Luis Vargas, M. Gutierrez-Reinoso, M. Barros-Rodríguez, R. Lima-Orozco, V. Andrade-Yucailla, M. García-Herreros
The feeding program carried out during the pre-partum and post-partum period in dairy cows is crucial to preserve their reproductive performance. The goal of the present study was to assess the effect of black wattle (Acacia mearnsii) supplementation on ovarian activity, uterine involution, and hormone profiles in dairy cows maintained at high-altitude conditions. A total of 10 Holstein dairy cows were divided into 2 groups: control (C; n=5) and supplemented group (AM; n=5). A dehydrated black wattle (AM; Acacia mearnsii) supplement was administered to AM group and adjusted to 22% crude protein per animal (Dehydrated Supplement; MS= 2.7 kg AM/day per cow, ~20% of total DM). Body weight (BW) was recorded individually in different periods (day 15 pre-partum, and day 19, 34, and 49 post-partum). Ovarian [follicles (FL), corpora lutea (CL)] and ovarian diameter (OD; mm)] and uterine structures [uterine horn thickness (HT; mm), cervix length (CrL; cm) and diameter (CD; cm), and uterine involution %] were assessed till day 34 postpartum. Moreover, hormone profiles such as thyroid-stimulating hormone (TSH; IU/mL), follicle-stimulating hormone (FSH; IU/mL), luteinizing hormone (LH; IU/mL), and progesterone (P4; ng/mL) were measured till day 49. Statistical differences were observed in BW when day 15 prepartum was compared to day 19 postpartum (p=0.05). However, no differences were observed between C and AM group regarding BW within each timepoint (p>0.05). No changes were observed in FL over time irrespective of the group studied (p<0.05). However, FL was significantly different when C and AM group were compared within each time-point (p< 0.05). Although no significant differences were observed on day 5 post- partum regarding the number of CL, significant differences were observed on day 25 and day 34 postpartum between groups (p> 0.05). OD increased from day 5 to Day 35 post-partum irrespective of the group assessed and differ between groups within time-points (p< 0.05). Differences were observed in TSH within day 19 and day 49 post-partum (p< 0.05). FSH was statistically different between groups on day 19 post-partum (p< 0.05). LH was more variable than TSH and FSH among and within time-points and groups (p< 0.05). Differences were observed between groups regarding P4 within day 19 and 49 post-partum (p< 0.05). In conclusion, overall although no differences were observed in BW between groups or time-points, the supplementation with AM showed differential patterns in reproductive tract structures/dimensions and hormone levels in dairy cows maintained at high-altitude conditions
{"title":"EFFECTS OF BLACK WATTLE (Acacia mearnsii) SUPPLEMENTATION ON OVARIAN ACTIVITY,\u0000 UTERINE INVOLUTION, AND HORMONE PROFILES IN DAIRY COWS AT HIGH-ALTITUDE\u0000 CONDITIONS","authors":"Luis Vargas, M. Gutierrez-Reinoso, M. Barros-Rodríguez, R. Lima-Orozco, V. Andrade-Yucailla, M. García-Herreros","doi":"10.18548/aspe/0010.09","DOIUrl":"https://doi.org/10.18548/aspe/0010.09","url":null,"abstract":"The feeding program carried out during the pre-partum and post-partum period in\u0000 dairy cows is crucial to preserve their reproductive performance. The goal of the\u0000 present study was to assess the effect of black wattle (Acacia mearnsii) supplementation\u0000 on ovarian activity, uterine involution, and hormone profiles in dairy cows maintained\u0000 at high-altitude conditions. A total of 10 Holstein dairy cows were divided into 2\u0000 groups: control (C; n=5) and supplemented group (AM; n=5). A dehydrated black wattle\u0000 (AM; Acacia mearnsii) supplement was administered to AM group and adjusted to 22% crude\u0000 protein per animal (Dehydrated Supplement; MS= 2.7 kg AM/day per cow, ~20% of total DM).\u0000 Body weight (BW) was recorded individually in different periods (day 15 pre-partum, and\u0000 day 19, 34, and 49 post-partum). Ovarian [follicles (FL), corpora lutea (CL)] and\u0000 ovarian diameter (OD; mm)] and uterine structures [uterine horn thickness (HT; mm),\u0000 cervix length (CrL; cm) and diameter (CD; cm), and uterine involution %] were assessed\u0000 till day 34 postpartum. Moreover, hormone profiles such as thyroid-stimulating hormone\u0000 (TSH; IU/mL), follicle-stimulating hormone (FSH; IU/mL), luteinizing hormone (LH;\u0000 IU/mL), and progesterone (P4; ng/mL) were measured till day 49. Statistical differences\u0000 were observed in BW when day 15 prepartum was compared to day 19 postpartum (p=0.05).\u0000 However, no differences were observed between C and AM group regarding BW within each\u0000 timepoint (p>0.05). No changes were observed in FL over time irrespective of the\u0000 group studied (p<0.05). However, FL was significantly different when C and AM group\u0000 were compared within each time-point (p< 0.05). Although no significant differences\u0000 were observed on day 5 post- partum regarding the number of CL, significant differences\u0000 were observed on day 25 and day 34 postpartum between groups (p> 0.05). OD increased\u0000 from day 5 to Day 35 post-partum irrespective of the group assessed and differ between\u0000 groups within time-points (p< 0.05). Differences were observed in TSH within day 19\u0000 and day 49 post-partum (p< 0.05). FSH was statistically different between groups on\u0000 day 19 post-partum (p< 0.05). LH was more variable than TSH and FSH among and within\u0000 time-points and groups (p< 0.05). Differences were observed between groups regarding\u0000 P4 within day 19 and 49 post-partum (p< 0.05). In conclusion, overall although no\u0000 differences were observed in BW between groups or time-points, the supplementation with\u0000 AM showed differential patterns in reproductive tract structures/dimensions and hormone\u0000 levels in dairy cows maintained at high-altitude conditions","PeriodicalId":36778,"journal":{"name":"Spermova","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42656652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The aim of the present study was to evaluate the effect of equilibration process on canine sperms after vitrification using coconut water extender with addition of soy lecithin and sucrose as nonpermeable cryoprotectants. Twelve ejaculates were collected separately by digital manipulation from 12 adult dogs. Only the second fraction of the ejaculate was used in this study, which was evaluated about volume, concetration, viability, total and progressive motility, kinetic parameters and morphology. After evaluation, semen was diluted with a coconut water extender (50% coconut water (v/v), 25% (v/v) distilled water and 25% (v/v) 5% anhydrous monosodium citrate solution) with addition of soy lecithin and fructose at 1% (v/v) and 0.25 M sucrose until final concentration of 100x106 spermatozoa/ml. Samples were divided into three aliquots and each of them was processed at different regimens: without equilibration, 5ºC for 30 minutes and 5ºC for 60 minutes and then vitrified by dropping 30 μl of sperm suspension directly into liquid nitrogen. Sperm pellets were devitrified at least one week later as three of them were dropped into 0.3 mL of CaniPlus AI (Minitüb, Germany), which was previously warmed in a water bath at 37ºC for 2 minutes. Sperm concentration and motility parameters were assayed using a computer-aided sperm analysis (CASA) system, viability-by supravital staining technique and morphology parameters were evaluated in Haemacolor® stained semen samples. In conclusion, our results demonstrate that when vitrification and coconut water extender with addition of 1% soy lecithin and 0.25 M sucrose as cryoprotectants were used, presence of equilibration time of 60 minutes returned the best canine sperm quality results.
{"title":"EFFECT OF EQUILIBRATION PROCESS ON CANINE SPERMATOZOA AFTER VITRIFICATION USING\u0000 COCONUT WATER EXTENDER WITH ADDITION OF SOY LECITHIN AND SUCROSE AS NONPERMEABLE\u0000 CRYOPROTECTANTS","authors":"A. Antonov, B. Ivanova","doi":"10.18548/aspe/0010.05","DOIUrl":"https://doi.org/10.18548/aspe/0010.05","url":null,"abstract":"The aim of the present study was to evaluate the effect of equilibration process\u0000 on canine sperms after vitrification using coconut water extender with addition of soy\u0000 lecithin and sucrose as nonpermeable cryoprotectants. Twelve ejaculates were collected\u0000 separately by digital manipulation from 12 adult dogs. Only the second fraction of the\u0000 ejaculate was used in this study, which was evaluated about volume, concetration,\u0000 viability, total and progressive motility, kinetic parameters and morphology. After\u0000 evaluation, semen was diluted with a coconut water extender (50% coconut water (v/v),\u0000 25% (v/v) distilled water and 25% (v/v) 5% anhydrous monosodium citrate solution) with\u0000 addition of soy lecithin and fructose at 1% (v/v) and 0.25 M sucrose until final\u0000 concentration of 100x106 spermatozoa/ml. Samples were divided into three aliquots and\u0000 each of them was processed at different regimens: without equilibration, 5ºC for 30\u0000 minutes and 5ºC for 60 minutes and then vitrified by dropping 30 μl of sperm suspension\u0000 directly into liquid nitrogen. Sperm pellets were devitrified at least one week later as\u0000 three of them were dropped into 0.3 mL of CaniPlus AI (Minitüb, Germany), which was\u0000 previously warmed in a water bath at 37ºC for 2 minutes. Sperm concentration and\u0000 motility parameters were assayed using a computer-aided sperm analysis (CASA) system,\u0000 viability-by supravital staining technique and morphology parameters were evaluated in\u0000 Haemacolor® stained semen samples. In conclusion, our results demonstrate that when\u0000 vitrification and coconut water extender with addition of 1% soy lecithin and 0.25 M\u0000 sucrose as cryoprotectants were used, presence of equilibration time of 60 minutes\u0000 returned the best canine sperm quality results.","PeriodicalId":36778,"journal":{"name":"Spermova","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49541248","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Daniel Gandarillas, Rosario Rios, Abel Quispe, Edith Torres, Angelica Puma
The objective was to evaluate the maturation time to obtain oocytes in Metaphase II (MII) stages and the rate of division and blastocysts after fertilization in alpacas. For this, the ovaries were obtained from benefited animals. Experiment 1: 441 oocytes were distributed in four times (26, 32, 38 and 42 hours) and matured in TCM-199. At the end of the maturation time, the oocytes were fixed in an ethanol solution: acetic acid (3:1) and stained with aceto-orcein (1%). Experiment 2: 959 oocytes were cultured under the same conditions as experiment 1 and fertilized with sperm obtained by artificial vagina and selected by the Swim Up method. At the end of the fertilization time, the presumed zygotes were cultured in SOFaa medium for 7 days. In experiment 1, 39.6 ± 8.0, 49.1 ± 18.6, 45.9 ± 9.1 and 38.2 ± 9.9% of oocytes were obtained in Metaphase-II for 26, 32, 38 and 42 h of maturation, respectively, without statistical difference (p>0.05). In experiment 2, the cleavage rate was 45.7 ± 3.5, 51.3 ± 10.2, 52.8 ± 5.6 and 50.9 ± 3.1 with no statistical difference between treatments (P>0.05) and the blastocyst rate was 20.5 ± 3.4, 30.5 ± 5.8, 21.2 ± 2.5 and 22.8 ± 3.5 for 26, 32, 38 and 42 hours, respectively, with a statistical difference between 32 hours and 26, 38 and 42 hours (p<0.05). The results obtained suggest that 32 hours of maturation are required to obtain 30.5% of blastocysts
{"title":"Evaluation of four times of maturation in vitro in alpaca oocytes on the rate of\u0000 maturation and embryonic development","authors":"Daniel Gandarillas, Rosario Rios, Abel Quispe, Edith Torres, Angelica Puma","doi":"10.18548/aspe/0010.02","DOIUrl":"https://doi.org/10.18548/aspe/0010.02","url":null,"abstract":"The objective was to evaluate the maturation time to obtain oocytes in Metaphase\u0000 II (MII) stages and the rate of division and blastocysts after fertilization in alpacas.\u0000 For this, the ovaries were obtained from benefited animals. Experiment 1: 441 oocytes\u0000 were distributed in four times (26, 32, 38 and 42 hours) and matured in TCM-199. At the\u0000 end of the maturation time, the oocytes were fixed in an ethanol solution: acetic acid\u0000 (3:1) and stained with aceto-orcein (1%). Experiment 2: 959 oocytes were cultured under\u0000 the same conditions as experiment 1 and fertilized with sperm obtained by artificial\u0000 vagina and selected by the Swim Up method. At the end of the fertilization time, the\u0000 presumed zygotes were cultured in SOFaa medium for 7 days. In experiment 1, 39.6 ± 8.0,\u0000 49.1 ± 18.6, 45.9 ± 9.1 and 38.2 ± 9.9% of oocytes were obtained in Metaphase-II for 26,\u0000 32, 38 and 42 h of maturation, respectively, without statistical difference (p>0.05).\u0000 In experiment 2, the cleavage rate was 45.7 ± 3.5, 51.3 ± 10.2, 52.8 ± 5.6 and 50.9 ±\u0000 3.1 with no statistical difference between treatments (P>0.05) and the blastocyst\u0000 rate was 20.5 ± 3.4, 30.5 ± 5.8, 21.2 ± 2.5 and 22.8 ± 3.5 for 26, 32, 38 and 42 hours,\u0000 respectively, with a statistical difference between 32 hours and 26, 38 and 42 hours\u0000 (p<0.05). The results obtained suggest that 32 hours of maturation are required to\u0000 obtain 30.5% of blastocysts","PeriodicalId":36778,"journal":{"name":"Spermova","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47292361","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
P. Baruselli, Bruna Catusi, L. Angelo de Abreu, B. Gonzales
It is well established that the timing of the first pregnancy in heifers has a major influence on their lifetime reproductive performance. However, zebuine heifers typically show delayed puberty with the first calving often occurring at around 36–48 months of age. Several factors affect the reproductive efficiency in heifers, such as weight, age, body condition, uterine development and genetics. The thickness of subcutaneous fat efficiently predicts the risk for the establishment of puberty and pregnancy in 14-month-old Nelore heifers submitted to FTAI. It was found that heifers with greater subcutaneous fat thickness were more likely to achieve puberty and to become pregnant at FTAI. Through adequate nutritional management combined with genetic selection for sexual precocity, it is possible to obtain also satisfactory reproductive efficiency in precocious primiparous. When the basic requirements (mainly nutrition and genetics) are met, the anticipation of the conception of heifers can be successfully established, improving the productivity and profitability of breeding livestock.
{"title":"FACTORS AFFECTING HASTEN OF REPRODUCTIVE AGE IN TAURINE AND ZEBUINE HEIFERS","authors":"P. Baruselli, Bruna Catusi, L. Angelo de Abreu, B. Gonzales","doi":"10.18548/aspe/0010.10","DOIUrl":"https://doi.org/10.18548/aspe/0010.10","url":null,"abstract":"It is well established that the timing of the first pregnancy in heifers has a\u0000 major influence on their lifetime reproductive performance. However, zebuine heifers\u0000 typically show delayed puberty with the first calving often occurring at around 36–48\u0000 months of age. Several factors affect the reproductive efficiency in heifers, such as\u0000 weight, age, body condition, uterine development and genetics. The thickness of\u0000 subcutaneous fat efficiently predicts the risk for the establishment of puberty and\u0000 pregnancy in 14-month-old Nelore heifers submitted to FTAI. It was found that heifers\u0000 with greater subcutaneous fat thickness were more likely to achieve puberty and to\u0000 become pregnant at FTAI. Through adequate nutritional management combined with genetic\u0000 selection for sexual precocity, it is possible to obtain also satisfactory reproductive\u0000 efficiency in precocious primiparous. When the basic requirements (mainly nutrition and\u0000 genetics) are met, the anticipation of the conception of heifers can be successfully\u0000 established, improving the productivity and profitability of breeding\u0000 livestock.","PeriodicalId":36778,"journal":{"name":"Spermova","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47081617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jesus Polo, Angela Brijaldo, M. Londoño, D. Velasco, J. Cardozo, Fabian Rueda
Seminal plasma proteins (SPP) are fundamental for oocyte fertilization by sperm cells. In bovine, the structure and function of SPP have been widely described in several studies, where the spermadhesin family has been highlighted. Spermadhesin proteins are closely related to sperm motility and viability along with protecting the sperm cells against oxidative stress. Spermadhesin-1, also known as acidic Seminal Fluid Proteins (aSFP), exhibits a redox activity that protects the sperm cells from reactive oxygen species (ROS), an important feature that could be taken in advantage to improve the postthaw seminal quality of sperm cells after cryopreservation processes. Therefore, the aim of this research was to produce heterologous aSFP using Escherichia coli as a cell factory. For this purpose, the DNA sequence of aSFP was inserted into a pDAss plasmid, containing a six-histidine tag (6xhis). The obtained construct was used to transform BL21 (DE3) E. coli competent cells. The expression trials were developed at three different temperatures (37°,25°, and 16°C) and two different concentrations of Isopropyl-β-D-1-thiogalactopiranoside (IPTG) as the expression inductor (0.5 y 1.0 mM). The aSFP-H6 produced was purified by affinity chromatography, and the peptide sequence was verified through mass spectrometry. Results evidenced the best conditions for the recombinant aSFP-H6 production (16°C and 1.0 mM of IPTG). In this sense, the viability and the conditions to produce heterologous bovine aSFP were described and could be further used to enhance cryopreservation bovine sperm mediums.
精浆蛋白（SPP）是精子细胞受精卵母细胞的基础。在牛中，SPP的结构和功能在几项研究中得到了广泛的描述，其中精子粘附素家族得到了强调。精子粘附素蛋白与精子活力和活力以及保护精子细胞免受氧化应激密切相关。精子粘附素-1，也被称为酸性精浆蛋白（aSFP），具有保护精子细胞免受活性氧（ROS）影响的氧化还原活性，这是一个重要特征，可以用来改善冷冻保存过程后精子细胞的精液质量。因此，本研究的目的是利用大肠杆菌作为细胞工厂生产异源aSFP。为此，将aSFP的DNA序列插入含有六组氨酸标签（6xhis）的pDAss质粒中。将获得的构建体用于转化BL21（DE3）大肠杆菌感受态细胞。表达试验在三种不同的温度（37°、25°和16°C）和两种不同浓度的异丙基-β-D-1硫代乳糖吡喃糖苷（IPTG）作为表达诱导剂（0.5 y 1.0mM）下进行。通过亲和层析纯化产生的aSFP-H6，并通过质谱法验证肽序列。结果证明了生产重组aSFP-H6的最佳条件（16°C和1.0mM IPTG）。从这个意义上讲，描述了产生异源牛aSFP的可行性和条件，并可进一步用于增强冷冻保存牛精子培养基。
{"title":"Heterologous expression of bovine spermadhesin-1 using escherichia coli as\u0000 biofactory","authors":"Jesus Polo, Angela Brijaldo, M. Londoño, D. Velasco, J. Cardozo, Fabian Rueda","doi":"10.18548/aspe/0010.08","DOIUrl":"https://doi.org/10.18548/aspe/0010.08","url":null,"abstract":"Seminal plasma proteins (SPP) are fundamental for oocyte fertilization by sperm\u0000 cells. In bovine, the structure and function of SPP have been widely described in\u0000 several studies, where the spermadhesin family has been highlighted. Spermadhesin\u0000 proteins are closely related to sperm motility and viability along with protecting the\u0000 sperm cells against oxidative stress. Spermadhesin-1, also known as acidic Seminal Fluid\u0000 Proteins (aSFP), exhibits a redox activity that protects the sperm cells from reactive\u0000 oxygen species (ROS), an important feature that could be taken in advantage to improve\u0000 the postthaw seminal quality of sperm cells after cryopreservation processes. Therefore,\u0000 the aim of this research was to produce heterologous aSFP using Escherichia coli as a\u0000 cell factory. For this purpose, the DNA sequence of aSFP was inserted into a pDAss\u0000 plasmid, containing a six-histidine tag (6xhis). The obtained construct was used to\u0000 transform BL21 (DE3) E. coli competent cells. The expression trials were developed at\u0000 three different temperatures (37°,25°, and 16°C) and two different concentrations of\u0000 Isopropyl-β-D-1-thiogalactopiranoside (IPTG) as the expression inductor (0.5 y 1.0 mM).\u0000 The aSFP-H6 produced was purified by affinity chromatography, and the peptide sequence\u0000 was verified through mass spectrometry. Results evidenced the best conditions for the\u0000 recombinant aSFP-H6 production (16°C and 1.0 mM of IPTG). In this sense, the viability\u0000 and the conditions to produce heterologous bovine aSFP were described and could be\u0000 further used to enhance cryopreservation bovine sperm mediums.","PeriodicalId":36778,"journal":{"name":"Spermova","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43314091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sperm morphology evaluation is an important parameter for determining the quality of semen and predicting fertility in rams. Different staining methods have been developed to detect the morphological status of sperm, but there is no optimized protocol, especially for animals yet. This study was designed to compare the results using SpermBlue®, Diff-Quick, and Coomassie Blue stain in the morphological evaluation of epididymal ram semen. In the study, samples collected from a Bafra ram (epididymal sperm/ram) known to have a good breeding history were diluted with Trisbased diluent and frozen. After thawing for each straw, three semen smears were made and stained with SpermBlue®, Diff-Quick, and Coomassie Blue. All morphological parameters were evaluated using a light microscope. 100 spermatozoa were examined randomly and classified according to their characteristics for each slide. In identifying morphological abnormalities, the staining protocols have compared amongst themselves, and no significant difference between DiffQuick and SpermBlue® staining methods was observed. However, significant differences were observed in midpiece abnormalities when SpermBlue® and Coomassie-Blue staining methods were compared, while significant difference was found in total abnormality in SpermBlue® and DiffQuick staining comparison (P <0.05). As a result, all staining methods evaluated can be easily optimized for laboratory conditions and used in the morphological analysis of ram semen.
{"title":"THE USE OF DIFFERENT STAINING METHODS IN THE EVALUATION OF FROZEN-THAWED CAUDA\u0000 EPIDIDYMAL RAM SPERM MORPHOLOGY","authors":"Cumali Kaya, M. Akar, Burcu Esin, M. Çevik","doi":"10.18548/aspe/0010.06","DOIUrl":"https://doi.org/10.18548/aspe/0010.06","url":null,"abstract":"Sperm morphology evaluation is an important parameter for determining the quality\u0000 of semen and predicting fertility in rams. Different staining methods have been\u0000 developed to detect the morphological status of sperm, but there is no optimized\u0000 protocol, especially for animals yet. This study was designed to compare the results\u0000 using SpermBlue®, Diff-Quick, and Coomassie Blue stain in the morphological evaluation\u0000 of epididymal ram semen. In the study, samples collected from a Bafra ram (epididymal\u0000 sperm/ram) known to have a good breeding history were diluted with Trisbased diluent and\u0000 frozen. After thawing for each straw, three semen smears were made and stained with\u0000 SpermBlue®, Diff-Quick, and Coomassie Blue. All morphological parameters were evaluated\u0000 using a light microscope. 100 spermatozoa were examined randomly and classified\u0000 according to their characteristics for each slide. In identifying morphological\u0000 abnormalities, the staining protocols have compared amongst themselves, and no\u0000 significant difference between DiffQuick and SpermBlue® staining methods was observed.\u0000 However, significant differences were observed in midpiece abnormalities when SpermBlue®\u0000 and Coomassie-Blue staining methods were compared, while significant difference was\u0000 found in total abnormality in SpermBlue® and DiffQuick staining comparison (P <0.05).\u0000 As a result, all staining methods evaluated can be easily optimized for laboratory\u0000 conditions and used in the morphological analysis of ram semen.","PeriodicalId":36778,"journal":{"name":"Spermova","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43571296","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The aim of this investigation was to evaluate the effect of the addition of GnRH at the beginning and end of a treatment with progesterone-releasing devices (P 4 ) and estradiol (E2) and the expression of estrus on pregnancy rates (PR) of lactating dairy cows inseminated at fixed time (FTAI). The estrus synchronization treatment consisted on the insertion of a P 4 device for 8 days and the administration of estradiol benzoate (EB) on the day of insertion and estradiol cypionate (ECP) at the time of device removal to induce ovulation. Experiment 1 was designed to determine whether the addition of GnRH at the start of the protocol improved the PR and if the division of the time of artificial insemination (AI) improved PR. The addition of GnRH on Day 0 did not improve PR (44% vs 40%) but the AI of cows showing estrus at 48 h and the administration of GnRH at 48 h and AI at 60 h of those not showing estrus resulted in higher PR than those cows that were FTAI at 48 h, regardless of estrus expression (53% vs 31%; P <0.01). Experiment 2 was designed to study follicular dynamics and assess whether the PR varies using different times of AI (48 h vs 60 h), according to the expression of estrus. The division of AI and adding GnRH at 48 h decreased the dispersion in the interval between AI and ovulation. There was an interaction (P<0.05) between the time of AI and expression of estrus. Pregnancy rates were higher in cows in estrus at 48 h that were AI at that time (47.8%) and those that were not in heat at 48 h and were AI at 60 h (53.4%) than those who were not in estrus at 48 h and were FTAI at that time (19.4%) and those that were in estrus at 48 h and were FTAI at 60 h (29.7%). In Experiment 3 PR was evaluated based on the addition or not of GnRH to cows not showing estrus at 48 h. Pregnancy rate was higher (P<0.05) in cows that were not in estrus at 48 h, that received GnRH at that time and were AI at 60 h than in cows not showing estrus and 48 h that were also AI at 60 h but without GnRH (56.0% vs 39.8%). Experiment 4 evaluated factors that may affect PR in cows treated with the protocol developed in this experiment. It was found that the heat stress of January, a body condition score <2.75 and the presence of only small follicles (<10 mm and without CL) adversely affect PR. In conclusion, the results of this series of experiments confirm the hypothesis that the inclusion of a dose of GnRH and the modification of the time of AI increase PR in lactating dairy cows treated with P 4 devices and estradiol that does not express estrus at the time of FTAI.
{"title":"Effect of estro expression and GnRH addition on pregnancy rate in lactating\u0000 holstein cows synchronized with estradiol and progesterone devices","authors":"Juan Tshopp, G. Bó","doi":"10.18548/aspe/0010.15","DOIUrl":"https://doi.org/10.18548/aspe/0010.15","url":null,"abstract":"The aim of this investigation was to evaluate the effect of the addition of GnRH\u0000 at the beginning and end of a treatment with progesterone-releasing devices (P 4 ) and\u0000 estradiol (E2) and the expression of estrus on pregnancy rates (PR) of lactating dairy\u0000 cows inseminated at fixed time (FTAI). The estrus synchronization treatment consisted on\u0000 the insertion of a P 4 device for 8 days and the administration of estradiol benzoate\u0000 (EB) on the day of insertion and estradiol cypionate (ECP) at the time of device removal\u0000 to induce ovulation. Experiment 1 was designed to determine whether the addition of GnRH\u0000 at the start of the protocol improved the PR and if the division of the time of\u0000 artificial insemination (AI) improved PR. The addition of GnRH on Day 0 did not improve\u0000 PR (44% vs 40%) but the AI of cows showing estrus at 48 h and the administration of GnRH\u0000 at 48 h and AI at 60 h of those not showing estrus resulted in higher PR than those cows\u0000 that were FTAI at 48 h, regardless of estrus expression (53% vs 31%; P <0.01).\u0000 Experiment 2 was designed to study follicular dynamics and assess whether the PR varies\u0000 using different times of AI (48 h vs 60 h), according to the expression of estrus. The\u0000 division of AI and adding GnRH at 48 h decreased the dispersion in the interval between\u0000 AI and ovulation. There was an interaction (P<0.05) between the time of AI and\u0000 expression of estrus. Pregnancy rates were higher in cows in estrus at 48 h that were AI\u0000 at that time (47.8%) and those that were not in heat at 48 h and were AI at 60 h (53.4%)\u0000 than those who were not in estrus at 48 h and were FTAI at that time (19.4%) and those\u0000 that were in estrus at 48 h and were FTAI at 60 h (29.7%). In Experiment 3 PR was\u0000 evaluated based on the addition or not of GnRH to cows not showing estrus at 48 h.\u0000 Pregnancy rate was higher (P<0.05) in cows that were not in estrus at 48 h, that\u0000 received GnRH at that time and were AI at 60 h than in cows not showing estrus and 48 h\u0000 that were also AI at 60 h but without GnRH (56.0% vs 39.8%). Experiment 4 evaluated\u0000 factors that may affect PR in cows treated with the protocol developed in this\u0000 experiment. It was found that the heat stress of January, a body condition score\u0000 <2.75 and the presence of only small follicles (<10 mm and without CL) adversely\u0000 affect PR. In conclusion, the results of this series of experiments confirm the\u0000 hypothesis that the inclusion of a dose of GnRH and the modification of the time of AI\u0000 increase PR in lactating dairy cows treated with P 4 devices and estradiol that does not\u0000 express estrus at the time of FTAI.","PeriodicalId":36778,"journal":{"name":"Spermova","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42760502","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The objective of this work was to carry out a review of how reproductive efficiency can be improved in dairies. Dairying in the world is demanding changes to which we have to constantly adapt if we want to remain in the system. The constant variation in the price of a liter of milk paid to the producer and in the cost of inputs that are affected by the demand and the economic policies of each country, have installed in the dairy producer the idea that milk production is a bad business due to the low profitability of the sector with respect to other less demanding agricultural activities such as agriculture. However, when analyzing production costs, there is a very wide variation between farms depending on the efficiency and effectiveness with which each establishment performs. The dairy chain has historically presented a dynamic conditioned by demand (internal and external) that ends up setting prices for the primary producer, thus conditioning the productive and technological path of the sector, giving rise to recurring conflicts of interest between producers and industrialists. The subsidy policies that are applied many times end up solving the specific situation of the producer, complicating the situation of the markets of other countries. The countries with the largest surpluses of milk, such as New Zealand, the United States of America, Germany, France, Australia and Ireland, have to sell a large part of their production to other markets, often complicating the producers of the importing countries, for which they must become increasingly efficient in order to compete with countries that subsidize primary production.
{"title":"Role of the advisor to improve reproductive efficiency and cash flow in intensive\u0000 dairy cattle production systems","authors":"J. Tschopp, G. Bó","doi":"10.18548/aspe/0010.21","DOIUrl":"https://doi.org/10.18548/aspe/0010.21","url":null,"abstract":"The objective of this work was to carry out a review of how reproductive\u0000 efficiency can be improved in dairies. Dairying in the world is demanding changes to\u0000 which we have to constantly adapt if we want to remain in the system. The constant\u0000 variation in the price of a liter of milk paid to the producer and in the cost of inputs\u0000 that are affected by the demand and the economic policies of each country, have\u0000 installed in the dairy producer the idea that milk production is a bad business due to\u0000 the low profitability of the sector with respect to other less demanding agricultural\u0000 activities such as agriculture. However, when analyzing production costs, there is a\u0000 very wide variation between farms depending on the efficiency and effectiveness with\u0000 which each establishment performs. The dairy chain has historically presented a dynamic\u0000 conditioned by demand (internal and external) that ends up setting prices for the\u0000 primary producer, thus conditioning the productive and technological path of the sector,\u0000 giving rise to recurring conflicts of interest between producers and industrialists. The\u0000 subsidy policies that are applied many times end up solving the specific situation of\u0000 the producer, complicating the situation of the markets of other countries. The\u0000 countries with the largest surpluses of milk, such as New Zealand, the United States of\u0000 America, Germany, France, Australia and Ireland, have to sell a large part of their\u0000 production to other markets, often complicating the producers of the importing\u0000 countries, for which they must become increasingly efficient in order to compete with\u0000 countries that subsidize primary production.","PeriodicalId":36778,"journal":{"name":"Spermova","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45957125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The incorporation of Genetics in bovine breeding herds through the use of Artificial Insemination at Fixed Time (TAI) has achieved an increase in profitability by allowing to obtain greater The objective of this review was to compare the performance of programs with natural service, treatment Synchronization before natural service, artificial insemination after oestrus detection and fixed-time artificial insemination (TAI) in beef cattle.
{"title":"Precision reproduction: role of the veterinarian on productivity in farming","authors":"G. Bó, E. Huguenine, P. Baruselli","doi":"10.18548/aspe/0010.17","DOIUrl":"https://doi.org/10.18548/aspe/0010.17","url":null,"abstract":"The incorporation of Genetics in bovine breeding herds through the use of\u0000 Artificial Insemination at Fixed Time (TAI) has achieved an increase in profitability by\u0000 allowing to obtain greater The objective of this review was to compare the performance\u0000 of programs with natural service, treatment Synchronization before natural service,\u0000 artificial insemination after oestrus detection and fixed-time artificial insemination\u0000 (TAI) in beef cattle.","PeriodicalId":36778,"journal":{"name":"Spermova","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43440166","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Moscoso, Jaime Reinoso, J. Samaniego, D. Argudo, J. Alvarado, D. Galarza
Glycerol (Gly) is the cryoprotective agent (CPA) most widely used to cryopreserve rooster semen. However, its cryoprotection, toxicity, and efficacy may vary in different breeds of roosters (e.g.,fighting rooster). In this sense, this investigation evaluated the effect of different concentrations of Gly added to the Lake-Ravie extender on the kinetic variables and plasma membrane integrity (PMI, equivalent to viability) of rooster spermatozoa. A total of 42 semen ejaculates from 6 Spanish fighting roosters (collected in 7 weekly sessions) by dorsal massage technique were used to conform 7-pools. Each pool was divided into 6-aliquots and then 6 treatments were formed according to Gly concentrations: 0 (control), 2% (Gly-2), 4% (Gly-4), 6% (Gly-6), 8% (Gly-8), and 10% (Gly10). The samples were frozen in static liquid nitrogen vapors, and their post-thaw sperm quality was analyzed using the CASA system (SCA-2018®) and Fluorescence (PI). Results after thawing showed that total (MT, %) and progressive (MP, %) motilities were higher (P<0.01) with the Gly-8 treatment compared to the control group and the Gly-4 and Gly-6 treatments. Regarding the postthaw kinetics, the oscillation (WOB, %) and the beat-cross frequency (BCF, Hz) were higher (P < 0.05) in the Gly-6, Gly-8, and Gly-10 treatments compared to their control. Finally, the PMI (%) was greater with the Gly-8 treatment compared to the Gly-2 treatment (P<0.05) and the control group (P<0.01). In conclusion, the addition of 8% glycerol to the freezing medium produced better sperm cryosurvival based on higher kinetics and integrity of the plasma membrane of fighting rooster spermatozoa.
{"title":"Cryopreservation of Fighting rooster spermatozoa: effect of different\u0000 concentrations of glycerol","authors":"A. Moscoso, Jaime Reinoso, J. Samaniego, D. Argudo, J. Alvarado, D. Galarza","doi":"10.18548/aspe/0010.04","DOIUrl":"https://doi.org/10.18548/aspe/0010.04","url":null,"abstract":"Glycerol (Gly) is the cryoprotective agent (CPA) most widely used to cryopreserve\u0000 rooster semen. However, its cryoprotection, toxicity, and efficacy may vary in different\u0000 breeds of roosters (e.g.,fighting rooster). In this sense, this investigation evaluated\u0000 the effect of different concentrations of Gly added to the Lake-Ravie extender on the\u0000 kinetic variables and plasma membrane integrity (PMI, equivalent to viability) of\u0000 rooster spermatozoa. A total of 42 semen ejaculates from 6 Spanish fighting roosters\u0000 (collected in 7 weekly sessions) by dorsal massage technique were used to conform\u0000 7-pools. Each pool was divided into 6-aliquots and then 6 treatments were formed\u0000 according to Gly concentrations: 0 (control), 2% (Gly-2), 4% (Gly-4), 6% (Gly-6), 8%\u0000 (Gly-8), and 10% (Gly10). The samples were frozen in static liquid nitrogen vapors, and\u0000 their post-thaw sperm quality was analyzed using the CASA system (SCA-2018®) and\u0000 Fluorescence (PI). Results after thawing showed that total (MT, %) and progressive (MP,\u0000 %) motilities were higher (P<0.01) with the Gly-8 treatment compared to the control\u0000 group and the Gly-4 and Gly-6 treatments. Regarding the postthaw kinetics, the\u0000 oscillation (WOB, %) and the beat-cross frequency (BCF, Hz) were higher (P < 0.05) in\u0000 the Gly-6, Gly-8, and Gly-10 treatments compared to their control. Finally, the PMI (%)\u0000 was greater with the Gly-8 treatment compared to the Gly-2 treatment (P<0.05) and the\u0000 control group (P<0.01). In conclusion, the addition of 8% glycerol to the freezing\u0000 medium produced better sperm cryosurvival based on higher kinetics and integrity of the\u0000 plasma membrane of fighting rooster spermatozoa.","PeriodicalId":36778,"journal":{"name":"Spermova","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43618744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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