Cryopreservation of mutton snapper ( Lutjanus analis) sperm

This study aimed to develop a protocol of semen cryopreservation of the mutton snapper Lutjanus analis. The interaction between three extenders ( pH 6.1; 7.8 and 8.2) , two concentrations of dimethyl sulfoxide ( DMSO, 5 and 10%) and three cooling rates ( -90; -60 and -30°C.min−1) on the sperm motility rate and motility time were analyzed by a factorial experiment. A sample of 30 fishes ( 1,261 ± 449 g) collected in the nature was kept in floating net cages. The semen was frozen by using cryogenic straws, in nitrogen vapour and transferred, later, to liquid nitrogen. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The highest sperm motility rate and motility time ( P < 0.05) was achieved by combining extender C ( pH 8.2) with DMSO ( 10%) and cooling rate of -60°C.min−1 ( P < 0.05) . The use of cryopreserved sperm presented fertilization rates higher than 59% validating the present protocol for mutton snapper.

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Sperm cryopreservation of lane snapper Lutjanus synagris(Linnaeus, 1758)

Brazilian Journal of Biology ◽

10.1590/1519-6984.20613 ◽

2015 ◽

Vol 75 (3) ◽

pp. 662-669 ◽

Cited By ~ 2

Author(s):

EG Sanches ◽

IR Oliveira ◽

PCS Serralheiro ◽

VR Cerqueira

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Equilibration Time ◽

Dilution Ratio ◽

Sperm Cryopreservation ◽

Ph Values ◽

Cryopreserved Sperm ◽

Time Density ◽

Motility Rate

AbstractThis study aims developing and evaluate a protocol of semen cryopreservation of the lane snapper Lutjanus synagris. Firstly, sperm motility rate, motility time, density and spermatocrit were appraised to characterize the sperm quality of the lane snapper. The effect of three extenders with distinct ionic compositions and pH values combined with seven concentrations of cryoprotector dimethylsulfoxide (0; 2.5; 5.0; 7.5; 10.0; 12.5 e 15.0%), five cooling rates (110, 90, 60, 45 e 30°C –min), nine equilibration time (1; 2,5; 5; 10; 15; 20; 25; 30 e 60 minutes) e five dilutions ratio (1:1; 1:3; 1:6; 1:10 e 1:20) on the sperm motility rate and motility time were analyzed. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The higher sperm motility rate and motility time (P<0.05) was achieved by combining extender with pH 8.2 with 10% concentration of dimethylsulfoxide and cooling rate 60°C –min, 1 minute of equilibration time and 1:3 (v/v) dilution ratio. The use of cryopreserved sperm presented fertilization rates >60% validating the present protocol for lane snapper. The cryoconserved sperm of lane snapper is a viable alternative, being possible to maintain appropriate sperm viability.

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Management of the storage of cryopreserved sperm on dairy cattle farms

Biotechnology in Animal Husbandry ◽

10.2298/bah1501085m ◽

2015 ◽

Vol 31 (1) ◽

pp. 85-100 ◽

Cited By ~ 2

Author(s):

R. Muiño ◽

A.I. Peña ◽

L.A. Quintela ◽

J. Becerra ◽

P. Herradón ◽

...

Keyword(s):

Multivariate Analysis ◽

Dairy Cattle ◽

Kinetic Parameters ◽

Liquid Nitrogen ◽

Sperm Motility ◽

Time Period ◽

Cryopreserved Sperm ◽

The Mean ◽

The Individual ◽

Cattle Farms

26 liquid nitrogen tanks were selected from different dairy cattle farms. Three sperm doses were introduced in a frequently used canister, while another three straws were deposited in another canister that did not contain any sperm doses, to determine whether the refilling with liquid nitrogen had been done appropriately. Then, 10 sperm doses belonging to the same freezing lot were stored in our laboratory under ideal conditions to be used as control doses. After certain time period, the doses were collected from the farms and were analysed to obtain data about their total sperm motility and the individual kinetic parameters of each sperm. Four sperm subpopulations (SP) with different patterns of motility were identified using a cluster multivariate analysis. The results show that the mean total sperm motility has hardly decreased for the doses stored in the frequently used canister (45.2 ? 6.9%) in comparison with the doses stored in the rarely used canister (46.9 ? 59.0). However, the decrease in total motility was greater when compared with the control doses (59.0%). As for the sperm SP, (SP4 rapid and progressive sperm) which contained 31% of the total of sperm (control doses), differed the most when control doses were compared to straw stored in farm tanks. The percentage of the latter was reduced to 10 % after being stored in the tanks of the farms for 7 mo. Such damage in SP 4 is progressive and cumulative and would probably reduce drastically compromising the fertility of the aforementioned sperm doses.

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57 EFFECT OF SEMINAL PLASMA REMOVAL ON SPERM CHARACTERISTICS AND MITOCHONDRIAL MEMBRANE FOLLOWING CRYOPRESERVATION OF SOUTH AFRICAN INDIGENOUS BUCK SEMEN

Reproduction Fertility and Development ◽

10.1071/rdv29n1ab57 ◽

2017 ◽

Vol 29 (1) ◽

pp. 136

Author(s):

L. P. Nethenzheni ◽

M. L. Mphaphathi ◽

P. V. M. Kalonji ◽

V. Monyelote ◽

N. C. Negota ◽

...

Keyword(s):

South African ◽

Liquid Nitrogen ◽

Sperm Motility ◽

Seminal Plasma ◽

Mitochondrial Membrane ◽

Membrane Damage ◽

Membrane Integrity ◽

Semen Sample ◽

Staining Solution ◽

Motility Rate

The removal of seminal plasma has a significant effect on semen characteristics. The aim of the study was to evaluate the effect of seminal plasma removal on sperm characteristics following semen dilution with Triladyl® or Bioxcell® extenders during cryopreservation. Semen samples were collected from 6 matured South African indigenous bucks for a period of 8 weeks by means of electro-ejaculation. Semen samples were pooled and then divided into 4 aliquots (Triladyl® -washed and non-washed or Bioxcell® -washed and non-washed) and diluted (1:4 vol/vol). Assessment of sperm motility characteristics was done by computer-aided sperm analysis (CASA) technology. Evaluation of mitochondria membrane integrity was done using JC-1 staining solution. Triladyl® and Bioxcell® washed semen sample groups were centrifuged at 1500 × g for 10 min, and seminal plasma was separated from sperm pellet using 1-mL plastic pipettes. After dilution of all semen sample groups, they were cooled by placing tubes into water (25°C) and then immediately placed in a 5°C fridge for equilibration for 2 h. At the end of equilibration period, all aliquot semen sample (final dilution concentration of 150 × 106 sperm/mL) groups were loaded into straws (0.25 mL) per treatment groups and placed horizontally 5 cm above the liquid nitrogen vapour for 10 min. At the end of freezing process, all semen straws per group were plunged directly into liquid nitrogen (−196°C) and stored until thawed. Frozen-thawed semen samples per treatment were analysed for sperm motility characteristics by CASA. JC-1 staining solution was also used during evaluation of mitochondria membrane integrity of frozen-thawed semen samples per treatment. Significant differences (P < 0.05) among mean values of semen parameters were determined by Tukey’s method. Sperm total motility rate of non-washed semen in Bioxcell® (85.0 ± 3.4) and Triladyl® (73.9 ± 13.8) was significantly reduced (P < 0.05) by cryopreservation compared with fresh (98.9 ± 1.2) semen sample. There was greater (P < 0.05) sperm mitochondrial membrane damage due to cryopreservation in non-washed semen group extended with Bioxcell® (50.2 ± 20.1) compared with semen extended with Triladyl® (31.3 ± 26.8) and fresh (16.6 ± 14.2) semen sample. Sperm total motility rate in Bioxcell® (68.2 ± 13.5) and Triladyl® (63.1 ± 15.1) groups on the non-washed semen were significantly reduced (P < 0.05) following cryopreservation, compared with fresh (98.3 ± 2.7) semen sample. The percentage of sperm with damaged mitochondria membrane in washed semen was significantly reduced (P < 0.05) in Triladyl® (21.6 ± 16.8) group compared with Bioxcell® (34.7 ± 14.9) group and fresh (35.0 ± 20.8) semen sample. In conclusion, seminal plasma removal reduced sperm motility rate in both extenders (Bioxcell® and Triladyl®) following post-thaw. In addition, sperm mitochondria membrane damage was higher in non-washed semen samples extended with Bioxcell® extender.

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Cryogenic Freezing of Silver Barb (Barbonymus gonionotus) Spermatozoa for Gene Pool Conservation

Progressive Agriculture ◽

10.3329/pa.v20i1-2.16863 ◽

2013 ◽

Vol 20 (1-2) ◽

pp. 117-124 ◽

Cited By ~ 1

Author(s):

MRI Sarder ◽

F Rahman ◽

MS Hossain ◽

MS Samad

Keyword(s):

Liquid Nitrogen ◽

Sperm Motility ◽

Gene Pool ◽

Egg Yolk ◽

Gradual Reduction ◽

Cryopreserved Sperm ◽

Thaw Period ◽

Silver Barb

Experiments were conducted to develop and standardize the protocols of cryopreservation of sperm of Barbonymus gonionotus. Three extenders Alsevers solution, urea-egg-yolk and egg-yolk citrate and four cryoprotectants methanol, ethanol, DMSO and DMA were used. Cryodiluents were prepared by adding 10% cryoprotectant to 90% extender (% v/v). Milt and cryodiluents were mixed at a ratio of 1 : 9 for Alsevers solution and 1 : 4 for urea egg-yolk and egg-yolk citrate solutions. Alsevers solution with 10% DMSO showed the best performance and produced 78 ± 2.55% sperm motility at the post-thaw period. Urea egg-yolk and egg-yolk citrate with 10% DMSO produced 76 ± 1.87% and 74 ± 1.87% post-thaw motility respectively. When cryopreserved sperm was stored in liquid nitrogen for 130 days, a gradual reduction in motility ranging from 31.25 to 37.50% was observed. This could be happened due to frequent opening of the nitrogen dewar that could cause thawing of the sample. In breeding trials, sperm preserved with Alsevers solution and urea egg-yolk plus 10% DMSO produced 14.28 ± 7.06% and 15.46 ± 5.50% hatching respectively. In contrast, sperm preserved with egg-yolk citrate and 10% DMSO produced poor hatching as 8.01 ± 2.15%.DOI: http://dx.doi.org/10.3329/pa.v20i1-2.16863 Progress. Agric. 20(1 & 2): 117 124, 2009

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Preservation of Morphological Integrity and Clot Retraction Activity of Human Platelets After Freezing

Thrombosis and Haemostasis ◽

10.1055/s-0038-1654822 ◽

1964 ◽

Vol 11 (01) ◽

pp. 222-229 ◽

Cited By ~ 8

Author(s):

Isaac Djerassi ◽

Albert Roy ◽

Jorge Alvarado ◽

Keyword(s):

Dimethyl Sulfoxide ◽

Liquid Nitrogen ◽

Human Platelets ◽

Slow Freezing ◽

Critical Conditions ◽

Plasma Medium ◽

Clot Retraction ◽

Controlled Freezing ◽

Direct Immersion

SummaryHuman platelets frozen at −195° C (liquid nitrogen) retain their morphological integrity and ability to promote clot retraction when 5% dimethyl-sulfoxide and 5% dextrose are added to the suspending plasma medium. Slow freezing was more effective than direct immersion in the liquid nitrogen. Although similar results may be achieved with dimethylsulfoxide alone with rigidly controlled freezing rates, the addition of sugars may permit freezing under less critical conditions.Dimethylsulfoxyd und 5% Dextrose dem Plasmamilieu hinzugefügt werden. Das langsame Einfrieren ist effektiver als das direkte Eintauchen in flüssigen Stickstoff. Obschon ähnliche Resultate mit Dimethylsulfoxyd allein unter exakter Kontrolle der Einfrierungsgeschwindig-keit erreicht werden können, erlaubt die Zugabe von Dextrose ein Einfrieren unter weniger kritischen Bedingungen.

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USING CRYOPRESERVED SPERM FOR CREATING STERLET BROOD STOCK

VESTNIK OF ASTRAKHAN STATE TECHNICAL UNIVERSITY SERIES FISHING INDUSTRY ◽

10.24143/2073-5529-2017-4-118-127 ◽

2017 ◽

pp. 118-127

Author(s):

Elena Nikolaevna Ponomareva ◽

Maria Mikhailovna Belaya ◽

Alexandra Andrianovna Krasilnikova ◽

Alexander Nickolaevich Nevalennyy

Keyword(s):

Liquid Nitrogen ◽

Control Group ◽

Test Results ◽

Brood Stock ◽

Active Response ◽

Acipenser Ruthenus ◽

Rostov Region ◽

The Central Nervous System ◽

Cryopreserved Sperm ◽

Sturgeon Fish

The research on the sterlet roe artificial insemination using cryopreserved sperm was carried out in the research base of the RAS Southern Scientific Centre (the Rostov region). Reproductive cells (including cryopreserved cells), larvae, sterlet fry ( Acipenser ruthenus Linnaeus, 1758) were taken as an object of research. A half of the roe (1.7 kg) taken from female starlet was inseminated by native sperm (control group); another half was inseminated by defrosted sperm of two males, which was stored in liquid nitrogen at -196ºC during 3 years (pilot group). Incubation lasted 5 days at water temperature 14.5-18.2ºC, with daily fluctuations of temperature 1.9ºC. Roe insemination in the control group made 90%, in the pilot group - 70%. Roe embryonic growth in the control group was faster, but embryogenesis duration in the pilot group met the standard time limits. Hatching prolarvae in the control group started one hour earlier, than in the pilot group; it made 75% and 60% of all incubated roe, correspondingly. Waste during the period of larvae maturing before they pass to mixed feeding was negligible - 2% in the control group and 3.4% in the pilot group. According to the test results, "open field" of reactivity of the central nervous system in the pilot group fry didn’t change from the control group fry, but more active response to stimuli was noted in the pilot group, which is very important for fry adaptation to the conditions in natural water basins. It was established that sterlet offspring obtained with use of defrosted sexual cells does not differ from the offspring obtained using native sperm and has higher morphometric characteristics. The test results prove the possibility and practicability of using sexual cells stored in liquid nitrogen for artificial restoration and formation of sturgeon fish broodstocks.

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Ridge-furrow with plastic film mulching system decreases the lodging risk for summer maize plants under different nitrogen fertilization rates and varieties in dry semi-humid areas

Field Crops Research ◽

10.1016/j.fcr.2021.108056 ◽

2021 ◽

Vol 263 ◽

pp. 108056

Author(s):

Changzhen Li ◽

Changjiang Li

Keyword(s):

Nitrogen Fertilization ◽

Plastic Film ◽

Summer Maize ◽

Plastic Film Mulching ◽

Fertilization Rates ◽

Film Mulching ◽

Maize Plants

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Effect of docosahexanoic acid on quality of frozen–thawed bull semen in BioXcell extender

Reproduction Fertility and Development ◽

10.1071/rd15089 ◽

2017 ◽

Vol 29 (3) ◽

pp. 490 ◽

Cited By ~ 3

Author(s):

Asmatullah Kaka ◽

Wahid Haron ◽

Rosnina Yusoff ◽

Nurhusien Yimer ◽

A. M. Khumran ◽

...

Keyword(s):

Lipid Peroxidation ◽

Liquid Nitrogen ◽

Sperm Motility ◽

Membrane Integrity ◽

Optimum Level ◽

Docosahexanoic Acid ◽

Normal Morphology ◽

Bull Semen ◽

Acrosome Integrity

This study was conducted to investigate the effect of docosahexanoic acid (DHA) supplementation in BioXcell extender on the quality of frozen–thawed bull semen. Twenty-four ejaculates were collected from three bulls (eight from each bull). Ejaculates with motility ≥70% and normal morphology ≥80% were extended into BioXcell extender to which 0 (control), 3, 5, 10 or 15 ng mL–1 DHA was added. The supplemented semen samples were incubated at 37°C for 15 min for DHA uptake by spermatozoa. Later, samples were cooled for 2 h at 5°C and packaged into 0.25-mL straws, frozen in liquid nitrogen for 24 h and subsequently thawed for evaluation. Results are presented as percentages ± s.e.m. Supplementation with DHA at 3 ng mL–1 significantly improved sperm functional parameters including sperm motility, normal morphology, viability, acrosome integrity and membrane integrity when compared with other supplemented groups and the control. Lipid peroxidation increased as the incorporation of DHA supplementation increased. In conclusion, 3 ng mL–1 concentration of DHA resulted in superior quality of frozen–thawed bull spermatozoa and is suggested as the optimum level of DHA to be added into BioXcell extender.

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