Effect of Shilajit on Freezing Rooster Semen

Mustafa Yiğit Nizam; Murat Selcuk

Article Details

Effect of Shilajit on Freezing Rooster Semen

Authors:

Mustafa Yiğit Nizam | Murat Selcuk

Publisher:

Afyon Kocatepe University

Main Topics:

Veteriner Bilimleri (Diğer)

Journal:

Kocatepe Veteriner Dergisi Vol.17 Issue 4, pp.393-400

Document Type:

Article

Publication Date:

3/2025

Document Language:

English

Document ID (Org No.):

2001327152

pISSN:

1308-1594

Keywords:

Abstract:

 Reactive oxygen species ROS are created in excess during the cryopreservation process, which speeds up the rate of lipid peroxidation LPO. This negatively impacts spermatozoa functions and reduces their capacity to fertilize. The spermatozoon plasma membrane consists of significant amounts of polyunsaturated fatty acids PUFA, which can be easily oxidized by ROS and produce harmful agents that are toxic to cells. The plasma membrane of rooster spermatozoa contains high concentrations of polyunsaturated fatty acids and very small amounts of mitochondria, cytoplasm, and cytoplasmic antioxidants. Cryopreservation of rooster semen has been associated with adverse effects, including increased lipid peroxidation, structural damage in the mitochondria and acrosomal area, changes in the integrity and permeability of the spermatozoon plasma membrane, and severe damage to DNA. In the study, semen taken from 20 Plymouth Rock roosters were pooled to eliminate individual differences. By adding 5 μgmL, 10 μgmL, 15 μgmL, 20 μgmL and 25 μgmL shilajit to Beltsville Poultry Semen Extender diluent, 5 experimental and 1 control groups were formed and frozen in 0.25 mL straws. After thawing in a water bath at 37oC, spermatologic parameters were analyzed with the CASA system. Viability evaluations were made with eosin  nigrosin stain and morphological evaluations were made with Hancock method. Sperm DNA integrity was examined with the COMET assay. As a result, it was concluded that the addition of 10, 15, 20 μgmL shilajit to rooster semen extender improves semen quality parameters and DNA integrity of semen after cryopreservation.

 Reactive oxygen species ROS are created in excess during the cryopreservation process, which speeds up the rate of lipid peroxidation LPO. This negatively impacts spermatozoa functions and reduces their capacity to fertilize. The spermatozoon plasma membrane consists of significant amounts of polyunsaturated fatty acids PUFA, which can be easily oxidized by ROS and produce harmful agents that are toxic to cells. The plasma membrane of rooster spermatozoa contains high concentrations of polyunsaturated fatty acids and very small amounts of mitochondria, cytoplasm, and cytoplasmic antioxidants. Cryopreservation of rooster semen has been associated with adverse effects, including increased lipid peroxidation, structural damage in the mitochondria and acrosomal area, changes in the integrity and permeability of the spermatozoon plasma membrane, and severe damage to DNA. In the study, semen taken from 20 Plymouth Rock roosters were pooled to eliminate individual differences. By adding 5 μgmL, 10 μgmL, 15 μgmL, 20 μgmL and 25 μgmL shilajit to Beltsville Poultry Semen Extender diluent, 5 experimental and 1 control groups were formed and frozen in 0.25 mL straws. After thawing in a water bath at 37oC, spermatologic parameters were analyzed with the CASA system. Viability evaluations were made with eosin  nigrosin stain and morphological evaluations were made with Hancock method. Sperm DNA integrity was examined with the COMET assay. As a result, it was concluded that the addition of 10, 15, 20 μgmL shilajit to rooster semen extender improves semen quality parameters and DNA integrity of semen after cryopreservation.

 Reactive oxygen species ROS are created in excess during the cryopreservation process, which speeds up the rate of lipid peroxidation LPO. This negatively impacts spermatozoa functions and reduces their capacity to fertilize. The spermatozoon plasma membrane consists of significant amounts of polyunsaturated fatty acids PUFA, which can be easily oxidized by ROS and produce harmful agents that are toxic to cells. The plasma membrane of rooster spermatozoa contains high concentrations of polyunsaturated fatty acids and very small amounts of mitochondria, cytoplasm, and cytoplasmic antioxidants. Cryopreservation of rooster semen has been associated with adverse effects, including increased lipid peroxidation, structural damage in the mitochondria and acrosomal area, changes in the integrity and permeability of the spermatozoon plasma membrane, and severe damage to DNA. In the study, semen taken from 20 Plymouth Rock roosters were pooled to eliminate individual differences. By adding 5 μgmL, 10 μgmL, 15 μgmL, 20 μgmL and 25 μgmL shilajit to Beltsville Poultry Semen Extender diluent, 5 experimental and 1 control groups were formed and frozen in 0.25 mL straws. After thawing in a water bath at 37oC, spermatologic parameters were analyzed with the CASA system. Viability evaluations were made with eosin  nigrosin stain and morphological evaluations were made with Hancock method. Sperm DNA integrity was examined with the COMET assay. As a result, it was concluded that the addition of 10, 15, 20 μgmL shilajit to rooster semen extender improves semen quality parameters and DNA integrity of semen after cryopreservation.

 Reaktif oksijen türleri ROS, kriyoprezervasyon sürecinde aşırı miktarda oluşmakta ve lipid peroksidasyonu LPO hızını artırmaktadır. Bu durum, spermatozoa fonksiyonlarını olumsuz etkilemekte ve fertilizasyon yeteneğini azaltmaktadır. Spermatozoon plazma membranı, önemli miktarda çoklu doymamış yağ asidi PUFA içermekte ve ROS tarafından kolayca oksitlenebilmekte ve hücrelere toksik olan zararlı maddeler üretebilmektedirler. Horoz spermatozoon plazma membranı, yüksek konsantrasyonlarda çoklu doymamış yağ asitleri ve çok az miktarda mitokondri, sitoplazma ve sitoplazmik antioksidan içermektedir. Horoz spermasının kriyoprezervasyonu, artmış lipid peroksidasyonu, mitokondri ve akrozomal bölgede yapısal hasar, spermatozoon plazma membranının bütünlüğü ve geçirgenliğinde değişiklikler ve DNAda ciddi hasar gibi olumsuz etkilerle ilişkilendirilmiştir. Çalışmada, 20 Plymouth Rock ırkı horozdan alınan sperma bireysel farklılıkları ortadan kaldırmak için bir araya getirildi. Beltsville Poultry Semen Extender BPSE Sulandırıcısına 5 μgmL, 10 μgmL, 15 μgmL, 20 μgmL ve 25 μgmL miktarlarda shilajit eklenerek 5 deney ve 1 kontrol grubu oluşturuldu ve 0.25 mL payetler içerisinde donduruldu. 37C su banyosunda çözdürüldükten sonra spermatolojik parametreler CASA sistemi ile belirlendi. Spermatozoa canlılık oranı eosin  nigrosin boyama metodu ile, morfolojik değerlendirmeler ise Hancock yöntemi ile yapıldı. Spermatozoonn DNA bütünlüğü COMET analiz yöntemi ile değerlendirildi. Sonuç olarak, horoz sperma sulandırıcısına 10, 15, 20 μgmL shilajit eklenmesinin, kriyoprezervasyon sonrası spermatolojik parametreleri ve DNA bütünlüğünü olumlu yönde etkilediği sonucuna varıldı.

Article Details

Effect of Shilajit on Freezing Rooster Semen

Authors:

Mustafa Yiğit Nizam | Murat Selcuk

Publisher:

Afyon Kocatepe University

Main Topics:

Veteriner Bilimleri (Diğer)

Journal:

Kocatepe Veteriner Dergisi Vol.17 Issue 4, pp.393-400

Document Type:

Article

Publication Date:

3/2025

Document Language:

English

Document ID (Org No.):

2001327152

pISSN:

1308-1594

Cryopreservation

Rooster

Shilajit

Spermkriyoprezervasyon

horoz

shilajit

sperma

Issues

Article Details

Authors: Mustafa Yiğit Nizam | Murat Selcuk

Publisher: Afyon Kocatepe University

Main Topics: Veteriner Bilimleri (Diğer)

Journal: Kocatepe Veteriner Dergisi Vol.17 Issue 4, pp.393-400

Document Type: Article

Publication Date: 3/2025

Document Language: English

Document ID (Org No.): 2001327152

pISSN: 1308-1594

Issues

Authors:

Mustafa Yiğit Nizam | Murat Selcuk

Publisher:

Afyon Kocatepe University

Main Topics:

Veteriner Bilimleri (Diğer)

Journal:

Kocatepe Veteriner Dergisi Vol.17 Issue 4, pp.393-400

Document Type:

Article

Publication Date:

3/2025

Document Language:

English

Document ID (Org No.):

2001327152

pISSN:

1308-1594