Efficiency of Different Diluents and Dilution Rates on the Fertilization Potential of Chicken Spermatozoa

Author(s):  
V. Beulah Pearlin ◽  
J. Mohan ◽  
J.S. Tyagi ◽  
M. Gopi ◽  
G. Kolluri ◽  
...  

Background: Artificial Insemination (AI) has been well established to obtain better fertility than natural mating. Semen extension and preservation play a major role in AI technique. The concentrated nature of chicken semen entails dilution of semen for easy handling and storage. With the availability of a wide range of extenders, exploring various extenders and dilution rates supplement the reproductive efficiency achieved through modern AI techniques.Methods: Pooled semen samples obtained from twenty White Leghorn breeder males were split equally into three portions, each for CARI poultry semen diluent, EK extender and Tselutin extender. At 1:2 dilution rate, the samples were subjected to sperm motility analysis from 0 to 96 h and AI was targeted at 0 and 24 h of semen collection. Fertility was assessed by candling of eggs collected from 2 to 8 days post insemination at 9th day of incubation. Further investigation of different dilution rates (1:2, 1:4, 1:6, 1:8, 1:10, 1:12, 1:14, 1:16, 1:18, 1:20) on fertility of fresh semen was done using the superior diluent.Result: Upon analysis, sperm motility showed no significant difference among the diluents at 0 h storage while CARI poultry semen diluent showed superior motility (%) at the subsequent storage periods, followed by EK and Tselutin extender. Higher fertility (p less than 0.05) was expressed in the diluents of CARI and EK at 0 h storage, whereas CARI poultry semen diluent showed superior fertility pattern (p less than 0.05) followed by EK and least by Tselutin extender at 24 h semen storage. Further investigation of different dilution rates using CARI diluent, exhibited fertility (p less than 0.05) of 90% and higher at 1:2 and 1:4 dilutions, followed by 1:6 and 1:8 at 2 to 6 days of fertile period.

2021 ◽  
Vol 19 (2) ◽  
pp. 237-244
Author(s):  
Nguyen Huu Duc ◽  
Pham Thu Giang ◽  
Tran Thi Binh Nguyen ◽  
Nguyen Thi Mai ◽  
Bui Dai Phong

The objective of this study was to determine the semen cryopreservation capacity of BBB bulls in Hanoi-Vietnam. Research conducted on the fresh semen collected from 05 BBB bulls. Results showed that semen color was normal (milky white, ivory white, ivory yellow), semen volume ranged from 6.35 mL to 7.48 mL (P <0.05), initial motility of semen ranged from 80.53% to 82.92% (P <0.05), sperm concentration in semen  ranged from 1.02 x 109 sperms/ml to 1.12 x 109 sperms/mL (P <0.05), abnormal sperm ratio ranged from 6.45% to 8.12% (P <0.05), alive sperm ratio ranged from 76.34% to 82.97% (P <0.05), sperm motility after thawing from straw semen ranged from 71.33% to 75.92% (P<0.05). In conclusion, successfully semen collection from 05 breeding BBB bulls at Hanoi Cattle Breeding Joint Stock Company, semen samples had normal color and good quantity and quality, suitable for production of frozen semen; and semen cryopreservation of straws of the 05 bull BBB semen mentioned at -196oC, sperm motility after freezing-thawing reached the economic and technical norms of 675/2014 of the Ministry of Agriculture and Rural Development.


2015 ◽  
Vol 36 (6Supl2) ◽  
pp. 4493
Author(s):  
Ronan Maciel Marcos ◽  
Giovano Neumann ◽  
Cesar Pereira Rebechi de Toledo ◽  
João Marcos Sena ◽  
Gilmar Baumgartner ◽  
...  

<p>This study describes the seminal and spermatic characteristics of fresh semen of <em>Steindachneridion melanodermatum </em>and investigates the effects of dilution, temperature, and storage period on its spermatic parameters. Sperm samples were collected from nine hormonally-induced males. The following parameters in fresh sperm were analyzed: seminal plasma osmolality (OSM), seminal pH, sperm motility (MOT), sperm velocity (SV) (including sperm curvilinear velocity (CVV), sperm straight-line velocity (SLV), and sperm average path velocity (APV)), total time of sperm motility (TEMP), sperm concentration (CONC), and index of sperm normality (NORM). Sperm samples from each male were diluted in a solution containing 5% fructose and 5% powdered milk, and stored at 10°C and 25°C. The same was carried out for sperm samples not subjected to dilution. From these samples, MOT, CVV, SLV, APV, SV, and TEMP were measured after 0 h, 5 h, 9 h, 18 h, 27 h, 36 h, 45 h, and 54 h. Males released 11.74 ± 5.38 mL of sperm, with an osmolality of 258.78 ± 29.36 mOsm.kg-1 and pH of 7.11 ± 0.31. The sperm presented a MOT of 99.86 ± 0.31% at a concentration of 1.03 × 1010 ± 3.65 × 109 spermatozoa.mL-1 with CVV of 185.58 ± 14.11 ?m.s-1, SLV of 49.15 ± 4.66 ?m.s-1, APV of 87.02 ± 4.13 ?m.s-1, SV of 106.52 ± 4.45 ?m.s-1, TEMP of 79.31 ± 5.62 s, NORM of 75.81 ± 5.71%. The results indicate that sperm motility, sperm velocity, and total time of sperm activation were affected by dilution, storage temperature, and storage period (p &lt; 0.05). Procedures for semen storage should be performed with undiluted sperm cooled at 10°C, or kept undiluted at 25°C for up to 27 h.</p>


2015 ◽  
Vol 49 (6) ◽  
Author(s):  
S Frydrychová ◽  
A Lustyková ◽  
E Václavková ◽  
J Lipenský ◽  
M Rozkot

The objective of this study was to investigate the effect of using different extenders <italic>viz.</italic> Androhep, Safecell Plus and SUS during cryopreservation on quality of frozen-thawed boar semen. Semen volume, sperm motility, sperm concentration, percentage of morphologically abnormal spermatozoa, total number of spermatozoa per ejaculate and activity of the enzyme aspartate aminotransferase (AST) were assessed in fresh semen collected from 39 fertile AI boars. Semen from each boar was divided into three portions and diluted 1:1.5 in extender Androhep, Safecell Plus and SUS and keep at 17°C for 15-h holding time before cryopreservation. Then sperm was cryopreserved. Straws were thawed in a water bath at 38°C for 40s and post-thaw sperm motility with AST activity was assessed. Significant difference in post-thaw sperm motility was found between extender Androhep and Safecell Plus (P<0.05). AST activity did not differ significantly between tested extenders (P>0.05). In conclusion, the results of the study indicate that using Safecell Plus extender during holding period before cryopreservation significantly affected post-thaw sperm motility.


2015 ◽  
Vol 36 (6Supl2) ◽  
pp. 4493
Author(s):  
Ronan Maciel Marcos ◽  
Giovano Neumann ◽  
Cesar Pereira Rebechi de Toledo ◽  
João Marcos Sena ◽  
Gilmar Baumgartner ◽  
...  

This study describes the seminal and spermatic characteristics of fresh semen of Steindachneridion melanodermatum and investigates the effects of dilution, temperature, and storage period on its spermatic parameters. Sperm samples were collected from nine hormonally-induced males. The following parameters in fresh sperm were analyzed: seminal plasma osmolality (OSM), seminal pH, sperm motility (MOT), sperm velocity (SV) (including sperm curvilinear velocity (CVV), sperm straight-line velocity (SLV), and sperm average path velocity (APV)), total time of sperm motility (TEMP), sperm concentration (CONC), and index of sperm normality (NORM). Sperm samples from each male were diluted in a solution containing 5% fructose and 5% powdered milk, and stored at 10°C and 25°C. The same was carried out for sperm samples not subjected to dilution. From these samples, MOT, CVV, SLV, APV, SV, and TEMP were measured after 0 h, 5 h, 9 h, 18 h, 27 h, 36 h, 45 h, and 54 h. Males released 11.74 ± 5.38 mL of sperm, with an osmolality of 258.78 ± 29.36 mOsm.kg-1 and pH of 7.11 ± 0.31. The sperm presented a MOT of 99.86 ± 0.31% at a concentration of 1.03 × 1010 ± 3.65 × 109 spermatozoa.mL-1 with CVV of 185.58 ± 14.11 ?m.s-1, SLV of 49.15 ± 4.66 ?m.s-1, APV of 87.02 ± 4.13 ?m.s-1, SV of 106.52 ± 4.45 ?m.s-1, TEMP of 79.31 ± 5.62 s, NORM of 75.81 ± 5.71%. The results indicate that sperm motility, sperm velocity, and total time of sperm activation were affected by dilution, storage temperature, and storage period (p < 0.05). Procedures for semen storage should be performed with undiluted sperm cooled at 10°C, or kept undiluted at 25°C for up to 27 h.


2016 ◽  
Vol 28 (2) ◽  
pp. 149
Author(s):  
M. M. Seshoka ◽  
M. L. Mphaphathi ◽  
K. S. Mafolo ◽  
M. Nkadimeng ◽  
Z. C. Raphalalani ◽  
...  

Traditionally, commercial hen egg yolk has been used in extenders or freezing media because of its easy availability. However, the use of quail egg yolk has not been used for preserving Nguni bull semen. The aim of the study was to compare the suitability of different quail egg yolk concentrations (5, 10, 15, and 20%) for cryopreserving Nguni bull semen. Semen was collected from 14 stud Nguni bulls with the aid of electro ejaculator. Collected semen samples were kept in a thermos-flask containing warm water at a temperature of 37°C and transported to the laboratory for further analyses. The sperm motility traits were evaluated using computer-aided sperm analysis prior extension. Semen samples were then randomly allocated into 5, 10, 15, and 20% of quail egg yolk and 20% concentration of chicken egg yolk (control) in citrate extender. The extender was supplemented with 12% of glycerol (Seshoka et al. 2012) as a cryoprotectant, and semen samples were diluted (1 : 2) and equilibrated for a period of 4 h at 5°C. After equilibration, semen samples were loaded into 0.25-mL straws, placed into a controlled rate programmable freezer, and stored in a LN tank (–196°C) until thawing. Frozen semen straws were thawed in a water bath at 37°C for 1 min. Thawed semen was evaluated for sperm motility traits using a computer-aided sperm analysis system. Data were analysed with ANOVA. A significant difference was recorded between fresh total sperm motility rate (99%) and frozen-thawed semen samples with either 5% (87.3%) quail or 20% (87.6%) chicken egg yolk citrate extender compared with 10% (92.6%), 15% (91.2%), or 20% (89.9%) quail egg yolk citrate extender. Moreover, fresh semen also resulted in a significantly higher progressive sperm motility rate (39.3%) as compared with frozen-thawed with 5% (26.2%) or 20% quail (28.5%) or 20% chicken (22.7%) egg yolk citrate extender. The results also demonstrated that the use of 10, 15, and 20% quail egg yolk in citrate extender yielded comparable results on total sperm motility with fresh semen as compared with 5% quail and 20% chicken egg yolk. In conclusion, quail egg yolk extender provided sufficient cryo-effectiveness to cryopreservation of Nguni bull semen.


2004 ◽  
Vol 24 (4) ◽  
pp. 385-391 ◽  
Author(s):  
Per Kjellstrand ◽  
Martin Erixon ◽  
Anders Wieslander ◽  
Torbjörn Lindén ◽  
Evi Martinson

Objective Bioincompatible glucose degradation products (GDPs) develop during heat sterilization of peritoneal dialysis (PD) fluids. However, degradation may also take place during storage. Consequently, storage may add to the bioincompatibility caused by heat sterilization. The aim of the present study was to investigate how different factors such as the sterilization procedure, pH, glucose concentration, and temperature influence GDP production during storage. Design Degradation in glucose solutions was followed by pH and UV absorbance at 228 nm and 284 nm over 2 years of storage. Different sterilization times, storage temperatures, pH, and glucose concentrations were included in the study. Peritoneal dialysis fluids were also used in the experiment. Bioincompatibility was estimated through inhibition of cell growth in L-929 fibroblasts, and GDPs through UV absorption and liquid chromatography. Results The most important factor determining the rate of GDP production during storage was temperature. The GDPs created by heat sterilization promoted further degradation of glucose during subsequent storage. A pH of around 3.2 protected glucose from degradation during both heat sterilization and storage. At a storage temperature of 20°C and a pH of 3.2, degradation was almost negligible. Heat sterilization produced considerable amounts of GDPs absorbing at 228 nm. During initial storage, these 228 nm-absorbing GDPs almost disappeared. After reaching a nadir, absorbance at 228 nm again started to increase. Contrary to this, absorbance at 284 nm [caused mainly by 5-hydroxymethyl-2-furaldehyde (5-HMF)] increased during the whole storage period. After 2 years at 40°C, the concentrations of GDPs produced during storage were of the same magnitude as those caused by heat sterilization. Inhibition of cell growth of L-929 fibroblasts correlated well with the part of the absorbance at 228 nm not caused by 5-HMF in glucose solutions that were heat sterilized under a wide range of conditions. This part of 228 nm absorbance (denoted 228corr) was caused almost entirely by 3,4-dideoxyglucosone-3-ene (3,4-DGE). Conclusions Temperature is the single most important factor for glucose degradation during storage. The concentrations of bioincompatible GDPs produced may, under improper conditions, be as high as those produced during sterilization. High concentrations of glucose and low pH protect glucose from being degraded during both sterilization and storage. A good estimate of 3,4-DGE concentration in the fluids can be obtained correcting the UV absorbance at 228 nm for the influence from 5-HMF (and, when appropriate, for lactate). The 228corr may thus be used as a simple quality control for the fluids.


1996 ◽  
Vol 62 (3) ◽  
pp. 599-604 ◽  
Author(s):  
C. Alexopoulos ◽  
C. Boscos ◽  
Ph. Saratsis ◽  
C. Saoulidis ◽  
S. Kyriakis

AbstractSemen collection from three boars was performed twice a week, and doses (100 ml each) of semen diluted with Beltsville Thaw Solution (BTS) extender were prepared containing 1 × 109, 3 × 109 and 5 × 109 spermatozoa. Diluted semen was then stored at 17°C for a maximum of 120 h. Percentage of motile spermatozoa (PMS) and type of spermatozoa motility (TSM) were determined by microscopic examinations performed at 24-h intervals, from zero time (beginning of storage at 17°C) up to 120 h of storage, and a sperm motility index (SMI) calculated. Sows (no. = 360) were divided into nine experimental groups (40 sows per group) and a combination of number of spermatozoa per dose and storage time was used for artificial insemination in each group. All the animals were inseminated twice at 12 and 36 h after detection of standing oestrus by a teaser boar. Return and farrowing data including litter size were recorded. PMS, TSM and SMI decreased significantly (P < 0·01) from 48 h up to 120 h of storage, this being more marked in semen doses of 5 × 109 spermatozoa. When semen stored for up to 24 h was used, no significant differences (P > 0·05) regarding return rates and farrowing rates were observed among the three groups inseminated with semen containing different numbers of spermatozoa per dose. Inseminations with semen doses of 1 × 109 and 3 × 109 spermatozoa stored for 48 to 72 h have significantly lower return rates (P < 0·05) and significantly higher farrowing rates (P < 0·05) than with doses of 5 × 109 spermatozoa. Inseminations with semen stored for more than 72 h gave relatively higher return rates and lower farrowing rates. Semen storage time and number of spermatozoa per dose appeared to reduce litter size only where semen doses of 5 X109 spermatozoa were inseminated after storage for more than 48 h.


2018 ◽  
Vol 52 (4) ◽  
pp. 299 ◽  
Author(s):  
S. SAMOUILIDIS (Σ. ΣΑΜΟΥΗΛΙΔΗΣ) ◽  
K. SAOULIDIS (Κ. ΣΑΟΥΛΙΔΗΣ) ◽  
A. FOUKOS (Α. ΦΟΥΚΟΣ) ◽  
P. YPSILANTIS (Π. ΥΨΗΛΑΝΤΗΣ) ◽  
A. DEMERTZIS (Α. ΔΕΜΕΡΤΖΗΣ) ◽  
...  

In this study, semen from 10 rabbits was frozen after it was diluted in two different extenders, A and B. Extender A consisted of Tris-buffer in which 20% of egg yolk and 3% of glycerol were added, while extender Β had the same composition, plus 3% of dimethylsulfoxide (DMSO). The semen of the buck exhibiting the best post-thaw motility in the two extenders, was used for the insemination of two corresponding groups (I and II) of 20 does each. The average sperm motility of fresh semen (77%) was significantly reduced (P<0.05) after it was frozen in the extender A (47%) and Β (54%). The post-thaw sperm motility in extender Β was significantly higher (P<0.05) than that in extender A. The percentage of the animals that gave birth in group II (60%) was increased compared to that of the animals in group I (50%), but not significantly (P>0.05). Also, no significant difference (P>0.05) was observed between the average litter size of group I (6,8±1,7) and group II (7,0±1,9). Thereafter, it is concluded that freezing rabbit semen in an extender that contains a combination of the cryoprotective agents glycerol and DMSO, in the proportion of 3% and 3%, respectively, results in a significant improvement of post-thaw sperm motility, while it doesn't affect significantly its fertility value neither the size of the litters.


1992 ◽  
Vol 19 (3) ◽  
pp. 227-232 ◽  
Author(s):  
Niall J. McGuinness ◽  
Christopher D. Stephens

Orthodontic study models form an essential part of the dental records of patients undergoing diagnosis and treatment. In order to ascertain the problems encountered by hospital orthodontic units in the utilization and storage of study models, a questionnaire was circulated in February 1991 to members of the Consultant Orthodontists Group. All respondents took pretreatment study models, while 9 per cent took their final study models at some time other than the end of active treatment; 85·5 per cent of respondents stored their study models in their units, but most were beginning to experience difficulties in this regard. There was a wide range for storage times, and only 10 per cent of employing authorities had a stated policy on the storage of study models. There was a highly significant difference (P&z.Ltc;0·001) between the time that models are stored at present, and the desired storage times. Most respondents appeared to be rather uncertain about the precise medico-legal requirements concerning model storage. The implications for audit and medico-legal matters are discussed in the light of these findings.


The evaluation of sperm quality in the laboratory is essential to improve efficiency in assisted reproduction. As in other species, for the dog there are reports that prolonged contact of sperm with some components of seminal plasma is associated with decreased motility and sperm viability. Thus, the centrifugation is a technique widely used to concentrate the spermatozoa and eliminate the supernatant. The purpose of this study was to evaluate the effect of double layer centrifugation on the percentages of total sperm motility and progressive sperm motility of the dog’s semen submitted to the dilution, single layer centrifugation, cooling and storage at 5 °C for 24 and 48 hours. For this purpose, ejaculates of 30 healthy male dogs were evaluated, by taking into account the comparison among the conventional sperm parameters (ejaculate volume, sperm concentration, total sperm motility and sperm progressive motility). The semen samples were examined in standard baseline condition of fresh semen (FS), after dilution (AD), after dilution and single layer centrifugation (SLC), after double layer centrifugation (DLC). According to the different time points, the semen samples were evaluated in baseline conditions, immediately after their collection at (T0), at 24 h (T24) and at 48 h (T48), to evaluate the effect of different treatments on the semen’s quality. Results showed a significant effect of double layer centrifugation on the improvement of total sperm motility and progressive sperm motility percentages of dogs. The use of cooling fresh semen soon after the double layer centrifugation will improve the semen quality up to 48h, with a special emphasis for the percentages of total sperm motility and sperm progressive motility, adding an alternative technical approach to reproductive performance in male breeding dogs.


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