{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:29:47Z","timestamp":1787027387095,"version":"build-2736575974"},"reference-count":71,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T00:00:00Z","timestamp":1766016000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>The current understanding of physiological parameters and redox balance in buffalo bull semen is limited and derived from various breeds. Moreover, the effects of cryopreservation in various buffalo breeds remain unclear.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>This study aimed to investigate the relationships between physiological parameters and compare fresh (F) and frozen-thawed (T) semen in Italian Mediterranean buffalos (IMB; 7, bulls). Buffalo ejaculates were collected using an artificial vagina and cryopreserved using a standard protocol. Both F and T were analyzed by CASA and flow cytometry Semen parameters assessed included motility, viability (using PI or SG counterstains for each assay), acrosome integrity (PSA), mitochondrial membrane potential (JC1), mitochondrial integrity (MT), intracytoplasmic (DHE) and mitochondrial (MX) superoxide production, and other intracellular reactive oxygen species (CR). The T samples were assessed immediately after thawing (T0) and following 3 h incubation at 37\u00b0C (T3).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Results showed significant correlations (\n                      <jats:italic>p<\/jats:italic>\n                      &amp;lt; 0.05) between total motility (TM) and progressive motility (PM) with mitochondrial membrane potential (MMP) and mitochondrial integrity (MI). The TM, PM, MI, and MMP positively correlated (\n                      <jats:italic>p<\/jats:italic>\n                      &amp;lt; 0.05) with total reactive oxygen species (ROS) production and negatively with superoxide production. Cryopreservation significantly decreased TM from 92.1 \u00b1 5.66% to 72.61 \u00b1 18.62% (\n                      <jats:italic>p<\/jats:italic>\n                      &amp;lt; 0.05), H\n                      <jats:sub>2<\/jats:sub>\n                      O\n                      <jats:sub>2<\/jats:sub>\n                      production from 23.02 \u00b1 7.42% to 11.49 \u00b1 8.85% (\n                      <jats:italic>p<\/jats:italic>\n                      &amp;lt; 0.05), and MMP from 83.29 \u00b1 11.20 to 56.98 \u00b1 15.87% (\n                      <jats:italic>p<\/jats:italic>\n                      &amp;lt; 0.05). After 3 h incubation, T semen exhibited increased mitochondrial damage and superoxide production, with decreased total ROS production.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>In conclusion, cryopreservation and subsequent incubation significantly affect mitochondrial functions, which strongly correlate with sperm motility. A deeper understanding of sperm energy metabolism and its relationship with redox regulation could allow for the optimization of current assisted reproductive technologies (ART), as these factors play a crucial role in sperm viability, motility, and fertilization capacity, which are critical for optimizing outcomes in ART procedures.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fvets.2025.1733446","type":"journal-article","created":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T15:07:27Z","timestamp":1766070447000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Mitochondrial function and reactive oxygen species dynamics in Italian Mediterranean buffalo semen following cryopreservation and post-thaw incubation"],"prefix":"10.3389","volume":"12","author":[{"given":"Giada","family":"Loddo","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Elena","family":"Gelain","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gianfranco","family":"Gabai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Asia","family":"D'Andrea","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisabetta","family":"Montanari","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chiara","family":"Milani","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisa","family":"Giaretta","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,12,18]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1137","DOI":"10.1016\/j.theriogenology.2005.07.012","article-title":"Blood testosterone levels in Italian Mediterranean buffalo bulls managed in two different breeding conditions","volume":"65","author":"Malfatti","year":"2006","journal-title":"Theriogenology."},{"key":"B2","doi-asserted-by":"publisher","first-page":"1171","DOI":"10.1016\/j.theriogenology.2013.02.014","article-title":"Minimum number of spermatozoa per dose in Mediterranean Italian buffalo (Bubalus bubalis) using sexed frozen semen and conventional artificial insemination","volume":"79","author":"Gaviraghi","year":"2013","journal-title":"Theriogenology."},{"key":"B3","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1016\/j.anireprosci.2016.06.010","article-title":"Sperm DNA integrity in frozen-thawed semen from Italian Mediterranean buffalo bulls and its relationship to in vivo fertility","volume":"172","author":"Serafini","year":"2016","journal-title":"Ani Rep Sci."},{"key":"B4","doi-asserted-by":"publisher","first-page":"81","DOI":"10.4142\/jvs.2017.18.1.81","article-title":"Alginate encapsulation preserves the quality and fertilizing ability of Mediterranean Italian water buffalo (Bubalus bubalis) and Holstein Friesian (Bos taurus) spermatozoa after cryopreservation","volume":"18","author":"Perteghella","year":"2017","journal-title":"J Vet Sci."},{"key":"B5","doi-asserted-by":"publisher","first-page":"116","DOI":"10.3390\/vetsci11030116","article-title":"Effect of the season on blood changes of oxidative stress index in the Italian Mediterranean buffalo (Bubalis bubalis)","volume":"11","author":"De Matteis","year":"2024","journal-title":"Vet Sci."},{"key":"B6","doi-asserted-by":"publisher","first-page":"570413","DOI":"10.3389\/fvets.2020.570413","volume":"7","author":"Minervino","year":"2020","journal-title":"Front Vet Sci."},{"key":"B7","doi-asserted-by":"publisher","first-page":"317","DOI":"10.1111\/j.1439-0531.2012.02152.x","article-title":"Dell'Aquila M, Barile V. 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