In this study, the quality of frozen bull semen was evaluated with the proAKAP4 level test. Sixty straws of frozen bull semen from various batches (n = 30) belonging to six bulls were used in the current study. The frozen bull semen samples were analysed in terms of proAKAP4 levels, sperm morphology and sperm movement parameters at hour 0 and hour 3 after thawing. The semen samples were divided into three groups according to the proAKAP4 levels: low concentration (<25 ng/10x106 spermatozoa), moderate concentration (25 to 39 ng/10x106 spermatozoa) and high concentration (≥40 ng/10x106 spermatozoa). A positive correlation was found between the proAKAP4 level and total motility (TM3), progressive motility (PM3), VSL3 and VCL3 values obtained after the third‐hour thermoresistance test (p < .05). There was a negative correlation between the percentage of sperm abnormal tail and the proAKAP4 level (p < .01). In addition, it was observed that the semen samples with proAKAP4 concentrations of 25 ng/106 spermatozoa and higher preserved the TM3 and PM3 motility characteristics. In conclusion, the proAKAP4 has the potential to become a biomarker protein to evaluate in the quality analysis of frozen‐thawed semen.

The objective was to determine effects of gallic acid (GA) and carnosic acid (CA), present in carob pods and rosemary extract respectively, on frozen-thawed ram spermatozoa. Thirty ejaculates were collected from five Merino rams, pooled, diluted in Tris-based extender and divided into five equal portions containing: 0.05 or 2 mM of GA; 0.05 or 0.2 mM of CA; or no additive (control). Extended semen was equilibrated at +4°C, loaded into straws, held 5 cm above liquid nitrogen for 12 min then plunged.Computer-aided sperm analysis was used to assess motility, whereas flow cytometry

The objective of this study was to determine effects of supplementing Tris-based semen extenders with either cholesterol-loaded cyclodextrin (CLC) or 7-dehydrocholesterol loaded cyclodextrin (7-DCLC) plus trehalose (T) for cryopreservation of ram semen. Semen was collected with an artificial vagina from five Merino rams (2–3 years of age) during the non-breeding season. Ejaculates were pooled, divided into eight equal portions, diluted with a standard Tris-based extender containing: no additive (control); T (50 mM); or T (50 mM) + 1.5, 2.5 or 3.5 mg of either 7-DCLC or CLC. Semen was chilled from 37°C to 4°C, placed in 0.25 ml French straws, held 5 cm above liquid nitrogen for 12 minutes, then plunged into liquid nitrogen. After thawing, a computer-aided semen analyzer system (CASA) was used to assess motility, whereas plasma membrane and acrosome integrity (PMAI) and high mitochondrial membrane potential (HMMP) were assessed with flow cytometry. Sperm supplemented with 2.5 mg and 3.5 mg CLC + T had the highest (P &lt; 0.05) total and progressive motility (65.2 ± 4.7 and 19.0 ± 1.0% respectively, mean ± SEM), albeit with no significant differences from sperm with 1.5 or 3.5 mg CLC + T. Sperm with 2.5 mg CLC + T had the highest (P &lt; 0.05) PMAI (59.3%; not different from 3.5 mg CLC + T) and highest (P &lt; 0.05) HMMP (64.6%; not different from 1.5 or 3.5 mg CLC + T). The lowest ALH value, 2.8 ± 0.3 µm was in the 2.5 mg 7-DCLC + T group; otherwise, there were no significant differences among groups for any other CASA end point. In conclusion, adding CLC + T to a tris-based extender optimized quality of frozen-thawed ram semen. Therefore, extenders including CLC + T have potential to improve quality of frozen-thawed ram sperm.

Conventional buffalo semen freezing studies are limited in Anatolian buffaloes, which are overly sensitive to exogenous stimulation. The present study's object was to determine the main features of Anatolian Buffalo semen obtained by artificial vagina method for the first time. A total number of 150 ejaculates were collected from three Anatolian Buffalo bulls (app. 4 years of age). The mean pH, volume and concentration of semen were found 6.63±0.15, 1.61±0.5 ml, 1629±222.67 x10 6 spermatozoa/ml, respectively. The sperm motion characteristics were determined by using a computerassisted sperm analysis system (CASA); the total and progressively motile sperm values were 57.

ÖZ Dietilheksil fitalat, sert yapılı plastiklere esneklik katmak amacıyla dünya genelinde yaygın bir şeklide kullanılan, çevresel toksik bir kimyasaldır. İçerisinde bulunduğu ve gevşek bağlarla tutunduğu ürünlerden kolayca ayrılarak çevreye yayılan dietilheksil fitalat sindirim, solunum ve deri teması yollarıyla insan ve hayvanların vücuduna geçmekte ve çeşitli toksik etkilere neden olmaktadır. Bu çalışmanın amacı, dietilheksil fitalatın boğa sperması üzerine in vitro toksik etkisini, bilgisayarlı sperm analiz sistemi ile araştırmaktır. Boğalardan suni vajen ile elde edilen spermalar, fosfat tampon solüsyonu ile 50x10 6 /ml spermatozoa olacak şekilde sulandırıldı ve dimetil sülfoksitte çözdürülen dietilheksil fitalatın 0, 1, 10, 100, 250 ve 500 µg/ml dozlarına maruz bırakılarak 1, 2, 3 ve 4 saat süreyle 37 °C su banyosu içerisinde inkübe edildi. Her inkübasyon süresinin sonunda kontrol ve deneme gruplarından alınan sperma örneklerinin motilite parametreleri analiz edildi. Yapılan değerlendirme sonucunda, dietilheksil fitalatın çalışmada kullanılan düşük dozlarında, doğrudan spermatozoa üzerine toksik etkisi tespit edilemezken, bu etkinin yüksek konsantrasyonlarda zamanla ve doza bağlı olarak ortaya çıktığı belirlendi. Özellikle 500 µg/ml dietilheksil fitalat maruziyetinin spermatozoon hareketlilik parametrelerinden, ortalama yol hızı ve doğrusal hızı 2. saatten itibaren, total motilite ve progresif motiliteyi ise 3. saatten itibaren kontrol gruplarına göre önemli derecede azalttığı belirlendi (p<0.05). İn vitro olarak elde edilen sonuçlar dietilheksil fitalatın erkek üreme organlarında toksik birikimine bağlı olarak boğalarda spermatogenezisi etkiliye bileceğini yönündedir. Effect of diethylhexyl phthalate on sperm motility parameters in bull ABSTRACT Diethylhexyl Phthalate is an environmentally toxic chemical commonly used in worldwide to make rigid plastics flexible. Due to its loosely bonded to plastic products, diethylhexyl phthalate easily leaches into the environment. After humans and animals exposure to diethylhexyl phthalate via digestion, respiration and skin contact, it causes various toxic effects in the body. The aim of this study was to investigate the in vitro toxic effect of diethylhexyl phthalate on bull semen motility via a computerized sperm analysis system. Semen collected from the bulls with artificial vagina were diluted with phosphate buffer solution to 50x10 6 /ml spermatozoa and incubated in a 37°C water bath for 1, 2, 3 and 4 h by exposing to 0, 1, 10, 100, 250 and 500 µg/ml doses of diethylhexyl phthalate dissolved in dimethyl sulfoxide. At the end of each incubation period, the motility parameters of semen samples taken from control and experimental groups were analyzed. As a result of the evaluation, the toxic effects of diethylhexyl phthalate on the spermatozoa were not determined at low doses used in this study, but this effect occurred at high concentrations by the time. It was observed that exposure of diethylhexyl phthalate particularly at the doses of 500 µg/ml significantly decr...