{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T05:01:23Z","timestamp":1789016483496,"version":"build-2803163510"},"reference-count":57,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2022,6,30]],"date-time":"2022-06-30T00:00:00Z","timestamp":1656547200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31802268"],"award-info":[{"award-number":["31802268"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["SWU117048"],"award-info":[{"award-number":["SWU117048"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012226","name":"Fundamental Research Funds for the Central Universities","doi-asserted-by":"publisher","award":["31802268"],"award-info":[{"award-number":["31802268"]}],"id":[{"id":"10.13039\/501100012226","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012226","name":"Fundamental Research Funds for the Central Universities","doi-asserted-by":"publisher","award":["SWU117048"],"award-info":[{"award-number":["SWU117048"]}],"id":[{"id":"10.13039\/501100012226","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Rice field eel (Monopterus albus), a protogynous hermaphrodite fish, is a good model for the research of sex determination and gonadal differentiation in teleosts. In this study, we cloned the full-length cDNA sequence of trh, which encoded a predicted protein with 270 amino acids. Trh mainly expressed in the brain, followed by the ovary, testis, muscle and pituitary, and had low levels in other peripheral tissues. During natural sex reversal, trh mRNA expression levels exhibited a significant increase at the late intersexual stage in the hypothalamus. In the gonad, trh mRNA expression levels showed a trend of increase followed by decrease, and only increased significantly at the middle intersexual stage. No matter static incubation or intraperitoneal (IP) injection, TRH had no significant effect on trh and thyroid-stimulating hormone \u03b2subunit (tsh\u03b2) mRNA expression levels, and serum T3, T4 and TRH release. After static incubation of ovarian fragments by TRH, the expression of gonadal soma derived factor (gsdf) was up-regulated significantly at both the doses of 10 and 100 nM. IP injection of TRH stimulated the expression of gsdf, and inhibited the expression of ovarian aromatase gene (cyp19a1a), accompanied by the increase of serum 11-KT levels. The results indicated that TRH may play a novel role in gonadal differentiation by the regulation of gonadal differentiation-related gene expression and sex steroid hormone secretion in rice field eel.<\/jats:p>","DOI":"10.3390\/ani12131691","type":"journal-article","created":{"date-parts":[[2022,6,30]],"date-time":"2022-06-30T20:53:02Z","timestamp":1656622382000},"page":"1691","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Molecular Cloning and Expression Analysis of Thyrotropin-Releasing Hormone, and Its Possible Role in Gonadal Differentiation in Rice Field eel Monopterus albus"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4476-1939","authenticated-orcid":false,"given":"Ke","family":"Feng","sequence":"first","affiliation":[{"name":"Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), College of Fisheries, Southwest University, Chongqing 400715, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jialin","family":"Su","sequence":"additional","affiliation":[{"name":"Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), College of Fisheries, Southwest University, Chongqing 400715, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengli","family":"Wu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), College of Fisheries, Southwest University, Chongqing 400715, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shengqi","family":"Su","sequence":"additional","affiliation":[{"name":"Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), College of Fisheries, Southwest University, Chongqing 400715, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weizhi","family":"Yao","sequence":"additional","affiliation":[{"name":"Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), College of Fisheries, Southwest University, Chongqing 400715, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.ygcen.2009.05.003","article-title":"TRH acts as a multifunctional hypophysiotropic factor in vertebrates","volume":"164","author":"Galas","year":"2009","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1737","DOI":"10.1016\/j.peptides.2010.05.017","article-title":"Molecular cloning and characterization of two putative appetite regulators in winter flounder (Pleuronectes americanus): Preprothyrotropin-releasing hormone (TRH) and preproorexin (OX)","volume":"31","author":"Buckley","year":"2010","journal-title":"Peptides"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"111229","DOI":"10.1016\/j.mce.2021.111229","article-title":"Response of the thyroid axis and appetite-regulating peptides to fasting and overfeeding in goldfish (Carassius auratus)","volume":"528","author":"Deal","year":"2021","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1038\/226321a0","article-title":"Characterization of ovine hypothalamic hypophysiotropic TSH-releasing factor","volume":"226","author":"Burgus","year":"1970","journal-title":"Nature"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1103","DOI":"10.1021\/bi00807a008","article-title":"Structure of porcine thyrotropin releasing hormone","volume":"9","author":"Nair","year":"1970","journal-title":"Biochemistry"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1111\/j.1365-2826.2010.01996.x","article-title":"Molecular evolution of the thyrotrophin-releasing hormone precursor in vertebrates: Insights from comparative genomics","volume":"22","author":"Wallis","year":"2010","journal-title":"J. Neuroendocrinol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1091","DOI":"10.1210\/en.2008-1795","article-title":"Thyrotropin-releasing hormone and the thyroid hormone feedback mechanism","volume":"150","author":"Chiamolera","year":"2009","journal-title":"Endocrinology"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Nakajima, Y., Yamada, M., Taguchi, R., Shibusawa, N., Ozawa, A., Tomaru, T., Hashimoto, K., Saito, T., Tsuchiya, T., and Okada, S. (2012). NR4A1 (Nur77) mediates thyrotropin-releasing hormone-induced stimulation of transcription of the thyrotropin \u03b2 gene: Analysis of TRH knockout mice. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0040437"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2795","DOI":"10.1210\/me.2007-0048","article-title":"Thyrotropin-releasing hormone receptor 1-deficient mice display increased depression and anxiety-like behavior","volume":"21","author":"Zeng","year":"2007","journal-title":"Mol. Endocrinol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1016\/0016-6480(88)90155-4","article-title":"The effects of TRH on plasma thyroid hormone levels of rainbow trout (Salmo gairdneri) and arctic charr (Salvelinus alpinus)","volume":"72","author":"Eales","year":"1988","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0303-7207(01)00392-6","article-title":"Molecular cloning of cDNA encoding thyroid stimulating hormone \u03b2 subunit of bighead carp Aristichthys nobilis and regulation of its gene expression","volume":"174","author":"Chatterjee","year":"2001","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1677\/jme.0.0320179","article-title":"Cloning of the cDNA for thyroid stimulating hormone beta subunit and changes in activity of the pituitary-thyroid axis during silvering of the Japanese eel, Anguilla japonica","volume":"32","author":"Han","year":"2004","journal-title":"J. Mol. Endocrinol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1006\/gcen.1995.1002","article-title":"Hypothalamic and thyroidal regulation of growth hormone in tilapia","volume":"97","author":"Melamed","year":"1995","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1006\/gcen.1998.7124","article-title":"Effects of thyrotropin-releasing hormone and its metabolites, cyclo(His-Pro) and TRH-OH, on growth hormone and prolactin synthesis in primary cultured pituitary cells of the common carp, Cyprinus carpio","volume":"111","author":"Kagabu","year":"1998","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1006\/gcen.1997.7031","article-title":"In vitro thyrotropin-releasing activity of corticotropin-releasing hormone-family peptides in coho salmon, Oncorhynchus kisutch","volume":"109","author":"Larsen","year":"1998","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.ygcen.2004.11.011","article-title":"Molecular cloning of prepro-thyrotropin-releasing hormone cDNAs from the common carp Cyprinus carpio and goldfish Carassius auratus","volume":"141","author":"Aoki","year":"2005","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/j.cbpb.2010.05.013","article-title":"Molecular characterization and gene expression of thyrotropin-releasing hormone in Senegalese sole (Solea senegalensis)","volume":"157","author":"Iziga","year":"2010","journal-title":"Comp. Biochem. Physiol. B"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1007\/s10695-017-0457-x","article-title":"Gene expression of thyrotropin- and corticotrophin-releasing hormones is regulated by environmental salinity in the euryhaline teleost Sparus aurata","volume":"44","author":"Mancera","year":"2018","journal-title":"Fish Physiol. Biochem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1080\/10495398.2011.547745","article-title":"Mapping, cDNA cloning and tissue expression of the porcine thyrotropin-releasing hormone receptor gene","volume":"22","author":"Jiang","year":"2011","journal-title":"Anim. Biotechnol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1016\/j.ygcen.2010.10.013","article-title":"Molecular cloning, gene structure, molecular evolution and expression analyses of thyrotropin-releasing hormone receptors from medaka (Oryzias latipes)","volume":"170","author":"Mekuchi","year":"2011","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.ygcen.2015.06.003","article-title":"\u201cBrain sex differentiation\u201d in teleosts: Emerging concepts with potential biomarkers","volume":"220","author":"Senthilkumaran","year":"2015","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1387\/ijdb.200072jf","article-title":"Stress and sex determination in fish: From brain to gonads","volume":"65","author":"Fernandino","year":"2021","journal-title":"Int. J. Dev. Biol."},{"key":"ref_23","unstructured":"Fisheries Bureau of Agriculture Ministry of China (2021). China Fishery Statistical Yearbook."},{"key":"ref_24","first-page":"1","article-title":"Rudimentary hermaphroditism in the symbranchoid eel, Monopterus albus","volume":"15","author":"Liu","year":"1944","journal-title":"Sinensia"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1016\/j.bbrc.2006.07.145","article-title":"Construction of a BAC library and identification of dmrt1 gene of the rice field eel, Monopterus albus","volume":"348","author":"Jang","year":"2006","journal-title":"Biochem. Bioph. Res. Comumun."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1016\/j.scib.2017.01.018","article-title":"High efficient gene targeting in rice field eel Monopterus albus by transcription activator-like effector nucleases","volume":"62","author":"Feng","year":"2017","journal-title":"Sci. Bull."},{"key":"ref_27","first-page":"775","article-title":"Sex determination and control in eels","volume":"Volume 11","author":"Wang","year":"2018","journal-title":"Sex Control in Aquaculture"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1721","DOI":"10.1007\/s10695-014-9962-3","article-title":"Reference gene selection for real-time RT-PCR normalization in rice field eel (Monopterus albus) during gonad development","volume":"40","author":"Hu","year":"2014","journal-title":"Fish Physiol. Biochem."},{"key":"ref_29","unstructured":"Green, M.R., and Sambrook, J. (2014). Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press. [4th ed.]."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1186\/1745-6150-3-20","article-title":"Splign: Algorithms for computing spliced alignments with identification of paralogs","volume":"3","author":"Kapustin","year":"2008","journal-title":"Bio. Direct"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1038\/s41587-019-0036-z","article-title":"SignalP 5.0 improves signal peptide predictions using deep neural networks","volume":"37","author":"Armenteros","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2947","DOI":"10.1093\/bioinformatics\/btm404","article-title":"Clustal W and Clustal X version 2.0","volume":"23","author":"Larkin","year":"2007","journal-title":"Bioinformatics"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1870","DOI":"10.1093\/molbev\/msw054","article-title":"MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets","volume":"33","author":"Kumar","year":"2016","journal-title":"Mol. Biol. Evol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real time quantitative PCR and the 2-\u25b3\u25b3Ct method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"729","DOI":"10.1016\/j.ygcen.2007.08.008","article-title":"Down-regulation of goldfish (Carassius auratus) prolactin gene expression by dopamine and thyrotropin releasing hormone","volume":"155","author":"Tse","year":"2008","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1007\/BF00306111","article-title":"Morphological identification of live gonadotropin, growth-hormone, and prolactin cells in goldfish (Carassius auratus) pituitary-cell cultures","volume":"276","author":"Goldberg","year":"1994","journal-title":"Cell Tissue Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2881","DOI":"10.1210\/en.2012-2220","article-title":"Epigenetic modifications during sex change repress gonadotropin stimulation of cyp19a1a in a teleost rice field eel (Monopterus albus)","volume":"154","author":"Zhang","year":"2013","journal-title":"Endocrinology"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.ygcen.2018.05.021","article-title":"Alternative splicing of GnRH2 and GnRH2-associated peptide plays roles in gonadal differentiation of the rice field eel, Monopterus albus","volume":"267","author":"Feng","year":"2018","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1016\/j.yhbeh.2010.12.008","article-title":"Thyrotropin releasing hormone (TRH) in goldfish (Carassius auratus): Role in the regulation of feeding and locomotor behaviors and interactions with the orexin system and cocaine- and amphetamine regulated transcript (CART)","volume":"59","author":"Abbott","year":"2011","journal-title":"Horm. Behav."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1677\/joe.1.06161","article-title":"Cloning and hypothalamic distribution of the chicken thyrotropin-releasing hormone precursor cDNA","volume":"186","author":"Vandenborne","year":"2005","journal-title":"J. Endocrinol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1126\/science.3079917","article-title":"Thyrotropin-releasing hormone precursor: Characterization in rat brain","volume":"231","author":"Lechan","year":"1986","journal-title":"Science"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1210\/mend-4-4-551","article-title":"Cloning and structure of human genomic DNA and hypothalamic cDNA encoding human preprothyrotropin-releasing hormone","volume":"4","author":"Yamada","year":"1990","journal-title":"Mol. Endocrinol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"292","DOI":"10.1016\/0014-5793(92)80307-3","article-title":"A cDNA from brain of Xenopus laevis coding for a new precursor of thyrotropin-releasing hormone","volume":"296","author":"Bulant","year":"1992","journal-title":"FEBS Lett."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1016\/j.ygcen.2006.09.009","article-title":"Molecular cloning of prepro-thyrotropin-releasing hormone cDNA from medaka (Oryzias latipes)","volume":"150","author":"Aoki","year":"2007","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"11731","DOI":"10.1016\/S0021-9258(19)38458-3","article-title":"Two precursors of thyrotropin-releasing hormone from skin of Xenopus laevis","volume":"265","author":"Kuchler","year":"1990","journal-title":"J. Biol. Chem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1070","DOI":"10.1002\/cne.24570","article-title":"Thyrotropin-releasing hormone (TRH) in the brain and pituitary of the teleost, Clarias batrachus and its role in regulation of hypophysiotropic dopamine neurons","volume":"527","author":"Singh","year":"2019","journal-title":"J. Comp. Neurol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.ygcen.2010.04.007","article-title":"Central administration of growth hormone-releasing hormone and corticotropin-releasing hormone stimulate downstream movement and thyroxine secretion in fall-smolting coho salmon (Oncorhynchus kisutch)","volume":"168","author":"Ojima","year":"2010","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"292","DOI":"10.1016\/0016-6480(75)90118-5","article-title":"The effects of mammalian thyrotropin-releasing hormone on the pituitary-thyroid axis of teleost fish","volume":"25","author":"Bromage","year":"1975","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1006\/gcen.1999.7253","article-title":"Immunohistochemical localization of corticotropin-releasing factor in the brain and corticotropin-releasing factor and thyrotropin-stimulating hormone in the pituitary of chinook salmon (Oncorhynchus tshawytscha)","volume":"114","author":"Matz","year":"1999","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1016\/j.ygcen.2018.04.023","article-title":"Thyroid hormone actions on male reproductive system of teleost fish","volume":"265","author":"Rodrigues","year":"2018","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.mce.2015.08.008","article-title":"Gonadal soma-derived factor (gsdf), a TGF-beta superfamily gene, induces testis differentiation in the teleost fish Oreochromis niloticus","volume":"415","author":"Kaneko","year":"2015","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.ygcen.2015.09.025","article-title":"The role of Amh signaling in teleost fish\u2014Multiple functions not restricted to the gonads","volume":"223","author":"Pfennig","year":"2015","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"352","DOI":"10.1016\/j.ygcen.2009.03.002","article-title":"Ovarian aromatase and estrogens: A pivotal role for gonadal sex differentiation and sex change in fish","volume":"165","author":"Guiguen","year":"2010","journal-title":"Gen. Comp. Endocrinol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"16667","DOI":"10.1038\/srep16667","article-title":"Molecular cloning and characterization of amh and dax1 genes and their expression during sex inversion in rice field eel Monopterus albus","volume":"5","author":"Hu","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1007\/s11033-016-3997-8","article-title":"Identification of gonadal soma-derived factor involvement in Monopterus albus (protogynous rice field eel) sex change","volume":"43","author":"Zhu","year":"2016","journal-title":"Mol. Biol. Rep."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1677\/JOE-08-0303","article-title":"Two cytochrome P450 aromatase genes in the hermaphrodite rice field eel Monopterus albus: mRNA expression during ovarian development and sex change","volume":"199","author":"Zhang","year":"2008","journal-title":"J. Endocrinol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"734536","DOI":"10.1016\/j.aquaculture.2019.734536","article-title":"Effects of environmental stress (sex steroids and heat) during sex differentiation in Japanese flounder (Paralichthys olivaceus): Insight from germ cell proliferation and gsdf-amh-cyp19a1a expression","volume":"515","author":"Yang","year":"2020","journal-title":"Aquaculture"}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/12\/13\/1691\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:40:58Z","timestamp":1760139658000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/12\/13\/1691"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,30]]},"references-count":57,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["ani12131691"],"URL":"https:\/\/doi.org\/10.3390\/ani12131691","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,30]]}}}