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The aim of the current study was to evaluate the effects of different concentrations of rainbow trout seminal plasma (RTSP) (0.1%, 1% and 10%) in extenders containing either egg yolk or lecithin for use in Awassi ram semen cryopreservation. Pooled sperm were diluted in a two-step dilution method to a final concentration of 1/5 (semen/extender) in egg yolk or lecithin extender containing no RTSP, 0.1%, 1% or 10% RTSP (v/v). Semen samples were assessed for sperm motility, plasma membrane integrity [hypoosmotic swelling test (HOST) and Hoechst 33258] and defective acrosomes [FITC-conjugated Pisum sativum agglutinin (PSA-FITC)] at the following five time points: after dilution with extender A; after equilibration; and post-thaw at 0 h, 3 h and 5 h. Malondialdehyde (MDA) was examined only after thawing. Freezing and thawing procedures (dilution, equilibration and post-thaw incubation at Oh, 3 h and 5 h) negatively affected the motility (P

The aim of the present study was to evaluate the correlation between the routine semen analyses and sperm DNA integrity, as assesed by TUNEL in fresh semen of rams in Kivircik and Awassi breeds in breeding season. Semen was collected by electro-ejaculation five times every other day. For that purpose, 50 fresh ejaculates from 6 Kivircik and 4 Awassi rams were evaluated. The mean percentages of semen volume, concentration, mass activity, motility, acrosomal defects, HOST and DNA fragmentation of Kivircik and Awassi rams were 1.3 ml, 1.9 x109, 3.3 (0-5 scale), 72.6%, 13.2%, 73.6%, 16.2% and 1.0 ml, 1.9 x109, 3.3 (0-5 scale), 72.5%, 11.1%, 77.9%, 15.9%; respectively (P>0.05). Sperm DNA fragmentation was correlated adversely with semen volume (r=-0.329, P0.05), mass activity (r=-0.349, P0.05) and HOS test (r=-0.513, P0.05), mass aktivite (r = -0.349, P0.05) ve HOS testi (r= -0.513, P

The aim of the current study was to evaluate the effect of egg yolk and different soybean lecithin concentrations on the efficiency of ram semen cryopreservation and to test the fertilizing ability of frozen-thawed ram semen. Ejaculates with a thick consistency, rapid wave motion (3-5), and >70% initial motility were pooled. Pooled semen were then divided into four groups and diluted at 1/5 (semen/extender) with 1%, 3%, 6% lecithin (L1, L3 and L6) or 20% egg yolk (EY20) using the two-step dilution method. As expected, the results of the current study showed that both motility and the rates of defective acrosomes in sperm were negatively affected by the cryopreservation procedure (P0.05). The results of in vitro fertilization, as assessed by the rate of blastocyst formation, were more successful in the EY20 group than those noted in different lecithin groups. In conclusion, freezing ram semen with an extender containing egg yolk could yield better post-thaw sperm parameters and embryonic development compared to lecithin containing extenders. Bu çalışmanın amacı, yumurta sarısı ve farklı lesitin konsantrasyonlarının koç spermasının dondurulabilirliği üzerine etkisini değerlendirmek ve dondurup çözdürülen koç spermasının fertilizasyon yeteneğini tespit etmektir. Kitle hareketi (3-5) ve >%70 motiliteye sahip ejakülatlar birleştirildi (pooling). Pooling yapılan sperma dört gruba bölündü ve %1, %3, %6 (L1, L3 ve L6) lesitin veya %20 yumurta sarısı (EY20) içeren sulandırıcılar ile 1/5 (sperma/sulandırıcı) oranında iki aşamalı sulandırma yöntemi kullanılarak sulandırıldı. Bu çalışmanın sonucunda; motilite ve akrozomal bozukluk oranlarının dondurma prosedüründen olumsuz yönde etkilendiği tespit edildi (P0.05). Blastosist oranları bakımından değerlendirilen in vitro fertilizasyon sonuçlarına göre, EY20 grubunun farklı lesitin gruplarından daha başarılı olduğu tespit edildi. Sonuç olarak, yumurta sarısı içeren sulandırıcı ile dondurulan koç spermasının lesitin gruplarına göre eritme sonrası spermatolojik özellikler ve embriyonik gelişim bakımından daha üstün olduğu tespit edildi.

The influence of different extender osmolality levels and the presence of different cryoprotectants on the post-thawed semen's characteristics and post-thawed plasma membrane integrity of ram spermatozoa were studied. Ram semen was frozen with TRIS-egg yolk based extender according to two-step dilution procedures. The final concentrations of the cryoprotectants: 6% glycerol, 6% 1,2-propanediol, 62.5 mM sucrose, and 62.5 mM trehalose were studied in three different extender B osmolality levels (350, 375, and 400 mOsm). The osmolality affected significantly the post-thawed semen's motility, defected acrosomes (DA), total morphological defect (TMD), along with the sperm's plasma membrane integrity (HOST). Type of cryoprotectant exerted significant effect (P

The aim of the study was to evaluate the effect Of the addition of 20% bull seminal plasma to semen extenders on post-thaw sperm motility, morphology, and fertility. Two Merino rains at 3-5 years of age, 43 ewes, and one Holstein bull were used. Fourteen ejaculates Were collected by the use of an artificial vagina, twice a day, at 60 min interval. Six semen ejaculates were extended with Tris-egg yolk extender containing 20% bull seminal plasma (TEY-BSP group), and 8 ejaculates were extended with Tris-egg yolk (TEY group), at a rate to obtain 150 x 10(6) spermatozoa/straw. The owes in oestrous were divided at random into two groups. Twenty-two ewes were inseminated with TEY-BSP and 21 ewes were inseminated with TEY. The mean percentage of motility, defected acrosome, and total morphological defect for the equilibrated and post-thaw semen parameters of the TEY, were 69.4%, 8.6%, and 17.0% and 42.8%, 49.9%, and 51.2%, respectively. The mean percentage of non-return rate (NRR) (n=43) was 30.2% and for the semen frozen with TEY-BSP (n=22) and TEY (n=21) - 31.8%, and 28.6%, respectively. There were no significant differences between TEY-BSP and TEY for NRR. In Conclusion, the addition of the bull seminal plasma to freezing extender has a Supportive effect on extended, equilibrated, and thawed semen. Moreover, the addition of bull seminal plasma to ram semen extender improved fertility.

Fertility of artificial insemination is relatively low when frozen semen is used in sheep (Ozturkler et al., 1999; Sanchez-Partida et al., 1992). Jones and Mann (1977) have suggested a relation between the lipid peroxidation formation in spermatozoa and quick decrease in motility and phospholipid loss in the membranes. Some researches agreed that dialyzed egg yolk, bull seminal plasma, milk and sodium sulphate could prevent spermatozoa from the post-thaw side effects of lipid peroxidation (Salamon and Maxwell, 1995; Aswort et al., 1994; Barrios et al., 2000). Jones and Mann (loc. cit.) claimed that egg yolk could decrease the peroxidation rate of ram spermatozoa when added to extenders but not as much as the bull seminal plasma. Researchers have found that bull spermatozoa can easily peroxidase due to the high level of unsaturated fatty acids bound to the phospholipids in the seminal plasma. The present study is planned to investigate the effects of bull seminal plasma in semen extenders during freezing of ram semen and its post-thaw motility and morphological characteristics.

In the present study, ten ejaculates were obtained from each of seven German shepherd dogs. Only the sperm-rich fractions of ejaculates from each dog once a week with a mean interval of 60 min, were used during five weeks. Semen quality was examined in total of 70 semen samples. After this examination, semen samples were centrifuged (5000 g x 10 minutes) and seminal plasma were separated. Plasma total protein, calcium, phosphorus, magnesium, sodium and potassium and alkaline phosphatase were determined. The semen quality and biochemical findings were compared between the first and the second ejaculate following very-short interval ejaculation. There were no significant differences in the percentage of motility, live spermatozoa, acrosomal, tail and total morphological defects of the second ejaculate compared with that of the first ejaculate. However, there were significant lower values for the volume and the spermatozoal concentration in the second ejaculate than that of the first ejaculate (p < 0.05). There were significantly lower values for the total protein, calcium, magnesium and alkaline phoshatase of the second ejaculate compared with that of the first ejaculate. Positive correlations were found between spermatozoal concentration and AP; motility and sodium concentration. The semen quality in the first and the second ejaculate in all dogs was normal for fertility. In conclusion, when the first ejaculate is inadequate, collection of second ejaculate may be useful for cryopreservation or insemination with fresh semen in dogs.

ÖZET Bu çalışmada dol tutmayan (Repeat Breeder) ineklerin kızgınlık sikluslarının geç luteal döneminde (12. veya 14. günler) prostaglandin F2a (PGF2a) enjeksiyonu ile oluşturulan kızgınlık senkronizasyonunun ineklerin ovary um fonksiyonları ve gebelik oranları üzerindeki etkisinin incelenmesi amaçlandı. Bu amaçla İzmir ili Menemen ilçesi ve çevresi başta olmak üzere reprodüktif özellikleri sürekli izlenen 2000 adet siyah alaca kültür ırkı süt ineği arasından döl tutmayan 60 inek araştırma için seçildi. Araştırma için seçilen bu inekler en az bir doğum yapmış, kızgınlık siklusları düzenli, genital kanallarının klinik muayenesinde herhangi bir bozukluk fark edilmeyen ve üst üste üç kez tohumlandığı halde gebe kalmayan hayvanlar arasından seçildi. Kızgınlık sikluslarının 12. veya 14. günlerinde tüm ineklere rektal muayene yapıldı. Rektal muayenede korpus luteumun bulunduğu ovaryumda palpe edilebilir folliküllere sahip olmayan 20 inek 1. grubu, muayene sırasında palpe edilebilir follikül tespit edilen (yaklaşık olarak 0.5 cm veya daha büyük çapta) ve elle follikülleri patlatılan 20 inek 2. grubu, muayene sırasında follikül tespit edilip herhangi el maniplasyonu yapılmayan 20 inek 3. grubu oluşturdu. Gruplardaki tüm ineklere rektal muayenenin hemen ardından tek doz kas içi PGF2a (Dinoprost 25 mg; Dinolitik, Eczacıbaşı) enjekte edildi. Enjeksiyondan sonra kızgınlık gösteren İnekler takip edilerek uygun zamanlama ile bir kez Holştayn ırkı boğa spermaları ile (6.25 nil. Payetler) tohumlandı. Tohumlamadan sonra tekrar kızgınlıkgösteren inekler kaydedildi. Kızgınlık göstermeyenler 45-60 gün sonra rektal yolla gebelik yönünden muayene edildi. Yapılan kızgınlık gözlemleri ve gebelik kontrollerinde birinci grupta %70, ikinci grupta %40 ve üçüncü grupta %25 oranında gebelik oluştuğu saptandı.En iyi birinci gruptaki ineklerin senkronize olduğu ve bu grupta daha fazla gebelik oluştuğu saptandı. Anahtar Kelimeler: Döl tutmama, luteal faz, prostaglandin F2a> follikül. SUMMARY Investigations of the Effect of Prostaglandin F2a Application on Ovarium Functions and Fertility of the Repeat Breeder Cows in the Luteal Phase In this study, it was aimed to investigate the ovarium functions and pregnancy rates of repeat breeder cows synchronized by prostaglandin F2a when they were in the late luteal phases of oestrus cycles (12th or 14th days). For this purpose, 60 out of 2000 repeat breeder black pied dairy cows bred in Menemen region of Izmir district were used as material. The criteria such as breed, age, nutritional situation, environmental factors and reproductive characteristics were the same for all selected animals with repeat breeder problem. Cows were selected among the animals which had normal cycles, pa rtu rated at least once, and had not shown abnormal clinical signs in their genital tract and had no pregnancy after three inseminations. Cows were divided into theree groups. First group consisted 20 cows which had not palpable follicles (appr. 0.5 cm in diameter or more) on their ovarium and had corpus luteum. The second group consisted 20 cows which had palpable follicles. These follicles were burnst manually by rectal palpation. The third group containing 20 cows had follicles but their follicles were not burnst. All cows were examined by rectal palpation between 12th and 14th days during luteal phase of their estrus cycle for determining the structures on the ovaries. A single dose of prostaglandin F2a (25 mg Dinoprost; Dinolytic, Eczacıbaşı) was injected to all cows in all groups by intra muscular route.In the trial group follicles larger than 0.5 cm in diameter were burst manually. All cows showing oestrus symptoms in all groups were inseminated with frozen black pied Holstein bulls' semen (0.25 ml. straws). AH inseminated cows were observed after insemination and returned ones were recorded. Cows without heat symptoms after 45- 60 days following insemination were controlled by rectal palpation for pregnancy diagnosis. 14 (70%), 8 (40%) and 5 (25%) cows in the first, second and third groups were determined as pregnant, respectively. After pregnancy diagnosis, best synchrorization rate was achieved in the first group and also more pregnancy was observed. Key Words: Repeat breeder, luteal phase, prostaglandin F2a> follicle. 41

ÖZET Bu çalışmada anöstrus döneminde bulunan koyunlara Florogeston asetat (FGA) ve Gebe Kısrak Serum Gonadotropin! (Pregnant Mare Serum Gonadotropin; PMSG) hormonları uygulayarak ovaryum aktivitelerinin uyarılması ve seksüel sikluslarının sinkronizasyonu ile bu koyunlarda değişik tohumlama metotlarının döl verimi üzerine etkilerinin karşılaştırılması amaçlandı. Materyal olarak Bursa Büyükorhan ilçesi Çeribaşı köyünde yetiştirici elinde bulunan 100 adet Merinos x Kıvırcık melezi koyun ve 4 adet Merinos x Kıvırcık melezi koç kullanıldı. Östrus sinkronizasyonunu sağlamak için 14 gün süreyle intravaginal yolla FGA emdirilmiş süngerler kullanıldı. Süngerler geri alındıktan hemen sonra her koyuna kas içi olarak (i. m.) 600 I.U. PMSG enjekte edildi. Florogeston asetat emdirilmiş vaginal sünger uygulanan 100 koyundan 99u östrus gösterdi. Çalışmada östrus gösteren koyunlardan dört grup oluşturuldu. Birinci grubu oluşturan 34 koyuna doğal aşım yaptırıldı. İkinci gruptaki 20 koyuna sulandırılmamış taze sperma ile, üçüncü gruptaki 19 koyuna sulandırılmış taze sperma ile, dördüncü gruptaki 20 koyuna da sulandırılarak ısısı +5°C'ye düşürülmüş sperma ile suni tohumlama yapıldı.Tohumlamalar servikal olarak tek tohumlama şeklinde yapıldı. Östrus gösteren koyunların altı tanesine genital kanal yangısı ve serviksin pozisyonunun uygun olmaması nedeniyle tohumlama yapılamadı. Tohumlamada kullanılan spermalar koçlardan suni vajen yardımıyla alındı. Alınan spermanın miktar ve pH yönünden makroskobik, kitle hareketi, motilite, yoğunluk yönünden de mikroskobik muayeneleri yapıldı. Alınan spermaların spermatolojik özelliklerini belirlemek için ölü/canlı spermatozoon oranını saptamak üzere Eosin-Nigrosin boyama tekniğinden, anormal spermatozoonların oranını saptamak amacıyla çini mürekkebinden, akrozom morfolojisini incelemek için de Giemsa boyama yönteminden yararlanıldı. Koçlardan alınan spermalar gerekli makroskopik ve mikroskobik muayeneleri yapıldıktan sonra Glikoz-Fosfat sulandırıcısı ile 1:1 oranında sulandırıldı. Doğal aşım uygulanan ve sulandırılmamış taze sperma, sulandırılmış taze sperma ve sulandırılarak ısısı +5°C'ye düşürülmüş sperma ile tohumlanan koyunlardan elde edilen gebelik oranlan % 64.7, % 50.0, % 47.3 ve % 40.0 olarak bulundu. Anahtar sözcükler : Koyun, sinkronizasyon, taze-, sulandırılmış-, soğutulmuş sperma, sun* i tohumlama. SUMMARY Effect of Oestrus Synchronization and Insemination Methods on Fertility in Ewes In the present study it is aimed to stimulate the ovarian activities and oestrus synchronization of ewes by applying Flourogestone acetate (FGA) and Pregnant Mare Serum Gonadotropin (PMSG) and to compare the effects of different insemination methods on fertility in the anoestrus period. A hundred Merino x Kıvırcık ewes and four Merino x Kıvırcık rams, bred in Çeribaşı village-Büyükorhan-Bursa, were used as material. FGA contained intravaginal sponges were used for 14 days for oestrus synchronization. 600 I.U. PMSG was injected to all ewes intramuscularly on the withdrawal day. Ninety nine of ewes showed oestrus. Ewes were divided into four groups. In the first group 34 ewes were mated naturally. In the second group 20 ewes were inseminated with undiluted fresh semen. In the third group 19 ewes were inseminated with diluted fresh semen. In the fourth group 20 ewes were inseminated with diluted and cooled (+5°C) semen. Inseminations were performed as single andcervically. Six ewes could not be inseminated because of inconvenient cervical position and genital inflammation. Semen was collected by means of artificial vagina. Volume and pH were examined macroscopically, massactivity, motility and sperm concentration were examined microscopically. Percentages of dead, abnormal and acrosome defected spermatozoids were determined by eosin-nigrosin, Indian ink and Giemsa's staining techniques, respectively. Examined semen was diluted of 1:1 ratio by Glucose-phosphate diluter. Conception rates were found as 64.7, 50.0, 47.3 and 40.0% for the four groups, respectively. Key Words : Sheep, synchronization, fresh-.diluted-, cooled semen, artificial insemination. 62

ÖZET Bu çalışmada çeşitli organizasyon biçimlerinin sığır sun'i tohumlama uy gulamaları ve dölverimine etkisi araştırıldı. Araştırmada Balıkesir - Manyas İlçesi, merkez ve köylerinden Holştayn ve Montafon ırkı toplam 460 inek ve düve kullanıldı. Araştırma materyali, yapılan genital organ muayenesinde anormal klinik bulgu saptanmayan inek ve düveler arasından seçildi. Tur, telefonla bildirim ve sinkronizasyon sistemlerin de sırasıyla 200, 200 ve 60 inek ve düve servikal olarak tohumlandı. Tohumlamayı izleyen 45 - 60. günler arasında rektal yolla gebelik saptandı. Sinkronizasyon çalışmasında, östruslarının düzenli olduğu önceden saptanan inek ve düvelere 11 gün ara ile iki enjeksiyon Prostaglandin F2 alfa (PGF2 a ) uygulandı. Tohumlamalar ikinci enjeksiyondan sonra kızgınlıklarına bakılmaksızın 80. saatte tek tohumlama şeklinde gerçekleştirildi. Tur geçeği 25 köyden geçen yaklaşık 180 km.lik bir yol birimiydi. Tur boyunca günde ortalama 15 materyal tohumlandı. Telefon bildirimi ile yapılan çalışmada yine Manyas merkez ilçe ve bağ lı 27 köyünde yetiştirici elinde bulunan inek ve düvelerde gerçekleştirildi. Bu sistemde kızgınlıkları sahipleri tarafından saptanan materyal merkeze bildirildi ve tohumlama için zaman verildi. Bu sistemde tohumlama saati pratik olarak sabah kızgınlık gösteren inek ve düvelerin öğleden sonra, öğleden sonra gös- 1terenlerinse ertesi sabah tohumlanmaları esas alındı. Tur, telefonla bildirim ve sinkronizasyon sistemleri ile tek tohumlamaya dayalı gebelik oranları sırasıyla % 54.5, % 75.0 ve % 60.0 olarak bulundu. Yapılan khi kare analizinde her üç yöntemde hayvanların gebelik oranları ara sında farklılık istatistiksel yönden önemli bulunmuştur (P< 0.05). Sonuç olarak, sığırlarda, günümüzde gelişmiş iletişim olanaklarından ya rarlanarak ve çağdaş biyoteknolojik olguların da bu alanda kullanılmasıyla da ha yüksek, başarılı dölverimi sonuçları elde edilebilecek organizasyon biçimle rinin gerekliliği görülmektedir. Anahtar sözcükler: Sun'i Tohumlama, Organizasyon, Fertilite. ZUSÂMMENFASSUNG Untersuchungen über den Einfluss verschiedener Organisationsformen der künstlichen Rinderbesamung auf die Fruchtbarkeit. in der vorliegenden Arbeit wurde der Einfluss verschiedener Organisationsfor men auf die künstliche Besamung und die Traechtigkeitsrate bei Rindern un- tersucht. In die Untersuchung wurden insgesamt 460 Kühe und Faersen aus der Pro- vinz Balıkesir - Manyas und den umgebenden Dörfern einbezogen. Das Un- tersuchungsmaterial wurde unter den Kühen und Faersen ausgewahlt, die bei der Genitaluntersuchung keine abnormale klinische Befunde aufwiesen. Mit- tels der Tour-, Telefonanrufs - und Brunstsynchronisationssysteme wurden der Reihe nach 200, 200 und 60 Kühe und Faersen cervikal besamt. 45 -60 Tage nach der Besamung wurde rektal die Traechtigkeit festgestellt. Zur Synchronisation erhielten die Kühe und Faersen, deren Brunst nach vo- rangegangenen Untersuchungen einen regelmaessigen Verlauf zeigte, im Ab- stand von 11 Tagen zwei Prostaglandin F2 a Injektionen. Die Besamung er- folgte im Anschluss an die zweite Injektion unabhaengig von einem Brunst- nachweis in der 80. Stunde in Form einer einzigen Applikation. Die Route verlief durch 25 Dörfer mit einer Laenge von etwa 180 km. An einem Tag komiten im Durchschnitt 15 Tiere besamt werden. 3Die Arbeit mit dem Telefonanrufssystem wurde auch in der Provinz Manyas und den 27 dazugehörigen Dörfern an in den Haenden von Züchtern befind- lichen Kühen und Faersen durchgeführt. Bei diesem System wurde die Brunst von den Tierbezitsern festgestellt, der Zentrale telefonisch mitgeteilt und der Besamungstermin festgelegt. Hierbei wurde praktischerweise der Zeitpunkt der Besamung bei Kühen und Faersen, die vormittags Brunstsympthome zeig- ten am Nachmittag und bei denen, die nachmittags Brunstsympthome zeigten, am naechsten Morgen festgelegt. Die erzielten Traechtigkeitsraten nach einma- liger Besamung betrugen beim Tour-, Telefonanrufs - und Brunstsynchronisati- onssystem der Reihe nach 54.5 %, 75.0 % und 60.0 %. Bei der chi2 Analyse zeigte die Traechtigkeitsrate aller drei Systeme einen statistisch signifikanten Unterschied (p< 0.05). Am Endeffekt zeigt sich die Notwendigkeit verschiedener Organisationsfor- men, urn bei Rindern mit Hilfe von entwickelten Kommunikationsmöglichkei- ten und der Anwendung moderner biotechnischer Applikationen höhere Tra echtigkeitsraten zu erzielen. Schliisselworter : Kiinstliche Besamung, Organisation, Fertilitaet. 41