Farklı süre ve sıcaklıklarda çözdürülen boğa spermalarının morfolojik fonksiyonlarının CASA cihazı ile değerlendirmesi

Thawing condition is one the most important factors affecting the re-animation of the spermatozoon in order to fertilise the oocyte. For that matter, we aimed to evaluate the morphological features of the head and midpiece of cryopreserved spermatozoa thawed at different temperatures and for various durations, with using CASA (Computer aided sperm analyser). Frozen semen samples belonging to the same batch, collected from three different bulls were grouped as; control group thawed for 20 seconds at 37 ℃; and experimental groups were thawed for 30, 40, 50 and 60 s at 25 ℃ and 37 ℃; for 10, 15, 20 and 25 s at 40 ℃; for 3, 6, 9, 12 s at 70 ℃. Morphometrical features of the samples were evaluated by using CASA system with nine repetitions. As a result, straws thawed at 25℃ for 40 s had the highest average length of head (6.22 ± 0.09 μm), and the width of midpiece (0.68 ± 0.01 μm). It was concluded that the thawing temperature and duration has affected/altered the morphometry of the sperm head and midpiece, although the results were not statistically significant (p> 0.005).Suni tohumlamada kullanılan dondurulmuş spermanın çözüm sonu parametrelerini belirleyen faktörlerin başında çözdürme koşulları gelmektedir. Yapılan araştırma ile Simental, Holstein ve Brown Swiss ırkı boğaların donmuş spermalarının, deneysel olarak belirlenen farklı sıcaklık ve sürelerde çözdürülmesi sonucu CASA cihazı ile spermatozoon başı ve orta kısmına ait parametreler bakımından değerlendirilmesi amaçlanmıştır. Çalışmada gruplar; kontrol grubu 37 ℃’de 20 saniye; deney grupları ise, 25 ℃’de 30, 40, 50 ve 60 sn.; 37 ℃’de 30, 40, 50 ve 60 sn.; 40 ℃’de 10, 15, 20 ve 25 sn.; 70 ℃’de 3, 6, 9, 12 sn. olarak belirlenmiştir. Üç farklı ırk boğaya ait, tek ejakülasyonda elde edilen dondurulmuş boğa spermaları, 9 tekrarda CASA parametreleri açısından değerlendirilmiştir. 25 ℃’de 40 sn.’de çözdürülen spermaların baş uzunluklarının (6.22 ± 0.09), orta kısım genişliğinin (0.68 ± 0.01) diğer sıcaklık ve sürelere göre daha yüksek ortalamaya sahip olduğu saptanmıştır. Çözdürme sıcaklık ve süresinin spermatozoon baş ve orta kısmına dair parametreleri değiştirdiği ancak elde edilen sonuçların istatiksel olarak anlamlı olmadığı belirlenmiştir (p> 0.005)

Koç spermasının çözüm sonu kalitesi üzerine farklı sulandırıcıların ve myo-inositolün etkileri

Bu çalışma koç spermasının çözüm sonu kalitesi, lipit peroksidasyonu ve antioksidan aktiviteleri üzerine farklı sulandırıcıların ve inositolün etkilerini değerlendirmek amacıyla yapıldı. Sperma 4 baş Karayaka koçundan suni vajen yardımıyla haftada üç kez alındı. Alınan sperma örneklerinden normospermi özellik gösterenler birleştirildi. Birleştirilen sperma örnekleri iki farklı dozda myo-inositol (5, 10 mM) içeren ve içermeyen (kontrol) üç farklı sulandırıcı (tris, yağsız süt tozu, sodyum sitrat) ile sulandırıldı. Örnekler dokuz ayrı çalışma grubuna ayrıldı: T-5I, T-10I, T (kontrol); M-5I, M-10I, M (kontrol); Na-5I, Na-10I, NaC (kontrol) sulandırılmış sperma içeren payetler 4°C’de 2 saat ekilibre edildi, sıvı azot buharında (-120°C’da 15 dakika) donduruldu ve sıvı azot (-196°C) içinde saklandı. Dondurulmuş spermalar su banyosunda 37°C’de 30 saniyede çözdürüldü. Sulandırıcılara eklenenen myo-inositol mikroskopik sperm ve oksidatif stres parametelerine önemli bir etkiye neden olmadı (P>0.05). T ve M sulandırıcıları, NaC sulandırıcısına göre donma-çözünme sonrası spermatozoon motilitesinde (%50.00±2.24% ve 55.00±3.42) ve HOS testte (%49.00±3.32% and 48.17±2.9) daha yüksek oranlar verdi (P0.05). MDA seviyesi T sulandırıcısında (1.22±0.07 nmol/ml), M ve NaC sulandırıcılarına göre daha düşük bulundu (P<0.05). GSH ve GSH-PX aktiviteleri için, T ve NaC sulandırıcıları M sulandırıcısına göre daha yüksek değerler verdi (P<0.01).The study was conducted to evaluate the eff ects of diff erent extenders and inositol additions on post-thaw semen quality, lipid peroxidation (LPO) and antioxidant activities. Semen was collected from four Karayaka rams from by artifi cial vagina three times a week. Semen samples showing normospermy quality were pooled. The pooled semen samples were extended in three extenders (Tris, T-, skimmed milk, M- and sodium citrate, NaC) with myo-inositol at two diff erent doses (5 mM, 10 mM) and no antioxidant (control). Nine experimental groups were assigned as follows: T-5I, T-10I, T (control); M-5I, M-10I, M (control); Na-5I, Na-10I, NaC (control). Straws containing extended semen were equilibrated at 4&deg;C for 2 h, frozen in vapor of (15 min at -120&deg;C) liquid nitrogen and stored in liquid nitrogen. Frozen semen was thawed in a water bath at 37&deg;C for 30 seconds. The use of all the extenders supplemented with diff erent doses of myo-inositol did not lead to any significant improvement in microscopic sperm and oxidative stress parameters (P&gt;0.05). Extenders of T and M resulted in higher sperm motility (50.00&plusmn;2.24% and 55.00&plusmn;0.42%) and HOST (49.00 3.32% and 48.17&plusmn;2.97%) rates, compared to NaC (37.00&plusmn;3.74% and 31.80&plusmn;2.96%, P&lt;0.01), following the freeze/thawing process. Extenders supplementated with myo-inositol not significantly aff ect malondialdehyde (MDA) levels and activities of catalase (CAT), superoxide dismutase (SOD), glutathione (GSH) and glutathione peroxidase (GSH-PX) in comparison to the control groups (P&gt;0.05), except for MDA level of T extender containing 10 mM inositol. MDA level was found lower (1.22&plusmn;0.07 nmol/ml) in T than those of the M and NaC (P&lt;0.05). For GSH and GSH-PX activities, T and NaC gave the higher values, compared to M, following the freeze/thawing process (P&lt;0.01)

The use of infrared thermography to detect the stages of estrus cycle and ovulation time in anatolian shepherd dogs

Abstract Background The aim of the study is to evaluate the effectiveness of thermographic monitoring, using the temperature changes of perianal and perivulvar areas for the determination of estrus in Anatolian Shepherd bitches. Fifteen bitches were used in the study. Blood and vaginal smear samples were collected and thermographic monitoring of perianal and perivulvar areas were carried out starting from proestrus to early diestrus. Also, external signs of estrus were investigated. Smear samples were evaluated by light microscopy after Diff-Quik staining method and superficial and keratinized superficial cells were determined as percentage (S + KS%). Progesterone and luteinizing hormone measurements were done by radioimmunoassay. The difference in temperature between perianal and perivulvar areas was evaluated through thermographic images by FLIR ResearchIR Software. Results According to the results obtained from the study, differences between progesterone and S + KS% were statistically significant (P  0,05). Serum luteinizing hormone levels did not sign any difference (P > 0,05). Conclusions As a result, thermographic monitoring alone is not enough for estrus detection in Anatolian Shepherd bitches. However, it can be used to assist the actual estrus detection technique in terms of providing some foreknowledge by evaluating the differences in temperature

Use of infrared thermography during ejaculation process and Its link with semen quality and freezability in dogs

This study aimed to describe the thermal variation of external reproductive tracts during ejaculation in relation to sperm quality in dogs. Forty-six adult fertile dogs were monitored using a thermal camera before, during and after the semen collection, taking into account penile and scrotal temperatures as reproductive thermal patterns while eye and perianal temperatures were recorded as complementary thermal patterns of behavioral response. The parameters were classified depending on age (≤4 years and >4 years), body weight (BW) (≤75 kg and >75 kg), sperm concentration (CON) (≤300 million and >300 million), total testicular volume (TTV) (≤600 cm3 and >600 cm3) and total ejaculation time (TET) (≤800 s and >800 s) of the animals from which semen was collected successfully. Heavier males (p < 0.05) that have more consistent testicles (p < 0.01) as well as quicker ejaculate responders (p < 0.001) and lower scrotal temperature had better semen (Δ motility) freezability. The lower eye temperature prior to the ejaculation (p < 0.01), lower scrotal temperature following ejaculation (p < 0.01), and conversely, higher penile temperature during the ejaculation (p < 0.001) had a higher sperm concentration. Furthermore, the sperm freezability was negatively correlated with total ejaculation time (r = -0.39, p < 0.05) and sperm abnormalities were lower in the ejaculate of dogs having a higher temperature of the scrotum, bulbus and penis. In conclusion, infrared monitoring throughout semen collection in dogs can provide information on behavioral reactions during human manipulation, as well as semen quality and testicular functionality