scholarly journals Efficiency of Tris-Based Extender Steridyl for Semen Cryopreservation in Stallions

Animals ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 1801
Author(s):  
Elena Nikitkina ◽  
Artem Musidray ◽  
Anna Krutikova ◽  
Polina Anipchenko ◽  
Kirill Plemyashov ◽  
...  
Keyword(s):  
Artificial Insemination ◽  
Egg Yolk ◽  
Dna Integrity ◽  
Normal Morphology ◽  
Progressive Motility ◽  
Frozen Semen ◽  
Fertilizing Ability ◽  
Motility Of Sperm ◽  
In Pregnancy ◽  
Stimulation Of

The fertilizing ability of stallion sperm after freezing is lower than in other species. The search for the optimal extender, combination of extenders, and the freezing protocol is relevant. The aim of this study was to compare lactose-chelate-citrate-yolk (LCCY) extender, usually used in Russia, and Steridyl® (Minitube) for freezing sperm of stallions. Steridyl is a concentrated extender medium for freezing ruminant semen. It already contains sterilized egg yolk. Semen was collected from nine stallions, aged from 7 to 12 years old. The total and progressive motility of sperm frozen in Steridyl was significantly higher than in semen frozen in LCCY. The number of spermatozoa with normal morphology in samples frozen in LCCY was 60.4 ± 1.72%, and with Steridyl, 72.4 ± 2.10% (p < 0.01). Semen frozen in Steridyl showed good stimulation of respiration by 2.4-DNP, which indicates that oxidative phosphorylation was retained after freezing–thawing. No differences among the extenders were seen with the DNA integrity of spermatozoa. Six out of ten (60%) mares were pregnant after artificial insemination (AI) by LCCY frozen semen, and 9/12 (75%) by Steridyl frozen semen. No differences among extenders were seen in pregnancy rate. In conclusion, Steridyl was proven to be a good diluent for freezing stallion semen, even though it was developed for ruminants.

scholarly journals Comparative analysis of various step-dilution techniques on the quality of frozen Limousin bull semen

2020 ◽  
Vol 13 (11) ◽  
pp. 2422-2428
Author(s):  
Ani Atul Arif ◽  
Tulus Maulana ◽  
Ekayanti Mulyawati Kaiin ◽  
Bambang Purwantara ◽  
Raden Iis Arifiantini ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Artificial Insemination ◽  
Skim Milk ◽  
Egg Yolk ◽  
Dna Integrity ◽  
Dilution Technique ◽  
Bull Semen ◽  
Frozen Semen ◽  
One Step

Background and Aim: Indonesia has two National Artificial Insemination centers and 17 Regional Artificial Insemination Centers. The frozen semen production techniques differed between the centers, including the type of diluent and semen dilution technique. The aim of the research was to compare the quality of frozen Limousin bull semen diluted using different techniques. Materials and Methods: Semen was collected from three sexually mature Limousin bulls using an artificial vagina. Immediately after collection, the semen was evaluated macroscopically and microscopically. Semen that had >70% motile sperm and <20% sperm abnormality was divided into three tubes and diluted with skim milk-egg yolk (SMEY) using three different dilution techniques: One-step dilution (100% SMEY with 8% glycerol) at room temperature ([RT] 20°C until 25°C) two-step dilution (50% SMEY without glycerol at RT, stored at 5°C; and 50% SMEY with 16% glycerol after 1 h stored at 5°C); and three-step dilution (50% SMEY without glycerol at RT, stored at 5°C; and 50% SMEY with 16% glycerol added twice at 1 h and 1.5 h after being stored at 5°C). The diluted semen was loaded into 0.25 mL mini straws, equilibrated, and frozen using a freezing machine. Sperm motility, viability, membranes, DNA integrity, and concentrations of malondialdehyde (MDA) and aspartate aminotransferase (AST) enzymes were evaluated after thawing. Results: The results showed that there were no significant differences in sperm motility and DNA integrity between dilutions (p>0.05). However, sperm viability and membrane intactness of one-step dilutions were higher than those of three-step dilutions. The concentrations of MDA and AST enzymes of sperm in one-step dilutions were lower than those of three-step dilutions (p<0.05). Conclusion: It was concluded that the one-step-dilution technique was better than three-step dilution for cryopreservation of Limousin bull semen.

scholarly journals Viability of Peranakan Etawah Liquid Semen Preserved in Tris Substituted with Various Energy Sources

2020 ◽  
Vol 25 (2) ◽  
pp. 68
Author(s):  
Nurul Afzan Hilda Zakiya ◽  
A H Yanti ◽  
T R Setyawati
Keyword(s):  
Energy Source ◽  
Artificial Insemination ◽  
Block Design ◽  
Egg Yolk ◽  
Energy Sources ◽  
Randomized Block Design ◽  
Frozen Semen ◽  
Semen Preservation ◽  
Semen Extender ◽  
Fresh Semen

The use of liquid semen for artificial insemination program of Etawah crossbreed goat (PE) is an alternative to replace frozen semen which is constrained by limited and expensive facilities. Production of liquid semen is faster than frozen semen, but the viability of liquid semen which preserved with a standard extender such as tris egg yolk is very short. The purpose of this study was to determine the viability of PE goat semen in egg yolk tris substituted with energy sources such as glucose, galactose, and mannose and to determine the most efficient energy source for semen preservation. This research was conducted from August to September 2018 at the Artificial Insemination Center in Lembang, West Java. This study was designed in a randomized block design (RBD) consist of three experimental groups divided into five groups. Fresh semen of PE goats were preserved using extender which energy source has been modified. Results showed that using glucose in PE goat semen extender produced the best motility among other groups (64.29 ± 9.2%). The highest viability was found in extender with fructose substitution (86.76 ± 2.3%). The longest viability of liquid semen was found in the extender with glucose substitution. It lasted for six days.

scholarly journals Decreased bull fertility: age-related changes in sperm motility and DNA fragmentation

2020 ◽  
Vol 151 ◽  
pp. 01010
Author(s):  
Berlin P. Pardede ◽  
Iman Supriatna ◽  
Yudi Yudi ◽  
Muhammad Agil
Keyword(s):  
Dna Fragmentation ◽  
Sperm Motility ◽  
Artificial Insemination ◽  
Fertility Rate ◽  
Blue Staining ◽  
Age Group ◽  
Progressive Motility ◽  
Frozen Semen ◽  
Group 3 ◽  
Group 2

This study aimed to analyze the effect of the age of bulls on sperm motility and DNA fragmentation and its impact on fertility. Ninety-six frozen semen straw from eight bulls were divided into four groups based on age (group-1: 5-6 years; group-2: 7-8 years; group-3: 9-10 years; group-4: 11-12 years). Total and progressive motility were detected by using computer-assisted semen analysis (CASA), while DNA fragmentation was detected by Toluidine blue staining. Over 500 artificial insemination services in the field were used for fertility rate analysis. The results of the analysis of total motility, progressive, and DNA fragmentation in all age groups still meet the minimum standard for artificial insemination programs. Analysis of progressive motility and DNA fragmentation showed significant differences in each age group (P<0.01), whereas analysis of total motility showed no significant differences in group-2 (7-8 years) and group-3 (9-10 years) (P>0.01). Increased age in bulls correlated significantly with increased sperm DNA fragmentation (P<0.01), decreased total and progressive motility (P<0,01), and potentially reduced the fertility rate (P<0.01). In conclusion, although the quality of frozen semen still meets the standards for artificial insemination programs, the age factor in bulls needs to be considered for achieving maximum fertility.

scholarly journals Current status and advances in ram semen cryopreservation

2018 ◽  
Vol 69 (2) ◽  
pp. 911 ◽  
Author(s):  
A. NTEMKA ◽  
I. A. TSAKMAKIDIS ◽  
E. KIOSSIS ◽  
A. MILOVANOVIĆ ◽  
C. M. BOSCOS
Keyword(s):  
Artificial Insemination ◽  
Storage Temperature ◽  
Egg Yolk ◽  
Current Status ◽  
Freezing Process ◽  
Freezing And Thawing ◽  
Semen Cryopreservation ◽  
Functional Changes ◽  
Fertilizing Ability ◽  
Semen Preservation

Ram semen cryopreservation contributes to genetic improvement through artificial insemination, eliminates geographical barriers in artificial insemination application and supports the preservation of endangered breeds thus the conservation of biodiversity. Sperm freezing process induces ultrastructural, biochemical and functional changes of spermatozoa. Especially, spermatozoa’s membranes and chromatin can be damaged, sperm membranes’ permeability is increased, hyper oxidation and formation of reactive oxygen species takes place, affecting fertilizing ability and subsequent early embryonic development. Aiming to improve ram frozen-thawed semen’s fertilizing capacity, many scientific investigations took place. Among them the composition of semen extenders, was a main point of interest. Semen preservation extenders regulate and support an environment of adequate pH and buffering capacity to protect spermatozoa from osmotic and cryogenic stress. Therefore, permeating (glycerol, dimethyl sulfoxide) and non-permeat ing (egg yolk, skimmed milk) cryoprotectants, sugars (glucose, lactose, trehalose, raffinose), salts (sodium citrate, citric acid) and antioxidants (amino acids, vitamins, enzymes) have been added and tested. Moreover, semen dilution rate, storage temperature, cooling rate and thawing protocol, are also some key factors that have been studied. The research results of this scientific topic are encouraging, not only about the freezing and thawing procedures, but also about the improvement of the additives’ properties. However, further research is needed to enhance the fertilizing ability of ram frozen-thawed semen, making its use practical in sheep reproductive management by the application of cervical artificial insemination.

scholarly journals Effect of preservation time on the quality of frozen semen in indigenous rams

2015 ◽  
Vol 44 (1) ◽  
pp. 10-15 ◽  
Author(s):  
BBA Mahmuda ◽  
Azizun Nesa ◽  
BF Zohara ◽  
MGS Alam ◽  
FY Bari
Keyword(s):  
Semen Quality ◽  
Egg Yolk ◽  
Volume Density ◽  
Normal Morphology ◽  
Mean Values ◽  
Frozen Semen ◽  
Significant Difference ◽  
The Mean ◽  
Fresh Semen

The study was carried out to observe the effects of preservation time on the quality of frozen semen of indigenous rams. Semen was collected using AV once a week from 4 rams. Tris based with 10% egg yolk and 7% glycerol extender was used to extend and freezing the semen. Fresh semen was evaluated for volume, density, mass motility and concentration, and mean values were observed as 0.8±0.2ml, 3.0±0.3, 3.2±0.7, 3.9±0.7×109/ml, respectively. Significant difference (p<0.05) was found for all the parameters among the rams. Mean values of motility, viability and normal morphology percentages were 83.3±4.3%, 88.2±4.4%, 84.2±3.5% in fresh semen while those of chilled semen at 40C were 74.7±2.3, 78.8±4.9 and 79.2±2.9%, respectively. For all the parameters, significant (p<0.05) difference was found among the rams. Frozen sperm motility was observed after thawing at 39-400C for 14-15 seconds. The mean motility, viability and normal morphology percentages after freezing for 24hrs, 7, 15 and 30 days of duration were 39.8±3.1, 41.1±4.3, 40.1±4.1 and 39.4±2.9%; 44.5±2.5, 45.3±2.8, 44.6±2.8 and 43.9±2.8%; 71.0±2.0, 71.7±1.5, 70.7±1.7 and 70.3±1.8%, respectively and values did not decrease significantly (p>0.05) with the increasing time of preservation. Non significantly decrease of the semen quality with advance of preservation time indicates the suitability of the protocol used for freezing of indigenous ram semen in Bangladesh.DOI: http://dx.doi.org/10.3329/bjas.v44i1.23113            Bang. J. Anim. Sci. 2014. 44 (1): 10-15

21 Effect of egg yolk extracted low-density lipoprotein on cryopreserved Nguni bull semen

2019 ◽  
Vol 31 (1) ◽  
pp. 136
Author(s):  
M. M. Tshabalala ◽  
K. A. Nephawe ◽  
M. L. Mphaphathi ◽  
C. M. Pilane ◽  
T. L. Nedambale
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Semen Parameters ◽  
Dna Integrity ◽  
Low Density ◽  
Bull Semen ◽  
Frozen Semen ◽  
Live Sperm

Egg yolk has been reported to have a beneficial effect on sperm quality following cryopreservation, and this led to its widespread use in semen extenders. However, egg yolk contains substances that inhibit respiration of sperm cells and diminish their motility rate. Moreover, it also contains low-density lipoproteins (LDL) that have a protective effect on sperm during the cryopreservation process. The objective of this study was improve cryopreservation of Nguni bull semen using egg yolk low-density protein. A total of 25 ejaculates were collected from 5 Nguni bulls aged 4 to 5 years using an electroejaculator during the natural breeding season. Collected raw semen samples were transported to the laboratory and evaluated for sperm motility, viability, plasma membrane, acrosome, and DNA integrity before dilution. Semen was randomly diluted with a sodium citrate extender supplemented with 20% egg yolk (control) and with 6, 8, 10, and 12% LDL concentrations. The diluted semen sample groups were equilibrated for 4h at 5°C. Following equilibration, semen was loaded into 0.25-mL straws and frozen in a controlled-rate programmable freezer. The groups of semen straws were then plunged into LN and transferred into LN tanks (−196°C) for storage. The frozen semen straws per treatment group were thawed at 37°C and evaluated for sperm motility, viability, plasma membrane, acrosome, and DNA integrity. Data were analysed with ANOVA using Stata V12 statistical software (StataCorp., College Station, TX), and treatment means were separated using Fisher’s protected t-test at the significant level of P&lt;0.05. Sperm motility and membrane integrity were significantly higher (P&lt;0.05) in frozen-thawed semen diluted with 8% LDL compared with the other concentrations. However, 6 and 8% LDL resulted in a significantly higher (P&lt;0.05) live sperm, DNA, and acrosome integrity. Frozen-thawed semen diluted with 10 and 12% LDL resulted in the lowest percentages of sperm motility, live sperm, plasma membrane, acrosome, and DNA integrity following cryopreservation. In conclusion, extender containing 8% LDL resulted in improved Nguni bull semen parameters such as sperm motility, viability, plasma membrane, acrosome, and DNA integrity following cryopreservation. Further studies are required to determine the fertilizing capacity of semen diluted and frozen with LDL in vitro and in vivo.

Natural bee honey and Conservation of drone semen in liquid nitrogen

2022 ◽  
pp. 17-22
Author(s):  
A. Gulov ◽  
A. Laskin
Keyword(s):  
Liquid Nitrogen ◽  
Honey Bees ◽  
Rock Type ◽  
Egg Yolk ◽  
Deep Freezing ◽  
Glass Capillaries ◽  
Fertilizing Ability ◽  
June July ◽  
Selection Of ◽  
Stimulation Of

Purpose: Conducting a honey diluent test for creeples of sperm of a drone honey bee.Materials and methods. The material for the research served a sperm of the milled drone drums of the "Prioksky" type of the Midway breed of bees. The selection of sperm was carried out in June-July 2020 g by the method of artificial stimulation of the turning of the endofalosha in half-armed drones aged 25-30 days. The rock type "Prioksky" of the middle Russian breed of bees. Before freezing, the sperm was stored in glass capillaries in the cooled state at 3 ° C for 2 months. The following composition of the diluent was tested - 10% honey, lactose, sucrose, egg yolk and dimethyl sulfoxide.Results. Studies have shown the viability of sperm at 64.0 ± 1.8% (41.5-83.7), and a total mobility of 2.2 ± 0.6% (0-11.5). To evaluate the fertilizing ability of sperm, carried out artificial insemination of 10 bee modules. In 4 seeded bees dykens, the presence of sperm in a seed-hearter with a concentration of sperm from 0.22-4.4 million / μl is revealed. In paired eggs of three other seeded matters, the presence of sperm and the complete absence of spermatozoa in the seed-receptionist are recorded.Conclusion. Tests of the honey diluent for deep freezing sperm of the drone honey bees in liquid nitrogen confirmed its cryophylactic properties.

scholarly journals The effect of freezing on cryosurvival of yak sperm

2018 ◽  
Vol 15 (2) ◽  
pp. 215-218
Author(s):  
S Deori
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Artificial Insemination ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
Mean Values ◽  
Frozen Semen ◽  
Sperm Abnormality ◽  
Semen Characteristics ◽  
Live Sperm

A study was carried out to study the effect of freezing on cryosurvival of yak semen. Artificial insemination in yak is still in infancy. Semen cryopreservation and use of artificial insemination can be applied in yak husbandry for conservation and rapid multiplication of superior germplasm. Semen was collected from four adult yak bulls using artificial vagina method managed under uniform conditions. A total of 40 ejaculates comprising of 10 ejaculates each bull were collected following twice a week schedule and evaluated for fresh semen characteristics. The fresh yak semen characteristics viz. ejaculate volume (ml), mass activity (0-4), initial sperm motility (%), sperm concentration (x 106/ml), live sperm (%), sperm abnormality (%) and intact acrosome (%) were 3.10 ± 0.18, 3.53 ± 0.96, 83.89 ± 2.87, 1180.22 ± 42.32, 77.63 ± 4.23, 8.45 ± 3.33 and 93.61 ± 3.78 respectively. The ejaculates were diluted (1:10) with Tris extender consisting of 6.4 ml glycerol and 20 ml of fresh egg yolk. Straws were equilibrated at 5°C for 4 hours followed by exposure to liquid nitrogen vapour for 10 minutes and finally transferred to liquid nitrogen container for storage. The cryosurvival rate was studied after 7 days of storage in liquid nitrogen. The frozen semen was thawed in warm water (37°C) for 30 seconds for evaluation. Mean values of postthaw sperm motility (%), live sperm (%) and intact acrosome (%) in yaks were 55.67 ± 4.67, 65.62 ± 3.23 and 89.26 ± 3.67 respectively. In conclusion, yak semen has a better cryosurvival while freezing in tris extender with 6.4 per cent glycerol and 20 per cent egg yolk following an equilibration period of 4h.SAARC J. Agri., 15(2): 215-218 (2017)

Cryopreservation and fertility of frozen thawed Chegu goat semen

2019 ◽  
Author(s):  
A. Sharma ◽  
P. Sood
Keyword(s):  
Endangered Species ◽  
Artificial Insemination ◽  
Arid Region ◽  
Egg Yolk ◽  
Conception Rate ◽  
Semen Parameters ◽  
Semen Cryopreservation ◽  
Livestock Species ◽  
Progressive Motility ◽  
Seminal Parameters

Goats are important livestock species of India. Chegu is a pashmina producing goat native to the cold arid region of Himachal Pradesh (H.P.), India. Semen cryopreservation from six Chegu elite males (aged 2.05±0.40 years; weighed 29.16±2.02 kg) was practiced using Tris Citrate Egg Yolk extender containing 10% EY and 6% Glycerol. Gross semen parameters includes volume (0.80.85±0.07 ml), color (Creamy white to yellowish), concentration (2238.5±231.0 x106 spermatozoa/ml) and mass motility (3.92±0.03).The significant changes (P less than 0.01) in post thaw seminal parameters (75.48±0.69 v/s 37.38±0.90; progressive motility), viability (75.79±0.95 v/s 48.25±1.78), morphological abnormalities (5.64±0.29 v/s 7.02±0.32) and HOST reactive spermatozoa (64.07±1.75 v/s 43.35±1.79) were observed in present study. Artificial insemination using frozen thawed semen having concentration (150 x106 spermatozoa/straw) from three different bucks was practiced in 40 synchronized goats with conception rate of 42.5 per cent. Non-significant variations amongst different bucks were observed with birth of 1.12 kids per doe and twinning rate of 11.8 per cent. It was concluded that semen cryopreservation along with artificial insemination can be practiced in Chegu goats to improve the population of this endangered species.

scholarly journals Effect of Monthly Variations on the Plasma Membrane, Acrosome and DNA Integrities of Spermatozoa in Friesian Holstein Bulls Born in Iraq

2020 ◽  
Vol 13 (2) ◽  
pp. 118-124
Author(s):  
Qusay Aboud ◽  
Saad Hatif
Keyword(s):  
Plasma Membrane ◽  
Artificial Insemination ◽  
Significant Decline ◽  
Dna Integrity ◽  
Frozen Semen ◽  
Negative Effect ◽  
Holstein Bulls ◽  
Monthly Variations ◽  
Artificial Vagina

This study was aimed to evaluate the influence of months of the year on the quality of semen in Holstein bulls. A study carried out at artificial insemination centre/ Abou-Ghareeb/ western of Baghdad. A total of 160 ejaculates were collected from 15 bulls born in Iraq via the artificial vagina. The age of the bulls ranged between 4 to 5 years and the study period were December to March and September. The semen samples were diluted with Tris base extender. The semen was packed in a straw according to the program of artificial insemination centre. Semen characteristics (plasma membrane, acrosome, and DNA integrities) were evaluated. The results revealed a significant decrease (P≤0.01) in the plasma membrane and acrosome integrity in September as compared with December, January, February, and March. There was a significant decline (P≤0.05) in DNA integrity in September as compared with December, January, February, and March in fresh and frozen semen. In conclusion, the September month had a negative effect on the plasma membrane, acrosome, and DNA percentage in all bulls.

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