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    Circulating miRNAs in maternal plasma as potential biomarkers of early pregnancy in sheep
    (Frontiers Media Sa, 2022) Hitit, Mustafa; Kose, Mehmet; Kaya, Mehmet Salih; Kirbas, Mesut; Dursun, Sukru; Alak, Ilyas; Atli, Mehmet Osman
    MicroRNA (miRNA) plays an important role in the control of gene expression and is implied in many biological functions, including embryo implantation and development. The aim was to assess plasma miRNA profiles during the peri-implantation and ascertain potential candidate miRNA markers for early pregnancy diagnosis in ovine plasma. The plasma samples were obtained from a total of 24 ewes on days 12 (pre-implantation; P12, n = 4), 16 (implantation; P16, n = 4) and 22 (post-implantation; P22, n = 4) after mating, and on their corresponding days of 12 (Pre-C; C12, n = 4), 16 (Imp-C; C16, n = 4) and 22 (Post-C; C22, n = 4) of the estrous cycle. The miRNA profiles in plasma were assessed by microarray technology. We detected the presence of 60 ovine-specific miRNAs in plasma samples. Of these miRNAs, 22 demonstrated a differential expression pattern, especially between the estrous cycle and early pregnancy, and targeted 521 genes. Two miRNAs (oar-miR-218a and oar-miR-1185-3p) were confirmed using RT-qPCR in the ovine plasma samples. Protein-protein interaction (PPI) network of target genes established six functional modules, of which modules 1 and 3 were enriched in the common GO terms, such as inflammatory response, defense response, and regulation of immune response. In contrast, module 2 was enriched in the developmental process involved in reproduction, embryo development, embryonic morphogenesis, and regulation of the developmental process. The results indicate that miRNAs profiles of plasma seemed to be modulated during the peri-implantation stage of pregnancy in ewes. Circulating miRNAs could be promising candidates for diagnosis in early ovine pregnancy.
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    Expression pattern of microRNAs in ovine endometrium during the peri-implantation
    (Elsevier Science Inc, 2022) Kose, Mehmet; Hitit, Mustafa; Kaya, Mehmet Salih; Kirbas, Mesut; Dursun, Sukru; Alak, Ilyas; Atli, Mehmet Osman
    MicroRNA (miRNA), acting as the transcriptional regulator of gene expression, has been widely demonstrated to be involved in many biological functions, including embryo implantation and devel-opment. The objective of the current study was to illuminate the expression pattern of microRNAs (miRNAs) in the endometrium during the peri-implantation in ewes. Intercaruncular endometrial samples was obtained from a total of 24 ewes on days of 12 (pre-implantation, n = 4), 16 (implantation, n = 4) and 22 (post-implantation, n = 4) of pregnancy following mating, and on their corresponding days of 12 (n = 4), 16 (n = 4) and 22 (n = 4) of the estrous cycle. The miRNA profiles were examined in the endometrium by microarray technology. We detected 116 ovine specifics miRNAs in the endometrium. Of these, nineteen were differentially expressed in early pregnancy. Four miRNAs (oar-miR-370-3p, oar-miR-411b-5p, oar-miR-379-3p and oar-miR-411a-3p) that had the most differential fold change were confirmed by RT-qPCR in ovine endometrium. The differentially expressed miRNAs targeted a total of 315 genes, resulting in 39 GO terms in molecular function, 353 in biological process, and 17 in the cellular component. The construction of the PPI network of target genes established two functional modules mostly enriched in the innate immune system, toll receptor cascades in module 1, whereas genes in module 2 were associated with GMCSF-mediated signaling events, insulin pathway, and mTOR signaling pathway. Based on the results, we may imply that miRNAs modulate ovine endometrium during the peri-implantation.(c) 2022 Elsevier Inc. All rights reserved.
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    Investigation of Accelerated Lambing Possibility of Anatolian Merino Sheep
    (Kafkas Univ, Veteriner Fakultesi Dergisi, 2014) Bulbul, Bulent; Kirbas, Mesut; Aktas, Ahmet Hamdi; Kose, Mehmet; Ataman, Mehmet Bozkurt; Coyan, Kenan; Kan, Mustafa
    Investigation of the possibilities to increase the number of lamb gained in a year via using the accelerated lambing method and, the profitability of a farm related to this is aimed with this study. As material, 525 Anatolian Merino ewes and 40 rams, aged at 2-4, in field conditions, and 199 ewes and 15 rams at Bahri Dagdas International Agricultural Research Institute were used. Ewes in the field condition and at the Institute were divided in to two groups as accelerated lambing and control and, 200 ewes in the field condition and 75 ewes at the Institute were remained as control to get one lamb per year while 325 in the field condition and 124 at the Institute were formed treatment group. The control ewes were bred in August and September, the traditional breeding season, in a 12 month interval while 3 lambings in 2 years were applied to the ewes in the treatment group and they were bred for one month again following the period of 5 months of pregnancy, 40 days of lactation and 20 days of weaning. Ram effect, ram effect + flushing and some different protocols were used for induction and synchronization of estrus for accelerated lambing in the treatment group. As a result, more fecundity and lamb productivity achieved by accelerated lambing than once a year lambing. Synchronization methods were found to be effective on fecundity and lamb productivity in accelerated lambing applications. However, lamb yield obtained by synchronization methods used in this study were not profitable.
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    Plasma profile of Pregnancy Associated Glycoproteins during Postpartum period in Konya Merino ewes lambing single and twin
    (Univ Zulia, Facultad Ciencias Veterinarias, 2023) Ucar, Eyyup Hakan; Kose, Mehmet; Kirbas, Mesut; Bozkaya, Faruk; Atli, Mehmet Osman
    This study investigated the plasma profile of Pregnancy Associated Glycoproteins (PAGs) after lambing in Konya Merino ewes using specific cattle pregnancy test kit. A total of 16 Konya Merino ewes were used as a material. Four groups were set up, ewes birthing a male lamb (SM group, n=4), a female lamb (SF group, n=4), twin male lambs (TM group, n=4) or twin female lambs (TF group, n=4). Blood plasma samples were taken on days 0 (lambing day), 3, 7, 10, 14 and then weekly until day 35. All samples were analysed using a bovine pregnancy test kit to detect of PAGs level. It was determined that there was a strong negative correlation between PAGs levels in peripheral blood and days after lambing (r2=0.969; P<0.01). However, no relationship was found between PAGs level and lamb gender or birth type. In conclusion, plasma PAGs level decreases rapidly in Konya Merino ewes regardless of lamb gender and birth type. Moreover, the results showed that the plasma profile of PAGs after lambing in Konya Merino ewes can be monitored with a bovine commercial ELISA-based pregnancy test kit, and the test results can be used in decisions and assessments based on the levels of PAG molecules.
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    Presynchronization in Sheep Ensures Synchronization of Next Estrus and Improves Fertility
    (Chulalongkorn Univ, 2023) Bulbul, Bulent; Kose, Mehmet; Kirbas, Mesut; Coyan, Kenan; Ataman, Mehmet Bozkurt; Bekci, Emine Aysun; Umutlu, Seyit
    In this study, fertility was evaluated either in the first hormonally synchronized or in the subsequent natural estrus among 202 sheep during the breeding season. Intramuscular (im) PGF(2 alpha )was administered 11 days apart in both the PG (n=50) and Pre-PG (n=50) groups, while im PGF(2 alpha )was given at the removal of an intravaginal progesterone sponge (after 11 days) in the Sponge (n=51) and Pre-Sponge (n=51) groups. Estrus was monitored following the last hormonal application (day 0) in all groups and starting from day 16 in the Pre-PG and Pre-Sponge groups for 5 days. Ewes were mated during the first hormonally synchronized estrus (PG and Sponge groups) or in the next (natural) estrus (Pre-PG and Pre-Sponge groups). The estrus rate and litter size in the Pre-Sponge group were higher than those in the other groups (P<0.05). In the Pre-Sponge group, pregnancy and lambing rates were higher than in the PG and Sponge groups, and fecundity was higher than in the PG group (P<0.05). It was concluded that progesterone and PGF(2 alpha )-based estrus synchronization may negatively affect fertility. Additionally, the next estrus is also synchronized after using progesterone and PGF(2 alpha )-based synchronization protocols. Fertility loss caused by the application of exogenous hormones can be mitigated by mating at the next natural estrus. Furthermore, the next natural estrus can be utilized in situations where hormone use is inappropriate but synchronization is desired.
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    Superovulation in cows synchronized with two different progesterone plus oestradiol protocols
    (Archiv Fur Tierzucht, 2013) Bulbul, Bulent; Kirbas, Mesut; Dursun, Sukru; Kose, Mehmet
    A total of 26 Brown Swiss cows were used to compare the synchronization and superovulatory response of follicle stimulating hormone treated cows that were synchronized with progesterone+oestradiol valerate or benzoate. Control cows (n=8) were superstimulated with follicle stimulating hormone using twice daily injections with decreasing doses from day 10-13 after determined reference oestrus. Cows in treatment groups were received either ear implant (n=9) containing norgestomet+oestradiol valerate or progesterone releasing intravaginal device (n=9) containing progesterone+oestradiol benzoate, at random stage of the oestrus cycle, for 9 days. Seven days after the implant and progesterone releasing intravaginal device insertion, follicle stimulating hormone was injected as described in the control group. There was no significant difference between the groups for superovulation responses. In conclusion, both protocols synchronized the oestrus cycle in follicle stimulating hormone treated cows and, any of the protocols evaluated in this study can be used as a pretreatment for superstimulation started on the seventh day of the implant or progesterone releasing intravaginal device insertion in Brown Swiss cows.