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1.
The objectives of this study were to evaluate the effects of varying dilutions, pH, temperature, osmolality, and cations on sperm motility parameters in waigieu seaperch, Psammoperca waigiensis. The maximum velocity of average in path (VAP), percentage of motile cells (MOT), and duration of sperm motility (DSM) were observed when semen was diluted in artificial seawater (ASW) at a ratio of 1:100 (144.9 ± 0.6 µm/sec, 95.6 ± 0.4%, and 230.3 ± 2.3 sec, respectively), at 30 C (142.0 ± 0.6 µm/sec, 93.6 ± 0.4%, and 238.3 ± 0.9 sec, respectively), and pH 8 (144.8 ± 0.6 µm/sec, 93.3 ± 0.4%, and 234.0 ± 1.5 sec, respectively). Maximum VAP, MOT, and DSM were obtained in each solution containing 0.6 M NaCl (143.8 ± 1.0 µm/sec, 91.3 ± 2.0%, and 230.6 ± 4.2 sec, respectively), 0.6 M KCl (135.1 ± 3.1 µm/sec, 91.1 ± 3.1%, and 230.3 ± 3.7sec, respectively), 0.2M CaCl2 (105.3 ± 4.7μm/sec, 47.9 ± 2.7%, and 120.7 ± 1.3 sec, respectively), 0.2 M MgCl2 (107.3 ± 3.0 m/s, 42.1 ± 3.3%, and 120.3 ± 4.8 sec, respectively), and osmolality of 400 mOsm/kg (145.1 ± 2.5 µm/sec, 93.0 ± 2.1%, and 346.5 ± 4.4s, respectively). We used these mediums as artificial insemination media for fertilizing matured eggs. The results showed that the fertilization and hatching rates in 0.6 M NaCl (75.3 ± 0.6% and 57.0 ± 2.4%, respectively), ASW (70.8 ± 1.2% and 51.2 ± 1.8%, respectively), or 400 mOsm/kg (72.9 ± 1.8% and 55.3 ± 1.6%, respectively) were higher than that in seawater (63.9 ± 1.2% and 39.2 ± 3.9%, respectively). In conclusion, using 0.6 M NaCl, ASW, or 400 mOsm/kg as an artificial insemination medium is effective for fertilizing of waigieu seaperch.  相似文献   

2.
Changes in semen quality and selected biochemical markers were analyzed during a week of spawning season of common carp Cyprinus carpio L. Semen was obtained twice, on May 30 and on June 7, and each time it was collected 24 h after hormonal stimulation using Ovopel [(d-Ala6, Pro9 NEt)-mGnRH + metoclopramide] in 1 pellet kg?1. The total volume of semen (ml), volume of semen per kg of body weight (ml kg?1 b.w.), sperm concentration (×109 ml?1), total number of sperm per kg of body weight (×109 kg?1 b.w.), pH of semen, pH of seminal plasma, seminal plasma osmotic pressure (mOsm kg?1) and the total protein content in seminal plasma (mg ml?1) were determined. A 10 mM Tris buffer containing 100 mM NaCl with 0.5 % BSA (pH 9.0, osmolality 200 mOsm kg?1) was used to activate sperm. The following computer-assisted sperm analysis (CASA) parameters were determined: percentage of motile sperm (MOT, %), progressively motile sperm (PRG, %), curvilinear velocity (VCL, μm s?1), straight-line velocity (VSL, μm s?1), movement linearity (LIN, %), wobbling index (WOB, %), amplitude of lateral head displacement (ALH, μm) and beat cross-frequency (BCF, Hz). The volume of semen per kg of BW, total number of sperm per kg of BW and semen pH were significantly lower at the second semen sampling compared to the first semen sampling. Volume of semen at the second sampling correlated positively with CASA parameters. A lack of differences among CASA parameters between both collection periods indicates good quality of carp sperm hormonally stimulated with Ovopel twice at a 1-week interval.  相似文献   

3.
The effectiveness of applying Ovaprim [(D-Arg6, Pro9NEt)-sGnRH + domperidone] and Ovopel [(D-Ala6, Pro9NEt)-mGnRH + metoclopramide] to male barbel Barbus barbus (L.) 6, 12 and 24 h after hormonal stimulation was analyzed. The control group (Control) during each time interval was stimulated with 0.9 % NaCl. Milt was collected from seven fish only once (n = 7) for Ovopel, Ovaprim and Control group in order to determine total volume of milt, volume of milt per kg of body weight, sperm concentration, total sperm production, seminal plasma osmotic pressure, pH of milt and pH of seminal plasma. Woynarovich’s solution (68 mM NaCl + 50 mM urea) with the addition of 0.5 % BSA (pH 7.7; 181 mOsm kg?1) was used as the activating liquid. Selected parameters of sperm motility (MOT %) and progressively motile sperm (%), curvilinear velocity (VCL, μm s?1), straight-line velocity (μm s?1), movement linearity (%), wobbling index (%), amplitude of lateral head displacement (μm) and beat cross frequency (Hz) were determined using the Computer-assisted sperm analysis system. A time of 6 h proved to be too short to obtain milt from barbel following hormonal stimulation with Ovaprim and Ovopel. Extending the time to 12 h, however, resulted in 100 % spermiation in males, regardless of hormonal preparation used for stimulation. The stimulation of spermiation in barbel is best performed using Ovopel 12 h upon application. Extending the latency period to 24 h following the application of this preparation results in a significant decrease in the volume of milt obtained, sperm count and motility parameters, including MOT and VCL, which may influence sperm fertilization ability.  相似文献   

4.
The effect of two commercial preparations containing different GnRH analogues with dopamine antagonists on quantitative and qualitative parameters of semen from chub Leuciscus cephalus L. collected in artificial conditions were examined. Semen was collected after the application of [(D‐Ala6, Pro9 NEt)‐mGnRH + metoclopramide] (Ovopel, n = 9), [(D‐Arg6, Pro9 Net)‐sGnR + domperidone] (Ovaprim, n = 9) and from the control group (0.9% NaCl, n = 9). Afterwards, semen volume, sperm concentration, total sperm production and semen pH were determined. Osmolality and pH of seminal plasma were also determined. Using the Computer‐assisted sperm analysis system (CASA), selected sperm parameters such as sperm motility (MOT %), progressively motile sperm (PRG, %), curvilinear velocity (VCL, μm s?1), straight‐line velocity (VSL, μm s?1), movement linearity (LIN, %), wobbling index (WOB, %), amplitude of lateral head displacement (ALH, μm) and beat cross frequency (BCF, Hz) were analysed. While Ovopel can also be used to stimulate chub spermation, the application of Ovaprim was much more effective for obtaining higher amounts of semen.  相似文献   

5.
The spermatozoa of oviparous fish, such as feral carp (Cyprinus carpio), are immotile in the presence of semen plasma or isotonic solutions, and to obtain good motility, they must be diluted with suitable medium. The objective of this study was to identify the best activating solution for feral carp sperm. Sperm motilities were compared in the new activating solution (a): (50 mM NaCl, 30 mM KCl, 30 mM Tris, pH = 8.5) and activating solution (b): (50 mM NaCl, 40 mM KCl, 30 mM Tris, pH = 8.5) based on effect of pH with everyone of Na+ and K+ ions versus four other activating solutions Billard’s saline solution, Poupard’s saline solution, distilled water and hatchery water that is routinely used for extending carp semen. Our results showed that maximum total motility period and percentage of motile sperm were seen in selected saline solution (a). The present study describes an activating solution that prolongs feral carp sperm motility.  相似文献   

6.
Three groups of captive-reared striped bass Morone saxatilis ages 1, 3 and 12 yr, were examined for age-related changes of sperm characteristics including short-term storage. All groups had similar ranges of the following parameters (mean× SEM): expressible milt (5.6× 0.5 mI/kg body weight (BW) to 7.5× 2.1 mL/kg BW), percentage of motile sperm (55× 6% to 60× 2%), duration of sperm motility (69× 3 sec to 72× 5 sec) and percentage of viable sperm (91× 2% to 93× 2%). Compared to the 1 and 12-yr-old fish, the 3-yr-old fish produced the greatest number of spermatozoa (1,190× 370× 109 spermatozoa/kg), sperm concentration (120× 8 × 109 spermatozoa/mL) and spermatocrit (74× 4%). In addition, during short-term storage at 4 C, extender-preserved sperm samples of the 3-yr-old group showed a significantly higher ( P < 0.05) percentage of motile sperm and duration of sperm motility, compared to the other two groups. This suggests that short-term storage may be affected by the age of the male fish. Sperm longevity of the 3-yr-old group was successfully maintained for as long as 15 d, longer than that of the 1-yr-old group (9 d) and 12-yr-old group (7 d). Overall, the 3-yr-old fish appeared to have superior sperm quality than the 1 or 12-yr-old fish based on higher sperm production and increased sperm longevity.  相似文献   

7.
Basic characteristics of the European smelt (Osmerus eperlanus) sperm are reported here for the first time. Smelt spermatozoa had a bullet-shaped head (1.42 μm length), a short midpiece and a long flagellum (27.72 μm). Two mitochondria were located along the flagella. The volume of smelt sperm was small (30-60 μl) and the duration of sperm motility was short (22 s in distilled water and 41 s in 20 mM sodium bicarbonate solution). Sodium chloride at concentrations ranging from 0-120 mM did not influence the percentage of motile spermatozoa but caused a steady increase in the duration of sperm movement. Potassium ions clearly reduced the percentage of motile sperm at a concentration of 10 mM. Spermatozoa were motile through a broad range of pH with an optimum from 7.5 to 8.5. Testicular spermatozoa had a different motility pattern compared to stripped spermatozoa (the latter exhibiting a reduction of motile spermatozoa by 30%, lower ALH and VCL and higher LIN and VSL). These results indicate that maturation of smelt spermatozoa occurring in sperm ducts is related not only to an increase of the percentage of motile spermatozoa but also to changes in the sperm motility pattern. Maintaining males with females resulted in stimulation of milt production. Our results indicate that European smelt sperm characteristics are similar to those of ayu (Osmeridae).  相似文献   

8.
The use of saline solutions was tested to determine their efficacy as replacements for ovarian fluid as sperm activators and to eliminate variability encountered with the use of Ovarian fluid. We tested fertilization rate of semen from eight males on Atlantic salmon Salmo salar eggs after five sperm-activating solutions and a non-activating saline were substituted for ovarian fluid. We used solutions shown acceptable for use with other salmonid species. The six solutions tested were a non sperm-activating phosphate-buffered saline (PBS, 7.2 g/L NaCl, 1.48 g/L Na2HPO4, 0.43 g/L K H2PO4), a Tris buffer (6.99 g/L NaCl, 3.63 g/L Tris and 2.42 g/L glycine), a Borax buffer (12.2 g boric acid/L in solution 1, 76 g disodium tetraborate/L combined 100:118, then 1 L combined with 3.7 L water and 18 g NaCl), and three solutions of 9.25 g/L (125 mM) NaCl buffered to pH 6.0, 7.5, and 8.9. The latter five solutions activated sperm immediately on contact, while PBS required additional water to activate sperm. The PBS solution was the least effective (mean percent eyed eggs 37.6%) for egg fertilization. The mean percent eyed eggs for the other five saline solutions (range 78% to 86%) were not significantly different. Sperm from one male provided significantly lower egg fertilization (39.6%) when compared with the other seven males (67.2–87.4% egg fertilization). Percent egg fertilization was not related to number of live sperm cells per egg. Our results show that osmotically-balanced sperm-activation solutions, even those with a pH range from 6.0 to 8.9 provide adequate media for fertilization of Atlantic salmon eggs. Fertilization in a deactivation saline and water reactivation of sperm resulted in low egg fertilization.  相似文献   

9.
The effect of sodium and potassium concentrations as well as optimal pH on the motility of common carp Cyprinus carpio L. sperm during short-term storage in artificial seminal plasma (ASP) was investigated. Sperm was collected from individual males (n?=?5) and each sample diluted tenfold (1:9) in ASP (sperm:extender) containing 2 mM CaCl2, 1 mM Mg2SO4 and 20 mM Tris at pH 8.0 and supplemented by the following concentrations of sodium and potassium (mM/mM): 0/150, 20/130, 40/110, 75/75, 110/40, 130/20 and 150/0. The osmolality of all ASP variants was set at 310 mOsm kg?1. Sperm motility was measured using a CASA system during 72 h of storage. Immediately after dilution, sperm motility was high (90%) both in each variant and in the control group (fresh sperm). After 72-h storage, the highest sperm motility was noted in ASP containing 110 mM NaCl and 40 mM KCl. No differences were found in the motility of samples preserved within the pH range of 7.0–9.0. Our data suggest that for the short-term storage of common carp sperm, whereas the pH of the solution does not play a crucial role, a specific potassium concentration of around 40 mM is required.  相似文献   

10.
舒德斌  郭柏福 《水产科学》2012,31(4):232-234
比较了史氏鲟精子在3种不同配比浓度稀释液的保存效果。试验结果表明,配方Ⅲ作为稀释液,8%甲醇作为抗冻剂,二步法超低温(-196℃)冷冻保存,5h后取出,38℃水浴解冻取得最好的冻后激活率,解冻后激活率为(52.3±3.5)%。解冻精子分别采用井水和激活液D(10mmol/L Tris+10mmol/L NaCl+25mmol/L Glu,pH 8.0)激活,进行人工授精。结果显示配方Ⅲ冻精采用激活液D激活授精获得最高受精率为68.56%,最高孵化率为52.91%。本次试验表明,1～2mmol/L范围内,低浓度K+比高浓度K+对史氏鲟精子保存有利;52～82mosmol/kg范围内,高渗稀释液有利于史氏鲟精子的保存;且激活授精方法是影响冻精受精率和孵化率的关键因素之一。  相似文献   

11.
Spermatozoa concentration, ionic composition, osmolality, glucose and total protein contents of seminal plasma and sperm motility were determined in Barbus sharpeyi (Cyprinidae, Teleosotei). Spermatozoa concentration ranged from 9.77 to 20.20 × 109 spermatozoa mL?1. Osmolality (mOsmol kg?1) and ionic contents (mM L?1) of the seminal plasma were 274.5±9.0, 70.0±3.4 Na+, 28.8±0.9 K+, 101.7±3.1 Cl?, 0.9±0.1 Mg2+ and 2.1±0.1 Ca2+ respectively. Total protein and glucose were 5.3±0.2 g L?1 and 76.7±4.3 mM L?1 respectively. Sperm motility was initiated in a hypo‐osmotic condition, composed of either an ionic (KCl or NaCl) or a non‐ionic (sucrose) activation medium. Duration of sperm motility was very short: <2 min after activation in distilled water. Percentage of motile spermatozoa was significantly higher in an activation medium containing NaCl compared with that of distilled water. An activating medium containing NaCl or KCl higher than 150 mM or sucrose higher than 275 mM totally inhibited the activation of sperm motility. Immediately after sperm activation, wave(s) propagated along the flagellum, but waves were restricted to the proximal part of the flagellum (close to the head) at 1 min post activation. Studied characteristics in the present study were compared with those of other cyprinids for understanding inter‐species differences.  相似文献   

12.
The effects of changes in osmotic pressure on the initiation of spermmotility in the river sculpin were studied. When the fresh milt was diluteddirectly with solutions of mannitol at various concentrations, 22% ofthe spermatozoa became motile in isotonic (300 mM) solution, 63%became motile in a solution of 200 mM, and more than 90% becamemotile in a solution of 100 mM and in 20 mM HEPES-NaOH. When the spermatozoawere transferred to various osmotic conditions after the termination ofswimming in the isotonic solution, 27.9% of spermatozoa became motilein the solution of 200 mM, while more than 80% of spermatozoa becamemotile in the solution of 100 mM and in the buffer solution. Aftertermination of swimming in the solutions of 300 mM and 200 mM, more than80% of spermatozoa became motile again when they were transferred toa solution of 100 mM and to the buffer solution. After the termination ofswimming in the solution of 100 mM, most of spermatozoa were immotile evenwhen they were transferred to the buffer solution. These results suggest theexistence of multiple osmotic switches in each sperm cell that initiate themotility; one reacts in response to the osmotic conditions above 200 mOsmkg-1, with the threshold varying for each cll, while anotherswitch, which is common to all the cells, reacts in response to osmoticpressure below 200 mOsm kg-1. The existence of the latterswitch was confirmed by the motility in the solution of 100 mM and in thebuffer solution in this study.  相似文献   

13.
Sperm of gilthead seabream, Sparus aurata, was diluted with solutions of different osmolarities and pH. The effect of the different diluents on sperm motility (intensity and percentage of motile sperm) was studied. Motility was induced as early as 10 s after mixing the sperm with diluents having an osmotic pressure higher than 500 mOsm/l. The intensity of motility decreased when the osmotic pressure was reduced, and was zero or significantly inhibited when the osmotic pressure of the diluent (300–380 mOsm/l) was close to that of the fish's seminal plasma (364.6±3.03 mOsm/l). The pH of the diluent did not have any effect on sperm motility (in a range of 6.8 to 8.9). A diluent which prevents spermatozoa motility (osmotic pressure 375 mOsm/l and pH 7.35) was successfully used to cryopreserve S. aurata sperm at −196°C. This diluent is considered promising for the long-term preservation of gilthead seabream sperm.  相似文献   

14.
中华倒刺鲃、白甲鱼和岩原鲤精子的生理特性比较   总被引：2，自引：0，他引：2  
对中华倒刺鲃(Spinibarbussinensis)、白甲鱼(Onychostomasimus)和岩原鲤(Procyprisrabaudi)3种鱼的鲜精在不同水溶液和不同浓度梯度的NaCl溶液中的活力以及精子形态、密度和精液pH值、浓度等进行了比较研究。结果表明:3种鱼的精液pH值都偏弱碱性,位于7·2~7·7之间;精液浓度依次为71·1%,76·1%和71%;精子头长依次为(3·89±0·53)μm,(2·98±0·08)μm和(6·42±0·59)μm,全长依次为(41·63±3·66)μm,(65·67±2·97)μm和(58·89±5·25)μm;精子密度依次为1·527×1010尾/mL,1·336×1010尾/mL和1·362×1010尾/mL。分析表明,3种鱼的精子大小与密度无相关性。在不同水溶液中,3种鱼的精子活力均以池塘水中最高;在不同浓度NaCl溶液中,3种鱼精子的最适浓度位于0·45%~0·55%之间,有效运动时间以中华倒刺鲃最高,为(85·23±12·02)s,其次是岩原鲤,为(50·45±6·89)s,白甲鱼最短,为(31·44±5·53)s。3种鱼的精子全长与精子活力存在负相关性,即精子越长,活力越低,精子越短,活力越高。  相似文献   

15.
Semen of the African catfish, Clarias gariepinus (Burchell, 1822), was investigated with respect to its cellular composition, sperm cell density, maturation grade, motility and fertility. Storage conditions were tested, whereby sperm viability was assessed by measurement of the motility after activation and by fertility tests. Testicular semen differed in its composition, i.e. the sperm density and numbers of spermatids, according to the maturity grade of the testis. Two semen types could be distinguished: semen type I was characterized by high sperm densities and low numbers of spermatids and semen type II had lower sperm densities and higher numbers of spermatids. Two semen types did not differ in motility and fertility (when adjusted for differences in sperm density). During storage, the sperm viability was influenced by the sodium concentration of the storage medium, temperature, membrane stabilizers as bovine serum albumen (BSA) or hen egg yolk, antibiotics and oxygen. Semen viability was maintained best when it was diluted at a ratio of 1:5 in storage solution (150 mmol L?1 NaCl, 2.5 mmol L?1 KCl, 1 mmol L?1 CaCl2, 1 mmol L?1 MgSO4, 20 mmol L?1 Tris (pH 8.5) and 0.5% BSA or 0.5% hen egg yolk) and stored at 4 °C. Oxygen gassing and addition of antibiotics (1 mg mL?1 gentamycine sulphate) to the storage solution affected the two semen types in different ways. Antibiotics had no effect on type I semen, but had a positive effect on type II semen. Oxygen gassing had a positive effect on type I semen but a negative effect on type II semen.  相似文献   

16.
The aims of this study were to describe the fresh spermatozoon ultrastructure using scanning and transmission electron microscopy and to improve the sperm cryopreservation methodology for the freshwater fish pirapitinga Brycon nattereri. Extenders (BTS? and NaCl), straw volumes (0.5 and 4.0 mL), thawing temperatures (30 and 60 °C) and activating agents (0.29% NaCl and 1% NaHCO3) were tested. Methylglycol was used as a cryoprotectant agent and sperm was frozen in nitrogen vapour (dry‐shipper). Post‐thawed sperm motility rate, motility quality (score 0=no movement; 5=rapidly swimming spermatozoa), duration of motility and spermatozoon morphology were evaluated. Fresh spermatozoon was 35.06 μm long, the head was ovoid (2.00 × 1.22 μm) with no acrosome, the midpiece was 2.15 μm long and the flagellum was 30.90 μm long with the typical 9+2 axoneme arrangement. Post‐thawed sperm motility rate (70–79% motile sperm), motility quality (score 3.1–3.7) and morphology (9.3–11.6% abnormal spermatozoa) were not affected by any of the parameters tested. The duration of sperm motility was longer when triggered in 1% NaHCO3 (392–1031 s) compared with 0.29% NaCl (144–338 s). Brycon nattereri sperm cryopreserved under the conditions described above yields over 70% motility and should last long enough to fertilize oocytes, even after 2 years of freezing.  相似文献   

17.
The high sperm density, together with the short spermatozoa swimming time, makes European eel sperm manipulation and assessment for quality difficult. Two diluting media (K15 and K30) previously designed for Japanese eel sperm were tested. After 24 h, European eel sperm showed significant reduction in the percentage of motile spermatozoa after activation and different motility parameters (VAP, angular velocity; VCL, curvilinear velocity; VSL, straight line velocity; BCF, beating cross frequency), concluding that these media are not suitable to preserve the sperm of this species. After a hormonal treatment to induce spermiation, sperm volume, density and motility were recorded at weekly samplings. The variation of the osmolality (325–330 mOsm kg−1), pH (8.4–8.6) and the ionic composition (concentration of Na+, K+, Mg2+ and Ca2+) of the seminal plasma were registered. Physio-chemical results were related with sperm quality throughout the treatment, to determine which must be the suitable characteristics of one extender for the sperm of this species, and to find the best conditions to obtain suitable cryopreservation media for European eel sperm. K+ concentration increased, while Ca2+ and Mg2+ concentrations showed a progressive reduction in correlation with the sperm quality improvement. Na+ showed a decreasing, but not significant tendency. P1 and P2 freezing media were designed considering the physio-chemical parameters as well as the ionic composition shown by the best quality sperm samples, and then compared with the previously described solutions, TNK and K30. Sperm quality was determined, checking the percentage of motile spermatozoa and motility parameters using computer-assisted sperm analysis (CASA) software. Samples were frozen after dilution (1:5, 1:20, 1:100) in different freezing media supplemented with 10% dimethyl sulfoxide (DMSO). After thawing, samples frozen with low dilution ratio (1:5) in TNK and P1 media showed higher, although not significant, spermatozoa survival (35.5 ± 14.5 and 36.6 ± 6.7%). The addition of l-α-phosphatidylcholine to the media seems to have a positive effect, as reported in the Japanese eel.  相似文献   

18.
Successful hybridization of sterlet (Acipenser ruthenus) × Siberian sturgeon (Acipenser baeri), sterlet × Russian sturgeon (Acipenser gueldenstaedti) and sterlet × European sturgeon (Acipenser sturio) was carried out for the first time using cryopreserved semen of sturgeon males and sterlet × sterlet crosses as controls. Sperm of all three species was diluted with a cryodiluent composed of 23.4 mM sucrose, 0.25 mM KCl, 30 mM Tris (pH 8.0) and 10% methanol. The samples were frozen in plastic straws in the vapor of liquid nitrogen at the height of 3 cm above the level of nitrogen for 3 min. Following thawing approximately 3000 sterlet eggs were fertilized with six straws of frozen-thawed sperm. The hatching rate with sterlet sperm was 30.6% while the hatching rate of A. ruthenus × A. baeri, A. ruthenus × A. gueldenstaedti and A. ruthenus × A. sturio hybrids was 50, 17.4 and 34%, respectively. Morphometric markers as well as random amplified polymorphic DNA (RAPD) assay was used to verify interspecific hybridization.  相似文献   

19.
In this study, the efficiency of a novel droplet vitrification technique along with different doses of fish antifreeze protein (AFP) type III on Persian sturgeon thawed spermatozoa quality (motility duration and motility percentage) was investigated. Semen of seven male individuals was pooled in equal volumes and diluted with 4°C Tris‐Hcl (100 mM), pH = 8 extenders containing 0, 5, 10, 15 μM of AFP type III in a ratio of 1:1 (semen/extenders). Treated semen was dropped into liquid nitrogen. Solidified droplets were stored for 2, 60 and 120 days and thawed by plunging them into a tube containing 5 mL Tris‐Hcl (100 mM), pH=8 with 1% BSA at 37°C. Motility duration in all treatments had no significant difference comparing to fresh sperm (P > 0.05), but their motility percentage was significantly lower. Treatment with 10 μM of AFP had significantly higher motility percentage (16.11 ± 0.5%) comparing to other treatments (P < 0.05). There was no significant difference between 0, 5, 15 μM of antifreeze protein treatments (P > 0.05), suggesting that antifreeze protein effectiveness are highly dose dependent, and dose of 10 μM is appropriate in Persian sturgeon spermatozoa droplet vitrification. Besides, the present technique obtained higher quality of spermatozoa comparing to its analogue techniques.  相似文献   

20.
A Preliminary Bacteriological Study of Refrigerated Channel Catfish Sperm   总被引：1，自引：0，他引：1  
Abstract.— This study was designed to simulate conditions encountered routinely during refrigerated storage of channel catfish sperm. Sperm samples were stored at 4 C in non-sterile and sterile Hanks' balanced salt solution (HBSS). Non-sterile HBSS was prepared with distilled water stored for 2 wk in a plastic carboy prior to use. Observations were made on the frequency and abundance of bacteria in samples, and on changes in sperm motility and quality. Sperm samples stored in non-sterile HBSS had a complete loss of motility within 72 h. Samples maintained in sterile HBSS showed an initial decrease in motility between 48 and 72 h, and a complete loss of motility within 10 d. Quality of the sperm in each buffer decreased as motility decreased; morphologic changes and reduced motility of sperm were coincident with increased bacterial numbers. Bacteria were cultured on tryptic soy agar and Pseudomonas F agar (PFA) by spread-plating 10–μL aliquots from each sample onto bacteriologic media and incubating for 5 d. The dominant bacteria observed were members of the genus Pseudomonas , representing 67% of the total bacteria identified. The dominant pseudomonad ( Pseudomonas sp.) cultured from sperm samples stored in sterile buffer produced caseinase, lecithinase, and was β-hemolytic, whereas the dominant bacteria ( P. putida ) cultured from samples stored in the non-sterile buffers did not. Highly motile pseudomonads, present in two samples stored in sterile buffer, colonized below the surface of the PFA media at 4 C. The attributes of the bacterial contaminants that likely contributed to the decrease in sperm quality were production of extracellular enzymes, consumption of oxygen, and a high level of motility. Potential sources of degradative bacteria were commensal flora of channel catfish and the water used in preparing the storage buffer.  相似文献   

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