{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T09:41:39Z","timestamp":1792489299984,"version":"4.5.0"},"reference-count":32,"publisher":"American Chemical Society (ACS)","issue":"43","license":[{"start":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T00:00:00Z","timestamp":1792454400000},"content-version":"vor","delay-in-days":365,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100008396","name":"Ministeriet for F? devarer, Landbrug og Fiskeri","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/100008396","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100022592","name":"Gr?nt Udviklings- og Demonstrations Program","doi-asserted-by":"publisher","award":["34009-19-1591"],"award-info":[{"award-number":["34009-19-1591"]}],"id":[{"id":"10.13039\/501100022592","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/na","name":"Organic RDD-2 program","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,10,29]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Biorefining of grasses and legume forages is carried out commercially to recover feed proteins for monogastric animals. The biorefining produces a fiber-rich pulp byproduct with additional valorization potential. Here, we explored whether pulp products from several biorefined plants could exhibit bioactivity against animal parasites. Among the tested plants, extracts derived from red clover possessed exceptionally high bioactivity against Ascaris suum larvae. GC-MS analysis revealed the antiparasitic compounds to be phytoestrogens, especially biochanin A and to a lesser extent formononetin. Higher concentrations of biochanin A and formononetin were found in biorefined pulp compared with unfractionated plants. These results underscore the dual benefit of biorefining: producing sustainable protein feed and valorizing residues for antiparasitic applications. These findings may offer a potential alternative to synthetic anthelmintics in addressing drug resistance challenges in livestock parasite management. Moreover, they pave the way for cost-effective, ecofriendly feed supplements, promoting circular economy principles in agriculture.<\/jats:p>","DOI":"10.1021\/acs.jafc.5c05338","type":"journal-article","created":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T11:27:02Z","timestamp":1760959622000},"page":"27213-27220","source":"Crossref","is-referenced-by-count":1,"title":["Biorefining of\nRed Clover (\n                    <i>Trifolium pratense<\/i>\n                    ) Yields a Phytoestrogen-Rich Byproduct with\nPotent Antiparasitic Activity"],"prefix":"10.1021","volume":"73","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2023-777X","authenticated-orcid":true,"given":"Aslak H. C.","family":"Christiansen","sequence":"first","affiliation":[{"name":"Aalborg University , , Fredrik Bajers Vej 7H , ,","place":["Aalborg, Denmark, DK-9220"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charlotte Smith","family":"Bonde","sequence":"additional","affiliation":[{"name":"University of Copenhagen , , Dyrl\u00e6gevej 100 , ,","place":["Frederiksberg, Denmark, DK-1870"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naud","family":"Megens","sequence":"additional","affiliation":[{"name":"Aalborg University , , Fredrik Bajers Vej 7H , ,","place":["Aalborg, Denmark, DK-9220"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8231-282X","authenticated-orcid":true,"given":"Andrew R.","family":"Williams","sequence":"additional","affiliation":[{"name":"University of Copenhagen , , Dyrl\u00e6gevej 100 , ,","place":["Frederiksberg, Denmark, DK-1870"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stig M.","family":"Thamsborg","sequence":"additional","affiliation":[{"name":"University of Copenhagen , , Dyrl\u00e6gevej 100 , ,","place":["Frederiksberg, Denmark, DK-1870"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1768-0279","authenticated-orcid":true,"given":"Mette","family":"Lu\u0308beck","sequence":"additional","affiliation":[{"name":"Aalborg University , , Fredrik Bajers Vej 7H , ,","place":["Aalborg, Denmark, DK-9220"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2025,10,20]]},"reference":[{"key":"2026102005060854000_cit1","doi-asserted-by":"publisher","first-page":"s2","DOI":"10.1017\/S1751731119003100","article-title":"Review: Fifty years\nof research on rumen methanogenesis: lessons learned and future challenges\nfor mitigation","volume":"14","author":"Beauchemin","year":"2020","journal-title":"Animal"},{"key":"2026102005060854000_cit2","doi-asserted-by":"publisher","first-page":"784","DOI":"10.1038\/s41893-021-00729-z","article-title":"Massive soybean expansion\nin South America since 2000 and implications\nfor conservation","volume":"4","author":"Song","year":"2021","journal-title":"Nature Sustainability"},{"key":"2026102005060854000_cit3","doi-asserted-by":"publisher","first-page":"605","DOI":"10.1002\/bbb.2098","article-title":"Green proteins: An energy-efficient solution for increased\nself-sufficiency in protein in Europe","volume":"14","author":"Njakou Djomo","year":"2020","journal-title":"Biofuels\nBioprod. Bioref."},{"key":"2026102005060854000_cit4","doi-asserted-by":"publisher","DOI":"10.1016\/j.anifeedsci.2021.114943","article-title":"Biorefined organic grass-clover protein\nconcentrate for growing pigs: Effect on growth performance and meat\nfatty acid profile","volume":"276","author":"St\u00f8dkilde","year":"2021","journal-title":"Anim. Feed Sci. Technol."},{"key":"2026102005060854000_cit5","doi-asserted-by":"publisher","first-page":"1757","DOI":"10.1111\/jpn.13406","article-title":"Biorefined grass-clover protein composition\nand effect on organic broiler performance and meat fatty acid profile","volume":"104","author":"St\u00f8dkilde","year":"2020","journal-title":"J. Animal Physiol. Animal Nutr."},{"key":"2026102005060854000_cit6","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1016\/j.anifeedsci.2018.08.004","article-title":"Protein\nvalue and degradation characteristics of pulp fibre fractions from\nscrew pressed grass, clover, and lucerne","volume":"244","author":"Damborg","year":"2018","journal-title":"Anim.\nFeed Sci. Technol."},{"key":"2026102005060854000_cit7","doi-asserted-by":"publisher","first-page":"769","DOI":"10.1016\/j.renene.2017.03.012","article-title":"Biogas potential\nof green biomass after protein extraction in an organic biorefinery\nconcept for feed, fuel and fertilizer production","volume":"129","author":"Santamari\u0301a-Ferna\u0301ndez","year":"2018","journal-title":"Renewable Energy"},{"key":"2026102005060854000_cit8","doi-asserted-by":"publisher","DOI":"10.1016\/j.anifeedsci.2020.114605","article-title":"Production\nof leaf protein concentrates in green biorefineries\nas alternative feed for monogastric animals","volume":"268","author":"Santamari\u0301a-Ferna\u0301ndez","year":"2020","journal-title":"Animal Feed Science and Technology"},{"key":"2026102005060854000_cit9","doi-asserted-by":"publisher","first-page":"1017","DOI":"10.1016\/j.ijpara.2018.06.001","article-title":"Ubiquitous parasites drive a 33%\nincrease in methane\nyield from livestock","volume":"48","author":"Fox","year":"2018","journal-title":"Int. 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Pharm. Res."},{"key":"2026102005060854000_cit17","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1016\/j.foodchem.2016.03.084","article-title":"Influence of solvents on the composition\nof condensed tannins in grape pomace seed extracts","volume":"207","author":"Bosso","year":"2016","journal-title":"Food Chem."},{"key":"2026102005060854000_cit18","doi-asserted-by":"publisher","first-page":"426","DOI":"10.1016\/j.jfoodeng.2013.01.014","article-title":"Techniques for extraction\nof bioactive compounds from plant materials:\nA review","volume":"117","author":"Azmir","year":"2013","journal-title":"Journal of Food Engineering"},{"key":"2026102005060854000_cit19","doi-asserted-by":"publisher","DOI":"10.1017\/S0022149X22000839","article-title":"Biochanin\nA in murine Schistosoma mansoni infection: effects on inflammation,\noxidative stress and fibrosis","volume":"97","author":"Elhenawy","year":"2023","journal-title":"J. Helminthol."},{"key":"2026102005060854000_cit20","doi-asserted-by":"publisher","first-page":"115","DOI":"10.1016\/j.exppara.2018.01.013","article-title":"Impact of treatment\nwith a Protein Tyrosine Kinase\nInhibitor (Genistein) on acute and chronic experimental Schistosoma\nmansoni infection","volume":"185","author":"Sobhy","year":"2018","journal-title":"Experimental Parasitology"},{"key":"2026102005060854000_cit21","doi-asserted-by":"publisher","first-page":"172","DOI":"10.1016\/j.vetpar.2017.07.005","article-title":"Screening of a small, well-curated natural\nproduct-based library identifies two rotenoids with potent nematocidal\nactivity against Haemonchus contortus","volume":"244","author":"Dilrukshi Herath","year":"2017","journal-title":"Veterinary\nParasitology"},{"key":"2026102005060854000_cit22","doi-asserted-by":"publisher","first-page":"35","DOI":"10.3390\/agriculture6030035","article-title":"Fertility\nof Herbivores Consuming Phytoestrogen-containing Medicago and Trifolium\nSpecies","author":"Reed","year":"2016","journal-title":"Agriculture"},{"key":"2026102005060854000_cit23","doi-asserted-by":"publisher","first-page":"394","DOI":"10.1016\/S1360-1385(99)01471-5","article-title":"Flavonoids and isoflavonoids\n- a gold mine for metabolic\nengineering","volume":"4","author":"Dixon","year":"1999","journal-title":"Trends in plant science"},{"key":"2026102005060854000_cit24","doi-asserted-by":"publisher","first-page":"694","DOI":"10.3389\/fpls.2014.00694","article-title":"Forage\npolyphenol oxidase and ruminant livestock nutrition","volume":"5","author":"Lee","year":"2014","journal-title":"Front. Plant Sci."},{"key":"2026102005060854000_cit25","doi-asserted-by":"publisher","first-page":"1528","DOI":"10.1017\/S1751731110000625","article-title":"Influence\nof damaging and wilting red clover on lipid metabolism during ensiling\nand in vitro rumen incubation","volume":"4","author":"Van Ranst","year":"2010","journal-title":"Animal"},{"key":"2026102005060854000_cit26","doi-asserted-by":"publisher","first-page":"475","DOI":"10.1186\/s13071-018-3012-4","article-title":"Antiparasitic\nactivity of chicory (Cichorium intybus) and its natural\nbioactive compounds in livestock: a review","volume":"11","author":"Pen\u0303a-Espinoza","year":"2018","journal-title":"Parasites Vectors"},{"key":"2026102005060854000_cit27","doi-asserted-by":"publisher","first-page":"211","DOI":"10.1111\/j.1439-037X.2004.00100.x","article-title":"Alfalfa phytoestrogen\ncontent: Impact\nof plant maturity and herbage components","volume":"190","author":"Seguin","year":"2004","journal-title":"Journal\nof Agronomy and Crop Science."},{"key":"2026102005060854000_cit28","doi-asserted-by":"publisher","first-page":"82","DOI":"10.1007\/s11250-020-02514-0","article-title":"Response of plantain\n(Plantago lanceolata\nL.) supplementation on nutritional, endo-parasitic, and endocrine\nstatus in lambs","volume":"53","author":"Reza","year":"2021","journal-title":"Trop. Anim. Health Prod."},{"key":"2026102005060854000_cit29","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1016\/j.vetpar.2010.06.017","article-title":"In vitro effects\nof Cassava (Manihot esculenta) leaf extracts on four development stages\nof Haemonchus contortus","volume":"173","author":"Marie-Magdeleine","year":"2010","journal-title":"Veterinary Parasitology"},{"key":"2026102005060854000_cit30","doi-asserted-by":"publisher","first-page":"251","DOI":"10.1007\/s11250-008-9182-x","article-title":"The effect\nof long-term feeding of fresh and ensiled cassava (Manihot esculenta)\nfoliage on gastrointestinal nematode infections in goats","volume":"41","author":"Sokerya","year":"2009","journal-title":"Tropical animal health and production"},{"key":"2026102005060854000_cit31","doi-asserted-by":"publisher","DOI":"10.1016\/j.jclepro.2022.134730","article-title":"Production\nof leaf protein concentrates from cassava: Protein distribution and\nanti-nutritional factors in biorefining fractions","volume":"379","author":"Gundersen","year":"2022","journal-title":"J. Cleaner Prod."},{"key":"2026102005060854000_cit32","doi-asserted-by":"publisher","first-page":"471","DOI":"10.1007\/s11101-021-09779-9","article-title":"Biologically\nactive compounds from forage plants","volume":"21","author":"Tava","year":"2022","journal-title":"Phytochemistry\nReviews"}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.5c05338","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/73\/43\/27213\/68909627\/acs.jafc.5c05338.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/73\/43\/27213\/68909627\/acs.jafc.5c05338.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T09:12:10Z","timestamp":1792487530000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/73\/43\/27213\/3643068\/Biorefining-of-Red-Clover-Trifolium-pratense"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,20]]},"references-count":32,"journal-issue":{"issue":"43","published-online":{"date-parts":[[2025,10,20]]},"published-print":{"date-parts":[[2025,10,29]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.5c05338","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,20]]}}}