Güneş YAMANER, Gökhan TUNÇELLİ, Momin MOMİN, Devrim MEMİŞ

Gökkuşağı Alabalığının (Oncorhynchus mykiss) Sperm Kalitesinde Bilgisayarlı Otomatik Sperm Analiz Sisteminde (CASA) Leja ve Makler Lamlarının Performansının Karşılaştırılması

Bu çalışmada gökkuşağı alabalığının (Oncorhynchus mykiss) spermine ait motilite ve motiliteye ait kinematik parametrelerin Bilgisayarlı Otomatik Sperm Analiz Sistemi (CASA) ile incelenmesinde farklı lamların kullanılmasının sonuçlar üzerindeki etkisi araştırılmıştır. Motilite ve parametreleri, oda sıcaklığında CEROS II (Hamilton-Thorne, Beverly, MA, USA) yazılım sistemine bağlı ışık mikroskobu (CX41, Olympus, Japan) ile incelenmiştir. Sperm örnekleri beş erkek balıktan abdominal masaj yöntemi ile balıkların üreme döneminde toplanmış ve Leja 2 (20 µl deep, Leja Products, Netherlands) ve Makler (10 µl deep, Sefi-Medical Instrument, Haifa, Israel) olmak üzere iki farklı lam kullanılarak analiz edilmişlerdir. Sperm örneklerinde toplam motilite (%), ve hız parametrelerinden eğrisel hız olarak ifade edilen VCL (µm/s) analiz edilmiştir. Dölleme çalışması için, bir dişi balıktan alınan yumurtalar (toplam 550 g, 7500 yumurta) beş eşit parçaya bölünmüştür. Bölünen yumurtaların her bir bölümü (1500 yumurta) analize tabi tutulan sperm örnekleri (5 ml) ile döllenmiştir. Dölleme, inkübasyon ve döllenme oranı prosedürleri, alabalık yetiştiriciliği için kullanılan rutin uygulama prosedürü altında gerçekleştirilmiştir. Döllenme yüzdesi her yumurta grubu için ˃%80 olarak bulunmuştur. İncelenen sperm örneklerinde Makler lamın kullanılması ile sperm motilite değerleri %26-45 arasında değişiklik göstermişken; Leja 2 lamı kullanılarak incelenen sperm örneklerinde her bir balıkta motilite değeri %90 ve üzeri bulunmuştur. Analiz esnasında iki farklı lam kullanılmasının motilite sonuçlarının belirlenmesini etkilediği bulunmuştur. Motilite sonuçları ve dölleme sonuçları karşılaştırıldığında ise Leja 2 lamı ile incelenen örneklerin motilite sonuçları döllenme oranı ile örtüştüğü ve daha kararlı sonuçlar verdiği görülmüştür. Motilite sonuçları ile yapılan istatistiksel değerlendirmede Leja 2 ve Makler lamlarından elde edilen sonuçlar arasında anlamlı bir farklılık tespit edilmiştir (p<0.05).

Anahtar Kelimeler:

Leja 2, CASA, Makler, Motilite parametreleri, Döllenme oranı, Gökkuşağı alabalığı

Comparison of Leja and Makler Chambers Performance in Rainbow Trout (Oncorhynchus mykiss) Sperm Quality in CASA System

In this study, the effect of the chamber used for the automated analysis of sperm motility and sperm kinematics parameters by CASA was evaluated of rainbow trout (Oncorhynchus mykiss) sperms. The assessment of motility parameters was carried out using CEROS II (Hamilton-Thorne, Beverly, MA, USA) connected to CX41 microscope (Olympus, Japan) at room temperature. Sperm samples were collected from five adult males by abdominal massage during the reproduction season and analyzed with two different chambers as follows specialty: Leja 2 cell chambered with 20 µl deep (Leja Products, Netherlands) and Makler chamber, round shape with 10 µl deep (Sefi-Medical Instrument, Haifa, Israel). Total sperm motility (Mot, %), and Velocity of Curvilinear (µm/s) were measured. For fertilization test, eggs from one female (550 g approximately 7500 eggs) were separated in equal five parts. Each part of eggs (approximately 1.500 eggs) was fertilized with each analyzed sperm (5ml). Fertilization, incubation procedure and calculation of fertilization rates have been kept as used routinely for rainbow trout culture procedure. The fertilization rates were found ˃80% for all used males. The motility percentage of samples analyzed by Leja has been found higher 90% while by makler changed between 26-45%. There is significantly effect on different chambers used in this study to determining the motility percentage. The high stability results and matched the fertilization success were detected in Leja 2-chamber. Statistical study with motility percentage showed a significant difference between Leja and Makler chambers (p<0,05).

Keywords:

CASA, Leja 2, Makler, Motility parameters, Fertilization rate, Rainbow trout,

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