scholarly journals The antioxidant effects of soybean lecithin- or low-density lipoprotein-based extenders for the cryopreservation of brown-bear (Ursus arctos) spermatozoa

2013 ◽  
Vol 25 (8) ◽  
pp. 1185 ◽  
Author(s):  
M. Alvarez-Rodríguez ◽  
M. Alvarez ◽  
L. Anel-López ◽  
C. Martínez-Rodríguez ◽  
F. Martínez-Pastor ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Ursus Arctos ◽  
Soybean Lecithin ◽  
Density Lipoprotein ◽  
Brown Bear ◽  
Egg Yolk ◽  
Type A ◽  
Low Density ◽  
Sperm Viability ◽  
Anti Oxidant

Egg yolk low-density lipoproteins (LDL) and soybean lecithin were evaluated as replacements for egg yolk in extenders used for the cryopreservation of brown-bear spermatozoa. The motility, viability and acrosomal status of post-thawed spermatozoa were analysed, and an egg-yolk extender was used as a control. The total antioxidant capacity of these extenders was tested. Soybean lecithin showed an effect that was dependent on the soybean concentration (2%, 3.5% and 5%) and source (Type A: 24% l-α-phosphatidylcholine, and Type B: 14–23% l-α-phosphatidylcholine). Only semen cryopreserved with 5% Type A soybean exhibited a sperm motility similar to that of semen cryopreserved in egg-yolk-based extender after thawing, although the sperm viability and acrosome status were not as high. Semen frozen in an extender containing LDL (10–15%) exhibited improved sperm viability in comparison with the control, but sperm motility was lower. The LDL-based extender exhibited a higher anti-oxidant activity than the egg-yolk extender and soy lecithin-based extenders. The extenders with higher anti-oxidant activity showed improvements in frozen sperm viability but lower semen motility. These results indicate that soybean lecithin did not have the same protective effect as egg yolk during the freezing of brown-bear spermatozoa but suggest that LDL (10–15%) could be a useful substitute for egg yolk in these extenders.

scholarly journals Efektivitas Low Density Lipoprotein dan Kuning Telur Ayam dan Puyuh pada Pengawetan Semen Ayam Merawang (EFFECTIVESS OF LOW DENSITY LIPOPROTEIN AND EGG YOLK FROM CHICKEN AND QUAIL ON MERAWANG SEMEN PRESERVATION)

2017 ◽  
Vol 18 (3) ◽  
pp. 345
Author(s):  
Magfira Magfira ◽  
Raden Iis Arifiantini ◽  
Ni Wayan Karniani Karja ◽  
Sri Darwati
Keyword(s):  
Sperm Motility ◽  
Cold Shock ◽  
Semen Quality ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Egg Yolk ◽  
Low Density ◽  
Sperm Abnormality ◽  
Semen Preservation ◽  
Chicken Egg Yolk

The successful of artificial insemination (AI) depends on the semen quality and extender. To minimize effect of cold shock during storage, extender is added with egg yolk. The objectives of this study were to compare the effectiveness of pure Low Density Lipoprotein (LDL) and egg yolk from domestic chicken and quail on motility and longevity of Merawang chicken sperm. The semen was collected by massage method from three Merawang roosters. Immediately after collection, semen was evaluated macroscopically and microscopically. Only semen demonstrated >70% motility and <20% sperm abnormality were used in this study. Semen divided into four aliquots and diluted with Lactate Ringer (LR) LDL chicken (RL-LDL-C), LR-LDL quail (LR-LDL-Q), LR- chicken Egg Yolk (LR-CEY), Ringer Lactate quail Egg Yolk (RL-QEY). Diluted semen than stored at 5oC. Sperm motility was examined twice a day and the longevity of sperm was determined every day until the sperm reach 0% motility. The motility of spermatozoa in the LR-LDL diluent differed from the sperm motility in the RL-QEY diluent at the 60th and 72th hour (P <0.05) poststorage. However, there was no difference in motility sperm in LR-LDL-C, RL-LDL-Q and RL-CEY. Additionally, there is no difference (P> 0.05) in spermatozoa longivity in the four diluents, with a range of longivities between 4.43 to 5.93 days. ABSTRAK Keberhasilan inseminasi buatan (IB) salah satunya bergantung pada kualitas semen dan pengencer yang digunakan. Dalam meminimalisir pengaruh cold shock saat penyimpanan, pengencer ditambahkan dengan kuning telur. Tujuan dari penelitian ini adalah untuk membandingkan efektivitas Low Density Lipoprotein (LDL) dan kuning telur yang berasal dari ayam kampung dan puyuh terhadap motilitas dan longivitas spermatozoa ayam. Koleksi semen dilakukan menggunakan metode pemijatan pada tiga ekor ayam merawang. Setelah semen dikoleksi, selanjutnya semen dievaluasi secara makroskopis dan mikroskopis. Semen yang menunjukkan motilitas 70% dan abnormalitas kurang dari 20% dibagi empat dan diencerkan menggunakan Ringer Laktat-LDLA (RL-LDLA), Ringer Laktat-(RL-LDLP), Ringer Laktatkuning telur ayam (RL-KTA), dan RL-kuning telur puyuh (RL-KTP). Semen yang telah diencerkan kemudian disimpan pada suhu 5oC. Motilitas spermatozoa diamati dua kali sehari sampai motilitas mencapai 0%. Motilitas spermatozoa dalam pengencer RL-LDLA berbeda dengan motilitas spermatozoa dalam pengencer RL-KTP pada jam ke-60 dan ke-72 (P<0.05) pascapenyimpanan. Akan tetapi tidak terdapat perbedaan motilitas spermatozoa dalam RL-LDLA, RL-LDLP dan RL-KTA. Longivitas spermatozoa dalam empat pengencer tidak terdapat perbedaan (P>0.05) dengan rentang longivitas antara 4,43 sampai 5,93 hari.

245 USE OF A TRIPLE STAIN (SYBR-14/PI/MC540) FOR VIABILITY AND CAPACITATION ASSESSMENT IN THAWED SEMEN FROM BROWN BEAR (URSUS ARCTOS)

2007 ◽  
Vol 19 (1) ◽  
pp. 239 ◽  
Author(s):  
V. Garcia-Macias ◽  
F. Martinez-Pastor ◽  
M. Alvarez ◽  
P. Paz ◽  
S. Borragan ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Ursus Arctos ◽  
Brown Bear ◽  
Emission Spectra ◽  
Egg Yolk ◽  
Membrane Lipid ◽  
Molecular Probes ◽  
Lower Percentage ◽  
Sperm Viability ◽  
Lipid Disorder

Application of new sperm assessment techniques would improve our capability to determine the effects of cryopreservation on sperm function. This becomes relevant when germplasm banks are established for endangered species, as in the case of the brown bear in Spain. Different triple stain techniques have been used in conjunction with flow cytometry to assess various sperm attributes including viability and acrosome status. However, fluorochromes with similar emission spectra may interfere with data resolution, making acquisition and interpretation of data difficult. The double stain combination of SYBR-14 and PI (propidium iodide; max λ 617) has been widely used to differentiate live from dead spermatozoa (spz), and, more recently, merocyanine 540 (MC; max λ 555) has been used to detect a sperm membrane lipid disorder associated with sperm capacitation. In the present study, we analyzed the suitability of combining SYBR-14/PI with MC for simultaneous determination of the viability and capacitation status of frozen–thawed spermatozoa of brown bears (n = 10; semi-free ranging; Cabarceno Park, Cantabria, Spain) obtained by electroejaculation under general anesthesia (7 mg kg-1 tiletamine + zolazepan and 2 mg kg-1 ketamine). Semen was diluted (Tes-Tris-fructose, 8% glycerol, 20% egg yolk, EDTA, and Equex paste), loaded in 0.25-mL straws, and frozen in a biofreezer at 20�C min-1 to -100�C. After storage in liquid nitrogen, samples were thawed at 65�C for 6 s, divided into 2 aliquots (1–2 million spz mL-1), extended with 300 �L PBS, and stained with SYBR14 (1.2 �L) and PI (3 �L; LIVE/DEAD� Sperm Viability Kit; Molecular Probes, Inc., Eugene, OR, USA). To evaluate the possible interaction of MC on sperm viability, half of the aliquots were counterstained with 1.5 �L of MC (diluted with 2.7 �M of DMSO); the other half were not counterstained (control). All tubes were incubated at 37�C for 30 min, and assessed by flow cytometry (FACSCalibur; BD Biosciences, San Jose, CA, USA). Data were analyzed with Bland-Altman. Results indicated that MC staining was mainly confined to dead spermatozoa (22.2 � 7.7%), whereas a lower percentage of live spermatozoa (5.7 � 1.6%; P &lt; 0.05) were also stained with MC. Possibly, the staining of dead spermatozoa with MC was due to capacitation changes induced by cryopreservation. The percentage of live spermatozoa was not different between samples counterstained with MC (68.9 � 9.2) and non-MC-stained control samples (68.6 � 8.8). Thus, we consider that MC does not influence SYBR14/PI discrimination of viable spermatozoa, and that the 3 stains can be used simultaneously. However, more studies are necessary to determine whether MC can be used to distinguish the capacitation status of brown bear thawed spermatozoa. This work was supported by CANTUR S.A. and CICYT (CGL 2004-0278/BOS).

Characterization of glycopeptides derived from the low-density lipoprotein of hen's egg yolk

1968 ◽  
Vol 46 (8) ◽  
pp. 983-988 ◽  
Author(s):  
J. Z. Augustyniak ◽  
W. G. Martin
Keyword(s):  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Egg Yolk ◽  
Low Density ◽  
Amide Nitrogen ◽  
Acetylneuraminic Acid ◽  
Hen’S Egg ◽  
Terminal Amino ◽  
Protein Moiety

Two glycopeptides (A and B) were isolated from pronase-digested vitellenin, the protein moiety of the low-density lipoprotein of hen's egg yolk. Aspartic acid was the only N-terminal amino acid of both glycopeptides but only A contained N-acetylneuraminic acid. A contained 55% hexose (mannose), 14% hexosamine, 12% N-acetylneuraminic acid, 0.71% amide nitrogen, and its molecular weight was 2.3 × 103. The corresponding values for B were 64, 17, 0.0, 0.75, and 2.0 × 103. Chemical analyses showed that B (and probably A) occurs in vitellenin with the heteropolysaccharide group bound N-glycosidically via the β-amide group of an asparaginyl residue. The indicated structure is R∙(NH)Asp∙Thr∙Ser∙(Ala, Gly, Val)∙Ile, where R, the heteropolysaccharide group, contains 2 hexosamine and 8 hexose residues.

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