{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"Research Square"}],"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T00:53:35Z","timestamp":1740099215454,"version":"3.37.3"},"posted":{"date-parts":[[2022,9,9]]},"group-title":"In Review","reference-count":35,"publisher":"Springer Science and Business Media LLC","license":[{"start":{"date-parts":[[2022,9,9]],"date-time":"2022-09-09T00:00:00Z","timestamp":1662681600000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2022,8,24]]},"abstract":"<title>Abstract<\/title>\n        <p>Histological techniques consist of traditional and very important tools for a better understanding of cellular morphology and physiology in ecotoxicological studies. Despite bioassays generally recommending the use of small test organisms, based on some advantages as their low maintenance costs and large replicates\u2019 number, the histological processing of small organisms is not commonly used due to demanding laboratorial handling. The present study proposes an accessible protocol for a histological method in fish larvae with a maximum length of 8 mm, as well as the use of histopathological biomarkers on Senegal sole larvae for ecotoxicological testing. Eight-day old larvae of <italic>Solea senegalensis<\/italic> obtained from breeders' natural spawns in cultivation conditions were submitted to histological processing by an adjusted protocol comprising fixation, agarose array pre-embedding, paraffin embedding, slides preparation, and image analysis by light optical microscope coupled to a digital camera. Low-cost and easy-to-use alternatives were applied in the technique execution. The histological steps were detailed and the multiple tissues and cellular structures of the <italic>S. senegalensis<\/italic> larvae were described and discussed. This can be considered a valuable background information for ecotoxicological studies where changes in cell morphology and physiology are expected.<\/p>","DOI":"10.21203\/rs.3.rs-1995398\/v1","type":"posted-content","created":{"date-parts":[[2022,9,9]],"date-time":"2022-09-09T22:49:17Z","timestamp":1662763757000},"source":"Crossref","is-referenced-by-count":0,"title":["Improvements in histological technique for the ecotoxicological assessment using small biological samples"],"prefix":"10.21203","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8398-2729","authenticated-orcid":false,"given":"LIGIA","family":"SANTANA","sequence":"first","affiliation":[{"name":"Federal University of Latin-American Integration: Universidade Federal da Integracao Latino-Americana"}]},{"given":"\u00c9vila P.","family":"Damasceno","sequence":"additional","affiliation":[{"name":"University of Aveiro: Universidade de Aveiro"}]},{"given":"Susana","family":"Loureiro","sequence":"additional","affiliation":[{"name":"University of Aveiro: Universidade de Aveiro"}]},{"given":"Amadeu M.V.M.","family":"Soares","sequence":"additional","affiliation":[{"name":"University of Aveiro: Universidade de Aveiro"}]},{"given":"Pedro","family":"Pous\u00e3o-Ferreira","sequence":"additional","affiliation":[{"name":"IPMA: Instituto Portugues do Mar e da Atmosfera"}]},{"given":"Denis M. S.","family":"Abessa","sequence":"additional","affiliation":[{"name":"UNESP: Universidade Estadual Paulista Julio de Mesquita Filho"}]},{"given":"Roberto","family":"Martins","sequence":"additional","affiliation":[{"name":"University of Aveiro: Universidade de Aveiro"}]},{"given":"Maria D.","family":"Pavlaki","sequence":"additional","affiliation":[{"name":"University of Aveiro: Universidade de Aveiro"}]}],"member":"297","reference":[{"issue":"3","key":"ref1","doi-asserted-by":"publisher","first-page":"229","DOI":"10.1016\/S1535-6108(02)00052-1","article-title":"Zebrafish as a cancer model system","volume":"1","author":"Amatruda JF","year":"2002","unstructured":"Amatruda JF, Shepard JL, Stern HM, Zon LI (2002) Zebrafish as a cancer model system. Cancer Cell 1(3):229\u2013231. https:\/\/doi.org\/10.1016\/S1535-6108(02)00052-1","journal-title":"Cancer Cell"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"885","DOI":"10.1016\/j.chemosphere.2019.06.183","article-title":"Effects of triclosan on early development of Solea senegalensis: from biochemical to individual level","volume":"235","author":"Ara\u00fajo MJ","year":"2019","unstructured":"Ara\u00fajo MJ, Quintaneiro C, Soares AMVM, Monteiro MS (2019) Effects of triclosan on early development of Solea senegalensis: from biochemical to individual level. Chemosphere 235:885\u2013899. https:\/\/doi.org\/10.1016\/j.chemosphere.2019.06.183","journal-title":"Chemosphere"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"1395","DOI":"10.1016\/j.scitotenv.2018.02.112","article-title":"Effects of UV filter 4-methylbenzylidene camphor during early development of Solea senegalensis Kaup 1858","volume":"628","author":"Ara\u00fajo MJ","year":"2018","unstructured":"Ara\u00fajo MJ, Rocha RJM, Soares AMVM, Bened\u00e9 JL, Chisvert A, Monteiro MS (2018) Effects of UV filter 4-methylbenzylidene camphor during early development of Solea senegalensis Kaup 1858. Sci Total Environ 628:1395\u20131404. https:\/\/doi.org\/10.1016\/j.scitotenv.2018.02.112","journal-title":"Sci Total Environ"},{"issue":"2","key":"ref4","first-page":"511","article-title":"Histological and histochemical observations in the stomach of the Senegal sole Solea senegalensis","volume":"16","author":"Arellano JM","year":"2001","unstructured":"Arellano JM, Storch V, Sarasquete C (2001) Histological and histochemical observations in the stomach of the Senegal sole Solea senegalensis. Histol Histopathol 16(2):511\u2013521. http:\/\/www.ehu.eslhistol-histopathol","journal-title":"Histol Histopathol"},{"issue":"1","key":"ref5","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1046\/j.1365-2761.1999.00134.x","article-title":"Histopathology in fish: proposal for a protocol to assess aquatic pollution","volume":"22","author":"Bernet D","year":"1999","unstructured":"Bernet D, Schmidt H, Meier W, Burkhardt-Holm P, Wahli T (1999) Histopathology in fish: proposal for a protocol to assess aquatic pollution. J Fish Dis 22(1):25\u201334. https:\/\/doi.org\/10.1046\/j.1365-2761.1999.00134.x","journal-title":"J Fish Dis"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"105351","DOI":"10.1016\/j.marenvres.2021.105351","article-title":"Biological responses and toxicopathic effects elicited in Solea senegalensis juveniles by waterborne exposure to benzo [a] pyrene","volume":"170","author":"Briaudeau T","year":"2021","unstructured":"Briaudeau T, Dos Santos LAA, Zorita I, Izagirre U, Marig\u00f3mez I (2021) Biological responses and toxicopathic effects elicited in Solea senegalensis juveniles by waterborne exposure to benzo [a] pyrene. Mar Environ Res 170:105351. https:\/\/doi.org\/10.1016\/j.marenvres.2021.105351","journal-title":"Mar Environ Res"},{"issue":"6","key":"ref7","doi-asserted-by":"publisher","first-page":"1993","DOI":"10.3390\/app10061993","article-title":"Mercury in juvenile Solea senegalensis: Linking bioaccumulation seafood safety and neuro-oxidative responses under climate change-related stressors","volume":"10","author":"Camacho C","year":"2020","unstructured":"Camacho C, Maulvault AL, Santos MT, Barbosa V, Foga\u00e7a FH, Pous\u00e3o-Ferreira P, Nunes ML, Rosa R, Marques A (2020) Mercury in juvenile Solea senegalensis: Linking bioaccumulation seafood safety and neuro-oxidative responses under climate change-related stressors. Appl Sci 10(6):1993. https:\/\/doi.org\/10.3390\/app10061993","journal-title":"Appl Sci"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"38","DOI":"10.1016\/j.cbpc.2017.11.003","article-title":"Comparative analysis of fixation and embedding techniques for optimized histological preparation of zebrafish","volume":"208","author":"Copper JE","year":"2018","unstructured":"Copper JE, Budgeon LR, Foutz CA, van Rossum DB, Vanselow DJ, Hubley MJ, Clark DP, Mandrell DT, Cheng KC (2018) Comparative analysis of fixation and embedding techniques for optimized histological preparation of zebrafish. Comp Biochem Physiol Part-C: Toxicol Pharmacol 208:38\u201346. https:\/\/doi.org\/10.1016\/j.cbpc.2017.11.003","journal-title":"Comp Biochem Physiol Part-C: Toxicol Pharmacol"},{"key":"ref9","unstructured":"EPA - Environmental Protection Agency (1998) Guidelines for Ecological Risk Assessment. EPA 630\/R-95-002F Washington D.C"},{"issue":"4","key":"ref10","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1007\/s11160-004-1632-6","article-title":"A review of the culture potential of Solea solea and S. senegalensis","volume":"13","author":"Imsland AK","year":"2003","unstructured":"Imsland AK, Foss A, Concei\u00e7ao LE, Dinis MT, Delbare D, Schram E, Rema P, White P (2003) A review of the culture potential of Solea solea and S. senegalensis. Rev Fish Biol Fish 13(4):379\u2013408. https:\/\/doi.org\/10.1007\/s11160-004-1632-6","journal-title":"Rev Fish Biol Fish"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"107207","DOI":"10.1016\/j.ecolind.2020.107207","article-title":"Biomarkers based tools to assess environmental and chemical stressors in aquatic systems","volume":"122","author":"Lomartire S","year":"2021","unstructured":"Lomartire S, Marques JC, Gon\u00e7alves AM (2021) Biomarkers based tools to assess environmental and chemical stressors in aquatic systems. Ecol Indic 122:107207. https:\/\/doi.org\/10.1016\/j.ecolind.2020.107207","journal-title":"Ecol Indic"},{"issue":"1","key":"ref12","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1016\/j.etap.2012.02.007","article-title":"The toxicity potential of pharmaceuticals found in the Douro River estuary (Portugal): evaluation of impacts on fish liver by histopathology stereology vitellogenin and CYP1A immunohistochemistry after sub-acute exposures of the zebrafish model","volume":"34","author":"Madureira TV","year":"2012","unstructured":"Madureira TV, Rocha MJ, Cruzeiro C, Rodrigues I, Monteiro RA, Rocha E (2012) The toxicity potential of pharmaceuticals found in the Douro River estuary (Portugal): evaluation of impacts on fish liver by histopathology stereology vitellogenin and CYP1A immunohistochemistry after sub-acute exposures of the zebrafish model. Environ Toxicol Pharmacol 34(1):34\u201345. https:\/\/doi.org\/10.1016\/j.etap.2012.02.007","journal-title":"Environ Toxicol Pharmacol"},{"issue":"3","key":"ref13","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1111\/raq.12091","article-title":"New developments and biological insights into the farming of Solea senegalensis reinforcing its aquaculture potential","volume":"8","author":"Morais S","year":"2016","unstructured":"Morais S, Arag\u00e3o C, Cabrita E, Concei\u00e7\u00e3o LE, Constenla M, Costas B, Dias J, Duncan N, Engrola S, Estevez A, Gisbert E, Ma\u00f1an\u00f3s E, Valente LMP, Y\u00fafera M, Dinis MT (2016) New developments and biological insights into the farming of Solea senegalensis reinforcing its aquaculture potential. Rev Aquac 8(3):227\u2013263. https:\/\/doi.org\/10.1111\/raq.12091","journal-title":"Rev Aquac"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"108722","DOI":"10.1016\/j.cbpc.2020.108722","article-title":"Toxic effects of environmentally realistic concentrations of diclofenac in organisms from two distinct trophic levels Hediste diversicolor and Solea senegalensis","volume":"231","author":"Nunes B","year":"2020","unstructured":"Nunes B, Daniel D, Canelas GG, Barros J, Correia AT (2020) Toxic effects of environmentally realistic concentrations of diclofenac in organisms from two distinct trophic levels Hediste diversicolor and Solea senegalensis. Comp Biochem Physiol Part-C: Toxicol Pharmacol 231:108722. https:\/\/doi.org\/10.1016\/j.cbpc.2020.108722","journal-title":"Comp Biochem Physiol Part-C: Toxicol Pharmacol"},{"issue":"9","key":"ref15","doi-asserted-by":"publisher","first-page":"1894","DOI":"10.1002\/tox.23310","article-title":"Toxicity of malathion during Senegalese sole Solea senegalensis larval development and metamorphosis: Histopathological disorders and effects on type B esterases and CYP1A enzymatic systems","volume":"36","author":"Ortiz-Delgado JB","year":"2021","unstructured":"Ortiz-Delgado JB, Funes V, Albend\u00edn G, Scala E, Sarasquete C (2021) Toxicity of malathion during Senegalese sole Solea senegalensis larval development and metamorphosis: Histopathological disorders and effects on type B esterases and CYP1A enzymatic systems. Environ Toxicol 36(9):1894\u20131910. https:\/\/doi.org\/10.1002\/tox.23310","journal-title":"Environ Toxicol"},{"issue":"1","key":"ref16","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1111\/j.1095-8649.2011.02942.x","article-title":"Morphological and histological study of larval development of the Senegal sole Solea senegalensis: an integrative study","volume":"79","author":"Padr\u00f3s F","year":"2011","unstructured":"Padr\u00f3s F, Villalta M, Gisbert E, Est\u00e9vez A (2011) Morphological and histological study of larval development of the Senegal sole Solea senegalensis: an integrative study. J Fish Biol 79(1):3\u201332. https:\/\/doi.org\/10.1111\/j.1095-8649.2011.02942.x","journal-title":"J Fish Biol"},{"key":"ref17","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1016\/j.envpol.2016.05.010","article-title":"Ecotoxicity and genotoxicity of cadmium in different marine trophic levels","volume":"215","author":"Pavlaki MD","year":"2016","unstructured":"Pavlaki MD, Ara\u00fajo MJ, Cardoso DN, Silva ARR, Cruz A, Mendo S, Soares AMVM, Calado R, Loureiro S (2016) Ecotoxicity and genotoxicity of cadmium in different marine trophic levels. Environ Pollut 215:203\u2013212. https:\/\/doi.org\/10.1016\/j.envpol.2016.05.010","journal-title":"Environ Pollut"},{"issue":"4","key":"ref18","doi-asserted-by":"publisher","first-page":"522","DOI":"10.3390\/w13040522","article-title":"Cadmium accumulation and kinetics in Solea senegalensis tissues under dietary and water exposure and the link to human health","volume":"13","author":"Pavlaki MD","year":"2021","unstructured":"Pavlaki MD, Morgado RG, Ferreira V, Rocha RJ, Soares AM, Calado R, Loureiro S (2021) Cadmium accumulation and kinetics in Solea senegalensis tissues under dietary and water exposure and the link to human health. Water 13(4):522. https:\/\/doi.org\/10.3390\/w13040522","journal-title":"Water"},{"key":"ref19","volume-title":"Microscopy and imaging science","author":"Plaul SE","year":"2017","unstructured":"Plaul SE, Andr\u00e9s Laube PF, Pacheco Marino SG, Santamar\u00eda Mart\u00edn CJ, Moyano DA, Barbeito CG, M\u00e9ndez-Vilas A (2017) Morphological techniques used in ichthyopathological diagnosis. In: M\u00e9ndez-Vilas A (ed) Microscopy and imaging science. practical approaches to applied research and education"},{"issue":"2","key":"ref20","doi-asserted-by":"publisher","first-page":"271","DOI":"10.7860\/JCDR\/2014\/7544.4079","article-title":"Health hazards of xylene: a literature review","volume":"8","author":"Rajan ST","year":"2014","unstructured":"Rajan ST, Malathi N (2014) Health hazards of xylene: a literature review. J Clin Diagn Res 8(2):271. https:\/\/doi.org\/10.7860\/JCDR\/2014\/7544.4079","journal-title":"J Clin Diagn Res"},{"issue":"3\u20134","key":"ref21","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1016\/S0044-8486(98)00496-7","article-title":"Histological and histochemical development of the digestive system of Solea senegalensis (Kaup 1858) larvae","volume":"171","author":"Ribeiro L","year":"1999","unstructured":"Ribeiro L, Sarasquete C, Dinis MT (1999) Histological and histochemical development of the digestive system of Solea senegalensis (Kaup 1858) larvae. Aquac 171(3\u20134):293\u2013308. https:\/\/doi.org\/10.1016\/S0044-8486(98)00496-7","journal-title":"Aquac"},{"issue":"6","key":"ref22","doi-asserted-by":"publisher","first-page":"448","DOI":"10.1136\/jclinpath-2018-205680","article-title":"Agar pre-embedding of small skin biopsies: real-life benefits and challenges in high throughput pathology laboratories","volume":"72","author":"Ridolfi M","year":"2019","unstructured":"Ridolfi M, Paudice M, Salvi S, Valle L, Gualco M, Perasole A, Anselmi L, Fiocca R, Mastracci L, Grillo F (2019) Agar pre-embedding of small skin biopsies: real-life benefits and challenges in high throughput pathology laboratories. J Clin Pathol 72(6):448\u2013451. http:\/\/dx.doi.org\/10.1136\/jclinpath-2018-205680","journal-title":"J Clin Pathol"},{"issue":"3","key":"ref23","doi-asserted-by":"publisher","first-page":"246","DOI":"10.1016\/j.ymeth.2006.03.001","article-title":"High-throughput zebrafish histology","volume":"39","author":"Sabaliauskas NA","year":"2006","unstructured":"Sabaliauskas NA, Foutz CA, Mest JR, Budgeon LR, Sidor AT, Gershenson JA, Joshi SB, Cheng KC (2006) High-throughput zebrafish histology. Meth 39(3):246\u2013254. https:\/\/doi.org\/10.1016\/j.ymeth.2006.03.001","journal-title":"Meth"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"429","DOI":"10.1016\/j.envpol.2017.10.061","article-title":"Ecological risk assessment of a contaminated stream using multi-level integrated biomarker response in Carassius auratus","volume":"233","author":"Samanta P","year":"2018","unstructured":"Samanta P, Im H, Na J, Jung J (2018) Ecological risk assessment of a contaminated stream using multi-level integrated biomarker response in Carassius auratus. Environ Pollut 233:429\u2013438. https:\/\/doi.org\/10.1016\/j.envpol.2017.10.061","journal-title":"Environ Pollut"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"150188","DOI":"10.1016\/j.scitotenv.2021.150188","article-title":"Can the toxicity of polyethylene microplastics and engineered nanoclays on flatfish (Solea senegalensis) be influenced by the presence of each other?","volume":"804","author":"Santana LMBM","year":"2022","unstructured":"Santana LMBM, Rodrigues AC, Campos D, Kaczerewska O, Figueiredo J, Silva S, Sousa I, Maia F, Tedim J, Abessa DMS, Pous\u00e3o-Ferreira P, Candeias-Mendes A, Soares F, Castanho S, Soares AMVM, Rocha RJM, Gravato C, Silva ALP, Martins R (2022) Can the toxicity of polyethylene microplastics and engineered nanoclays on flatfish (Solea senegalensis) be influenced by the presence of each other? Sci Total Environ 804:150188. https:\/\/doi.org\/10.1016\/j.scitotenv.2021.150188","journal-title":"Sci Total Environ"},{"issue":"11","key":"ref26","doi-asserted-by":"publisher","first-page":"1762","DOI":"10.1016\/j.marpolbul.2007.07.014","article-title":"Estuarine fish health assessment: evidence of wastewater impacts based on nitrogen isotopes and histopathology","volume":"54","author":"Schlacher TA","year":"2007","unstructured":"Schlacher TA, Mondon JA, Connolly RM (2007) Estuarine fish health assessment: evidence of wastewater impacts based on nitrogen isotopes and histopathology. Mar Pollut Bull 54(11):1762\u20131776. https:\/\/doi.org\/10.1016\/j.marpolbul.2007.07.014","journal-title":"Mar Pollut Bull"},{"issue":"1","key":"ref27","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1023\/A:1008212000208","article-title":"The use of histopathological indicators to evaluate contaminant-related stress in fish","volume":"6","author":"Schwaiger J","year":"1997","unstructured":"Schwaiger J, Wanke R, Adam S, Pawert M, Honnen W, Triebskorn R (1997) The use of histopathological indicators to evaluate contaminant-related stress in fish. J Aquat Ecosyst Stress Recovery 6(1):75\u201386. https:\/\/doi.org\/10.1023\/A:1008212000208","journal-title":"J Aquat Ecosyst Stress Recovery"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"132103","DOI":"10.1016\/j.chemosphere.2021.132103","article-title":"Biochemical genotoxic histological and ultrastructural effects on liver and gills of fresh water fish Channa punctatus exposed to textile industry intermediate 2 ABS","volume":"287","author":"Sharma K","year":"2022","unstructured":"Sharma K, Sharma P, Dhiman SK, Chadha P, Saini HS (2022) Biochemical genotoxic histological and ultrastructural effects on liver and gills of fresh water fish Channa punctatus exposed to textile industry intermediate 2 ABS. Chemosphere 287:132103. https:\/\/doi.org\/10.1016\/j.chemosphere.2021.132103","journal-title":"Chemosphere"},{"key":"ref29","volume-title":"Atlas of fish histology and histopathology","author":"Sharon G","year":"2012","unstructured":"Sharon G, Zilberg D (2012) Atlas of fish histology and histopathology. Central and Northen Arava Research and Development Centers 78"},{"issue":"3","key":"ref30","doi-asserted-by":"publisher","DOI":"10.17221\/8763-VETMED","article-title":"Morphologic evaluation of the gills as a tool in the diagnostics of pathological conditions in fish and pollution in the aquatic environment: a review","volume":"61","author":"Strzyzewska E","year":"2016","unstructured":"Strzyzewska E, Szarek J, Babinska I (2016) Morphologic evaluation of the gills as a tool in the diagnostics of pathological conditions in fish and pollution in the aquatic environment: a review. Vet Med 61(3). https:\/\/doi.org\/10.17221\/8763-VETMED","journal-title":"Vet Med"},{"issue":"4","key":"ref31","doi-asserted-by":"publisher","first-page":"614","DOI":"10.2144\/98254st02","article-title":"Agarose-embedded tissue arrays for histologic and genetic analysis","volume":"25","author":"Tsao-Wu GS","year":"1998","unstructured":"Tsao-Wu GS, Weber CH, Budgeon LR, Cheng KC (1998) Agarose-embedded tissue arrays for histologic and genetic analysis. Biotechniques 25(4):614\u2013618. https:\/\/doi.org\/10.2144\/98254st02","journal-title":"Biotechniques"},{"issue":"3","key":"ref32","doi-asserted-by":"publisher","first-page":"395","DOI":"10.4025\/actascibiolsci.v35i3.18032","article-title":"Fish histopathology and catalase activity as biomarkers of the environmental quality of the industrial district on the Amazon estuary Brazil","volume":"35","author":"Viana AP","year":"2013","unstructured":"Viana AP, Fr\u00e9dou FL, da Silva Montes C, Rocha RM (2013) Fish histopathology and catalase activity as biomarkers of the environmental quality of the industrial district on the Amazon estuary Brazil. Acta Sci Biol Sci 35(3):395\u2013401. https:\/\/doi.org\/10.4025\/actascibiolsci.v35i3.18032","journal-title":"Acta Sci Biol Sci"},{"issue":"3\u20134","key":"ref33","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1016\/S1382-6689(02)00021-2","article-title":"Aquatic toxicology: opportunities for enhancement through histopathology","volume":"11","author":"Wester PW","year":"2002","unstructured":"Wester PW, Van der Ven LTM, Vethaak AD, Grinwis GCM, Vos JG (2002) Aquatic toxicology: opportunities for enhancement through histopathology. Environ Toxicol Pharmacol 11(3\u20134):289\u2013295. https:\/\/doi.org\/10.1016\/S1382-6689(02)00021-2","journal-title":"Environ Toxicol Pharmacol"},{"issue":"3","key":"ref34","doi-asserted-by":"publisher","first-page":"297","DOI":"10.1177\/0192623314540229","article-title":"Nonlesions misdiagnoses missed diagnoses and other interpretive challenges in fish histopathology studies: a guide for investigators authors reviewers and readers","volume":"43","author":"Wolf JC","year":"2015","unstructured":"Wolf JC, Baumgartner WA, Blazer VS, Camus AC, Engelhardt JA, Fournie JW, Frasca S Jr, Groman DB, Kent ML, Khoo LH, Law JM, Lombardini ED, Ruehl-Fehlert C, Segner HE, Smith SA, Spitsbergen JM, Weber K, Wolfe MJ (2015) Nonlesions misdiagnoses missed diagnoses and other interpretive challenges in fish histopathology studies: a guide for investigators authors reviewers and readers. Toxicol Pathol 43(3):297\u2013325. https:\/\/doi.org\/10.1177\/0192623314540229","journal-title":"Toxicol Pathol"},{"key":"ref35","unstructured":"Statements & Declarations"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/www.researchsquare.com\/article\/rs-1995398\/v1","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.researchsquare.com\/article\/rs-1995398\/v1.html","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,10,11]],"date-time":"2022-10-11T08:44:52Z","timestamp":1665477892000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.researchsquare.com\/article\/rs-1995398\/v1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,9]]},"references-count":35,"URL":"https:\/\/doi.org\/10.21203\/rs.3.rs-1995398\/v1","relation":{},"subject":[],"published":{"date-parts":[[2022,9,9]]},"subtype":"preprint"}}