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<\/p>\n<\/p>\n
Y. Arboleda Mar\u00edn<\/strong>, D. A. Gonz\u00e1lez Gaviria<\/strong>, L. K. Lopera Mesa<\/strong>, P. C. Lalinde Acevedo<\/strong>, A. F. Mej\u00eda<\/strong>, W. D. Cardona Maya<\/strong> and H. Cardona Cadavid<\/strong>*<\/p>\n <\/a><\/p>\n <\/a> ▲<\/a><\/p>\n Reproductive evaluation, the selection of individual breeders, and reproductive biotechnologies are important tools in developing productive and reproductive rates. When choosing a male as a breeding future, determining its general health status and genotypic and phenotypic superiority are necessary. This study evaluated the conventional and functional seminal quality in ovine males. The semen of eleven ovines of different pure races was collected by electroejaculation. The following average values \u200b\u200bwere observed for each conventional parameter: scrotal circumference (CE, 34.3 cm), seminal volume (vol, 1.63 mL), concentration (C, 768.4 x 106<\/sup>\/mL), individual motility (MI, 80%), masal motility (MM, 4) and vigor (Vi, 3.7). On the other hand, functional analysis showed plasma membrane integrity (PMI) of 45.7%, mitochondrial membrane potential (MMP) of 38.5%, DNA fragmentation index (DFI) of 17.0%, lipoperoxidation of the membrane (LPO) of 32.7%, and the production of reactive oxygen species (ROS) of 28.56%. Key words:<\/strong> semen quality; flow cytometry; reproduction; sheep; semen<\/em><\/p><\/blockquote>\n <\/a> ▲<\/a><\/p>\n Ovine species in Colombia are kept by small producers as a source of food (meat and milk) of high nutritional value and income when marketing products such as skin, wool, and others (Barrios). In any productive activity, including sheep production, the evaluation, and control of aspects such as management, feeding, facilities, production, and reproduction, are determining factors for establishing the viability of sheep. The area of animal reproduction has been extensively studied. However, in practice, there are still many shortcomings that hinder the fertility and fertilization process, both in ovine species and in other domestic species: inadequate nutritional and sanitary conditions at holdings, poorly trained workers, and failures in artificial reproduction processes (Gonz\u00e1lez-Stagnaro, 2011).<\/p>\n The female and the male are essential and indispensable in a production system; however, it is essential to state that the male accounts for more than 80% of the economic viability of a farm (Barrios, 2007). Consequently, strong excellent genotypic and phenotypic selection of males is essential; therefore, it is necessary that the holding knows the reproductive status of each animal. The techniques most used today for collecting semen in domestic animals, mainly ruminants (bovines, sheep, and goats), are the artificial vagina and electroejaculation (Morillo, Salazar, & Castillo, 2012; Yamasaki Maza et al<\/em>., 2005). The use of FC in veterinary medicine has been increasing, though there is still much to evaluate (C\u00f3rdova-Izquierdo, Saltijeral Oaxaca, Mu\u00f1oz Mendoza, C\u00f3rdova-Jim\u00e9nez, & C\u00f3rdova-Jim\u00e9nez, 2006). An innovative idea has been to implement FC for sperm evaluation in male sheep. The objective of this study was to analyse the conventional and functional semen quality of sheep.<\/p>\n <\/a> ▲<\/a><\/p>\n Semen samples were collected from 11 sheep pure bred rams: Dorper (n<\/em>=4), Katahdin (n<\/em>=4), and Santa Ines (n<\/em>=3). The Ethics Committee approved this work for the Ethics Committee for Experimentation with Animals (CEEA) of the University of Antioquia in Act No. 109\/2014.<\/p>\n Each collection began with a physical examination of the whole animal, including the fore and hind limbs, physical examination of the foreskin, penis, scrotum, testicles, and epididymis, in addition to assessing body condition and the ruling out any vision problems. The seminal sample was obtained by using the electroejaculation method (Yamasaki Maza et al<\/em>., 2005). Once the sample was obtained, macroscopic evaluation was performed (volume, appearance, colour, and presence of foreign elements), and then it was divided into two fractions: one for conventional field tests or microscopic evaluation (mass motility (MM, scale 1 to 5), percentage of individual motility (MI), vigor (Vi, scale 1 to 5), sperm concentration (C, 106<\/sup>\/mL)) and the second was taken to the laboratory within 2 hours to quantify the mitochondrial membrane potential (MMP) using FC (Ospina Medina and Cardona Maya, 2015). This was performed using the dye 3,3\u2019-dihexyloxacarbocyanine iodide (DIOC6) (Mayorga-Torres et al<\/em>., 2013; 2015; Mayorga-Torres et al<\/em>., 2015), intracellular reactive oxygen species (ROS) (Mayorga-Torres et al<\/em>., 2013; 2015; Mayorga-Torres et al<\/em>., 2015), using 2\u2019,7\u2019-dichlorofluorescein diacetate (DCFH-DA) staining, plasma membrane integrity (PMI) (Mayorga-Torres et al<\/em>., 2015; B. J. Mayorga-Torres et al<\/em>., 2015; B. J. Mayorga-Torres et al<\/em>., 2013) using SYBR 14, lipoperoxidation (LPO) (Mayorga-Torres et al<\/em>., 2015; B. J. Mayorga-Torres et al<\/em>., 2015; B. J. Mayorga-Torres et al<\/em>., 2013) using BODIPY C11 dye, and DNA fragmentation index (DFI) (Mayorga-Torres et al<\/em>., 2015; B. J. Mayorga-Torres et al<\/em>., 2015; B. J. Mayorga-Torres et al<\/em>., 2013) using acridine orange.<\/p>\n The characteristics analysed in this study were described using descriptive statistics (Table 1) and Pearson\u2019s correlation (Table 2), all variables were continuous. The statistical program R-Project was used through the RCORR procedure and the HMISC library.<\/p>\n <\/a> ▲<\/a><\/p>\n No males with physical problems or lesions were found in the physical evaluation. The evaluation of the reproductive system organs was within the normal parameters. It should also be considered that the evaluated males belong to different breeds and different management systems.<\/p>\n Correlations between all analysed variables (Table 2) by conventional methods and FC were determined, with their respective confidence levels. A strongly positive correlation was obtained for essential variables in determining semen quality, such as Vi and MM, Vi and MI, MMP and MM, MMP and MI, PMI and MMP, and ROS and MMP, while strongly negative correlations were found for C and Vol, and LPO and CE, Table 2.<\/p>\n The values analysed by FC showed that an average of 38.5% of the cells evaluated present high MMP: 28.56% ROS production, 17% DNA fragmentation, 32.7% OLP, an average of 32.7% was obtained, and 45.7% of the analysed cells have intact MI.<\/p>\n <\/a> ▲<\/a><\/p>\n The average semen volume was 1.63 mL, which is higher than that reported by Avellaneda et al<\/em>. (2006) (0.6 mL) who also obtained the semen sample by electroejaculation, was higher than reported by C\u00f3rdoba-Izquierdo et al<\/em>. (2006) using an artificial vagina for natural mounting (0.57 mL), and similar to (Cabrera & Pantoja, 2008) obtained by artificial vagina (1.7 mL). It should be noted that the breeds used in the different studies cited are different from those examined in this study. Like sperm concentration, this characteristic presented a wide range of variation (0.25 to 4.5 mL and 133 to 2810 x 106<\/sup>\/ mL, respectively), possibly due to the heterogeneity presented in the study population regarding bred.<\/p>\n The average scrotal circumference was 34.3 cm, which is larger than the report by (Avellaneda et al<\/em>., 2006) (22.6 cm), despite a similar average age of males in both studies (31.25 vs<\/em>. 34.7 months).<\/p>\n The sperm concentration in the present study was 768.4 x 106<\/sup>\/mL, which was lower than that reported by (Avellaneda et al<\/em>., 2006) (1285.7 x 106<\/sup>\/ mL) and (Cabrera and Pantoja, 2008) (2735 x 106<\/sup>\/mL), but higher than that reported by (C\u00f3rdova-Izquierdo et al<\/em>., 2006) (206.4 x 106<\/sup>\/mL using an artificial vagina) and when obtaining the semen sample directly from the vagina after a natural mount (176.02 x 106<\/sup>\/mL) in the same study.<\/p>\n Regarding individual motility, this was 80% on average, which was higher than the report by (Avellaneda et al<\/em>., 2006) (58.87%) and (C\u00f3rdova- Izquierdo et al<\/em>., 2006) (64.5% using artificial vagina) and when obtaining the semen sample directly from the vagina after natural mating (60.4%) in the same study. Mass motility was 4.0 on average, similar to the report by (Cabrera and Pantoja, 2008).<\/p>\n Concerning the functional parameters, in humans as the most widely studied group, the reference limit for MMP has been estimated at 60% (Mayorga-Torres et al<\/em>., 2013; 2017; Ospina Medina and Cardona Maya, 2015), suggesting that it is low in the present study. It should be noted that MMP is a predictive characteristic in fertilisation and it is highly correlated with some of the most predictive conventional characteristics of semen quality, such as mass and individual motility (Table 2).<\/p>\n The production of a high ROS average was 28; this value is considered optimal in humans (Mayorga-Torres et al<\/em>., 2013; 2017). Likewise, ROS production is related to DFI, although a low negative correlation was observed in the present study (Table 2).<\/p>\n The results obtained for DFI show an average of 17.0% of DNA fragmentation, a high figure compared to that reported as optimal in humans (<15%) (Mayorga-Torres et al<\/em>., 2013; 2017), though this can be considered to still represent semen of acceptable quality and may indicate a high percentage of sperm viability. Regarding the LPO analysis, an average of 32.7% of cells with lipid oxidation was obtained; this is a functional parameter of interest because it is theoretically related to motility, although a low positive correlation is not significant in this study (Table 2). The PIM analysis shows that on average, 45.7% of the analysed cells have an intact MI which, as a parameter correlated with sperm viability, should be higher (> 50%) (Mayorga-Torres et al<\/em>., 2013; 2017) as reported as optimal in humans.<\/p>\n Of the five variables analysed in FC, only one, the production is ROS, and it was within the values reported as optimal for humans. Still, this cannot assure that the evaluated males have low fertility since they are different species, and it would therefore be necessary to determine the optimal values in specific ovine species.<\/p>\n In conclusion, the values obtained in the conventional evaluation may vary according to the method used to obtain the semen sample and the breed of the males. Therefore, more studies are needed to estimate the minimum acceptable values to conclude that a male has good fertility.<\/p>\n Given that this study has a number of uncontrolled variables, several of the results may not be significant; therefore, a larger study with controlled variables such as breed, environment, age, time of feedings, and several feedings should be considered to relate these variables with fertility.<\/p>\n <\/a> 1. AVELLANEDA, Y., F. RODR\u00cdGUEZ, H. GRAJALES, R. MART\u00cdNEZ and R. VASQUEZ (2006): Determinaci\u00f3n de la pubertad en corderos en el tr\u00f3pico alto colombiano por caracter\u00edsticas corporales, calidad del eyaculado y valoraci\u00f3n de testosterona. Livest. Res. Rural. Dev. 18, 10. <\/a> ▲<\/a><\/p>\n Reproduktivna procjena, odabir pojedina\u010dnih rasplodnih mu\u017ejaka i reproduktivne biotehnologije va\u017eni su alati u unaprje\u0111enju produktivnih i reproduktivnih stopa. Pri odabiru mu\u017ejaka za rasplod, potrebno je odre\u0111ivanje op\u0107eg zdravstvenog statusa te genotipske i fenotipske superiornosti. Klju\u010dne rije\u010di:<\/strong> kvaliteta sjemena, proto\u010dna citometrija, reprodukcija, ovce, sjeme<\/em><\/p><\/blockquote>\n","protected":false},"excerpt":{"rendered":" Y. Arboleda Mar\u00edn, D. A. Gonz\u00e1lez Gaviria, L. K. Lopera Mesa, P. C. Lalinde Acevedo, A. F. Mej\u00eda, W. D.<\/p>\n","protected":false},"author":8,"featured_media":0,"menu_order":8,"comment_status":"closed","ping_status":"open","template":"","format":"standard","meta":{"footnotes":""},"categories":[21],"tags":[1488,922,1170,722,1227],"issuem_issue":[1438],"ppma_author":[1489,1490,1491,1492,728,1493,1494],"class_list":["post-5081","article","type-article","status-publish","format-standard","hentry","category-original-scientific-articles","tag-kvaliteta-sjemena","tag-ovce","tag-protocna-citometrija","tag-reprodukcija","tag-sjeme","issuem_issue-veterinarska-stanica-54-1"],"yoast_head":"\n
\nYurani ARBOLEDA MARIN<\/strong>, Daniela Alejandra GONZ\u00c1LEZ GAVIRIA<\/strong>, Leidy Katerine LOPERA MESA<\/strong>, Semillero de Investigaci\u00f3n y Grupo de Estudio en Caprinos y Ovinos (GECO-GaMMA), Facultad de Ciencias Agrarias, Universidad de Antioquia. Medell\u00edn, Colombia; Paula Andrea LALINDE ACEVEDO<\/strong>, Walter D. CARDONA MAYA<\/strong>, Grupo Reproducci\u00f3n, Facultad de Medicina, Universidad de Antioquia, Medell\u00edn, Colombia; Andr\u00e9s Felipe MEJIA<\/strong>, Empresa de Biotecnolog\u00eda de la Reproducci\u00f3n Ge3, Medell\u00edn, Colombia; Henry CARDONA CADAVID<\/strong>*, (Corresponding author, e-mail: henry.cardona@udea.edu.co), Grupo de Investigaci\u00f3n en Gen\u00e9tica, Mejoramiento y Modelaci\u00f3n Animal (GaMMA), Universidad de Antioquia, Medell\u00edn, Colombia<\/div>\n![\"\"](\"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2021\/03\/pdf.png\")
<\/a>https:\/\/doi.org\/10.46419\/vs.54.1.1<\/a><\/div>\n<\/p>\n\nAbstract<\/a>Introduction<\/a>Material and methods<\/a>Results<\/a>Discussion<\/a>References<\/a>Sa\u017eetak<\/a><\/span><\/div>\n<\/div>\n\n
Abstract<\/h2>\n
\n
\nAdditionally, a high and positive correlation was found for variables concerning seminal quality: Vi and MM, Vi and MI, MMP and MM, MMP and MI, PMI and MMP, as well as ROS and MMP. In contrast, two strongly negative correlations were obtained (C vs<\/em>. Vol and LPO vs<\/em>. CE). These are the first estimates of functional seminal quality evaluation of sheep in Colombia. In addition, this study provides a baseline for conventional and functional seminal parameters in the region.<\/p>\nIntroduction<\/h2>\n
\n
\nTo date, the census of sheep exploitation in the country is very low, since there are only two million animals, distributed mainly in six regions: La Guajira (42.4%), Magdalena (10.9%), C\u00f3rdoba (7%), and Cesar (7.9%) and Boyac\u00e1 (6.7%) (Instituto Colombianao Agropecuario, 2018). However, at present, the business of sheep farming is increasing due to the increase in meat consumption and the profitability that entrepreneurs show in this activity, so much so that in recent years the development of productions and agribusiness in the sheep sector has promoted everything related to the import of animals, the incorporation of technology, the acquisition of equipment and implements and technical training.
\nThis has enabled the high demand for products to be partly met in Colombia and in some markets of neighbouring countries (Restrepo).<\/p>\n
\nReproductive examination of a male consists of a physical examination, in which scrotal circumference, body development, vision, good poise, body condition, and overall reproductive organ health are evaluated, in addition to a seminal quality test to know the male\u2019s fertilising capacity.<\/p>\n
\nSemen can be evaluated by conventional methods in which the quality is defined according to macroscopic characteristics, usually flow cytometry (FC) (Izquierdo et al<\/em>., 2016) to quantify functional seminal parameters that allow for better evaluation of the capacity for fertilisation (Laguado, 2007).<\/p>\nMaterial and methods<\/h2>\n
\nStudy area and population<\/h3>\n
Sample collection and evaluation<\/h3>\n
\nFinally, the scrotal circumference (SC) was measured.<\/p>\nStatistical analysis<\/h3>\n
![\"\"](\"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2022\/06\/table01-semen-quality.png\")
![\"\"](\"https:\/\/veterinarska-stanica-journal.hr\/wp-content\/uploads\/2022\/06\/table02-semen-quality.png\")
ROS: Reactive oxygen species; *: P<0.05; **: P<0.01.<\/figcaption><\/figure>\nResults<\/h2>\n
\nDiscussion<\/h2>\n
\n
\nReferences<\/strong> [… show]<\/span><\/a><\/span><\/p>\n ▲<\/a><\/p>\n
\n2.\tBARRIOS, C. (2007): Gu\u00eda pr\u00e1ctica de ovinocultura enfocada hacia la producci\u00f3n de carne. Bogot\u00e1, Colombia: Bacom LTDA.
\n3.\tCABRERA, V. and A. PANTOJA (2008): Influencia de los dilutores tris y ovine freezing sobre la integridad de la membrana citoplasm\u00e1tica durante la congelaci\u00f3n de semen de ovinos en pajillas de 0.5 ml. Revista de Investigaciones Veterinarias del Per\u00fa, 19, 152-159. 10.15381\/rivep.v19i2.1162
\n4.\tC\u00d3RDOVA-IZQUIERDO, A., J. SALTIJERAL OAXACA, R. MU\u00d1OZ MENDOZA, M. C\u00d3RDOVA-JIM\u00c9NEZ and C. C\u00d3RDOVA-JIM\u00c9NEZ (2006): Efecto del m\u00e9todo de obtenci\u00f3n de semen de ovino sobre la calidad. REDVET. Revista Electr\u00f3nica de Veterinaria 7, 1-4.
\n5.\tGONZ\u00c1LEZ-STAGNARO, C. (2011): Buenas Pr\u00e1cticas Ganaderas (BPG) en el manejo de la reproducci\u00f3n en reba\u00f1os Doble Prop\u00f3sito. Innovaci\u00f3n & Tecnolog\u00edaen la Ganader\u00eda Doble Prop\u00f3sito.
\n6.\tInstituto Colombianao Agropecuario (2018): Censo Pecuario Nacional \u2013 2018. https:\/\/www.ica.gov.co\/areas\/pecuaria\/servicios\/epidemiologia-veterinaria\/censos-2016\/censo-2018.aspx.
\n7.\tIZQUIERDO, A. C., A. E. I. REYES, R. ESPINOSA, J. E. G. L. CERVANTES CASTRO, J. F. I. DE LOURDES, et al. (2016): Aplicaci\u00f3n de la citometr\u00eda de flujo en veterinaria a. Revista Complutense de Ciencias Veterinarias 10, 60-73. 10.5209\/RCCV.54514
\n8.\tLAGUADO, J. (2007): Aplicaciones de la citometr\u00eda de flujo en microbiolog\u00eda, veterinaria y agricultura. Revista MVZ C\u00f3rdoba 12, 1077-1095. 10.21897\/rmvz.430
\n9.\tMAYORGA TORRES, J. M., B. PE\u00d1A, A. P. CADAVID, W. D. CARDONA MAYA (2015): La importancia cl\u00ednica del ADN esperm\u00e1tico en el an\u00e1lisis seminal cotidiano. Revista chilena de obstetricia y ginecolog\u00eda 80, 265-268. 10.4067\/S0717-75262015000300012
\n10.\tMAYORGA-TORRES, B. J., M. CAMARGO, A. AGARWAL, S. S. DU PLESSIS, A. P. CADAVID and W. D. CARDONA MAYA (2015): Influence of ejaculation frequency on seminal parameters. Reprod. Biol. Endocrinol. 13, 47. 10.1186\/s12958-015-0045-9
\n11.\tMAYORGA-TORRES, B. J., W. CARDONA-MAYA, A. CADAVID and M. CAMARGO (2013): Evaluacion de los parametros funcionales espermaticos en individuos infertiles normozooespermicos. Actas Urol. Esp., 37, 221-227. 10.1016\/j.acuro.2012.06.008
\n12.\tMAYORGA-TORRES, B. J. M., M. CAMARGO, A. P. CADAVID, S. S. DU PLESSIS and W. D. CARDONA MAYA (2017): Are oxidative stress markers associated with unexplained male infertility? Andrologia 49. 10.1111\/and.12659
\n13.\tMORILLO, M., S. SALAZAR and E. CASTILLO (2012): Evaluaci\u00f3n del potencial reproductivo del macho bovino. Instituto Nacional de Investigaciones Agr\u00edcolas, 30-44.
\n14.\tOSPINA MEDINA, L., and W. D. CARDONA MAYA (2015): Evaluaci\u00f3n del potencial de membrana mitocondrial en espermatozoides humanos capacitados. Medicina & Laboratorio 21, 375-382. 10.36384\/01232576.135
\n15.\tRESTREPO, M. (2018): Plan estrat\u00e9gico para el desarrollo gremial 2010-2018. Bogot\u00e1, Colombia: Asoovinos.
\n16.\tYAMASAKI MAZA, A., P. PEDRAZA VILLAG\u00d3MEZ, M. M. PERALTA LAILSON, G. YONG ANGEL, J. E. ROTHSCHUH VILLANUEVA and D. YAMASAKI MAZA (2005): Dise\u00f1o y construcci\u00f3n de electroeyaculador para ovinos y caprinos. REDVET. Revista Electr\u00f3nica de Veterinaria, 6.
\n<\/em><\/p>\n<\/div>\n\n
Kvaliteta sjemena u ovaca<\/h2>\n
\nYurani ARBOLEDA MARIN<\/strong>, Daniela Alejandra GONZ\u00c1LEZ GAVIRIA<\/strong>, Leidy Katerine LOPERA MESA<\/strong>, Semillero de Investigaci\u00f3n y Grupo de Estudio en Caprinos y Ovinos (GECO-GaMMA), Facultad de Ciencias Agrarias, Universidad de Antioquia. Medell\u00edn, Colombia; Paula Andrea LALINDE ACEVEDO<\/strong>, Walter D. CARDONA MAYA<\/strong>, Grupo Reproducci\u00f3n, Facultad de Medicina, Universidad de Antioquia, Medell\u00edn, Colombia; Andr\u00e9s Felipe MEJIA<\/strong>, Empresa de Biotecnolog\u00eda de la Reproducci\u00f3n Ge3, Medell\u00edn, Colombia; Henry CARDONA CADAVID<\/strong>, Grupo de Investigaci\u00f3n en Gen\u00e9tica, Mejoramiento y Modelaci\u00f3n Animal (GaMMA), Universidad de Antioquia, Medell\u00edn, Colombia<\/div>\n
\n
\nOvaj rad procjenjuje konvencionalnu i funkcionalnu kvalitetu sjemena u ovnova.
\nSjeme jedanaest ovnova razli\u010ditih \u010distokrvnih pasmina prikupljeno je elektro ejakulacijom.
\nZamije\u0107ene su sljede\u0107e prosje\u010dne vrijednosti za svaki konvencionalni parametar: opseg skrotuma (CE, 34,3 cm), volumen sjemena (vol, 1,63 mL), koncentracija (C, 768,4 x 106<\/sup>\/mL), pojedina\u010dna pokretljivost (MI, 80 %), ukupna pokretljivost (MM, 4) i vitalnost (Vi, 3,7). S druge strane, funkcionalna analiza pokazala je integritet stani\u010dne membrane (PMI) od 45,7 %, potencijal mitohondrijske membrane (MMP) od 38,5 %, indeks fragmentacije DNK (DFI) od 17,0%, lipoperoksidaciju membrane (LPO) od 32,7 % i proizvodnju reaktivnih kisikovih spojeva (ROS) od 28,56 %. Uz to je otkrivena i visoka i pozitivna korelacija za varijable o kvaliteti sjemena: Vi i MM, Vi i MI, MMP i MM, MMP i MI, PMI i MMP, kao i ROS i MMP. Suprotno tome, dobivene su dvije vrlo negativne korelacije (C nasuprot Vol i LPO nasuprot CE). To su prve procjene ocjenjivanja funkcionalne kvalitete sjemena ovaca u Kolumbiji. Uz to, ova studija osigurava po\u010detnu vrijednost za sve konvencionalne i funkcionalne parametre sjemena u regiji.<\/p>\n