{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T17:19:07Z","timestamp":1788455947690,"version":"build-2803163510"},"reference-count":43,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Zhejiang Province Agriculture, Rural Areas, and Nine Directions Project","award":["2025SNJF010"],"award-info":[{"award-number":["2025SNJF010"]}]},{"name":"Zhejiang Province Agriculture, Rural Areas, and Nine Directions Project","award":["2025SNJF097"],"award-info":[{"award-number":["2025SNJF097"]}]},{"name":"Zhejiang Provincial Research Institute Special Project","award":["2026YSZX01-5"],"award-info":[{"award-number":["2026YSZX01-5"]}]},{"name":"National Freshwater Aquatic Germplasm Resource Center","award":["FGRC18537"],"award-info":[{"award-number":["FGRC18537"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Procambarus clarkii and Macrobrachium nipponense are two freshwater shrimp species used in rice-farming systems. To guide their cultivation in saline\u2013alkaline waters, two 96 h experiments were conducted on juvenilels of each species using 540 juveniles per species (initial body length: 1.51 \u00b1 0.16 cm for P. clarkii and 0.88 \u00b1 0.03 cm for M. nipponense): combined salinity\u2013pH stress (0\/15.0\/25.0\u2030 at pH 7.4\/9.0\/10.0) and NaHCO3 alkalinity stress (0\/10\/20 mmol\/L). Under salinity\u2013pH stress, survival of P. clarkii was 100%, 72.23%, and 53.33%, respectively; for M. nipponense, it was 93.33%, 61.13%, and 23.33%. Under alkalinity stress, survival of P. clarkii was 92.22%, 82.22%, and 61.11%; for M. nipponense, it was 91.11%, 65.55%, and 46.67%. In both species, superoxide dismutase, acid phosphatase, and alkaline phosphatase decreased while malondialdehyde increased with stress intensity. Metabolomics revealed species-specific reprogramming: P. clarkii upregulated protective lipids (e.g., glycerophosphoethanolamines, fatty acid esters), whereas M. nipponense showed depletion of critical osmolytes (glycerophosphocholines) and accumulation of apoptosis-related ceramides. KEGG analysis confirmed that energy metabolism, amino acid homeostasis, and ABC transporters were primary pathways affected. In conclusion, extreme stress significantly reduced survival, suppressed immunity, and triggered metabolic reprogramming in both species, with P. clarkii showing substantially higher tolerance. For cultivation, P. clarkii is preferred in moderately high saline\u2013alkaline waters, while M. nipponense requires low-alkalinity freshwater with strict monitoring. These findings support species-specific management of saline\u2013alkaline rice\u2013shrimp systems.<\/jats:p>","DOI":"10.3390\/ani16172778","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T16:21:21Z","timestamp":1788452481000},"page":"2778","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Comparative Physiological and Metabolomic Responses of Procambarus clarkii and Macrobrachium nipponense to Combined Salinity\u2013pH and NaHCO3 Alkalinity Stress"],"prefix":"10.3390","volume":"16","author":[{"given":"Meili","family":"Chi","sequence":"first","affiliation":[{"name":"Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries (Zhejiang Freshwater Fishery Environmental Monitoring Station), Ministry of Agriculture and Rural Affairs, Huzhou 313001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shun","family":"Cheng","sequence":"additional","affiliation":[{"name":"Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries (Zhejiang Freshwater Fishery Environmental Monitoring Station), Ministry of Agriculture and Rural Affairs, Huzhou 313001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenping","family":"Jiang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries (Zhejiang Freshwater Fishery Environmental Monitoring Station), Ministry of Agriculture and Rural Affairs, Huzhou 313001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junzhi","family":"Luo","sequence":"additional","affiliation":[{"name":"Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries (Zhejiang Freshwater Fishery Environmental Monitoring Station), Ministry of Agriculture and Rural Affairs, Huzhou 313001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3929-649X","authenticated-orcid":false,"given":"Shili","family":"Liu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries (Zhejiang Freshwater Fishery Environmental Monitoring Station), Ministry of Agriculture and Rural Affairs, Huzhou 313001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6795-0252","authenticated-orcid":false,"given":"Jianbo","family":"Zheng","sequence":"additional","affiliation":[{"name":"Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries (Zhejiang Freshwater Fishery Environmental Monitoring Station), Ministry of Agriculture and Rural Affairs, Huzhou 313001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chao","family":"Zhu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries (Zhejiang Freshwater Fishery Environmental Monitoring Station), Ministry of Agriculture and Rural Affairs, Huzhou 313001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miao","family":"Peng","sequence":"additional","affiliation":[{"name":"Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries (Zhejiang Freshwater Fishery Environmental Monitoring Station), Ministry of Agriculture and Rural Affairs, Huzhou 313001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fei","family":"Li","sequence":"additional","affiliation":[{"name":"Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries (Zhejiang Freshwater Fishery Environmental Monitoring Station), Ministry of Agriculture and Rural Affairs, Huzhou 313001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,3]]},"reference":[{"key":"ref_1","first-page":"64","article-title":"Distribution and industrial development of crayfish in the world","volume":"16","author":"Mu","year":"2007","journal-title":"J. Shanghai Ocean Univ."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Chen, P.C., Shih, C.H., Chu, T.J., Lee, Y.C., and Tzeng, T.D. (2017). Phylogeography and genetic structure of the oriental river prawn Macrobrachium nipponense (Crustacea: Decapoda: Palaemonidae) in East Asia. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0173490"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"35749","DOI":"10.1038\/s41598-025-20622-5","article-title":"Effect of salinity exposure on the antioxidant system of \u201cTaihu No. 3\u201d Macrobrachium nipponense","volume":"15","author":"Zhou","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Huang, Y.Q., Gao, P.C., Yu, D.D., Sun, Z., Yang, X., Lai, Q.F., and Chi, H. (2025). A comparative analysis on the biochemical composition and nutrition evaluation of crayfish (Procambarus clarkii) cultivated in saline-alkali and fresh water. Foods, 14.","DOI":"10.3390\/foods14111997"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Wang, P.P., Bakari, J.S., Han, Y.Q., Hu, H.H., Liu, Z.L., Zhang, Y.W., Chen, Z.G., Huang, C.G., Wang, M.M., and Chen, H.G. (2026). Effects of thermal stress on growth and reproduction of Procambarus clarkii and aquaculture best practices. Animals, 16.","DOI":"10.3390\/ani16030495"},{"key":"ref_6","unstructured":"Colin, J.B., Richard, J.G., and Jonathan, M.W. (2012). Extreme environments: Hypersaline, alkaline, and ion-poor waters. Fish Physiology, Elsevier."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Guo, K., Zhao, Z.G., Zhang, R., Luo, L., Wang, S.H., and Xu, W. (2025). Integrated untargeted metabolomics and physiological analysis reveal the response mechanisms of Procambarus clarkii larvae to carbonate alkalinity exposure. Comparative Biochemistry and Physiology. Toxicology & pharmacology: CBP, Elsevier.","DOI":"10.2139\/ssrn.5229041"},{"key":"ref_8","first-page":"1694","article-title":"Acute toxicity of high pH stress and its effect on enzymes activity and histological structure of gill and hepatopancreas in Procambarus clarkii","volume":"40","author":"Tao","year":"2016","journal-title":"J. Fish. China"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Yu, H., Zou, S.B., Yuan, H.W., Liu, M., Ni, M., and Yuan, J.L. (2025). Physiology and molecular response mechanisms in the gills of Macrobrachium rosenbergii under acute NaHCO3 alkaline stress. Antioxidants, 14.","DOI":"10.3390\/antiox14101266"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/j.aquaculture.2017.03.016","article-title":"Effects of alkalinity and total hardness on growth and survival of postlarvae freshwater prawns, Macrobrachium rosenbergii (De Man 1879)","volume":"473","author":"Brown","year":"2017","journal-title":"Aquaculture"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1007\/s10126-025-10522-0","article-title":"Acute alkaline stress activates glucose metabolism for energy supply and induces the immune response in the oriental river prawn, Macrobrachium nipponense","volume":"27","author":"Li","year":"2025","journal-title":"Mar. Biotechnol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Jin, S., Xu, M., Gao, X., Jiang, S., Xiong, Y., Zhang, W., Qiao, H., Wu, Y., and Fu, H. (2024). Effects of alkalinity exposure on antioxidant status, metabolic function, and immune response in the hepatopancreas of Macrobrachium nipponense. Antioxidants, 13.","DOI":"10.3390\/antiox13010129"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"820","DOI":"10.1007\/s10126-022-10151-x","article-title":"RNA-seq provides novel insights into response to acute salinity stress in oriental river prawn Macrobrachium nipponense","volume":"24","author":"Fan","year":"2022","journal-title":"Mar. Biotechnol."},{"key":"ref_14","unstructured":"Bao, L. (2000). The Effects of pH, Salinity, and Dietary Phosphorus on the Growth of Macrobrachium nipponense. [Master\u2019s Thesis, Hebei University]."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Jin, S.B., Zhou, R., Gao, X.B., Xiong, Y.W., Zhang, W.Y., Qiao, H., Wu, Y., Jiang, S.F., and Fu, H.T. (2024). Identification of the effects of alkalinity exposure on the gills of oriental river prawns, Macrobrachium nipponense. BMC Genom., 25.","DOI":"10.1186\/s12864-024-10659-7"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Jin, S.B., Zhang, Y.F., Fu, H.T., Zhang, W.Y., Qiao, H., Xiong, Y.W., and Jiang, S.F. (2025). Transcriptome profiling analysis reveals changes in the antioxidant defense system, morphology, and gene expression in the gills of Macrobrachium nipponense caused by alkalinity exposure. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26094321"},{"key":"ref_17","first-page":"419","article-title":"Effects of salinity fluctuation on the molt, growth, and energy budget of juvenile Fenneropenaeus chinensis","volume":"19","author":"Ding","year":"2008","journal-title":"Chin. J. Appl. Ecol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1007\/s10126-024-10314-y","article-title":"Temporal transcriptomic profiling reveals dynamic changes in gene expression of giant freshwater prawn upon acute saline-alkaline stresses","volume":"26","author":"Chen","year":"2024","journal-title":"Mar. Biotechnol."},{"key":"ref_19","first-page":"419","article-title":"Effect of temperature and salinity on the survival and growth of juvenile redclaw (Cherax quadricarinatus)","volume":"10","author":"Austin","year":"1995","journal-title":"Freshw. Crayfish"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1111\/jwas.12762","article-title":"Influence of salinity on survival, growth, hemolymph osmolality, gill sodium potassium ATPase activity, and sodium potassium chloride co-transporter expression in the redclaw crayfish Cherax quadricarinatus","volume":"52","author":"Rida","year":"2020","journal-title":"J. World Aquac. Soc."},{"key":"ref_21","first-page":"44","article-title":"Effects of acute salinity on antioxidation and non-specific immunity of Procambarus clarkia","volume":"10","author":"Qiao","year":"2020","journal-title":"Chin. Fish. Qual. Stand."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"741409","DOI":"10.1016\/j.aquaculture.2024.741409","article-title":"Effects of bicarbonate on antioxidant, immune, osmolytic and metabolic capabilities of Scylla paramamosain","volume":"594","author":"Che","year":"2024","journal-title":"Aquaculture"},{"key":"ref_23","first-page":"278","article-title":"Effects of immunopoiysaccharide on LSZ, ALP, ACP and POD activities of Penaeus chinensis serum","volume":"30","author":"Liu","year":"1999","journal-title":"Oceanol. Limnol. Sin."},{"key":"ref_24","first-page":"833","article-title":"Effects of high-salt stress on daily weight gain, osmoregulation and immune related enzyme activities in Litopenaeus vannamei postlarvae","volume":"43","author":"Zhao","year":"2019","journal-title":"J. Fish. China"},{"key":"ref_25","first-page":"1","article-title":"Effect of salinities on activities of antioxidant enzymes and immune-ralated enzymes of the white shrimp Litopenaeus vannamei","volume":"12","author":"Lu","year":"2019","journal-title":"Hebei Fish."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e13735","DOI":"10.1111\/obr.13735","article-title":"Fatty acid esters of hydroxy fatty acids: A potential treatment for obesity-related diseases","volume":"25","author":"Li","year":"2024","journal-title":"Obes. Rev."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1016\/j.tifs.2022.11.005","article-title":"Exploring foods as natural sources of FAHFAs\u2014A review of occurrence, extraction, analytical techniques and emerging bioactive potential","volume":"129","author":"Olajide","year":"2022","journal-title":"Trends Food Sci. Technol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1038\/nri3859","article-title":"Eicosanoid storm in infection and inflammation","volume":"15","author":"Dennis","year":"2015","journal-title":"Nat. Rev. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Heckmann, L.H., Sibly, R.M., Timmermans, M.J., and Callaghan, A. (2008). Outlining eicosanoid biosynthesis in the crustacean Daphnia. Front. Zool., 5.","DOI":"10.1186\/1742-9994-5-11"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1039\/D1NP00038A","article-title":"Structure\u2013function relationship of terpenoid glycosyltransferases from plants","volume":"39","author":"Kurze","year":"2022","journal-title":"Nat. Prod. Rep."},{"key":"ref_31","first-page":"53","article-title":"Pharmacology of inhibitor-protected beta-lactam agents","volume":"3","author":"Kryukova","year":"2020","journal-title":"Novejshie Zarubezhnye I Otechestvennye Lek. Prep. Farmakoter. Farmakodinamika Farmakokinet."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Theyagarajan, K., Sruthi, V.P., Mohanapriya, D., Thenmozhi, K., and Senthilkumar, S. (2024). Biosensor platform for testing active pharmaceutical ingredients. Health and Environmental Applications of Biosensing Technologies, Elsevier.","DOI":"10.1016\/B978-0-443-19039-1.00007-9"},{"key":"ref_33","unstructured":"Hall, L.W., and Trim, C.M. (2016). Principles of sedation, anticholinergic agents, and principles of premedication. Veterinary Anaesthesia, Elsevier. [11th ed.]."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"6977","DOI":"10.1038\/s41467-023-42464-3","article-title":"Discovery of isoflavone phytoalexins in wheat reveals an alternative route to isoflavonoid biosynthesis","volume":"14","author":"Polturak","year":"2023","journal-title":"Nat. Commun."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1007\/s13659-022-00357-w","article-title":"Beauty of the beast: Anticholinergic tropane alkaloids in therapeutics","volume":"12","author":"Kyu","year":"2022","journal-title":"Nat. Prod. Bioprospecting"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1721","DOI":"10.1039\/D2NP00030J","article-title":"Biosynthesis of pyrroloindoline-containing natural products","volume":"39","author":"Sun","year":"2022","journal-title":"Nat. Prod. Rep."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4428","DOI":"10.1038\/s41467-018-06528-z","article-title":"Efficient biosynthesis of heterodimeric C3-aryl pyrroloindoline alkaloids","volume":"9","author":"Tian","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1016\/j.lfs.2010.08.003","article-title":"Enzymes involved in osmolyte synthesis: How does oxidative stress affect osmoregulation in renal cells?","volume":"87","author":"Rosas","year":"2010","journal-title":"Life Sci."},{"key":"ref_39","first-page":"165","article-title":"Analysis of expression patterns of PcABCG5 Gene in Penaeus chinensis under saline-alkali stress","volume":"45","author":"Li","year":"2024","journal-title":"Prog. Fish. Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"106569","DOI":"10.1016\/j.aquatox.2023.106569","article-title":"Integration of transcriptomic and metabolomic reveals carbonate alkalinity stress responses in the hepatopancreas of Litopenaeus vannamei","volume":"260","author":"Duan","year":"2023","journal-title":"Aquat. Toxicol."},{"key":"ref_41","first-page":"101546","article-title":"Identification of the changes of immune response, morphology, and gene expressions in hepatopancreas of Macrobrachium nipponense under the treatment of alkalinity exposure","volume":"55","author":"Jin","year":"2025","journal-title":"Comp. Biochem. Physiol. Part D. Genom. Proteom."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Lambert, C., Schmidt, K., Karger, M., Stadler, M., Stradal, T.E.B., and Rottner, K. (2025). Cytochalasans and their impact on actin filament remodeling. Biomolecules, 15.","DOI":"10.3390\/biom15071030"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1016\/j.chempr.2020.11.010","article-title":"Biosynthetically inspired divergent syntheses of merocytochalasans","volume":"7","author":"Long","year":"2021","journal-title":"Chem"}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/17\/2778\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T16:32:04Z","timestamp":1788453124000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/17\/2778"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,3]]},"references-count":43,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ani16172778"],"URL":"https:\/\/doi.org\/10.3390\/ani16172778","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,3]]}}}