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1
Evans, G., F. K. Hollinshead, and W. M. C. Maxwell. "Preservation and artificial insemination of sexed semen in sheep." Reproduction, Fertility and Development 16, no. 4 (2004): 455. http://dx.doi.org/10.1071/rd04032.
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Sperm-sexing technology using flow cytometry is in advanced stages of development for the sperm of several species. The sorting process could compromise sperm viability and sperm require specific handling procedures both before and after sorting to maintain the integrity and function of the sorted sperm. Standard freezing protocols have been modified for post-sorting cryopreservation of sperm and frozen sperm have been successfully thawed, sorted, refrozen and subsequently used to produce offspring. The relatively low numbers of available sorted sperm have, in some cases, led to modification of artificial insemination techniques to maximise efficiency of use. Multiple ovulation and embryo transfer, or in vitro fertilisation and associated technology, may lead to the more efficient use of sexed sperm.
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Zegeye, Zemenu Birhan, Nóra Vass, and Andualem Tomano. "Application of laparoscopic artificial insemination in conventional Lacaune sheep farm using frozen-thawed semen." Acta Agraria Debreceniensis, no. 2 (December 1, 2020): 133–38. http://dx.doi.org/10.34101/actaagrar/2/7113.
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The complex anatomical structure of the ewe reproductive tract accompanied with low quality of frozen ram semen for artificial insemination, resulted in a challenge with regard to using superior genotypes for reproductive ovine performance. Hence, improved genetics in ovine management has not been efficiently and widely used especially in undeveloped countries. Therefore, intrauterine semen deposition by laparoscopic insemination should be adopted in the current sheep production systems. Thus, this study aimed to assess the pregnancy rate and lambing rate of ewe inseminated by laparoscopic insemination techniques using frozen-thawed semen. The research used imported frozen semen from two rams of the Lacaune breed. Ewes were grouped according to age in years (1, 2 and 4). Before insemination, the semen was examined microscopically for its motility and viability and thereafter the laparoscopic artificial insemination technique was performed to 19 Lacaune breed ewes using frozen-thawed semen. The overall pregnancy and prolificacy rates were 31.57% and 42.10% respectively. Out of 2 ewes in the 1-year age group that were inseminated, only 1 ewe lambed representing 50%. However, from 16 ewes inseminated of four-year age group, 5 ewes lambed representing 31.25%. Significant difference based on age group was not evaluated due disproportionate of the data, (such that the data included 2 ewes in one-year-old age, 1 ewe in 2-year-old age and 16 ewes in 4-year-old age). Based on the ram semen, 33.33% and 30% of the inseminated ewes were pregnant from ram A and ram B semen respectively. However, in the case of prolificacy rate, 44.44% and 40 % of the ewes lambed from using semen of ram A and B, respectively. There was no significant difference (p>0.05) in pregnancy and prolificacy rates based on semen from the two rams. In conclusion, in this research study, ram semen had no significant effect on pregnancy and prolificacy rates using laparoscopic AI on Lacaune sheep. This could be due to the fact that the rams had very good quality semen. Evaluation of ram semen, accompanied with appropriate ewe selection based on age and rightful deposition of semen could lead to better and more consistent results. Overall this could contribute to the successful application of laparoscopic artificial insemination in Lacaune sheep production systems for enhanced productivity.
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Hunton, J. R., Sally E. Flecker, and W. M. C. Maxwell. "Pregnancy rates following intra-uterine insemination with pellet or straw-frozen ram semen." Journal of Agricultural Science 109, no. 1 (August 1987): 189–91. http://dx.doi.org/10.1017/s0021859600081144.
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There has been considerable research on techniques for artificial insemination of sheep with frozen ram semen (Maxwell, 1984). Acceptable pregnancy rates were reported following ‘two-step’ dilution and freezing of semen in P.V.C. ‘ministraws’ (Colas, 1975; Colas & Guerin, 1981); however, other workers have obtained poor fertility following cervical insemination with semen frozen in straws (Maxwell, etal. 1980; Tervit elal. 1984).
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Haresign, W., S. R. Read, R. M. Curnock, and H. C. B. Reed. "A note on the use of laparoscopy for intrauterine insemination of frozen-thawed semen in the ewe." Animal Science 43, no. 3 (December 1986): 553–56. http://dx.doi.org/10.1017/s0003356100002762.
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One of the major limitations to the widespread use of artificial insemination (AI) in the United Kingdom (UK) sheep industry is that with frozen-thawed semen current insemination techniques result in lowered fertility. Consequently, only fresh or liquid-chilled ram semen can be used if commercially acceptable conception rates are to be achieved.
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Maxwell, W. M. C., and L. J. Hewitt. "A comparison of vaginal, cervical and intrauterine insemination of sheep." Journal of Agricultural Science 106, no. 1 (February 1986): 191–93. http://dx.doi.org/10.1017/s0021859600061906.
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Practical systems for the artificial insemination of sheep have been available for many years (reviewed by Maxwell, 1984). The traditional method is to separate from the flock ewes exhibiting a natural oestrus as identified by ‘teaser’ rams, and inseminate these ewes with freshly collected and diluted semen. The ewes are generally inseminated by suspension of the hindquarters over an elevated rail and deposition of the semen within the first fold of the cervix using a plastic pipette, speculum and headlamp (the cervical insemination method, Salamon, 1976).
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Sarradin, Pierre, Sandrine Melo, Céline Barc, Céline Lecomte, Olivier Andréoletti, Frédéric Lantier, Jean-Louis Dacheux, and Jean-Luc Gatti. "Semen from scrapie-infected rams does not transmit prion infection to transgenic mice." REPRODUCTION 135, no. 3 (March 2008): 415–18. http://dx.doi.org/10.1530/rep-07-0388.
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Scrapie is the most common transmissible spongiform encephalopathy (TSE) in livestock. Natural contamination in sheep flocks is presumed to occur by maternal transmission to offspring. However, horizontal prion transmission from animal to animal exists and may be significant in sustaining and spreading contagion in the field. Artificial insemination is widely used in modern farming, and as large amounts of prion protein have been found in sheep sperm membrane, epididymal fluid and seminal plasma, horizontal transmission by this route was hypothesized since no clear information has been obtained on possible sexual transmission of TSE. We therefore tested the contamination levels of semen from scrapie-infected rams at different stages of incubation, including the clinical phase of the disease. We report here that under our experimental conditions ram semen did not transmit infectivity to scrapie-susceptible transgenic mice overexpressing the V136R154Q171 allele of the sheep prion (PRNP) gene. These results suggest that artificial insemination and natural mating have a very low or negligible potential for the transmission of scrapie in sheep flocks.
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Haresign, W., R. M. Curnock, and H. C. B. Reed. "Conception rates following cervical and intrauterine insemination in the ewe." Proceedings of the British Society of Animal Production (1972) 1986 (March 1986): 65. http://dx.doi.org/10.1017/s0308229600015749.
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The current technique for artificial insemination (AI) of ewes during the breeding season necessitates the synchronisation of oestrus with progestagen sponges and PMSG, and involves depositing semen into the posterior cervix at a fixed time after sponge removal. Extensive field trials over a number of years in the UK have indicated that a single insemination using fresh diluted semen 56 h after sponge removal generally results in a conception rate of 70%, while conception rates following a single insemination of frozen-thawed semen 57 h after sponge removal have ranged from 19% to 34% (mean 28%). Giving two inseminations of frozen-thawed semen at 50 h and 60 h after sponge removal increased the overall mean conception rate, but only up to 48%. This inability to achieve acceptable conception rates with frozen-thawed semen nullifies many of the potential benefits of AI in sheep flocks.It is well established that the cervix presents a major barrier to sperm transport in the ewe, particularly when oestrus has been synchronised with progestagens and PMSG.
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Findlater, R. C. F., W. Haresign, and R. M. Curnock. "Effect of timing of intrauterine insemination with frozen-thawed semen on conception rates in ewes." Proceedings of the British Society of Animal Production (1972) 1987 (March 1997): 27. http://dx.doi.org/10.1017/s0308229600034681.
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The widespread use of artificial insemination (AI) in the United Kingdom sheep industry has been limited by the poor conception rates obtained after cervical insemination of frozen-thawed semen. The major problem in this respect is the impairment of sperm transport through the cervix, particularly when AI is used in conjunction with oestrus synchronisation.Previous studies (Killeen and Caffery, 1982; Maxwell, 1984) have indicated that a laparo-scopic technique for intrauterine insemination in ewes may overcome such limitations. At the moment, however, sufficient data on the optimum time of insemination and sperm doses required to maximise fertility in British breeds are not available. The present study was conducted to establish the optimum time of intrauterine insemination using frozen-thawed semen.
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FUKUI, Yutaka, Hirohide KOHNO, Tetsuro TOGARI, Mami HIWASA, and Kentaro OKABE. "Fertility after Artificial Insemination Using a Soybean-Based Semen Extender in Sheep." Journal of Reproduction and Development 54, no. 4 (2008): 286–89. http://dx.doi.org/10.1262/jrd.20004.
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Fukui, Y., H. Kohno, T. Togari, T. Matsuoka, and H. Imai. "10 EFFECTS OF INSEMINATION TIME, BREED, AND INSEMINATOR ON FERTILITY OF EWES INTRAUTERINALLY INSEMINATED WITH FROZEN - THAWED SEMEN IMPORTED FROM NEW ZEALAND." Reproduction, Fertility and Development 19, no. 1 (2007): 123. http://dx.doi.org/10.1071/rdv19n1ab10.
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Artificial insemination, especially with the use of frozen semen, is one of the important tools for embryo transfer program in sheep. The present study investigated the effects of insemination times, breeds, and two inseminators on the fertility of ewes intrauterinally inseminated with frozen–thawed ram semen imported from New Zealand. At 8 sheep farms located in Hokkaido, Japan, during the breeding season (October to December) in 2005, a total of 64 mature (1- to 6-year old) Suffolk (32 heads) and Polled Dorset (32 heads) ewes were used. The ewes were treated with controlled intravaginal drug release (CIDR containing 0.3 g progesterone; Pharmacia & Upjohn, Ltd., Auckland, New Zealand) for 12 days and an injection of 500 IU equine chorionic gonadotropin one day before CIDR removal. The fixed-time intrauterine inseminations (early: 43–46 h; late: 47–50 h) after CIDR removal were performed using the frozen–thawed semen from a Suffolk and Polled Dorset ram by two inseminators. The effects of breeds (Suffolk and Polled Dorset), fixed-time insemination times (early and late phases), and two inseminators on pregnancy (number of pregnant ewes/number of ewes inseminated, 60 days after insemination) and lambing (number of lambed ewes/number of ewes inseminated) rates were analyzed by chi-square test. The prolificacy was compared by Student's t-test, and differences were also analyzed by Tukey's omega procedure. The effect of the different farms on fertility was not examined due to the small numbers of ewes per farm. Pregnancy (60.0 and 72.4%, respectively) and lambing (60.0 and 71.4%, respectively) rates were not significantly different between Suffolk and Polled Dorset ewes. The inseminators also did not affect pregnancy (62.6 and 68.8%) and lambing (62.6 and 67.7%) rates. For the insemination times, the lambing rate tended to be higher (P ≤ 0.10) in the early insemination than in the late insemination (76.7% and 53.6%, respectively). The present results show acceptable fertility in ewes inseminated with Suffolk and Polled Dorset frozen semen imported from New Zealand. The early intrauterine insemination (43–46 h after CIDR removal) tended to result in higher fertility than the late insemination (47–50 h after CIDR removal). From 38 lambed ewes, 60 newborn lambs were produced, and this has provided new blood lines of Suffolk and Polled Dorset sheep in Japan.
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Critser, John K., and Jeanne V. Linden. "Therapeutic insemination by donor I: A review of its efficacy." Reproductive Medicine Review 4, no. 1 (March 1995): 9–17. http://dx.doi.org/10.1017/s0962279900001022.
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Of all the assisted reproductive technologies in current use, artificial insemination has by far the longest history. While the earliest verifiable reports using this technique date to the eighteenth century for nonhuman artificial insemination and to the nineteenth century for human artificial insemination, systematic use of this approach to assist reproduction did not occur until the early part of this century. During the early 1900s, in Russia, Ivanov developed methods for semen collection from and insemination of horses. These techniques were later modified to apply to other agriculturally important species so that by the 1930s, millions of horses, cattle and sheep were being bred using artificial insemination. The adaptation of widespread use of artificial insemination (primarily in cattle) in agriculture extended to Britain in the early 1940s and to the USA in the 1950s. Corresponding implementation of artificial insemination in human reproductive medicine closely followed these innovations in the animal husbandry field.
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Vlasenko, S., O. Zhulinska, and O. Yeroshenko. "Clinical and laboratory prognostic indicators for fertility in sheep." Naukovij vìsnik veterinarnoï medicini, no. 1(149) (May 30, 2019): 6–14. http://dx.doi.org/10.33245/2310-4902-2019-149-1-6-14.
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With the use of technology of artificial insemination in sheep farms are not yet defined criteria for assessing the full value of the preparation of females for insemination, and hence – the possibility of prediction and correction of their fertilization, which prevents the rational use of cryopreserved semen and ensuring the maximum reception of the offspring. We have proved the prognostic importance of fertilization of the morphofunctional state of the vulva and the vagina and the quality of mucus in sheep breeds during estrus. The material of the study was 327 sheep of ascanian breeding, which during the sexual intercourse before insemination examined the vulva, vagina and evaluated the estrus slime (number, color, consistency, presence of impurities, elasticity, electrical resistance, type of crystallization, protein content). The results of ultrasound diagnosis of pregnancy were determined by the fertility of sheep with different integral compositions of clinical and laboratory parameters. It has been established that in sheep, in which fertility in the first sexual cycle reached 65.1%, most often found a pink, moderately edematous vulva with clear mucus. In females with pale mucous membrane and insignificant amount of mucus, the effectiveness of inseminates declined to 53.3-58.0%. In a significant amount of estral mucus, the proportion of infertile animals increased twofold. At the same time, the selection of liquid, but cloudy, or thick mucus is a sign of an unfavorable prognosis, in which fertility decreases by 1.8-2.1 times (p <0.001). Dense, white, paste-like isolates were observed in a small number of sheep, mostly bright at the beginning of the anestral season. Low fertility in the first sexual hunting (35.5%) and a high multiplicity of repeated inseminations (29.0%) indicate that sheep with thick estral slime are only beginning to enter the sexual season, and this quality of secrecy indicates an inadequate estrogenization of the body. It was also found that in the infertile sheep during sexual hunting, the protein content of cervical mucus was 4.8 times higher, and the elasticity of mucus was reduced by 2.9 times. The most prevalent was the prognosis of average fertilization (53.3-58.0%), which was recorded in 62.9% of experimental sheep. The prognosis of high fertilization, which resulted in 62.5-65.1% of oseminins, was found in 27.8% of females. At the same time, the number of females with a fertility forecast at 40% was only 3.1%, and the prevalence of an unfavorable prognosis, in which fertility was the smallest (30.0-35.5%), reached 6.2%. Key words: sheep, askanian breeding, estrus, fertility prognosis, estral mucus, vulva, vagina, artificial insemination.
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Lucidi, P., B. Barboni, and M. Mattioli. "Ram-induced ovulation to improve artificial insemination efficiency with frozen semen in sheep." Theriogenology 55, no. 9 (June 2001): 1797–805. http://dx.doi.org/10.1016/s0093-691x(01)00522-2.
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Kumar, Davendra, and Krishnappa Balagnur. "Recent advances in cryopreservation of semen and artificial insemination in sheep: A review." Indian Journal of Small Ruminants (The) 25, no. 2 (2019): 134. http://dx.doi.org/10.5958/0973-9718.2019.00043.6.
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Consalter, Angélica, Andressa F. Silva, Edwards Frazão-Teixeira, Luis F. Matos, Francisco C. R. de Oliveira, Juliana S. Leite, Franciele B. F. Silva, and Ana M. R. Ferreira. "Toxoplasma gondii transmission by artificial insemination in sheep with experimentally contaminated frozen semen." Theriogenology 90 (March 2017): 169–74. http://dx.doi.org/10.1016/j.theriogenology.2016.12.004.
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HIWASA, Mami, Hirohide KOHNO, Tetsuro TOGARI, Kentaro OKABE, and Yutaka FUKUI. "Fertility after Different Artificial Insemination Methods Using a Synthetic Semen Extender in Sheep." Journal of Reproduction and Development 55, no. 1 (2009): 50–54. http://dx.doi.org/10.1262/jrd.20062.
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Ahangari, Y. J., M. Nowrozi, and A. Soleimani. "The effect of dilution rates and freezing methods on post-thawing motility of Baluchi ram spermatozoa." Proceedings of the British Society of Animal Science 2002 (2002): 130. http://dx.doi.org/10.1017/s1752756200007869.
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For the effective use of Artificial insemination technique in sheep industry, investigation on the methods of ram semen dilution and freezing is necessary. Ahangari (1992) showed that various rates of dilution from 1:1 to 1:4 did not affect p>0.05 post thawing survival of Cambridge ram spermatozoa. Fiser et al (1987) achieved a 73% c.f. 67% pregnancy rate using thawed semen, previously frozen to -100 and -79 respectively. The objective of this study was to investigate the effect of two rates of dilution of semen and two methods of freezing on post-thawing motility of ram spermatozoa.
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Morrier, A., F. Castonguay, and J. L. Bailey. "Glycerol addition and conservation of fresh and cryopreserved ram spermatozoa." Canadian Journal of Animal Science 82, no. 3 (September 1, 2002): 347–56. http://dx.doi.org/10.4141/a01-045.
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Fresh extended ram semen has a short fertile lifespan whereas acceptable fertility with cryopreserved semen is achieved only by laparoscopy, which limits widespread artificial insemination in sheep. Although glycerol is considered essential for freezing spermatozoa, it is often included in extenders for short-term storage at above-freezing temperatures. To test the hypothesis that glycerol reduces the function of fresh sperm, ram semen was divided into two aliquots and diluted with commercial extenders that were identical, except that one contained 7% glycerol (n = 6). In a second experiment, ram semen was prepared for cryopreservation by a one-step dilution with a 7% glycerol extender or gradually, with a two-step protocol, to test the hypothesis that the method and time of glycerol addition affects sperm quality after freezing and thawing (n = 7). For both experiments, semen was diluted in a synthetic oviductal fluid (SOF-m) and sperm quality was assessed by computer-assisted motility, viability and chlortetracycline fluorescence (CTC) patterns (an indicator of capacitation status). The presence of glycerol did not affect the quality of fresh sperm (P > 0.27). For cryopreserved sperm, the method of glycerol addition also did not affect thawed sperm. However, a decrease in sperm motility and viability, and different distribution of CTC patterns occurred due to the duration of time in extender and in SOF-m (P ≤ 0.0002). Cryo-capacitation was also observed. In conclusion, the presence of glycerol in the extender did not reduce ram sperm quality during conservation of the semen at 5°C or when it was used to completely and rapidly dilute the semen before cooling for cryopreservation. Key words: Sheep, Triladyl, Biladyl, chlortetracycline, artificial insemination, spermatozoa.
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Madrigali, A., A. Rota, D. Panzani, S. Castellani, M. Shawahina, A. Hassan, F. Di Iacovo, C. Rossignoli, and F. Camillo. "Artificial Insemination in Sheep with Fresh Diluted Semen: Comparison Between Two Different Semen Extenders and Management Protocols." Tropical Animal Science Journal 44, no. 3 (August 21, 2021): 255–60. http://dx.doi.org/10.5398/tasj.2021.44.3.255.
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Masoudi, Reza, Ahmad Zare Shahneh, Armin Towhidi, Hamid Kohram, Abbas Akbarisharif, and Mohsen Sharafi. "Fertility response of artificial insemination methods in sheep with fresh and frozen-thawed semen." Cryobiology 74 (February 2017): 77–80. http://dx.doi.org/10.1016/j.cryobiol.2016.11.012.
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NTEMKA, A., I. A. TSAKMAKIDIS, E. KIOSSIS, A. MILOVANOVIĆ, and C. M. BOSCOS. "Current status and advances in ram semen cryopreservation." Journal of the Hellenic Veterinary Medical Society 69, no. 2 (July 18, 2018): 911. http://dx.doi.org/10.12681/jhvms.18014.
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Ram semen cryopreservation contributes to genetic improvement through artificial insemination, eliminates geographical barriers in artificial insemination application and supports the preservation of endangered breeds thus the conservation of biodiversity. Sperm freezing process induces ultrastructural, biochemical and functional changes of spermatozoa. Especially, spermatozoa’s membranes and chromatin can be damaged, sperm membranes’ permeability is increased, hyper oxidation and formation of reactive oxygen species takes place, affecting fertilizing ability and subsequent early embryonic development. Aiming to improve ram frozen-thawed semen’s fertilizing capacity, many scientific investigations took place. Among them the composition of semen extenders, was a main point of interest. Semen preservation extenders regulate and support an environment of adequate pH and buffering capacity to protect spermatozoa from osmotic and cryogenic stress. Therefore, permeating (glycerol, dimethyl sulfoxide) and non-permeat ing (egg yolk, skimmed milk) cryoprotectants, sugars (glucose, lactose, trehalose, raffinose), salts (sodium citrate, citric acid) and antioxidants (amino acids, vitamins, enzymes) have been added and tested. Moreover, semen dilution rate, storage temperature, cooling rate and thawing protocol, are also some key factors that have been studied. The research results of this scientific topic are encouraging, not only about the freezing and thawing procedures, but also about the improvement of the additives’ properties. However, further research is needed to enhance the fertilizing ability of ram frozen-thawed semen, making its use practical in sheep reproductive management by the application of cervical artificial insemination.
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Geenty, K. G., F. D. Brien, G. N. Hinch, R. C. Dobos, G. Refshauge, M. McCaskill, A. J. Ball, et al. "Reproductive performance in the Sheep CRC Information Nucleus using artificial insemination across different sheep-production environments in southern Australia." Animal Production Science 54, no. 6 (2014): 715. http://dx.doi.org/10.1071/an11323.
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The present paper covers reproductive performance in an artificial-insemination (AI) program of the Sheep CRC Information Nucleus with 24 699 lambs born at eight locations in southern Australia across five lambings between 2007 and 2011. Results from AI with frozen semen compared well with industry standards for natural mating. Conception rates averaged 72%, and 1.45 lambs were born per ewe pregnant for Merino ewes and 1.67 for crossbreds. Lamb deaths averaged 21% for Merino ewes and 15% for crossbreds and 19%, 22% and 20% for lambs from ewes that were mated to terminal, Merino and maternal sire types, respectively. Net reproductive rates were 82% for Merino ewes and 102% for crossbreds. From 3198 necropsies across 4 years, dystocia and starvation-mismothering accounted for 72% of lamb deaths within 5 days of lambing. Major risk factors for lamb mortality were birth type (single, twin or higher order), birthweight and dam breed. Losses were higher for twin and triplet lambs than for singles and there was greater mortality at relatively lighter and heavier birthweights. We conclude that reproductive rate in this AI program compared favourably with natural mating. Lamb birthweight for optimum survival was in the 4–8-kg range. Crossbred ewes had greater reproductive efficiency than did Merinos.
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Jaśkowski, Bartłomiej M., Magdalena Woźna, and Marek Gehrke. "Sex-sorted sperm in farm animals: scale of use, the effectiveness of insemination, risk factors." Medycyna Weterynaryjna 73, no. 5 (2017): 272–79. http://dx.doi.org/10.21521/mw.5696.
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The aim of the study was to present the scale of use, risk factors and possibilities, which sorter semen gives in biotechnics used in reproduction of cattle. Modern sorters allow for the evaluation of 6 million X and Y spermatozoa per hour. Sex-sorted semen, which is commercially used, contains 2.1 x 106 of spermatozoa. It is used mostly in AI of milk heifers, mainly in large cattle herds. Sorted semen containing Y spermatozoa is sold less often in the world than the one with X spermatozoa. The percentage of the desired sex of the young is higher than 90. The pregnancy rate after application of sorted semen is about 20–25% lower than after insemination of non-sorted semen and depends on a number of factors. The main factors are: breed of female, service number, the herd of origin, the depth of semen deposition, the bull producing semen, ambient temperature and technical parameters during sperm sorting. A number of methods have been developed to improve conception rate, including timed artificial insemination (TAI) and synchronization of heat and ovulation. Results of donor inseminations with the use of sorter semen are presented, with the lower percentage of embryos suitable for the transfer and embryos of the highest quality highlighted. Previous studies do not indicate a reduction of the conception rate after the transfer of embryos obtained in vitro and in vivo after fertilization using sorted semen. It remains difficult to justify a significant increase in the frequency of stillbirths of bulls after using sorted sperm. Similarly, 16% of stillbirths of bulls were observed after embryo transfer, when donors were inseminated with sorter semen. The percentage of stillbirths of bulls after embryo transfer with the use of conventional semen is 9%. The sorted semen is not often used for inseminations in pigs, sheep and goats.
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Fogarty, N. M., W. M. C. Maxwell, J. Eppleston, and G. Evans. "The viability of transferred sheep embryos after long-term cryopreservation." Reproduction, Fertility and Development 12, no. 2 (2000): 31. http://dx.doi.org/10.1071/rd00020.
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The survival to term of 414 sheep embryos, thawed and transferred after conventional cryopreservation and storage for 13 years, was evaluated. A concurrent experiment involving the transfer of vitrified embryos to 91 ewes and artificial insemination of 51 ewes with frozen–thawed semen from sires of the long-term cryopreserved embryos provided forms of control treatments. The donor ewes had a mean ovulation rate of 10.9, and 7.1 embryos per ewe were cryopreserved. Each recipient ewe received two embryos and pregnancy was assessed at Day 18, Day 54 and term. The pregnancy rate was lower in the long-term embryo group than the artificial insemination group at Day 18 (P<0.01) and Day 54 (P<0.05), although the difference at term (31% v. 49%) was not significant, with the vitrified embryo group being similar to the long-term group. Embryo survival to birth was 21%, with the majority of loss (80%) occurring by Day 18. The later stage of development and higher grade of transferred embryos and the older age of donor ewes resulted in a significantly higher (P<0.01) pregnancy rate at Day 54 and term, and percentage of lambs born and weaned. Other effects of donor ewes (genotype, superovulation treatment, number of ovulations and embryos cryopreserved) were not significant. Implications for the design of genetic evaluation and germplasm conservation programmes using embryo cryopreservation technology are discussed.
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AMIRIDIS (Γ.Σ. ΑΜΟΙΡΙΔΗΣ), G. S., B. KUEHHOLZER, U. BESENFELDER, A. LYMBEROPOULOS (Α. ΛΥΜΠΕΡΟΠΟΥΛΟΣ), and E. VAINAS (Ε. ΒΑΪΝΑΣ). "Laparoscopie collection and transfer of Chios sheep embryos." Journal of the Hellenic Veterinary Medical Society 50, no. 3 (January 31, 2018): 244. http://dx.doi.org/10.12681/jhvms.15717.
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The efficiency of two different laparoscopic techniques for embryo collection was examined. Two groups (A and B) of 12 Chios breed ewes were used as donors and 20 crossbred Chios ewes as recipients. Oestrus was synchronised by intravaginal sponges, the donors were supervoulated by 8,8 mg of ovine FSH and laparoscopic intrauterine artificial insemination was conducted with extended fresh ram semen. Embryo collection was carried out the 6th day after AI. The uteri of the animals of group A were flushed after catheterisation of the ipsilateral oviduct. In the animals of group Β a flushing catheter was inserted close to the tip of the horn. Fifty nine and 46 embryos were collected from groups A and Β corresponding to recovery rate of 51.75% and 39% respectively (P>0.05). Laparoscopic transfer of 43 fresh embryos resulted to 11 (55%) pregnancies with (39.5%) surviving fetuses.
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PAPADOPOULOS (Σ. ΠΑΠΑΔΟΠΟΥΛΟΣ), S., E. THEODOSIADOU (Α. ΘΕΟΔΟΣΙΑΔΟΥ), D. KANTAS (Δ. ΚΑΝΤΑΣ), and E. VALASI (Ε. ΒΑΛΑΣΗ). "The application of in vitro fertilization techniques for the evaluation of ram fertility." Journal of the Hellenic Veterinary Medical Society 66, no. 2 (January 31, 2018): 63. http://dx.doi.org/10.12681/jhvms.15588.
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The prediction of field fertility of a given ram by using in vitro tests would be of great importance for the reproductive management in sheep flocks. There are many in vitro procedures available for evaluating semen quality and fertilizing ability, and the method chosen depends on the objective of evaluating the sperm and the available resources. The in vitro evaluation of semen fertilizing ability was firstly developed for the artificial insemination (AI) purposes and secondly for the application of in vitro fertilization (IVF) technique. The IVF techniques allow the assessment of fertility in terms of ability to penetrate and fertilize in vitro mature oocytes and ultimately to yield component embryos following IVF and culture. In this review are briefly presented in vitro studies performed in an attempt to establish an accurate laboratory test for the evaluation and, even more, the prediction of field fertility in sheep.
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Buckrell, B. C., C. Buschbeck, C. J. Gartley, T. Kroetsch, W. McCutcheon, J. Martin, W. K. Penner, and J. S. Walton. "Further development of a transcervical technique for artificial insemination in sheep using previously frozen semen." Theriogenology 42, no. 4 (September 1994): 601–11. http://dx.doi.org/10.1016/0093-691x(94)90377-u.
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Wildeus, S., D. L. Wright, and D. O'Brien. "103 Use of Liquid Semen Artificial Insemination in Katahdin Sheep in a Commercial Farm Setting." Journal of Animal Science 95, suppl_1 (December 1, 2016): 51. http://dx.doi.org/10.2527/ssasas2017.0103.
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Mansur, Md Abdullah Al, Md Golam Shahi Alam, Pankaj Kumar Jha, Md Asaduzzaman Rimon, Nazmun Naher, and Farida Yeasmin Bari. "Conception rate following intra-cervical artificial insemination using frozen semen at field level in indigenous sheep of Bangladesh." Asian Journal of Medical and Biological Research 4, no. 1 (June 7, 2018): 55–62. http://dx.doi.org/10.3329/ajmbr.v4i1.36822.
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This study was undertaken to study the AI conception rate using frozen semen at field level. Five farms in Mymensingh, Bangladesh were selected for AI Trial in field ewes. Four rams were selected for semen collection, evaluation, and frozen semen production and further to study conception rate followed by intra-cervical AI in both natural and synchronized ewes. Conception rate were confirmed by non-returned rate and ultrasound scanning at 30-40 days of post insemination. The volume, colour, mass activity, sperm motility, viability, concentration, HOST +ve (%) and normal spermatozoa percentages were 0.8±0.3 ml, 3.9±0.3, 4.4±0.6, 81.3±5.0%, 90.0±4.0%, 3519.0±545.6x106/ml, 87.4±3.3% and 85.6±1.8%, respectively. The sperm concentration of ram R#6 was significantly higher (P<0.05) (4120.5±93.5x106/ml) compared with other rams. The mean motility and viability of pre-dilution, 120 minutes of addition of Part-A, 240 minutes of addition of Part-B and post-thaw were (83.8±4.8%, 81.3±2.5%, 80.0±4.1% and 41.3±9.5%) and (93.3±1.0%, 90.0±1.4%, 88.8±1.0% and 58.3±8.7%), respectively. There were no significant difference (P˃0.05) between pre-dilution and post-dilution sperm motility and viability percentage however, post-thaw sperm motility and viability significantly (P<0.05) decreased compare with the motility and viability of pre-dilution and post-dilution values. Motility and viability percentages of frozen semen did not decrease significantly (P> 0.05) with the increase of preservation time. The mean motility and viability at 24 hrs, day 7, day 15 and day 30 were 41.3±9.5%, 41.5±8.5%, 41.8±9.9% and 40.5±10.2%; and 59.0±10.1%, 58.5±7.7%, 59.0±8.8% and 57.8±8.3%, respectively. The conception rates in natural and synchronized estrous were 26.7% and 25%, respectively. There was no significant difference in conception rates between the natural and synchronized oestrous in field level. However, the present non-return rate and conception rate indicate the suitability of produced frozen semen application in the field level.Asian J. Med. Biol. Res. March 2018, 4(1): 55-62
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Pérez-Garnelo, S. S., C. Borque, N. Madrid-Bury, M. Delclaux, C. Talavera, E. Martínez, A. T. Palasz, and J. De La Fuente. "199 BASIC CHARACTERISTICS AND CRYOBANKING OF BARBARY SHEEP (AMMOTRAGUS LERVIA) SEMEN." Reproduction, Fertility and Development 17, no. 2 (2005): 249. http://dx.doi.org/10.1071/rdv17n2ab199.
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Barbary sheep (Ammotragus lervia) are considered vulnerable species by the World Conservation Union (IUCN). The purpose of this study was to describe the basic characteristics of fresh semen, test the efficacy of commercial extender Triladyl, and collect necessary data that may help to create a frozen semen bank for the Barbary sheep in Spain. A total of 21 ejaculates were collected by rectal-probe electroejaculation from one dominant (D) and three minor (M) adult males housed in the Madrid Zoo. After ejaculation, semen volume, concentration, and mass motility were assessed. Remaining raw semen was diluted at 37°C with TRIS-based extender Triladyl (Minitüb, Tiefenbach, Germany) and 20% egg yolk to a final concentration of 200 × 106 sperm per mL. Diluted samples were kept at 5°C for 4 h and then loaded into 0.25-mL French straws, frozen at 5 cm above liquid nitrogen (LN2) for 10 min and then plunged into LN2. Samples were thawed in a water bath at 37°C for 30 s. Post-thaw semen survival was evaluated by sperm motility (%M), quality of movement (Q), normal acrosome status (%NAS), normal sperm morphology (%NOR), membrane integrity (hypo-osmotic test; %HOST), and sperm viability (eosin-nigrosin vital staining; %V), and were compared with the same parameters in the fresh semen. Data between D and M males were analyzed by one way ANOVA. Mean volume of ejaculates, total sperm concentration and mass motility of raw semen were respectively; 5.2 ± 1.56 mL, 2800.0 ± 1290.5 × 106 and 3.4 ± 0.4 for the D male, and 3.5 ± 3.2 mL, 251.2 ± 103.9 × 106, and 1.88 ± 1.4 for M males (P < 0.05). Remaining semen parameters evaluated in raw semen showed no differences between D and M males. However, post-thaw semen quality was significantly (P < 0.05) reduced in all analyzed parameters except %NAS and %NOR in M males groups as compared to the D male (Table 1). It can be concluded that Barbary sheep raw semen collected by electroejaculation is of sufficient quality to be used in an artificial insemination program and can be successfully frozen in commercially available Triladyl extender. However, the post-thaw viability of semen may considerably depend on the male reproductive status in the flock. Table 1. Characteristics of fresh and cryopreserved Barbary sheep semen This work was supported by CAM 07B/007/1999 (Analysis of ejaculate traits and development of methods of semen preservation in wild ungulates from the Madrid Zoo).
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Oláh, J., S. Kusza, S. Harangi, J. Posta, A. Kovács, A. Pécsi, C. Budai, and A. Jávor. "Seasonal changes in scrotal circumference, the quantity and quality of ram semen in Hungary." Archives Animal Breeding 56, no. 1 (October 10, 2013): 102–8. http://dx.doi.org/10.7482/0003-9438-56-010.
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Abstract. In this study, the quantitative and qualitative traits of semen were studied in seven rams of different breeds (Prolific Merino, Cokanski Tsigai, Barbados Blackbelly, Bábolna Tetra, Awassi, Ile de France and Suffolk), bred in Hungary. The semen parameters (density, volume, pH, mass motility, % motility, thawing and heat resistance), freezability of semen and the factors influencing these parameters were evaluated with respect to breed and season. The fresh and post-thawing quality of semen varied greatly with the breed and the season. The postthawing motility of semen cells was outstandingly high for Awassi rams in three seasons. During the test period, the smallest scrotal circumference was measured for Barbados Blackbelly, except for the summer when it increased by 12.5 cm. The reintroduction of artificial insemination could lead to a significant advancement of the sheep sector in Hungary. To promote this, we have provided useful and new information for breeders and organisations.
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Purdy, Phillip H. "174 What Quality and Fertility Should We Expect When Using Semen Cryopreservation and AI with Livestock? a Comparison Across Species." Journal of Animal Science 99, Supplement_1 (May 1, 2021): 113. http://dx.doi.org/10.1093/jas/skab054.186.
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Abstract Assisted reproductive technologies (ART) can be used across most agricultural species and will result in some degree of fertility when employed correctly. Still, conversations with agricultural producers and scientists (corporate, academic, governmental) repeatedly reveal that they do not know what success rates they should anticipate when using some ARTs, specifically semen cryopreservation and artificial insemination, with agricultural species (beef and dairy cattle, goats, pigs, poultry, sheep). These perceptions hinder ART application within the agricultural and scientific communities. Understanding these expected results is a critical component that is used to guide the USDA National Animal Germplasm Program laboratory operations for collecting, freezing and using germ plasm (semen, eggs, embryos, DNA, tissues, organs, cells), has consequently resulted in growth of the national collection, and provided tools, technologies, and educational opportunities for agricultural producers with documented success. Therefore, the intent of this presentation is to provide an overview of what results should be expected when using semen cryopreservation and artificial insemination across livestock species, explain the factors that influence successful use of these ARTs, which should encourage a more broad acceptance of their use with all agricultural species, and discuss opportunities for research and optimization that will improve fertility when using these technologies.
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Hodge, Marnie J., Sara de las Heras-Saldana, Sally J. Rindfleish, Cyril P. Stephen, and Sameer D. Pant. "Characterization of Breed Specific Differences in Spermatozoal Transcriptomes of Sheep in Australia." Genes 12, no. 2 (January 30, 2021): 203. http://dx.doi.org/10.3390/genes12020203.
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Reduced reproductive efficiency results in economic losses to the Australian sheep industry. Reproductive success, particularly after artificial insemination, is dependent on a number of contributing factors on both ewe and ram sides. Despite considerable emphasis placed on characterising ewe side contributions, little emphasis has been placed on characterising ram side contributions to conception success. Over 14,000 transcripts are in spermatozoa of other species, which are transferred to the ova on fertilisation. These transcripts conceivably influence early embryonic development and whether conception is successful. Semen was collected (n = 45) across three breeds; Merino, Dohne, and Poll Dorset. Following collection, each ejaculate was split in two; an aliquot was assessed utilising Computer Assisted Semen Analysis (CASA) and the remaining was utilised for RNA extraction and subsequent next-generation sequencing. Overall, 754 differentially expressed genes were identified in breed contrasts and contrast between ejaculates of different quality. Downstream analysis indicated that these genes could play significant roles in a broad range of physiological functions, including maintenance of spermatogenesis, fertilisation, conception, embryonic development, and offspring production performance. Overall results provide evidence that the spermatozoal transcriptome could be a crucial contributing factor in improving reproductive performance as well as in the overall productivity and profitability of sheep industries.
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Godfrey, Robert W., Sabrina Keiper, and Sue Lakos. "78 Evaluation of extended hair sheep ram semen stored as liquid at 5°C." Journal of Animal Science 98, Supplement_2 (November 1, 2020): 74–75. http://dx.doi.org/10.1093/jas/skz397.175.
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Abstract This study was designed to evaluate the quality of extended hair sheep ram semen stored as a liquid at 5°C. St Croix White (STX; n = 6) and Dorper x STX (DRPX, n = 5) rams were collected weekly for 3 wk using estrus ewes fitted with an intravaginal collection vial. Semen was kept at 32°C during transport to the lab and during processing. Semen was evaluated for percent motility (MOT), viability (LIVE) using eosin-nigrosin stain and concentration using a hemocytometer counting chamber. Semen was extended to a final concentration of 250 x 106/mL in a one-step dilution with a skim UHT milk extender with 10% egg yolk by volume and packaged into 0.5 mL straws. Straws were stored in a styrofoam box in a refrigerator at 5°C for 96 h, or in an Equitainer® set up using the manufacturer’s instructions, for 24 h at which time they were transferred to the styrofoam box in the refrigerator for 72 h. Semen was evaluated for MOT and LIVE at -1, 0, 24, 48 72 and 96 h relative to cooling. Semen traits were analyzed using GLM of SAS for repeated measures with ram, time, breed and cooling method in the model. There was no difference (P > 0.10) in MOT or LIVE over time between breeds or cooling method. One STX ram did not produce any samples that survived extension; one STX and four DRPX rams produced at least one sample that did not survive extension and all were removed from analysis. The MOT decreased (P < 0.0001) from 81.7 ± 2.9 % at -1 h to 52.2 ± 2.9% at 96 h. The LIVE decreased (P < 0.0001) from 83.1 ± 3.6% at -1 h to 50.4 ± 3.6% at 96 h. These results show that ram semen stored as a liquid at 5°C can maintain motility and viability for 96 h. Further studies will be conducted to evaluate cooling of extended ram semen and fertility after artificial insemination.
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Aboneev, V., V. Marchenko, and E. Aboneeva. "Some features of experimental researches in sheep breeding: to help of young scientists." Glavnyj zootehnik (Head of Animal Breeding), no. 6 (June 1, 2020): 58–64. http://dx.doi.org/10.33920/sel-03-2006-08.
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Organizing and conducting scientific and farm experiment is one of the most important components of any scientific and research work. The features of the modern period of animal husbandry development, the state of the feed base in all categories of farms, the availability of qualified specialists, the level and culture of conducting selection and technological techniques, create the main conditions for reliable performance of experimental researches. In sheep breeding the success of research work on the selection plan depends to a large extent on careful individual accounting of the results of insemination and lambing of ewes. Currently, in different categories of sheep farms crossbreeding is used as one of the methods for faster improvement and increasing the productivity of animals. At the same time, it is not always clear from the materials of many researches how the experimental groups of breeding stock were formed, what rules of selection were observed during artificial insemination, how accounting was carried out during insemination and lambing of ewes, what justifies the calculation of some indicators of economic effectiveness of the research without taking into account important economically useful features. The recommendations on how to eliminate possible errors when conducting research and production experiments by young scientists have been provided in the article. In particular, the main principles of forming experimental groups of sheep by age, productivity and origin have been shown. The necessity of housing all sex and age groups of experimental animals with complete feeding has been noted. The most important condition is the same number of ewes inseminated every day with the semen of each mating stud ram. It has been recommended to use vasectomy teaser rams for the authenticity of the origin of the obtained off spring. Special attention has been paid to the need to take into account all valuable economic useful features studied in the course of scientific and production experiments to calculate some indicators of economic effectiveness when rearing off spring of different origin.
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Pau, Salvatore, Laura Falchi, Mauro Ledda, Luisa Bogliolo, Federica Ariu, and Maria Teresa Zedda. "Surgery on cervical folds for transcervical intrauterine artificial insemination with frozen-thawed semen enhances pregnancy rates in the sheep." Theriogenology 126 (March 2019): 28–35. http://dx.doi.org/10.1016/j.theriogenology.2018.11.019.
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Wusiman, Abulizi, Yan-Ping Wang, Kang Ren, Guang-Bin Zhou, Xiang-Wei Fu, Lun Suo, Zhi-Qiang Fan, Liang Wang, and Shi-En Zhu. "Semen Storage at 23, 4 or -196°C and its Application to Artificial Insemination in Small-tail Han Sheep." Asian Journal of Animal and Veterinary Advances 7, no. 4 (March 15, 2012): 299–308. http://dx.doi.org/10.3923/ajava.2012.299.308.
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Wulster-Radcliffe, Meghan C., and Gregory S. Lewis. "Development of a new transcervical artificial insemination method for sheep: effects of a new transcervical artificial insemination catheter and traversing the cervix on semen quality and fertility." Theriogenology 58, no. 7 (October 2002): 1361–71. http://dx.doi.org/10.1016/s0093-691x(02)01042-7.
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Mittleider, T., S. Collins, P. Gibbons, and J. Gibbons. "12 Artificial insemination and embryo transfer results in ewes during a long daylength period." Reproduction, Fertility and Development 33, no. 2 (2021): 113. http://dx.doi.org/10.1071/rdv33n2ab12.
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Sheep are polyestrous, short-day breeders (∼11h of daylength), and exhibit oestrus approximately every 16–17 days during the breeding season, usually in late September to late December in the Northern Hemisphere. Progressive sheep producers often use assisted reproductive techniques such as laparoscopic AI and ovarian hyper-stimulation, embryo collection, and embryo transfer (ET) to increase genetic gain, and strive to have early December lambs to target specific show markets, which dictates AI or ET during the late summer. This field trial compared pregnancy rates following AI or ET in July and August (∼14h of daylength) in southwest Virginia (36–38′12″ N). Ewes (AI, n=83; ET recipients, n=33) were synchronized using a modified Ovsynch protocol involving intravaginal progesterone implants for 14 days, prostaglandin F2α (intramuscular) 48h before expected oestrus, and PG600 (IM) and gonadotrophin-releasing hormone (intramuscular) 52 to 54h and 16h before AI, respectively. Ewes were subjected to AI (frozen/thawed semen) regardless of whether they displayed signs of oestrus, and ewes selected as embryo recipients were subjected to a similar protocol but instead received 1 or 2 embryos (based upon the number of viable embryos produced per embryo donor) 6 days following the AI of the embryo donors. Ovarian hyper-stimulation of the embryo donors (n=13) was enabled by twice-daily FSH injections [totalling 350–455IU of Folltropin V (10–13mL)] for the 4 days before AI. Six days following AI, embryos were recovered surgically from the embryo donors (n=13) and yielded an average (±s.e.m.) of 6.6±1.2 total ova, 4.7±1.1 transferable quality embryos, and 1.9±0.8 unfertilized ova per collection. Pregnancy was detected using transrectal ultrasonography at ∼30 days of gestation and the pregnancy rates were analysed using Chi-squared. There was a tendency (P=0.092) for more pregnancies to be established following ET (22/33; 66%) compared with AI (41/83; 49%). There was no statistical relationship between AI ewes or ET recipient ewes that became pregnant relative to whether they displayed signs of oestrus or not. Embryo transfer was a more successful approach to produce pregnancies in ewes compared with AI during long daylength periods in this field trial. Further, ova from hyper-stimulated embryo donor ewes experienced a very high fertilization rate. Future studies will evaluate the ova capability directly via laparoscopic aspiration of ovarian follicles and IVF and evaluation of hyper- and non-hyper-stimulated follicular waves (using transrectal ultrasonography) and endocrine dynamics during different long and short daylength periods. Extending the opportunity to generate embryos and offspring regardless of daylength will assist aggressive sheep producers in reaching their reproductive, financial, and genetic goals.
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Mittleider, T., S. Collins, P. Gibbons, and J. Gibbons. "12 Artificial insemination and embryo transfer results in ewes during a long daylength period." Reproduction, Fertility and Development 33, no. 2 (2021): 113. http://dx.doi.org/10.1071/rdv33n2ab12.
Full textAbstract:
Sheep are polyestrous, short-day breeders (∼11h of daylength), and exhibit oestrus approximately every 16–17 days during the breeding season, usually in late September to late December in the Northern Hemisphere. Progressive sheep producers often use assisted reproductive techniques such as laparoscopic AI and ovarian hyper-stimulation, embryo collection, and embryo transfer (ET) to increase genetic gain, and strive to have early December lambs to target specific show markets, which dictates AI or ET during the late summer. This field trial compared pregnancy rates following AI or ET in July and August (∼14h of daylength) in southwest Virginia (36–38′12″ N). Ewes (AI, n=83; ET recipients, n=33) were synchronized using a modified Ovsynch protocol involving intravaginal progesterone implants for 14 days, prostaglandin F2α (intramuscular) 48h before expected oestrus, and PG600 (IM) and gonadotrophin-releasing hormone (intramuscular) 52 to 54h and 16h before AI, respectively. Ewes were subjected to AI (frozen/thawed semen) regardless of whether they displayed signs of oestrus, and ewes selected as embryo recipients were subjected to a similar protocol but instead received 1 or 2 embryos (based upon the number of viable embryos produced per embryo donor) 6 days following the AI of the embryo donors. Ovarian hyper-stimulation of the embryo donors (n=13) was enabled by twice-daily FSH injections [totalling 350–455IU of Folltropin V (10–13mL)] for the 4 days before AI. Six days following AI, embryos were recovered surgically from the embryo donors (n=13) and yielded an average (±s.e.m.) of 6.6±1.2 total ova, 4.7±1.1 transferable quality embryos, and 1.9±0.8 unfertilized ova per collection. Pregnancy was detected using transrectal ultrasonography at ∼30 days of gestation and the pregnancy rates were analysed using Chi-squared. There was a tendency (P=0.092) for more pregnancies to be established following ET (22/33; 66%) compared with AI (41/83; 49%). There was no statistical relationship between AI ewes or ET recipient ewes that became pregnant relative to whether they displayed signs of oestrus or not. Embryo transfer was a more successful approach to produce pregnancies in ewes compared with AI during long daylength periods in this field trial. Further, ova from hyper-stimulated embryo donor ewes experienced a very high fertilization rate. Future studies will evaluate the ova capability directly via laparoscopic aspiration of ovarian follicles and IVF and evaluation of hyper- and non-hyper-stimulated follicular waves (using transrectal ultrasonography) and endocrine dynamics during different long and short daylength periods. Extending the opportunity to generate embryos and offspring regardless of daylength will assist aggressive sheep producers in reaching their reproductive, financial, and genetic goals.
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Vallejo, D. A., J. D. Londoño, Y. A. Yepes, V. Tamayo, A. F. Mejia, and J. G. Maldonado. "Pregnancy rates in hair sheep after Ovsynch synchronization and a combined intracervical fixed-time artificial insemination and 10-day mating period." November-2019 12, no. 11 (November 2019): 1779–83. http://dx.doi.org/10.14202/vetworld.2019.1779-1783.
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Aim: This study aimed to evaluate the pregnancy rates in hair ewes using an Ovsynch synchronization protocol under a breeding system that combines fixed-time insemination plus a 10-day mating period as an alternative. Materials and Methods: Through an experimental study (n=27), ewes were randomly located into one of three treatments: (1) Pre-synch (n=9): Prostaglandin F2α (PGF2α)+Gonadotropin-releasing hormone (GnRH)+PGF2α+GnRH; (2) Ovsynch (n=9): GnRH+PGF2α+GnRH; and (3) control: Ewes bred by natural mating (NM) (n=9). Ewes were fixed-time inseminated (fixed-time artificial insemination [FTAI]) with fresh semen, collected just before the insemination time through vaginoscopy at 16 h after the second GnRH (gonadorelin) injection. Each experimental group was placed separately during 15 days and, after this time, fertile rams were allowed back with ewes for a 10-day mating period. Control group ewes remained with the rest of the herd suitable for breeding and were bred under NM. Pregnancy diagnosis was performed by ultrasound at 28-, 56-, and 84-day post-breeding to differentiate between FTAI and NM pregnancies. Total (FTAI±NM) pregnancy rates at 56-day post-breeding were used to compared Pre-synch, Ovsynch, and control. For this purpose, two-tailed proportions comparison z-test was used with a 95% confidence level, for testing as the null hypothesis whether two proportions were equal. Results: Pregnancy rates were higher in control ewes (66.4%) than FTAI (46.6%). When pregnancy rates after a 10-day mating period (40%) were added, the final rate (86.6%) was significantly (p<0.05) higher in Ovsynch-based protocols. The pregnancy rate was significantly lower in FTAI ewes compared to FTAI +10-day mating group (p<0.05). The overall pregnancy rate was 88.0, 85.7, and 67.0 (p>0.05) for Pre-synch, Ovsynch, and control ewes, respectively. Conclusion: These results provide evidence on the benefits of combined FTAI protocols for improving the reproductive efficiency of sheep.
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Crilly, J. P., L. Söderquist, A. Holmström, and N. D. Sargison. "Proof of concept of ovine artificial insemination by vaginal deposition of frozen-thawed semen under UK sheep-farming conditions: TABLE 1:." Veterinary Record 178, no. 21 (April 15, 2016): 532.1–532. http://dx.doi.org/10.1136/vr.103417.
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Dunne, L. D., M. G. Diskin, and J. M. Sreenan. "The effect of reducing feed intake following insemination on embryo survival in cattle." Proceedings of the British Society of Animal Science 1999 (1999): 3. http://dx.doi.org/10.1017/s1752756200001587.
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In farm animals early embryonic loss is recognised as a major cause of reproductive wastage. In sheep and pigs there is evidence that high energy intakes prior to ovulation and in early pregnancy depress systemic progesterone with a detrimental effect on embryo survival. The objective of this study was to investigate the effect of short term nutritional changes pre- and post-insemination on embryo survival and systemic progesterone in cattle. Preliminary results have been presented previously (Dunne etal., 1997).Oestrus was synchronised in 247 beef cross heifers aged 18-24 months using two injections of prostaglandin (PG) administered 10 days apart. At the oestrus following the first PG injection heifers were allocated to either a Low (L, 0.6 M) or High (H, 2.3M) pasture allowance for the 10 day period prior to artificial insemination (AI). AI was carried out at the oestrus following the second PG injection using semen from a single Limousin bull. On the day following AI, heifers were randomly reallocated to either a L or H pasture allowance until embryo recovery at day 14 to 16 or pregnancy diagnosis at Day 30.
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Herrid, M., M. Jackson, N. Suchowerska, S. Stockwell, K. Hutton, R. Davey, J. Olejnik, S. Hope, and J. R. Hill. "251. Production of donor-derived live lambs following testis germ cell transplantation." Reproduction, Fertility and Development 20, no. 9 (2008): 51. http://dx.doi.org/10.1071/srb08abs251.
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Testes germ cell transplantation in livestock has the potential for amplification of transgenic genotypes and for use as an alternative to artificial insemination. This study investigated a workable protocol for testis germ cell transplantation in sheep between animals of the same breed and different breeds. Testes of two groups of recipients at the stage of pre-pubertal (transition from gonocytes to spermatogonia, n = 2) or peri-pubertal (spermatogenesis initiated, n = 2) were treated with a single dose of 9 grey (Gy) or 15 Gy with a 6MV photon beam irradiation, respectively. In the first experiment, using pre-pubertal irradiated animals, testis germ cell transplantation between the same breed was performed at 16 weeks post irradiation. The left testes of recipient rams were injected with donor cells labelled with fluorescent dye PHK26, while the right testes were given unlabelled cells. The left testes of recipients were removed by castration after 2 weeks following transplantation to evaluate the location of the transferred cells, while the right testes were kept in place for long-term assessment of sperm output. In cryosections of the left testes, PKH26 positive cells were found both on the basement membrane as single cells or in the interstitial area. In the second experiment, animals irradiated at the peri-pubertal stage, received donor cells at 5 weeks post irradiation and animals were kept intact for semen production. For a period of two years after transplantation, semen samples were collected routinely from two groups of rams and analysed using microsatellite markers. Two recipients (50%) demonstrated the presence of donor DNA in their ejaculates. In order to investigate the fertility of the donor-origin sperm in recipient ejaculates, 99 ewes were artificially inseminated with semen from two positive rams. Four lambs (8%) have been identified as being sired by donor-derived sperm produced in the recipient ram that received a Merino to Merino transplantation, while no donor-derived offspring was obtained from the recipient with Border Leicester to Merino transplantation. This study represents the first report of the production of live progeny following testis germ cell transplantation in sheep.
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Pereyra-Bonnet, F., A. Gibbons, M. Cueto, R. Fernández-Martín, and D. Salamone. "307 TRANSGENIC OVINE EMBRYOS BY ARTIFICIAL INSEMINATION, IN VITRO FERTILIZATION AND INTRACYTOPLASMIC SPERM INJECTION." Reproduction, Fertility and Development 21, no. 1 (2009): 250. http://dx.doi.org/10.1071/rdv21n1ab307.
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Nowadays, transgenesis in animals constitutes an important tool for pharmacological protein production and livestock improvement. In 1971 Brackett first described that heterologous DNA can be introduced into a mammalian oocyte using sperm cells as vectors. We evaluated the capacity of AI, IVF and ICSI to produce transgenic embryos, in ovine, using sperm that had been exposed to a pCX-EGFP plasmid in Long and Short incubation protocols. The pCX-EGFP plasmid contains an enhanced green fluorescent protein gene (egfp) under the chimeric cytomegalovirus-IE-chicken β-actin enhancer-promoter control. Sperm/pCX-EGFP incubation was carried out by Long Incubation (2 h at 17°C in 200 μL of SFM medium: 100 mL contains glucose 1.2 g, Na citrate 1.0 g, EDTA 0.4 g, Citric acid 0.3 g, Trizma 0.6 g) and Short Incubation (5 min at 0–5°C in 10–100 μL of extender medium: 100 mL contains Na Citrate 2.8 g and EDTA 4 mg). For AI, Merino sheep (n = 17) were superovulated and inseminated with fresh semen (200 millions sperm/sheep) from eight Merino rams. The embryos were recovered by flushing the uterine horns by standard procedures. In IVF and ICSI, slaughterhouse oocytes were fertilized with frozen/thawed sperm. IVF was carried out in Brackett-Oliphant medium with 5 mm of caffeine, 20 IU mL–1 of heparin with 20 million sperm mL–1 during 5 h in an atmosphere of 5% CO2 in air. In ICSI, the spermatozoon was immobilized by breaking its tail and injected into MII oocytes. Immediately the oocytes were activated by incubation in TALP-HEPES with 5 μm ionomycin for 4 min, cultured in TCM199 for 3 h and transferred to a droplet of 1.9 mm DMAP for 3 h. Maturation and cultivation conditions were determined by standard operating procedures. All embryos were exposed to blue light (488 nm) to determine the percentage of morulae/blastocysts showing green fluorescence. Results are shown in Table 1. Statistical analysis was done by Fisher test. AI and IVF were able to produce a high percentage of morula and blastocyst stage, but were unable to produce transgenic embryos. In contrast, regardless of the sperm/plasmid incubation protocol, high percentages of transgenic morulae and blastocysts were always obtained by ICSI and the highest rate was achieved with Short Incubation (P < 0.05). In order to demonstrate ICSI-Short incubation embryo viability, two-day-old non-selected fluorescence embryos (n = 45), were transferred into the oviducts of five surrogate mothers. Pregnancy was diagnosed at day 25 (2/5; 40%), and one normal female lamb was recently born (1/5; 20%). In conclusion, our results show that in ovine, ICSI seems to be the only method for producing transgenic embryos using sperm cells as vectors. In addition the offspring born confirm the viability of these embryos. Table 1.Development and fluorescence expression of ovine embryos
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Garcia-Alvarez, O., A. Maroto-Morales, M. D. Perez-Guzman, and A. J. Soler. "115 RELATIONSHIP BETWEEN IN VIVO FERTILITY OF MANCHEGA EWES LAPAROSCOPICALLY INSEMINATED WITH FROZEN - THAWED RAM SEMEN AND HETEROLOGOUS CALF IN VITRO FERTILIZATION." Reproduction, Fertility and Development 19, no. 1 (2007): 175. http://dx.doi.org/10.1071/rdv19n1ab115.
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Cryopreservation may damage the fertilizing ability of spermatozoa. For this reason, it is important to assess the sperm before artificial insemination. In vitro fertilization (IVF) tests are the most suitable for assessing overall sperm function during fertilization. The aim of this work was to study whether a relationship exists between heterologous calf in vitro fertilization and in vivo fertility in Manchega sheep, with the purpose of predicting in vivo fertility of males before using their semen. Frozen–thawed sperm from 4 Manchega rams was used to laparoscopically inseminate 481 ewes of the same breed. Sperm was cryopreserved in a TRIS-yolk-glycerol extender. A minimum of 10 females were laparoscopicly inseminated per ram. These same straws were used for IVF (4 replicates per male). Domestic calf ovaries were collected at a slaughterhouse. Inmature cumulus–oocyte complexes (COCs) were aspirated and matured in vitro in TCM-199 with 10 ng mL-1 EGF and 100 �M cysteamine. After 24 h, zona-intact mature oocytes were incubated in synthetic oviduct fluid supplemented with 10% estrous sheep serum. Thawed spermatozoa were co-incubated with the oocytes (one million per well) for 40 h, and the cleavage rate was asessed. A regresion analysis was performed. The in vivo fertility ranged from 25.00 to 62.50%. Two rams had a fertility under 30.00% and the other ones over 55.00%. The in vitro fertility ranged from 42.50 to 58.50%. The in vivo fertility was not related to the in vitro fertility (P = 0.17). This could be due to the low number of males used in this work. Heterologous calf in vitro fertilization tests cannot be used to predict in vivo fertility of ram semen since no relationship was found between both variables. Nevertheless, these results are preliminary and we are working with more rams and replicates to obtain more information. This work was funded by INIA and Consejeria de Agricultura de Castilla-La Mancha. Garcia-Alvarez enjoyed a studentship from the INIA
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Galarza, D. A., M. Ladrón de Guevara, P. Beltrán-Breña, M. J. Sánchez-Calabuig, A. López-Sebastián, J. Santiago-Moreno, and D. Rizos. "137 Assessment of fertilizing ability of Merino ram semen cold stored up to 48h by heterologous IVF of bovine oocytes." Reproduction, Fertility and Development 31, no. 1 (2019): 194. http://dx.doi.org/10.1071/rdv31n1ab137.
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The use of cold-stored ram semen has been applied in sheep AI programs, because it preserves its fertilizing ability similar to fresh. Besides, the heterologous IVF has been successfully employed to assess semen fertilizing ability in several species. Hence, we aimed to evaluate the fertilizing ability of ram semen cold stored up to 48h at 5°C by assessing heterologous IVF using bovine oocytes. Fifteen pools of 3 normospermic Merino ram (2-7 years) ejaculates were collected using artificial vagina, diluted to 200×106 spermatozoa mL−1 with ultra-heat-treatment-based extender (skim milk-6% egg yolk) and cold stored up to 48h. In vitro matured zona-intact bovine oocytes were subjected to heterologous IVF using fresh semen (FS, n=707), semen cold stored to 24h (CS24, n=832) or semen cold stored to 48h (CS48, n=611). In parallel, homologous IVF (control, n=1356) and parthenogenesis (parth control non-fertilized oocytes, n=334) were performed. Ram non-selected and selected (BoviPure, Nidacon International, Mölndal, Sweden) semen parameters were evaluated by computer-assisted semen analysis. Sperm-oocyte interaction was assessed at 2.5h post-insemination (hpi) by evaluating the number of bound spermatozoa, whereas penetration and polyspermy were evaluated after 12 hpi. Presumptive zygotes were fixed and stained with Hoechst 33342 at 18, 20, 22, 24 and 26 hpi to assess pronuclear formation using phase contrast and confocal microscopy. Cleavage rate was evaluated in all groups at 48 hpi. Data obtained from 5 replicates were analysed using one-way ANOVA. Data was expressed as mean±standard error of the mean. In terms of sperm storage time, non-selected semen showed a significant decrease (P<0.05) for CS24 and CS48 compared with FS on progressive motility [SPM (%): 52.30±4.1 and 36.9±5.5v. 71.3±1.6] and straight-line velocity (mm s−1: 132.2±6.1 and 109.7±6.3v. 176.7±4.3), respectively. However, selected semen showed a decrease (P<0.05) only for CS48 when compared with CS24 or FS on SPM (35.6±3.9v. 56.1±6.91 and 59.3±2.6) and straight-line velocity (83.5±4.4v. 105.3±6.5 and 110±2.0), respectively. No differences were observed between heterologous IVF groups in all parameters evaluated. Homologous IVF showed a higher percentage of penetration only when compared with heterologous FS group (44.4±6.8v.12.5±4.5%; P<0.01). The polyspermy was higher in heterologous CS24 group when compared with homologous IVF (11.4±3.4v. 3.8±2.2; P<0.05). The homologous IVF group, as expected, showed the higher percentage of pronuclear formation at 18 hpi compared with heterologous IVF with FS (67.3±5.8v. 35.2±5.6%), CS24 (72.1±4.5 v. 37.2±5.7%) and CS48 (63.0±6.0 v. 27.0±5.6%), respectively (P<0.001). Likewise, cleavage rate was higher in homologous group compared with heterologous IVF and parthenogenetic groups for FS (78.3±2.6.8v. 46.3±3.2 and 7.0±2.3%), CS24 (78.4±2.6 v. 48.3±3.2 and 4.9±2.0%), and CS48 (78.4±3.3 v. 43.3±3.5 and 4.3±1.2%), respectively (P<0.001). In conclusion, Merino ram semen cold stored up to 48h maintains its fertilization ability to the same extend as fresh and can be used for sheep crossbreeding programs.
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Panarace, M., G. Jauregui, J. Smith, D. Ferguson, J. Hill, and M. J. Medina. "314 ULTRASONOGRAPHIC ASSESSMENT OF FETAL DEVELOPMENT IN AI AND ET SHEEP FETUSES." Reproduction, Fertility and Development 20, no. 1 (2008): 237. http://dx.doi.org/10.1071/rdv20n1ab314.
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The environment to which the mammalian embryo is exposed during the preimplantation period of development has a profound effect on the physiology and viability of the conceptus (Gardner et al. 2005 Reprod. Fertil. Dev. 17, 361–370); therefore superovulation has been implicated in abnormal methylation and imprinting in the resultant embryo. In addition, the first week after mating is now known to be a critical time for embryo programming in sheep. However, the effect of altering the maternal environment by superovulation followed by embryo transfer (ET) into non-superovulated recipient ewes has not been thoroughly investigated in livestock animals. The objective of the present study was to determine if fetal and placental growth curves are affected by the mating type (AI or ET). In the ET group, 10 ewes underwent a superovulation program prior to artificial insemination; then 29 embryos were transferred singly into recipients. In the AI group, 25 ewes were inseminated with semen from the same sire as used for the ET group. All ewes used were Merino breed. At Day 30 after AI or Day 24 after ET, ewes were scanned for pregnancy using a 5-MHz transrectal probe (Honda, Tokyo, Japan). First, crown rump length (CRL, mm) was measured at Days 30 and 44, while abdominal circumference (ABD, mm) and femur length (FEM, mm) were measured monthly from Day 58 to term using a transabdominal 3-MHz convex probe. Data were analyzed by ANOVA for repeated measures. At Day 30, CRL was not different among AI and ET fetuses, but at Day 44, AI fetuses had significantly longer CRL than ET fetuses (P < 0.001, Table 1). Also, at the first measurement of FEM and ABD (Day 58) mating types were not significantly different, but measurements at Day 85 showed AI fetuses had larger FEM, P < 0.05, and ABD, P < 0.01. From Day 114 onward, there was no difference in fetal size between AI and ET fetuses. Univariate analysis of variance indicated no significant mating type or gender effects on gestation length, placenta weight, cotyledon count, weight of cotyledons as a proportion of total placenta, or weight of lambs at birth, prior to lamb-marking and weaning. We conclude that in the ET group, the first 90 days of the embryonic/fetal growth curve could subtly be affected by superovulation treatment; however, it did not affect weight and survival rate of this type of fetus. Table 1. Mating type effects on crown rump length (CRL, mm), femur length (FEM, mm), and abdominal circumference (ABD, mm), mean ± SE
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PAPADOPOULOS, S., C. DELIGIANNIS, E. K. THEODOSIADOU, D. KANTAS, TH LAINAS, P. GOULAS, G. C. FTHENAKIS, and I. VALASI. "Fertility rate of short-term progestagen pretreated ewes in relation to breed: A field study." Journal of the Hellenic Veterinary Medical Society 68, no. 1 (January 29, 2018): 35. http://dx.doi.org/10.12681/jhvms.15554.
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The effect of short-term instead of long-term progestagen treatment on fertility of Karagouniko and Chios ewes, after natural mating or artificial insemination, was investigated. Two experiments were performed during the transition period from anοestrous to the breeding season. In the 1st experiment (natural mating, NM), Karagouniko and Chios ewes were randomly allocated into 3 groups, that were KLM (long-term progestagen treatment; n=35), KSM (short-term progestagen treatment; n=34), KSP (short-term progestagen treatment; prostaglandin; n=35) and CLM (n=40), CSM (n=35), CSP (n=38), respectively. In the 2nd experiment (intracervical artificial insemination, AI) Karagouniko and Chios ewes were randomly allocated into 3 groups, that were KLA (long-term progestagen treatment; AI at 54h; n=50), KSA1 (short-term progestagen treatment; AI at 54h; n=20), KSA2 (short-term progestagen treatment; AI at 48h; n=28) and CLA (n=40), CSA1 (n=16), CSA2 (n=20), respectively. At sponges’ removal (d0) all ewes received 400 IU eCG. Ten rams served NM, while for AI fresh diluted semen was used. Pregnancy diagnosis was performed, 45-50 days later. Ιn the 1st experiment, blood samples were collected, daily for 5 days, starting on d0, for serum progesterone assessment. Conception rate in Karagouniko ewes after NM was higher (P<0.05) in KSM (35.29%) compared to KLM (17.14%) group, but did not differ with KSP (28.57%) group, while after AI it was higher (P<0.05) in KLA (42.00%) or KSA1 (40.00%) compared to KSA2 (14.29%) group. In Chios ewes no significant differences were observed between groups either after NM [CLM (45.00%), CSM (36.84%), CSP (34.29%)] or after AI [CSA1 (50.00%), CSA2 (50.00%), CLA (45.00%)]. No significant differences were observed after NM or after AI in the litter size in both breeds. These results indicate that short-term progestagen treatment for oestrus synchronization could be applied in indigenous Greek sheep breeds, resulting in equal (Chios) or improved fertility (Karagouniko) than the common long-term one. Also, the fertility rate in ewes subjected to short-term progestagen treatment depends on the time of AI in relation to breed.
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Polge, Christopher. "Lionel Edward Aston Rowson, O.B.E. 28 May 1914 — 26 July 1989." Biographical Memoirs of Fellows of the Royal Society 46 (January 2000): 483–97. http://dx.doi.org/10.1098/rsbm.1999.0097.
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L.E.A. Rowson was always known as Tim to his family, friends and colleagues alike. He was a veterinary surgeon who made important contributions to research in reproductive physiology in farm animals and its application to animal breeding. In particular, he was a pioneer of artificial insemination (AI) and embryo transfer, which have become two of the most important technologies for livestock improvement in modern times.He was appointed Director of the first AI centre for cattle breeding in Britain, established at Cambridge in 1942, and played a leading role in the application and rapid growth of this technology. In 1952 he contributed to the development of successful methods for the freezing and long-term storage of bull semen at very low temperatures. This had far-reaching consequences for the future of AI and cattle breeding worldwide.For thirty years he also worked at the Animal Research Station in Cambridge on methods for embryo transfer in sheep and cattle and their use in research and breeding. This culminated in the 1970s with the development of effective methods for collection and transfer of cattle embryos by non-surgical means. The birth of the first calf after transfer of a deepfrozen embryo in 1973 was another landmark, and these advances led quite quickly to the commercial application of embryo transfer in cattle breeding.Tim Rowson is generally regarded as the founder of embryo transfer in farm animals, but important contributions were made by many collaborators. He always considered that he was privileged during his early years to have worked with Dr John (later Sir John) Hammond, F.R.S., who maintained that the function of applied science was to synthesize the detailed knowledge gained from fundamental research into a constructive whole so that it could be used for a specific purpose. Tim was a true disciple of this philosophy and always tried to relate a fundamental approach to a practical outcome.