":"e371","article-title":"Effect of Protein Source and Probiotic on the Intestinal Tract of Pacific White Shrimp Litopenaeus Vannamei","volume":"44","author":"Moraes","year":"2018","journal-title":"Bol. Inst. Pesca"},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1111\/anu.12900","article-title":"Litopenaeus Vannamei Fed Diets with Different Replacement Levels of Fish Meal by Fish Silage: A Molecular Approach on Intestinal Microbiota","volume":"25","author":"Shao","year":"2019","journal-title":"Aquac. Nutr."},{"key":"ref_96","doi-asserted-by":"crossref","unstructured":"Banu, H., Swain, H.S., Das, P.C., Velmani, V., and Kumari, R. (2024). Comparative Microbial Community Occurrence Pattern, Growth Attributes, and Digestive Enzyme Indices of Puntius Gonionotus (Bleeker, 1850), Pangasianodon Hypophthalmus (Sauvage, 1878) and Heteropneustus Fossilis (Bloch, 1794) under Freshwater Biofloc Based Polyculture System. BMC Microbiol., 24.","DOI":"10.1186\/s12866-024-03473-4"},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"103329","DOI":"10.1016\/j.aqrep.2025.103329","article-title":"Biofloc-Based Bacterial Community Forms an Ecological Barrier for Resisting Vibrio Parahaemolyticus Infection in Shrimp","volume":"46","author":"Wang","year":"2026","journal-title":"Aquac. Rep."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"737061","DOI":"10.1016\/j.aquaculture.2021.737061","article-title":"Vibrio Parahaemolyticus Alters the Community Composition and Function of Intestinal Microbiota in Pacific White Shrimp, Penaeus Vannamei","volume":"544","author":"Zhang","year":"2021","journal-title":"Aquaculture"},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"e12986","DOI":"10.1111\/raq.12986","article-title":"Shrimp Intestinal Microbiota Homeostasis: Dynamic Interplay Between the Microbiota and Host Immunity","volume":"17","author":"Zhou","year":"2025","journal-title":"Rev. Aquac."}],"container-title":["Current Issues in Molecular Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1467-3045\/48\/9\/904\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,6]],"date-time":"2026-09-06T04:24:47Z","timestamp":1788668687000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1467-3045\/48\/9\/904"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,4]]},"references-count":99,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["cimb48090904"],"URL":"https:\/\/doi.org\/10.3390\/cimb48090904","relation":{},"ISSN":["1467-3045"],"issn-type":[{"value":"1467-3045","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,4]]}}}