157 EFFECT OF DENSITY GRADIENT SELECTION ON EMBRYO RECOVERY RATES OF FRESH AND COOLED STALLION SEMEN

2013 ◽  
Vol 25 (1) ◽  
pp. 227
Author(s):  
F. Papa ◽  
M. Carmo ◽  
P. Papa ◽  
J. Dell'Aqua ◽  
M. Alvarenga
Keyword(s):  
Plasma Membrane ◽  
Density Gradient ◽  
Recovery Rate ◽  
Ovulation Induction ◽  
Membrane Integrity ◽  
Recovery Rates ◽  
Plasma Membrane Integrity ◽  
Embryo Recovery ◽  
Fresh Semen ◽  
Sperm Recovery

The aim of this study was to improve the spermatic kinetic parameters from stallions with poor quality of fresh and chilled semen by the use of density gradient Equipure™ (Nidacon, Mölndal, Sweden). Semen from 5 Quarter Horse stallions aged 8 and 16 years with history of low embryo recovery rates were used. The kinetics sperm evaluation was performed by computerized semen analysis (CASA) and plasma membrane integrity with fluorescent probes. The average motility parameters for fresh semen before selection were total motility (MT) 60%, progressive motility (PM) 30%, and plasma membrane integrity (IMP) 60% and for cooled semen (24 h at 5°C) were: MT 50%, MP 18%, and 50% IMP. For the group of fresh and cooled semen with no density gradient selection (NS), mares were inseminated with 1 billion viable sperm diluted in skim milk extender in the uterine body, 24 h after ovulation induction with 1 mg of deslorelin. For EquipureTM selection group (SE) semen was concentrated through Spermfilter membrane™ and resuspended in 5 mL of BotuSemenTM (BotupharmaTM, Brazil). In a 15 mL conic tube 5 mL of EquipureTM was added and another part containing 5 mL of the resuspended sperm was slowly added over the EquipureTM column. The 15 mL conic tube was centrifuged at 400g for 20 min. After centrifugation, the sperm pellet was carefully aspirated and to the pellet was resuspended in 4 mL of BotucrioTM. The sperm recovery rate with EquipureTM was 40%. Deep uterine AI was performed 24 h after ovulation induction with 1 mg of deslorelin. Data were analyzed by ANOVA followed by Tukey’s test (P < 0.05). The analysis of semen after EquipureTM selection resulted in average: 75, 35, 65, and 40%, respectively, for MT, MP, and IMP index and sperm recovery. The embryo recovery rate from the 5 stallions showed the following results: Stallion 1 (fresh semen): 12 mares (NS)/4 embryos (33%) and for group SE 22 mares/16 embryos, (72%; P ≤ 0.05). Stallion 2 (fresh semen): 9 mares (NS)/4 embryos, (44%) and for group SE, 12 mares/8 embryos (66%; P > 0.1). Stallion 3 (fresh semen): 4 mares (NS)/1 embryo (25%) and 7 mares for SE/4 embryos (57%; P > 0.1). Stallion 4 (chilled semen): 4 mares (NS)/0 embryo 0% and for SE group 8 mares/6 embryos 75%; P ≤ 0.001). Stallion 5 (chilled semen): 10 mares (NS)/2 embryos (20%) and for SE 6 mares/3 embryos (50%; P > 0.1). For the overall results, 39 inseminations were performed on the no selected group and 11 embryos recovered (28%) for the selected group 55 inseminations and 37 embryos recovered (67%; P < 0.01). The results clearly showed that selection by sperm density gradient EquipureTM was a very effective technique that allowed an improvement on semen quality and fertility. The authors acknowledge support from FAPESP and Botupharma.

65 ARE “BAD FREEZER” STALLIONS ALSO “BAD COOLER” STALLIONS?

2015 ◽  
Vol 27 (1) ◽  
pp. 125
Author(s):  
C. Ramires Neto ◽  
M. M. B. Castro-Chaves ◽  
Y. F. R. Sancler-Silva ◽  
R. C. Uliani ◽  
P. V. L. Oliveira ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Seminal Plasma ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Cooling Process ◽  
Freezing Resistance ◽  
Plasma Membrane Integrity ◽  
Progressive Motility ◽  
Frozen Semen ◽  
Fresh Semen

Several factors can interfere with sperm cryopreservation resistance, especially the genetic factors and those related to the plasma membrane composition of the sperm and seminal plasma. However, it is still unclear if the same factors that confer freezing resistance will perform the same role during the cooling process. Thus, the aim of this study was to determine the relation between the resistance to freezing and cooling processes in stallions. Two ejaculates from each of 75 stallions were used. All animals showed good quality of fresh semen (total motility higher than 60% and plasma membrane integrity higher than 50%). After collection, the semen was diluted 1 : 1 with commercial skim milk-based extender (Botu-SemenTM, Botupharma, Brazil) and then a part was designed to cooling and the another to freezing. The cooled semen was divided into 2 groups: Group PS, in which the semen was diluted with Botu-SemenTM at a concentration of 50 × 106 sperm mL–1, and Group SPS, which was subjected to a centrifugation at 600 × g for 10 min and resuspended with Botu-SemenTM at 50 × 106 sperm mL–1. Semen samples from both groups were placed in the same cooling passive system for a period of 24 h/5°C. To accomplish the freezing process, the semen sample was subjected to centrifugation at 600 × g for 10 min. The supernatant was discarded, and the pellet was re-suspended in a Botu-CrioTM. The straws were frozen according to the manufacture. The sperm parameters from fresh semen, cooled semen for 24 h with and without seminal plasma, and frozen semen were evaluated for kinetics by computer-assisted semen analysis and for plasma membrane integrity (IMP%) by epi-fluorescence microscopy. The animals were classified in relation to their resistance to cooling and freezing processes as follow: “bad coolers” – reduction in sperm total motility and in plasma membrane integrity higher than 35% after 24 h of cooling in samples with seminal plasma; “good coolers” – reduction in sperm total motility and in plasma membrane integrity lower than 35% after 24 h of cooling in samples with seminal plasma; “bad freezer” – sperm total motility lower than 40% and progressive motility lower than 20% in seminal sample after thawing; “good freezer” – sperm total motility higher than 60% and progressive motility higher than 30% in seminal sample after thawing. The comparison between the resistance to cooling and freezing processes was performed by Fisher's exact test. The level of significance was 5%. No difference (P < 0.05) between the resistance to cooling and freezing processes was observed. The percentage of stallions “good freezer” and “good cooler” was 54%, “good freezer” and “bad cooler” was 22.6%, “bad freezer” and “good cooler” was 12%, and “bad freezer” and “bad cooler” was 10.6%. Within stallions classified as “good freezer” and “bad cooler,” 52.9% also were “good cooler” when the seminal plasma was removed before the cooling process, and 47.1% remained as “bad cooler.” The result of this study demonstrates that there is a strong relation between the resistance to cooling and freezing processes in stallions. In stallions categorized as “bad cooler,” the seminal plasma presents a major influence on the quality and longevity of cooled semen.

Cryoprotective Effect of Glycerol Concentrations on Indian Red Jungle Fowl (Gallus Gallus Murghi) Spermatozoa

2018 ◽  
Vol 11 (2) ◽  
pp. 80-88 ◽  
Author(s):  
Bushra Allah Rakha ◽  
Muhammad Sajjad Ansari ◽  
Shamim Akhter ◽  
Elisabeth Blesbois
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Gallus Gallus ◽  
Recovery Rates ◽  
Plasma Membrane Integrity ◽  
Red Jungle Fowl ◽  
Frozen Semen ◽  
Acrosome Integrity

Semen cryopreservation protocols for wild avian species need to be optimised in order to achieve optimum post-thaw sperm quality and fertility. The present study was designed to evaluate the cryoprotective effect of different glycerol concentrations (11%, 15% and 20%) on post-thaw quality, recovery rates, absolute livability index and fertility of Indian Red Jungle Fowl (Gallus gallus murghi) semen. Semen was collected from eight mature cocks and cryopreserved for storage at −196 °C. Frozen semen was thawed at 37 °C for 30 s and assessed for motility, plasma membrane integrity, viability and acrosome integrity at 0, 2 and 4 h incubation at 37 °C. Percentages of motility, plasma membrane integrity, viability and acrosome integrity were recorded higher (P<0.05) post-thaw at 0, 2 and 4 h at 37 °C with 20% glycerol compared to 15% and 11% glycerol. Likewise, recovery rates (%) of aforementioned parameters after cryopreservation and absolute livability index were observed highest (P<0.05) with 20% glycerol. By comparing values of R2 after multivariate regression analysis, least negative effects of hours of incubation were observed on semen quality in extenders with 20% glycerol followed by 15% and 11% glycerol. The fertility outcomes (number of fertile eggs, fertility [%], number of hatched chicks, percent hatch and hatchability of fertilised eggs) were recorded higher (P<0.05) with 20% glycerol followed by 15% and 11% glycerol. It is concluded that the concentration of 20% glycerol gives the best cryoprotection for quality and fertility of Indian Red Jungle Fowl semen.

The effects of Trolox on the quality of sperm from captive squirrel monkey during liquefaction in the extender ACP-118™

Zygote ◽  
2018 ◽  
Vol 26 (4) ◽  
pp. 333-335 ◽  
Author(s):  
D. V. C. Almeida ◽  
J. S. Lima ◽  
D. L. Leão ◽  
K. G. Oliveira ◽  
R. R. Santos ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Squirrel Monkey ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Plasma Membrane Integrity ◽  
Time Periods ◽  
Semen Extender ◽  
Fresh Semen

SummaryThe aim of this study was to evaluate the effect of incubating semen for different periods (90, 270 or 450 min) with or without Trolox® (100 or 150 µM) on the quality of sperm from Saimiri collinsi. Sperm motility, vigour, and plasma membrane integrity (PMI) were evaluated in both fresh semen and semen incubated for different time periods, i.e. 90, 270 or 450 min of incubation. Supplementation of semen extender with Trolox® 100 µM improved sperm motility, vigour and PMI for up to 270 min of incubation.

scholarly journals PENGARUH METODE PENCUCIAN SPERMATOZOA SAPI ACEH TERHADAP MOTILITAS, PERSENTASE HIDUP, DAN INTEGRITAS MEMBRAN PLASMA UTUH SPERMATOZOA (The Infuence of Aceh Bull Spermatozoa Washing Method on Spermatozoa Motility and Plasma Membrane Integrity of Intact Spermatozoa)

2015 ◽  
Vol 9 (2) ◽  
Author(s):  
Listin Handayani ◽  
Dasrul Dasrul ◽  
Muslim Akmal ◽  
Cut Nila Thasmi ◽  
Hamdan Hamdan ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Quality Parameters ◽  
Control Group ◽  
Plasma Membrane Integrity ◽  
Spermatozoa Motility ◽  
Sperm Washing ◽  
Fresh Semen

This study aimed to determine the effect of sperm washing by swim up and centrifugation in isotonic medium on sperm quality of aceh bull. In this study, fresh semen from healthy male aceh bull aged 3-4 months was collected using artificial vagina. Immediately after semen collection, fresh semen quality was examined macroscopically and microscopically. Subsequently, sperm washing was performed by centrifugation and swim up in sperm washing medium. Group 1 (P0) as control group, cement washed with isotonic solution (andromed medium: saline solution) with ratio of 1:8. 2. Group 2 (P1), cement was separated by centrifugation method, group 3 (P2), all cement was separated by swim up method then examined the sperm quality sperm washing results. Each treatment was repeated 5 times. Quality parameters measured were the percentage of spermatozoa motility, sperm viability, and plasma membrane integrity intact spermatozoa. Data were analyzed with analysis of variance one-way pattern, followed by Duncan's multiple test. The results showed the mean ± SD percentage of sperm motility of each treatment group (P0; P1; P2) respectively amounted to 72.00±3.74, 66.40±4.77, and 73.60±3.29%. The percentage of viability was 72.00 ±3.74%, 66.40±2.88%, 71.80±2.17%. The percentage of plasma membrane integrity is intact spermatozoa was 68.20±1.79%, 57.20±3.77%, 69.00±2.00%. Results of this study showed that the percentage of motility, live spermatozoa and plasma membrane integrity intact after separation by swim-up method were significantly different (P <0.05) compared with no separation.Key words: spermatozoa quality, aceh bulls, centrifugation, swim up

Comparision of DNA fragmentantion and plasma membrane integrity between chilled and frozen semen of bulls

2014 ◽  
Author(s):  
Mello Papa Patricia de ◽  
Carlos Ramires Neto ◽  
Priscilla Nascimento Guasti ◽  
Rosiara Rosaria Dias Maziero ◽  
Yame F R Sancler-Silva ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Integrity ◽  
Plasma Membrane Integrity ◽  
Frozen Semen

scholarly journals Plasma membrane integrity in health and disease: significance and therapeutic potential

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Catarina Dias ◽  
Jesper Nylandsted
Keyword(s):  
Plasma Membrane ◽  
Therapeutic Potential ◽  
Calcium Influx ◽  
Membrane Integrity ◽  
Cell Types ◽  
Physical Parameters ◽  
Membrane Repair ◽  
Plasma Membrane Integrity ◽  
Repair Mechanisms ◽  
And Function

AbstractMaintenance of plasma membrane integrity is essential for normal cell viability and function. Thus, robust membrane repair mechanisms have evolved to counteract the eminent threat of a torn plasma membrane. Different repair mechanisms and the bio-physical parameters required for efficient repair are now emerging from different research groups. However, less is known about when these mechanisms come into play. This review focuses on the existence of membrane disruptions and repair mechanisms in both physiological and pathological conditions, and across multiple cell types, albeit to different degrees. Fundamentally, irrespective of the source of membrane disruption, aberrant calcium influx is the common stimulus that activates the membrane repair response. Inadequate repair responses can tip the balance between physiology and pathology, highlighting the significance of plasma membrane integrity. For example, an over-activated repair response can promote cancer invasion, while the inability to efficiently repair membrane can drive neurodegeneration and muscular dystrophies. The interdisciplinary view explored here emphasises the widespread potential of targeting plasma membrane repair mechanisms for therapeutic purposes.

scholarly journals Plasma membrane integrity: implications for health and disease

BMC Biology ◽  
2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Dustin A. Ammendolia ◽  
William M. Bement ◽  
John H. Brumell
Keyword(s):  
Plasma Membrane ◽  
Membrane Damage ◽  
Membrane Integrity ◽  
Whole Body ◽  
Plasma Membrane Integrity ◽  
Plasma Membrane Damage ◽  
Cellular Environment ◽  
Health And Disease ◽  
Repair Pathways ◽  
The Impact

AbstractPlasma membrane integrity is essential for cellular homeostasis. In vivo, cells experience plasma membrane damage from a multitude of stressors in the extra- and intra-cellular environment. To avoid lethal consequences, cells are equipped with repair pathways to restore membrane integrity. Here, we assess plasma membrane damage and repair from a whole-body perspective. We highlight the role of tissue-specific stressors in health and disease and examine membrane repair pathways across diverse cell types. Furthermore, we outline the impact of genetic and environmental factors on plasma membrane integrity and how these contribute to disease pathogenesis in different tissues.

Assessment of the Functional Integrity of Hepatocytes: A Brief Review

1993 ◽  
Vol 21 (3) ◽  
pp. 324-329
Author(s):  
Jeffrey R. Fry ◽  
Alison H. Hammond
Keyword(s):  
Plasma Membrane ◽  
Trypan Blue ◽  
Membrane Integrity ◽  
Blue Dye ◽  
Plasma Membrane Integrity ◽  
Functional Integrity ◽  
Cellular Integrity

A variety of approaches to assessment of cellular integrity exist, based on tests of integrity of the plasma membrane, tests of metabolic competence, and asessment of morphology. By definition, such approaches address different aspects of cellular integrity and hence are not interchangeable indices of cellular integrity. Accordingly, it would be most appropriate to characterise hepatocyte preparations on the basis of more than just trypan blue dye exclusion (a test of plasma membrane integrity) as is customary. A scheme for the choice of the most appropriate mix of tests of cellular integrity is presented.

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