{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T01:14:29Z","timestamp":1785892469789,"version":"3.56.0"},"reference-count":53,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,2,8]],"date-time":"2022-02-08T00:00:00Z","timestamp":1644278400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Diputaci\u00f3n general de arag\u00f3n","award":["A07 17R"],"award-info":[{"award-number":["A07 17R"]}]},{"DOI":"10.13039\/501100003329","name":"Ministry of Economy, Industry and Competitiveness","doi-asserted-by":"publisher","award":["CICYT AGL-2017-83799-R"],"award-info":[{"award-number":["CICYT AGL-2017-83799-R"]}],"id":[{"id":"10.13039\/501100003329","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100014440","name":"Ministerio de ciencia, innovaci\u00f3n y universidades","doi-asserted-by":"publisher","award":["MICINN PRE2018-085198"],"award-info":[{"award-number":["MICINN PRE2018-085198"]}],"id":[{"id":"10.13039\/100014440","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biology"],"abstract":"<jats:p>Testicular ultrasound is a non-invasive technique that could be very useful for predicting ram seminal quality. Recent software developments allow macroscopic and microscopic evaluation of testicular parenchyma. Thus, the aim of this study was to evaluate the testicular echotexture using ultrasound-video analysis and investigate its possible correlation with semen quality. Nine rams were evaluated for one year using a portable ultrasound scanner and the echotexture was analyzed with ECOTEXT\u00ae software. The number of black (Ec1), white (Ec2), and grey pixels (Ec3), tubular density (TD), lumen area (LA), and lumen diameter (LD) were analyzed. Semen was collected by an artificial vagina the same day and the sperm concentration, morphology, motility, viability, phosphatidylserine (PS) translocation, reactive-oxygen-species (ROS) levels, DNA damage and capacitation state were evaluated. Ec2 and Ec3 correlated positively with \u201cbad quality\u201d sperm parameters (the percentage of spermatozoa with high ROS levels, with PS translocation and proximal cytoplasmic droplets), and negatively with motility. In contrast, TD and LA showed a positive correlation with \u201cgood quality\u201d parameters (motility or normal morphology) and a negative correlation with spermatozoa with high ROS levels, with DNA fragmentation, and proximal or distal cytoplasmic droplets. Thus, echotexture analysis by ultrasound-video analysis could be a valuable tool for assessing ram fertility.<\/jats:p>","DOI":"10.3390\/biology11020261","type":"journal-article","created":{"date-parts":[[2022,2,9]],"date-time":"2022-02-09T01:03:40Z","timestamp":1644368620000},"page":"261","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Testicular Ultrasound Analysis as a Predictive Tool of Ram Sperm Quality"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3929-5064","authenticated-orcid":false,"given":"Melissa","family":"Carvajal-Serna","sequence":"first","affiliation":[{"name":"Grupo BIOFITER-Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular y Celular, Instituto Universitario de Investigaci\u00f3n en Ciencias Ambientales de Arag\u00f3n (IUCA), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6733-9373","authenticated-orcid":false,"given":"Sara","family":"Miguel-Jim\u00e9nez","sequence":"additional","affiliation":[{"name":"Grupo BIOFITER-Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular y Celular, Instituto Universitario de Investigaci\u00f3n en Ciencias Ambientales de Arag\u00f3n (IUCA), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2312-6402","authenticated-orcid":false,"given":"Rosaura","family":"P\u00e9rez-Pe","sequence":"additional","affiliation":[{"name":"Grupo BIOFITER-Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular y Celular, Instituto Universitario de Investigaci\u00f3n en Ciencias Ambientales de Arag\u00f3n (IUCA), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1997-4262","authenticated-orcid":false,"given":"Adriana","family":"Casao","sequence":"additional","affiliation":[{"name":"Grupo BIOFITER-Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular y Celular, Instituto Universitario de Investigaci\u00f3n en Ciencias Ambientales de Arag\u00f3n (IUCA), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/0378-4320(96)01505-9","article-title":"Seasonal breeding in sheep: Mechanism of action of melatonin","volume":"42","author":"Malpaux","year":"1996","journal-title":"Anim. Reprod. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1111\/j.1439-0531.2008.01141.x","article-title":"Seasonality of reproduction in mammals: Intimate regulatory mechanisms and practical implications","volume":"43","author":"Chemineau","year":"2008","journal-title":"Reprod. Domest. Anim."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1095\/biolreprod30.5.1039","article-title":"Profiles of luteinizing hormone, follicle-stimulating hormone, testosterone and prolactin in rams of diverse breeds: Effects of contrasting short (8L:16D) and long (16L:8D) photoperiods","volume":"30","author":"Schanbacher","year":"1984","journal-title":"Biol. Reprod."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"623","DOI":"10.1530\/jrf.0.0880623","article-title":"Seasonal cycles in the blood plasma concentration of FSH, inhibin and testosterone, and testicular size in rams of wild, feral and domesticated breeds of sheep","volume":"88","author":"Lincoln","year":"1990","journal-title":"J. Reprod. Fertil."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1530\/jrf.0.0790609","article-title":"The response of rams and ewes of three breeds to artificial photoperiod","volume":"79","author":"Poulton","year":"1987","journal-title":"J. Reprod. Fertil."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1095\/biolreprod42.1.63","article-title":"Continuous exposure of Suffolk ewes to an equatorial photoperiod disrupts expression of the annual breeding-season","volume":"42","author":"Jackson","year":"1990","journal-title":"Biol. Reprod."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/S0921-4488(99)00143-1","article-title":"Seasonal variation in semen characteristics of Chios and Friesian rams in Greece","volume":"37","author":"Karagiannidis","year":"2000","journal-title":"Small Rumin. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/S0921-4488(97)00073-4","article-title":"Influence of season and age on sexual maturation parameters of Texel, Suffolk and Ile-de-France rams: 1. Testicular size, semen quality and reproductive capacity","volume":"28","author":"Mandiki","year":"1998","journal-title":"Small Rumin. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/0093-691X(81)90112-6","article-title":"The effect of season on the scrotal circumference and sperm motility and morphology in rams","volume":"16","author":"Mickelsen","year":"1981","journal-title":"Theriogenology"},{"key":"ref_10","first-page":"383","article-title":"The effect of melatonin on sperm quality and testicular size of normospermic and pathospermic rams in the anoestrous season","volume":"85","author":"Coyan","year":"1998","journal-title":"Wien. Tierarztl. Monatsschr."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1959","DOI":"10.1016\/S0093-691X(02)01259-1","article-title":"Ultrasonographic appearance of clinically healthy testicles and epididymides of rams","volume":"59","author":"Gouletsou","year":"2003","journal-title":"Theriogenology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.smallrumres.2016.12.022","article-title":"Ultrasonographic examination of the scrotal contents in rams","volume":"152","author":"Gouletsou","year":"2017","journal-title":"Small Rumin. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1606","DOI":"10.1177\/1535370214543063","article-title":"Correlations among ultrasonographic and microscopic characteristics of prepubescent ram lamb testes","volume":"239","author":"Giffin","year":"2014","journal-title":"Exp. Biol. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"794","DOI":"10.3181\/0812-RM-364","article-title":"A Study of Morphological and Haemodynamic Determinants of Testicular Echotexture Characteristics in the Ram","volume":"234","author":"Giffin","year":"2009","journal-title":"Exp. Biol. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1258\/ebm.2011.011050","article-title":"Suitability of epididymal and testicular ultrasonography and computerized image analysis for assessment of current and future semen quality in the ram","volume":"237","author":"Ahmadi","year":"2012","journal-title":"Exp. Biol. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"49","DOI":"10.4103\/2305-0500.275635","article-title":"Seasonal changes in testicular ultrasonogram pixel-intensity and their association with semen characteristics in rams","volume":"9","author":"Hedia","year":"2020","journal-title":"Asian Pac. J. Reprod."},{"key":"ref_17","first-page":"936","article-title":"Lack of relationship between testicular echotexture and seminal characteristics in adult Texel rams","volume":"9","author":"Urt","year":"2018","journal-title":"Agric. Sci."},{"key":"ref_18","first-page":"110","article-title":"Testicular and epididymal ultrasonography in Santa Ines lambs raised in Brazil","volume":"11","author":"Andrade","year":"2014","journal-title":"Anim. Reprod."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1111\/rda.13213","article-title":"Changes in testicular size, echotexture, and arterial blood flow associated with the attainment of puberty in Dorper rams raised in a subtropical climate","volume":"54","author":"Camela","year":"2019","journal-title":"Reprod. Domest. Anim."},{"key":"ref_20","unstructured":"Echegaray, A., Marcantonio, S., Maraboli, C., Munoz, I., Escartin, N., and Gnemmi, G. (2018). New echotexture parameters to evaluate the testicular parenchyma in bulls. Reproduction in Domestic Animals, Proceedings of the 22nd Annual Conference of the European Society for Domestic Animal Reproduction (ESDAR), Cordoba, Spain, 27\u201329 September 2018, Wiley Blackwell."},{"key":"ref_21","unstructured":"Escartin, N., Munoz, I., Gil, A., and Echegaray, A. (2018). New echotexture parameters to evaluate the testicular parenchyma in rams. Reproduction in Domestic Animals, Proceedings of the 22nd Annual Conference of the European Society for Domestic Animal Reproduction (ESDAR), Cordoba, Spain, 27\u201329 September 2018, Wiley Blackwell."},{"key":"ref_22","unstructured":"Lafuente, A., Lafuente, M., Escartin, N., Munoz, I., and Echegaray, A. (2018). New echotexture parameters to evaluate the testicular parenchyma in stallions. Reproduction in Domestic Animals, Proceedings of the 22nd Annual Conference of the European Society for Domestic Animal Reproduction (ESDAR), Cordoba, Spain, 27\u201329 September 2018, Wiley Blackwell."},{"key":"ref_23","unstructured":"Munoz, I., Escartin, N., Zamora, A., and Echegaray, A. (2018). New echotexture parameters to evaluate the testicular parenchyma in boars. Reproduction in Domestic Animals, Proceedings of the 22nd Annual Conference of the European Society for Domestic Animal Reproduction (ESDAR), Cordoba, Spain, 27\u201329 September 2018, Wiley Blackwell."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"S133","DOI":"10.1017\/S1751731118001167","article-title":"Review: Testicular vascular cone development and its association with scrotal thermoregulation, semen quality and sperm production in bulls","volume":"12","author":"Kastelic","year":"2018","journal-title":"Animal"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1016\/j.rvsc.2013.04.023","article-title":"Evaluation of testicular artery blood flow by Doppler ultrasonography as a predictor of spermatogenesis in the dog","volume":"95","author":"Zelli","year":"2013","journal-title":"Res. Vet. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.anireprosci.2018.08.027","article-title":"Effect of seasonality on testicular blood flow as determined by color Doppler ultrasonography and hormonal profiles in Shiba goats","volume":"197","author":"Samir","year":"2018","journal-title":"Anim. Reprod. Sci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.theriogenology.2018.09.032","article-title":"Monthly changes in testicular blood flow dynamics and their association with testicular volume, plasma steroid hormones profile and semen characteristics in rams","volume":"123","author":"Hedia","year":"2019","journal-title":"Theriogenology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1111\/rda.13279","article-title":"Effects of testicular hemodynamic and echogenicity changes on ram semen characteristics","volume":"53","author":"Ntemka","year":"2018","journal-title":"Reprod. Domest. Anim."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1136\/vr.146.1.10","article-title":"Ultrasonography of spontaneous lesions of the genital system of three rams, and their influence on semen quality","volume":"146","author":"Ahmad","year":"2000","journal-title":"Vet. Rec."},{"key":"ref_30","first-page":"260","article-title":"Significance of Non-conventional Parameters in the Evaluation of Cooling-induced Damage to Ram Spermatozoa Diluted in Three Different Media","volume":"45","author":"Mendoza","year":"2010","journal-title":"Reprod. Domest. Anim."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"413","DOI":"10.3389\/fvets.2021.650572","article-title":"Influence of Non-conventional Sperm Quality Parameters on Field Fertility in Ovine","volume":"8","author":"Mendoza","year":"2021","journal-title":"Front. Vet. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1530\/jrf.0.0880343","article-title":"Use of fluorescent probes to assess membrane integrity in mammalian spermatozoa","volume":"88","author":"Harrison","year":"1990","journal-title":"J. Reprod. Fertil."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2089","DOI":"10.2527\/jas.2005-766","article-title":"Determination of intracellular reactive oxygen species and high mitochondrial membrane potential in Percoll-treated viable boar sperm using fluorescence-activated flow cytometry1","volume":"84","author":"Guthrie","year":"2006","journal-title":"J. Anim. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/0012-1606(84)90084-8","article-title":"Determination of the time course of capacitation in mouse spermatozoa using a chlortetracycline fluorescence assay","volume":"104","author":"Ward","year":"1984","journal-title":"Dev. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1071\/R96046","article-title":"Capacitation status and fertility of fresh and frozen-thawed ram spermatozoa","volume":"9","author":"Gillan","year":"1997","journal-title":"Reprod. Fertil. Dev."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Hafez, E.S.E., and Hafez, B. (2000). Reproduction in Farm Animals, Wiley Blackwell. [7th ed.].","DOI":"10.1002\/9781119265306"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"714","DOI":"10.2527\/1998.763714x","article-title":"Evaluation of sexual behavior of hair sheep rams in a tropical environment","volume":"76","author":"Godfrey","year":"1998","journal-title":"J. Anim. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"384","DOI":"10.11606\/issn.2318-3659.v50i5p384-395","article-title":"Correlations between testicular hemodynamic and sperm characteristics in rams","volume":"50","author":"Batissaco","year":"2013","journal-title":"Braz. J. Vet. Res. Anim. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2554","DOI":"10.1017\/S1751731120001330","article-title":"The continuous presence of ewes in estrus in spring influences testicular volume, testicular echogenicity and testosterone concentration, but not LH pulsatility in rams","volume":"14","author":"Abecia","year":"2020","journal-title":"Animal"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.smallrumres.2012.11.036","article-title":"Seasonal changes of scrotal circumference, blood plasma testosterone concentration and semen characteristics in Racka rams","volume":"111","author":"Egerszegi","year":"2013","journal-title":"Small Rumin. Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0301-6226(12)80009-1","article-title":"Physiology and genetics of testicular size in sheep: A review","volume":"32","author":"Matos","year":"1992","journal-title":"Livest. Prod. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1530\/jrf.0.0710351","article-title":"Changes in photoperiod and nutrition and their effect on testicular growth of rams","volume":"71","author":"Lindsay","year":"1984","journal-title":"Reproduction"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/j.theriogenology.2003.10.004","article-title":"Seasonal variation in testicular volume and sexual behavior of Chios and Serres rams","volume":"62","author":"Avdi","year":"2004","journal-title":"Theriogenology"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1016\/j.tvjl.2013.06.001","article-title":"Preliminary assessment of the quantitative relationships between testicular tissue composition and ultrasonographic image attributes in the ram","volume":"198","author":"Ahmadi","year":"2013","journal-title":"Vet. J."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1016\/0093-691X(96)00190-2","article-title":"Changes in circulating hormone concentrations, testes histology and testes ultrasonography during sexual maturation in beef bulls","volume":"46","author":"Evans","year":"1996","journal-title":"Theriogenology"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/j.smallrumres.2003.07.007","article-title":"Seasonal variation in semen characteristics, scrotal circumference and libido of Persian Karakul rams","volume":"53","author":"Kafi","year":"2004","journal-title":"Small Rumin. Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1177\/074873001129002051","article-title":"Biology of mammalian photoperiodism and the critical role of the pineal gland and melatonin","volume":"16","author":"Malpaux","year":"2001","journal-title":"J. Biol. Rhythm."},{"key":"ref_48","unstructured":"Senger, P.L. (2003). Pathways to Pregnancy and Parturition, Current Conceptions, Inc.. [2nd ed.]."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1016\/j.theriogenology.2004.12.008","article-title":"Relationship between semen quality and pixel\u2013intensity of testicular ultrasonograms after scrotal insulation in beef bulls","volume":"64","author":"Arteaga","year":"2005","journal-title":"Theriogenology"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.anireprosci.2017.01.015","article-title":"Prospective ultrasonographic and endocrine predictors of spermatogenic onset in ram lambs","volume":"179","author":"Bartlewski","year":"2017","journal-title":"Anim. Reprod. Sci."},{"key":"ref_51","first-page":"493","article-title":"Role of sperm apoptosis and oxidative stress in male infertility: A narrative review","volume":"19","author":"Asadi","year":"2021","journal-title":"Int. J. Reprod. Biomed."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"616821","DOI":"10.4161\/oxim.1.1.6843","article-title":"Antioxidant Systems and Oxidative Stress in the Testes","volume":"1","author":"Aitken","year":"2008","journal-title":"Oxid. Med. Cell Longev."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1002\/jcu.20439","article-title":"Long-term effect of varicocele repair on intratesticular arterial resistance index","volume":"36","author":"Balci","year":"2008","journal-title":"J. Clin. Ultrasound"}],"container-title":["Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-7737\/11\/2\/261\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:15:54Z","timestamp":1760134554000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-7737\/11\/2\/261"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,8]]},"references-count":53,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["biology11020261"],"URL":"https:\/\/doi.org\/10.3390\/biology11020261","relation":{},"ISSN":["2079-7737"],"issn-type":[{"value":"2079-7737","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,8]]}}}