263. Substitution of skim milk with bovine serum albumin in a stallion semen diluent

2008 ◽  
Vol 20 (9) ◽  
pp. 63
Author(s):  
Z. Gibb ◽  
C. G. Grupen ◽  
L. H. A. Morris ◽  
G. Evans ◽  
W. M. C. Maxwell
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Sperm Quality ◽  
Skim Milk ◽  
Lower Percentage ◽  
Animal Science ◽  
Flow Cytometric ◽  
Alternative Protein ◽  
Protective Proteins

Skim milk has long been utilised as a source of protective proteins in stallion semen diluents. However, skim milk is also thought to contain components that are toxic to sperm and reduces the clarity of sperm suspensions, which impedes sperm assessments. This may also reduce the effectiveness of staining procedures used to process sperm for flow cytometric sex-sorting. The aim of this study was to ascertain the optimal concentration of bovine serum albumin (BSA) to replace skim milk in a traditional stallion semen diluent, Kenney's Modified Tyrode's (KMT) Medium1, for handling and processing stallion sperm before flow cytometric sex-sorting. Two ejaculates were collected from each of three pony stallions. Each ejaculate was divided into five aliquots and diluted in either KMT with skim milk or KMT supplemented with 0, 0.25, 0.5 or 1% BSA. Diluted samples were further divided into two aliquots and either stored at 15°C for 18 h before incubation and assessment, or incubated and assessed immediately upon arrival. Samples were incubated at 34°C and evaluated at 0, 45 and 90 min for objective motility and acrosome integrity. No interactions were observed between any treatments over time. There was a lower percentage of intact and a higher percentage of detached acrosomes for sperm incubated in KMT containing 0% BSA than all other treatments. A greater proportion of sperm incubated in KMT with skim milk had partial acrosome damage compared with other treatments. There was no difference in % total motility for sperm incubated in KMT with skim milk, and KMT containing 0.5 and 1% BSA (Table 1). These results indicate that BSA may be suitable as an alternative protein source in stallion semen diluents. Further studies are required to compare sex-sorting rates and sperm quality after sex-sorting, incubation and staining in skim milk compared with BSA-based media. (1) Padilla, A W and Foote, R H 1991, ‘Extender and centrifugation effects on the motility patterns of slow-cooled stallion spermatozoa’, Journal of Animal Science, vol. 69, no. 8, pp. 3308–331

scholarly journals PENGGANTIAN BOVINE SERUM ALBUMIN PADA PENGENCER CEP-2 DENGAN SERUM DARAH SAPI DAN PUTIH TELUR TERHADAP KUALITAS SEMEN CAIR SAPI LIMOUSIN SELAMA PENDINGINAN (The Substitution of Bovine Serum Albumin with Cattle Blood Serum and Egg White in CEP-2 Diluent on the Quality of Limousin Bull Liquid Semen during Refrigerating)

2016 ◽  
Vol 10 (2) ◽  
pp. 98-102
Author(s):  
Trinil Susilawati ◽  
Feri Eka Wahyudi ◽  
Inna Anggraeni ◽  
Nurul Isnaini ◽  
Muhammad Nur Ihsan
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Blood Serum ◽  
Sperm Motility ◽  
Bovine Serum ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Egg White ◽  
Cattle Blood

This study aims to determine the effect of the substitution of bovine serum albumin (BSA) with cattle blood serum and egg white in the diluent of Cauda epididymal Plasma 2 (CEP-2) on sperm quality of Limousin cattle during cooling at 3-5 C. The research material used was rejected Limousin bull sperm (motility of 50-60%) from Artificial Insemination Centre Singosari, Malang. This research was a laboratory experiment using a randomized block design which was composed of six treatments with 10 replications, those were T0 as controls ((90% CEP-2 with BSA + 10% egg yolk); T1 (83.84% CEP-2 + 6.16% cattle blood serum + 10% egg yolk); T2 (81.84% CEP-2 + 8.16% cattle blood serum + 10% egg yolk); dan T3 (90% CEP-2 + 0,4% egg white + 10% egg yolk); T4 (90% CEP-2 + 0.8% egg white + 10% egg yolk); and T5 (90% CEP-2 without BSA + 10% egg yolk). Parameters measured were the percentage of motility, viability, and abnormality of sperms. Results of research after 48 hours of storage showed that the percentage of sperm motility in T0, T1, T2, T3, T4, and T5 were 40.50±5.90, 36±36.16, 34.00±6.58, 40.50±3.69, 38.50±3.37, and 38.50±4.12, respectively, while the percentage of sperms viability were 75.16±4.30, 70.50±2.88, 73.80±2.80, 74.80±3.30, 75.13±3.13, and 74.03±4.13, respectively, and the percentage of sperms abnormality were 10.14±2.34, 10.62±1.34, 11.33±2.00, 10.94±2.82, 10.02±1.95, and 10.78±1.96, respectively. In conclusion, CEP-2 diluent with or without the addition of 19% egg yolk in BSA and the substitution of BSA with 0.4-0.8% egg white can maintain semen quality to hour of 48 in cold storage.This study aims to determine the effect of the substitution of bovine serum albumin (BSA) with cattle blood serum and egg white in the diluent of Cauda epididymal Plasma 2 (CEP-2) on sperm quality of Limousin cattle during cooling at 3-5 C. The research material used was rejected Limousin bull sperm (motility of 50-60%) from Artificial Insemination Centre Singosari, Malang. This research was a laboratory experiment using a randomized block design which was composed of six treatments with 10 replications, those were T0 as controls ((90% CEP-2 with BSA + 10% egg yolk); T1 (83.84% CEP-2 + 6.16% cattle blood serum + 10% egg yolk); T2 (81.84% CEP-2 + 8.16% cattle blood serum + 10% egg yolk); dan T3 (90% CEP-2 + 0,4% egg white + 10% egg yolk); T4 (90% CEP-2 + 0.8% egg white + 10% egg yolk); and T5 (90% CEP-2 without BSA + 10% egg yolk). Parameters measured were the percentage of motility, viability, and abnormality of sperms. Results of research after 48 hours of storage showed that the percentage of sperm motility in T0, T1, T2, T3, T4, and T5 were 40.50±5.90, 36±36.16, 34.00±6.58, 40.50±3.69, 38.50±3.37, and 38.50±4.12, respectively, while the percentage of sperms viability were 75.16±4.30, 70.50±2.88, 73.80±2.80, 74.80±3.30, 75.13±3.13, and 74.03±4.13, respectively, and the percentage of sperms abnormality were 10.14±2.34, 10.62±1.34, 11.33±2.00, 10.94±2.82, 10.02±1.95, and 10.78±1.96, respectively. In conclusion, CEP-2 diluent with or without the addition of 19% egg yolk in BSA and the substitution of BSA with 0.4-0.8% egg white can maintain semen quality to hour of 48 in cold storage.

scholarly journals Allogeneic Mesenchymal Stem Cell Treatment Induces Specific Alloantibodies in Horses

2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
Sean D. Owens ◽  
Amir Kol ◽  
Naomi J. Walker ◽  
Dori L. Borjesson
Keyword(s):  
Bone Marrow ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Research Projects ◽  
Allogeneic Mscs ◽  
Humoral Immune ◽  
Flow Cytometric ◽  
Stem Cell Treatment ◽  
Mesenchymal Stem Cell Treatment

Background.It is unknown whether horses that receive allogeneic mesenchymal stem cells (MSCs) injections develop specific humoral immune response. Our goal was to develop and validate a flow cytometric MSC crossmatch procedure and to determine if horses that received allogeneic MSCs in a clinical setting developed measurable antibodies following MSC administration.Methods.Serum was collected from a total of 19 horses enrolled in 3 different research projects. Horses in the 3 studies all received unmatched allogeneic MSCs. Bone marrow (BM) or adipose tissue derived MSCs (ad-MSCs) were administered via intravenous, intra-arterial, intratendon, or intraocular routes. Anti-MSCs and anti-bovine serum albumin antibodies were detected via flow cytometry and ELISA, respectively.Results.Overall, anti-MSC antibodies were detected in 37% of the horses. The majority of horses (89%) were positive for anti-bovine serum albumin (BSA) antibodies prior to and after MSC injection. Finally, there was no correlation between the amount of anti-BSA antibody and the development of anti-MSC antibodies.Conclusion.Anti allo-MSC antibody development was common; however, the significance of these antibodies is unknown. There was no correlation between either the presence or absence of antibodies and the percent antibody binding to MSCs and any adverse reaction to a MSC injection.

Effects of Bovine Serum Albumin on Boar Sperm Quality During Liquid Storage at 17°C

2015 ◽  
Vol 50 (2) ◽  
pp. 263-269 ◽  
Author(s):  
X-G Zhang ◽  
G-J Yan ◽  
J-Y Hong ◽  
Z-Z Su ◽  
G-S Yang ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Sperm Quality ◽  
Liquid Storage ◽  
Boar Sperm

Viability of Staphylococci in Various Diluents

1986 ◽  
Vol 7 (7) ◽  
pp. 370-372 ◽  
Author(s):  
Razmick Essiain ◽  
D.J Flournoy
Keyword(s):  
Peritoneal Dialysis ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Tap Water ◽  
Skim Milk ◽  
Deionized Water ◽  
Dialysis Fluid ◽  
Peritoneal Dialysis Fluid ◽  
Better Than

AbstractStandardized suspensions of 78 staphylococci and micrococci were stored in different diluents and tested periodically for 3 weeks to determine their ability to maintain viability. In 92% of the tests, diluent suspensions yielded viable organisms for at least 21 days. Organism survival was maintained in the following order, from best to worst: skim milk, tap water, 0.9% NaCl, deionized water, Dianeal (peritoneal dialysis) fluid, 0.2% bovine serum albumin and 5% glycerol. In 81% of the instances, oxacillin-susceptible staphylococci survived better than oxacillin-resistant staphylococci.

150 α6 Integrin for evaluating sperm quality in bulls with different capacities of invitro embryo production

2020 ◽  
Vol 32 (2) ◽  
pp. 201
Author(s):  
E. G. A. Perez ◽  
D. L. D'Ercole ◽  
M. H. Macedo ◽  
A. A. F. Rodrigues ◽  
R. F. Gonçalves
Keyword(s):  
Bovine Serum Albumin ◽  
Fluorescence Intensity ◽  
Bovine Serum ◽  
Sperm Quality ◽  
Flow Cytometric Analysis ◽  
Fluorescein Isothiocyanate ◽  
Embryo Production ◽  
Logarithmic Amplifier ◽  
Flow Cytometric ◽  
The Mean

The α6 integrin, an adhesion molecule, is expressed on bovine sperm, but major questions about the role of integrins in sperm-oocyte fusion remain unsolved. In this work, we show the results of characterisation of sperm α6 integrin from 4 bulls with different capacities of invitro embryo production using flow cytometric analysis. The bull capacities judged by the rate of blastocyst formation after invitro embryo production with semen from 5 ejaculates per animal were 44.3, 17.1, 13.2, and 15.0% for bulls 1, 2, 3, and 4, respectively (P<0.05). For flow cytometric analysis, surface expression of α6 integrin was evaluated using a fluorescence-activated cell sorter (FACScan, Coulter Electronics) using a 520-nm excitation from an argon laser at 150 mW for excitation. Frozen-thawed sperm were centrifuged at 700×g for 10min and washed once in warm phosphate buffered saline (PBS). Briefly, the spermatozoa (5×105 cells per sample) were resuspended in 100μL of PBS containing 1% bovine serum albumin. After washing 3 times, the live cells were incubated at room temperature for 1h with 100μL (1:500) of α6 monoclonal antibody (Chemicon) in PBS (0.1M, pH 7.4) with 1% bovine serum albumin. After washing three times, the cells were incubated at 4°C for 1h in the dark with the fluorescein isothiocyanate-conjugated F(ab)2 fragment of affinity isolated goat anti-mouse antibody (Invitrogen). The cells were washed three times, resuspended in 100μL of PBS, and analysed. For each sample, 10 000 cells were recorded at a flow rate of 200-300 cells s−1 using forward scatter (cell size) and side angle of light scatter (cell density), the first using a logarithmic amplifier and the second using a linear amplifier. The fluorescence data were collected using the logarithmic amplifier. The percentage of positive cells and the mean fluorescence channel on a 1023-channel scale were calculated using Epics Profile II software (Epics II Software). To define the forward and side-scatter regions corresponding to sperm, binding of fluorescein isothiocyanate-conjugated Pisum sativum agglutinin to the acrosome was used for setting the bitmap on the dot plot. Initially, the percentage of spermatozoa stained with α6 antibody was calculated on a per-individual basis. Subsequently, the mean±standard deviation was calculated for each group. The Mann-Whitney U test was used to compare the differences in the expression of α6 between groups. Expression of α6 integrin was higher in bull 1 (control) than in bulls with low capacities of invitro embryo production. The spermatozoa from bull 1 was distributed in a single broad peak with a mean fluorescence intensity of 36.16±4.17%. The increased distance of the fluorescence intensity in bull 1 compared with the weak fluorescence peaks in the others bulls reflects the increased width and distance of the population distribution. In conclusion, α6 integrin may be used as a biomarker to evaluate sperm quality. This study was supported by FAPESP grants 2010/01077-9, 2011/18085-7, and 2016/00976-6.

Effects of α-Linolenic Acid and Bovine Serum Albumin on Frozen-thawed Boar Sperm Quality during Cryopreservation

2016 ◽  
Vol 40 (4) ◽  
pp. 33-37 ◽  
Author(s):  
Won-Hee Lee ◽  
◽  
Yong Hwangbo ◽  
Sang-Hee Lee ◽  
Hee-Tae Cheong ◽  
...  
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Linolenic Acid ◽  
Bovine Serum ◽  
Sperm Quality ◽  
Boar Sperm

scholarly journals The quality of Etawah crossbreed sperm after sexing with different combination of Bovine Serum Albumin concentrations

2021 ◽  
Vol 888 (1) ◽  
pp. 012025
Author(s):  
S D Rasad ◽  
N Solihati ◽  
K Winangun
Keyword(s):  
Plasma Membrane ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Sperm Quality ◽  
Duncan Multiple Range Test ◽  
Randomized Design ◽  
Completely Randomized Design ◽  
Range Test

Abstract This research aimed to determine the quality of Etawah crossbreed sperm after sexing with different combinantion of Bovine Serum Albumin (BSA) concentrations. The parameter of this research were motility, viability, intact plasma membrane (IPM) and intact acrosome cap (IAC) (%). The Completely randomized design (CRD) was applied in this experiment involving 4 treatments of the combination of BSA concentrations (T1=3%:6%, T2 = 4%:8%, T3=5%:10%, T4=6%:12% at upper and lower fraction) and each treatments was repeated 5 times at post chilled and post sexing. Data were analyzed by using analysis of variance (ANOVA), followed by Duncan Multiple Range Test. The result showed that thecombination of BSA concentrations affected (P<0.05) motility, viability, IPM and IAC. The highest value of sperm quality in upper and lower fraction was obtained from the combination BSA of 5%:10% (motility 78.60 ± 2.61% and 73.80 ± 2.49%; viability 282 ± 14.30 and 252.8 ± 12.97 hours; IPM-value 79.60 ± 1.98% and 74.70 ± 1.82% and IAC 81.00 ± 1.46% and 76.90 ± 1.29%). Based on the results it can be concluded that the quality of Etawah crossbreed sperm after sexing is affected by the combination of BSA concentrations.

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