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

2013 ◽  
Vol 20 (1-2) ◽  
pp. 117-124 ◽  
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 Alsever’s 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 Alsever’s solution and 1 : 4 for urea egg-yolk and egg-yolk citrate solutions. Alsever’s 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 Alsever’s 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

77 ADDITION OF CHOLESTEROL IN FRESH GOAT SPERM IMPROVES CRYOSURVIVAL RATES

2013 ◽  
Vol 25 (1) ◽  
pp. 186
Author(s):  
B. G. Silva ◽  
E. A. Moraes ◽  
C. S. Oliveira ◽  
W. D. Ferrari Junior ◽  
W. C. G. Matos ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Phase Contrast ◽  
Room Temperature ◽  
Petri Dish ◽  
Egg Yolk ◽  
Irreversible Damage ◽  
Hypoosmotic Swelling Test ◽  
Contrast Microscopy ◽  
Working Solution

Cryopreservation causes irreversible damage to goat sperm membranes, measured by a loss of motile and functional normal cells, compared with fresh sperm. The objective of this study was to determine if the addition of cholesterol-loaded cyclodextrin (CLC) to goat semen improved sperm cryosurvival. The CLC was prepared as described by Purdy and Graham (2004 Cryobiology 48, 36–45) with some modifications: 200 mg of cholesterol were dissolved in 1 mL of chloroform and 1 g of methyl-beta-cyclodextrin was dissolved in 2 mL of methanol. A 0.45-mL aliquot of the cholesterol solution was added to the cyclodextrin solution, after which the mixture was poured into a glass Petri dish and the solvents allowed to evaporate on a warm plate for 24 h. The resulting crystals were removed from the dish and stored at 22°C. A working solution of the CLC was prepared by adding 50 mg of CLC to 1 mL TALP at 37°C. Thirty ejaculates from 5 bucks were collected, diluted 1 : 1 in Tris diluent, divided into 7 equal aliquots, and centrifuged at 800g for 10 min. The sperm pellets were resuspended in Tris diluent, to which 0, 0.75, 1.5, 3.0, 4.5, 6.0, or 7.5 mg of CLC/120 million sperm were added. All treatments were incubated for 15 min at room temperature and then cooled to 4°C over 2 h. The samples were then diluted with Tris-egg-yolk diluent containing 2% glycerol, and the sperm were packaged into 0.5-mL straws, frozen in static liquid-nitrogen vapour for 20 min, and plunged into liquid nitrogen. Straws were thawed in 37°C water for 30 s, extended in Tris, and analyzed using optic microscopy. To test thermal resistance, after thawing, 0.5 mL of semen from each treatment were placed in 1.5-mL Eppendorf tubes in a water bath at 37°C for 3 h. At 0, 60, 120, and 180 min, subsamples were evaluated for sperm progressive motility. A hyposmotic test was also conducted by adding 10 µL of sperm to 2 mL of each solution and incubating them for 1 h/37°C. Sequentially, 20 µL of sperm was diluted in hypoosmotic solution (150 mOsm), and the samples were evaluated using phase-contrast microscopy. A total of 100 spermatozoa were counted in at least 5 different fields, and sperm tails were classified as either noncoiled or coiled. Data were analyzed using ANOVA, and treatment means were separated, using the SNK test at 5% probability. The sperm motility (50.4, 33.8, and 22.5%) was significantly higher for sperm treated with 0.75 mg of cholesterol after 0, 60, and 120 min of incubation after thawing, when compared with other treatments. No treatment differences in the hypoosmotic swelling test were observed. The addition of 0.75 mg of cholesterol to fresh goat semen improved sperm motility after cryopreservation for up to 3 h. Supported by FACEPE and CAPES.

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)

scholarly journals FREEZING CAPABILITY OF PASUNDAN BULL SPERM USING TRIS-EGG YOLK, TRIS-SOY, AND ANDROMED® DILUENTS

2017 ◽  
Vol 11 (1) ◽  
pp. 45-49
Author(s):  
Abdullah Baharum ◽  
R. Iis Arifiantini ◽  
Tuty Laswardi Yusuf
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
Freezing Process ◽  
Sperm Abnormalities ◽  
Bull Sperm ◽  
Motility Of Sperm ◽  
Artificial Vagina

The aims of this study were to investigate the freezing capability of Pasundan bull spermatozoa in Tris-egg yolk (TEY), Tris-soy (TS), and AndroMed® as diluents. Semen were collected twice a week from four Pasundan bulls aged 3-5 years old using an artificial vagina and evaluated macro- and microscopically. Semen had ≥70% sperm motility, ≥800x106/mL sperm concentration, and less than 20% sperm abnormalities were divided into three parts and each of them diluted with TEY, TS, or AndroMed®. After an equilibration step at 5°C for four hours, diluted semen were packaged in 0.25 mL straw, frozen in liquid nitrogen for ten minutes and kept in liquid nitrogen container until examination. Motility test on fresh, diluted, equilibrated, and after-thawed semen was done using Androvision®. The results showed that after thawing motility of sperm diluted in AndroMed® (58.64±0.72%) was higher than in TEY (49.45±1.22%) and TS (39.34±6.33%). Sperm motility of Pasundan bulls diluted in these three diluents reduced around 33.27±2.45% during freezing process.

scholarly journals Management of the storage of cryopreserved sperm on dairy cattle farms

2015 ◽  
Vol 31 (1) ◽  
pp. 85-100 ◽  
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.

Erratum to: 56 SINGLE LAYER CENTRIFUGATION THROUGH PURESPERM® 80 IMPROVES QUALITY OF CRYOPRESERVED DOG SPERMATOZOA

2013 ◽  
Vol 25 (1) ◽  
pp. 175
Author(s):  
L. Alcaráz ◽  
M. Hidalgo ◽  
M. J. Galvez ◽  
D. Acha ◽  
I. Ortiz ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Semen Quality ◽  
Semen Analysis ◽  
Gradient Centrifugation ◽  
Single Layer ◽  
Egg Yolk ◽  
Final Concentration ◽  
Viable Sperm

Density gradient centrifugation with PureSperm® (PureSperm® 40 + PureSperm® 80; Nidacon International, Mölndal, Sweden) has been satisfactorily used to enhance the quality of dog semen samples; however, no studies have been performed on the effect of single layer centrifugation (SLC) with PureSperm® on frozen–thawed dog semen. The aim of this study was to investigate if SLC with PureSperm® 80 can improve the post-thaw semen quality of dog. Semen from 5 dogs was collected by digital manipulation. Two ejaculates from each dog were centrifuged with Tris-based extender, supernatant was removed, and sperm pellet was suspended to a final concentration of 300–400 × 106 sperm mL–1 with CaniPROTM Freeze A plus 20% egg yolk at 22°C. Extended semen was cooled to 5°C within an hour and then diluted to a final concentration of 150–200 × 106 sperm mL–1 in CaniPROTM Freeze B plus 20% egg yolk at 5°C, loaded in 0.5-mL plastic straws and frozen horizontally in ranks placed 4 cm above the surface of liquid nitrogen vapors for 10 min, after which they were directly placed in liquid nitrogen. After 24 to 48 h of storage, straws were thawed in a water bath at 37°C for 30 s. After thawing, semen samples were divided in 2 aliquots: one of them was used as control and the other one was processed by SLC PureSperm® 80. Assessment of sperm motility (assessed by computerized-assisted semen analysis), morphology (Diff-Quick staining), and viability [triple fluorescent stain of propidium iodine/isothiocyanate-labeled peanut (Arachis hypogaea) agglutinin/Rhodamine 123] were evaluated in control and treated semen samples. Data were studied by ANOVA. Results are expressed as mean ± SEM. Significant (P < 0.001) differences were found between SLC-treated and control semen for sperm motility (percentage of total motile spermatozoa: 93.65 ± 0.05 v. 83.79 ± 0.13; percentage of progressive motile spermatozoa: 79.38 ± 6.66 v. 54.61 ± 16.11), morphology (86.45 ± 0.01 v. 83.51 ± 0.01), and viability (percentage of viable sperm with an intact acrosome: 58.32 ± 0.04 v. 36.50 ± 0.17; percentage of viable sperm with an acrosome reaction: 2.81 ± 0.01 v. 9.74 ± 0.21). Based on our results, we can conclude that SLC with PureSperm® 80 is an alternative and successful method for improving the quality of frozen–thawed dog spermatozoa, selecting good-quality spermatozoa (motile, morphologically normal, viable, and acrosome intact spermatozoa) from the rest of the semen sample.

scholarly journals Cryopreservation of mutton snapper ( Lutjanus analis) sperm

2013 ◽  
Vol 85 (3) ◽  
pp. 1083-1092 ◽  
Author(s):  
EDUARDO G. SANCHES ◽  
IDILI R. OLIVEIRA ◽  
PEDRO C. DA SILVA SERRALHEIRO ◽  
VINICIUS R. CERQUEIRA
Keyword(s):  
Dimethyl Sulfoxide ◽  
Cooling Rate ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Nitrogen Fertilization ◽  
Factorial Experiment ◽  
Fertilization Rates ◽  
Cryopreserved Sperm ◽  
Net Cages ◽  
Motility Rate

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.

scholarly journals Comparison between different dilution rates on canine semen freezing using Tris-buffer with the addition of egg-yolk and glycerol

2005 ◽  
Vol 57 (6) ◽  
pp. 764-771
Author(s):  
A.R. Silva ◽  
R.C.S. Cardoso ◽  
L.D.M. Silva
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Sperm Morphology ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
Tris Buffer ◽  
Normal Sperm ◽  
Manual Stimulation

Standardized sperm concentration and volume:volume extension were compared as dilution rates for canine semen freezing. Six proven stud dogs were submitted to two seminal collections by manual stimulation. Semen was evaluated and extended in tris plus egg-yolk and glycerol according to two different dilution rates. The first one was based on a standardized sperm concentration of 200x10(6) spermatozoa/ml and the second was a volume:volume extension at a proportion of one part semen to one part extender. Semen was frozen, stored in liquid nitrogen and thawed after one week. Sperm motility and vigor were appraised after each stage of the process and at 15 and 30min post-thawing. Sperm morphology was analyzed after collection and thawing. No differences were observed between treatments after thawing regarding sperm motility and vigor, normal sperm morphology rate or longevity. Both dilution rates can be efficiently used for canine semen freezing.

scholarly journals Kriopreservasi sperma ikan kawan Poropontius tawarensis menggunakan Dimetil sulfoxida (DMSO)

DEPIK ◽  
2019 ◽  
Vol 8 (3) ◽  
pp. 158-166
Author(s):  
Cut Ruhul Muthmainnah ◽  
Zainal A. Muchlisin ◽  
Kartini Eriani ◽  
Iwan Hasri ◽  
Nur Fadli ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Egg Yolk ◽  
Dna Integrity ◽  
Hatching Rate ◽  
Fish Eggs ◽  
Integrity Test ◽  
Critical Material ◽  
Fish Sperm ◽  
Dmso Concentration

Abstract. Kawan fish (Poropuntius tawarensis) is an endemic fish found in Danau Laut Tawar, Central Aceh, Indonesia. This species has been threatened by ecological partubation, unfrindly fishing practices and pollution.  Cryopreservation is one of the ways to maintain the presence of these fish. Cryoprotectant (CP) is a critical material in the cryopreservation and DMSO is a common CP used in cryopreservation. Hence, the aim of the present study was to determine the optimum DMSO concentration for kawan fish sperm. The completely randomized design with 6 treatments and 3 replications were used in this study. The tested treatment was the difference of DMSO concentration, namely; 0, 3%, 6%, 9%; 12%, and 15% DMSO was combined with 5% egg yolk. The ratio of sperm to diluent is 1: 20. The cryotubes containing diluented sperm were evaporated at 5 cm from the surface of liquid nitrogen for 10 min, then stored in a liquid nitrogen container at -1960C for 2 weeks, then thawed and analyzed for the quality. The results showed that fresh sperm of kawan fish had motility of 48.67%, pH 7, milky white, with moderate consistency. The assessment of mass movements shows that the sperm has good quality. The ANOVA test showed that the addition of DMSO in diluents gavee significant effect on sperm motility, fertility and hatchability rates of fish eggs (P 0.05). The highest percentage of sperm motility and fertilization rates of fish eggs were found at concentration of 6%, respectively with the value of 46.67% and 45.67%, respectively. The highest percentage of hatching rate was also found in similar concentration of DMSO with the value of 19.33%. %. The DNA integrity test using the electrophoresis gel method showed that there was damage to DNA fish sperm after freezing, the the lower damage was found at 9% and 12% DMSO. It is concluded that the optimum concentration of DMSO for kawan fish sperm is at 6% of DMSO. Key words: kawan fish (Poropuntius tawarensis), cryopreservation, DMSO, DNA integrity Abstrak. Ikan kawan (Poropuntius tawarensis) merupakan ikan endemik yang terdapat di Danau Laut Tawar, Aceh Tengah, Indonesia. Menurut IUCN (International Union for Conservation of Nature), ikan ini termasuk ikan yang terancam punah oleh sebab kerusakan lingkungan, penangkapan tidak ramah lingkungan dan polusi. Salah satu cara untuk menjaga keberadaan ikan tersebut adalah dengan penerapan metode kriopreservasi sperma. Oleh karena itu, penelitian ini bertujuan untuk menentukan konsentrasi DMSO optimum dan melihat kerusakan DNA yang terjadi pada sperma ikan kawan(Poropontius tawarensis) pasca pembekuan.Metode yang digunakan dalam penelitian ini adalah Rancangan Acak Lengkap (RAL) dengan 6 perlakuan dan 3 ulangan.Perlakuan yang diuji adalah perbedaan konsentrasi DMSO dengan konsentrasi 0; 3%; 6%; 9%; 12% dan 15%.DMSO tersebut dikombinasikan dengan 5% kuning telur. Perbandingan sperma dengan pengencer adalah 1 : 20. Semua cryotube yang berisi sperma dan pengencer diuapkan pada jarak 5 cm dari permukaan nitrogen cair selama 10 menit, selanjutnya, disimpan dalam kontainer nitrogen cair bersuhu -1960C untuk disimpan selama 2 minggu. Hasil penelitian menunjukkan bahwa sperma segar ikan kawan memiliki nilai motilitas sebesar 48,67%, pH 7, berwarna putih susu, dengan konsistensi sedang. Penilaian gerakan massa menujukkan bahwa sperma tersebut berkualitas baik. Hasil uji ANOVA menunjukkan bahwa penambahan DMSO dalam pengencer berpengaruh nyata terhadap motilitas, fertilitas dan daya tetas telur ikan kawan (Poropontius tawarensis) (P0,05) setelah pembekuan. Selanjutnya, uji lanjut Duncan menunjukkan bahwa persentase motilitas sperma dan pembuahan telur ikan kawan tertinggi terdapat pada penambahan DMSO dengan konsentrasi 6%, masing-masing sebesar 46,67% dan 45,67%. Persentase penetasan telur tertinggi juga dijumpai pada perlakuan 6% DMSO, dengan nilai 19,33%. Hasil uji integritas DNA menggunakan metode elektrofresis gel menunjukkan bahwa terdapat kerusakan pada DNA sperma ikan pasca pembekuan, Kerusakan yang terendah terdapat pada konsentrasi DMSO 9% dan 12%. Namun secara umum, disimpulkan bahwa konsentrasi optimum untuk kriopreservasi ikan kawan adalah 6% DMSO.Kata kunci: ikan kawan (Poropuntius tawarensis), kriopreservasi, DMSO, integritas DNA

67 DELIVERING CHOLESTANOL OR DESMOSTEROL TO BULL SPERM MEMBRANES IMPROVES CRYOSURVIVAL

2008 ◽  
Vol 20 (1) ◽  
pp. 114
Author(s):  
E. A. M. Amorim ◽  
J. K. Graham ◽  
M. Meyers ◽  
B. Spizziri
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Plasma Membranes ◽  
Egg Yolk ◽  
Positive Control ◽  
Epifluorescence Microscopy ◽  
Osmotic Tolerance ◽  
Nitrogen Vapor ◽  
Bull Sperm ◽  
And Function

Altering the lipid composition of sperm plasma membranes affects sperm cryosurvival. Cryopreservation induces many stresses on the spermatozoa, including destabilization of the plasma membrane, which results in the loss of sperm motility and function. Treating bull spermatozoa with cholesterolloaded cyclodextrin (CLC) prior to cryopreservation increases sperm cryosurvival rates. This study compared the effect of adding other sterols, which should incorporate into the membrane and increase membrane fluidity at low temperatures, thereby increasing cryosurvival. Ejaculates from four bulls were divided into two experiments (E). In E1, ejaculates were extended with Tris, and then subdivided into four treatments: No additive (control), 1.5 mg CLC/120 million sperm (positive control), and 1.5 mg/120 million sperm in cyclodextrin pre-loaded with either cholestanol or desmosterol. Spermatozoa were incubated for 15 min at 22�C after which both the ability of fresh spermatozoa to bind to the zona pellucida (ZP) and chicken egg perivitelline membrane (EPM) and their osmotic tolerance were evaluated. In E2, sperm were diluted to 120 million cells mL–1 in a Tris diluent and treated as described for E1. Then, samples were diluted 1:1 (v:v) in Tris with 20% Egg Yolk (EY) and cooled to 5�C. After dilution 1:1 (v:v) with Tris containing 10% EY and 16% glycerol, samples were allowed to equilibrate for 15 min, and then were packaged into 0.5-mL straws, frozen in static liquid nitrogen vapor for 20 min, and plunged into liquid nitrogen for storage. Straws were thawed and the motility and zona-binding ability were determined using a Hamilton Thorne Motility Analyzer (Hamilton Thorne Biosciences, Beverly, MA, USA) and epifluorescence microscopy, respectively. Treatment differences for sperm motility, osmotic tolerance, and zona binding were determined using analysis of variance. Treating spermatozoa with CLC resulted in more fresh bull spermatozoa binding to the EPM and ZP compared to cholestanolor desmosterol-loaded cyclodextrin-treated spermatozoa or control cells (P < 0.05). No differences were observed between EPM and ZP binding (P > 0.05). The percentages of total and progressively motile spermatozoa were higher for fresh samples treated with cholesterol-, cholestanol-, or desmosterol-loaded cyclodextrin than for control cells (P < 0.05) when spermatozoa were exposed to anismotic conditions, and then returned to isosmolality. After cryopreservation, the percentages of motile spermatozoa and number of spermatozoa binding to ZP were similar for spermatozoa treated with CLC (56% and 115 sperm/ZP) and cholestanol (53% and 108 sperm/ZP) compared to spermatozoa treated with desmosterol (42% and 86 sperm/ZP; P < 0.05). All treatments provided higher motility and binding efficiency than control spermatozoa (32% and 62 sperm/ZP; P < 0.05). Therefore, adding cholesterol or cholestanol to bull sperm membranes improved cell cryosurvival. Studies to determine if cholestanol affects sperm capacitation need to be conducted.

scholarly journals Comparison of growth performance between cryopreserved and fresh sperm-originated fry of Barbonymus gonionotus

1970 ◽  
Vol 7 (1) ◽  
pp. 145-149 ◽  
Author(s):  
F Rahman ◽  
MRI Sarder ◽  
MA Rouf
Keyword(s):  
Growth Performance ◽  
Egg Yolk ◽  
Citrate Solution ◽  
Control Groups ◽  
Significant Difference ◽  
Cryopreserved Sperm ◽  
Survival And Growth ◽  
And Control ◽  
Silver Barb ◽  
Glass Aquarium

This experiment was conducted to compare the growth performance of silver barb fry produced from cryopreserved sperm with those produced from fresh sperm. Cryopreserved sperm used for fry production was preserved with three extenders, Alsever's solution, urea egg-yolk, egg-yolk citrate solution and one cryoprotectant, DMSO. Cryodiluents were prepared by mixing the cryoprotectants at 10% concentration of the extender (% v/v). Fry produced with fresh sperm was considered as control. For comparing the growth, 60 fry of 15 day-old for each treatment of both cryopreserved and control groups were stocked to glass aquarium (50 cm x 30 cm x 28 cm) and reared them for ten weeks. Growth of fry in terms of length and weight increment in both cryopreserved and control groups were measured weekly. The growth pattern was more or less similar for all the treatments and there was no significant difference (P>0.05) between them. The survival rate of fry produced from cryopreserved sperm was 82.3% and that from fresh sperm 88%, and there was no significant difference (P>0.05) between two groups. It is therefore, concluded that the use of cryopreserved sperm does not impair survival and growth of fry. Keywords: Silver barb; Cryopreservation; Growth; Survival DOI: 10.3329/jbau.v7i1.4977 J. Bangladesh Agril. Univ. 7(1): 145-149, 2009

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