Sex-preselected buffalo (Bubalus bubalis) calves derived from artificial insemination with sexed sperm

doi:10.1016/j.anireprosci.2010.01.001

Animal Reproduction Science

Volume 119, Issues 3–4, June 2010, Pages 169-171

https://doi.org/10.1016/j.anireprosci.2010.01.001 Get rights and content

Abstract

Flow cytometry sorting of X- and Y-chromosome bearing sperm has been emerging as a promising technology to alter the sex ratio in progenies of mammals in the resent years. The objective of this study was to evaluate the efficiency of AI by using the sexed sperm to produce sex-preselected calves in buffalo species. A total of 43 buffalo cows were inseminated with X-sorted sperm, 30 of which were confirmed pregnant 3 mo following AI. In terms of conception rate, significant difference was observed between AI with sexed sperm derived from different bulls (P < 0.05), but not between sexed and non-sexed sperm (P > 0.05), nor between heifers and parous buffalo cows (P > 0.05). A total of 29 sex-preselected calves, 24 females and 5 males, developed to term and were viable on delivery. Results of this study indicate the feasibility of the application of the sexing technology to accelerate the genetic improvement in swamp buffalo.

Introduction

Buffalo is an important domestic animal in agriculture and animal production industry in many Asian and Mediterranean countries. Over 22.7 million buffaloes, most of which are swamp type, are distributed in the subtropical provinces in southern China (FAO, 2003). As the consumption of high quality dairy product increase, there is a need to expand the genetic merit of river type buffalo, whose milk production is much higher than that of swamp buffalo. So far it has been demonstrated that the application of sexed bovine sperm using AI is effective in altering the sex ratio and rapidly expanding dairy herds carrying high genetic value animals (Johnson, 2000, Garner, 2006). Flow sorting of X- and Y-chromosome bearing buffalo sperm and subsequent use in IVF has been previously reported (Lu et al., 2006, Lu et al., 2007), which validated the feasibility of this sex-preselecting technology in buffalo species. The practical application of sexed sperm in buffalo breeding would be of great interest both in biological and economic terms.

Over the last several decades, AI has been shown to be one of the most successful breeding strategies to quickly disseminate genes from the best available males for improvement of production traits. Previous reports revealed that, following Ovsynch protocol and AI with sexed sperm into the utero-tubal junction, a conception rate of 42.8% was observed in Mediterranean Italian buffaloes (Presicce et al., 2005). However, buffalo in households of China and many Asian countries are usually raised in very small herds (2–5 buffaloes per family) and the Ovsynch protocol is rarely practiced in buffalo species. Artificial insemination following spontaneous estrous should be a more viable way to rapidly disseminate the sexed buffalo sperm at present. In this study, semen was collected from river type buffalo (Nili-Ravi and Murrah) and sorted for X-sperm enriched sample. This was inseminated by AI into the deep uterine horn of spontaneous estrous buffalo. Conception rate was recorded to determine the efficiency of the use of this technology for buffalo.

Section snippets

Animals

One Murrah and one Nili-Ravi bull of proven fertility were used in semen collection by artificial vagina and unsexed sperm used in the experiment was collected from 10 bulls of proven fertility. All bulls were raised in Guangxi Livestock and Poultry Breeding Station, China. The swamp type buffalo cows and F1 crossbreeds of river type and swamp type used for AI were raised in households in Lingshan county, Guangxi, China.

Sperm sorting

Only ejaculates with >75% morphologically normal sperm and >65% progressive

Conception rates in AI with sexed and non-sexed buffalo sperm

Flow cytometry sorting of X- and Y-sperm followed by insemination for production of sex-preselected offspring has been successful in various animals (Cattle: (Seidel et al., 1999a); Sheep: (Cran et al., 1997, Hollinshead et al., 2002); Horse: (Buchanan et al., 2000); Pig: (Johnson, 1991, Grossfeld et al., 2005)). However, the low dose of sexed sperm using in AI, combined with the negative effect of sorting procedures on sperm, usually resulted in a low conception rate. In cattle, the conception

Acknowledgements

We acknowledge the staff at Animal Husbandry and Veterinarian Service Station in Lingshan County for assistance in carrying out the AI. This research was jointly supported by National High-tech R & D Program (2008AA101004), National Science and Technology Supporting Program (No. 2006BAD04A18), Guangxi Science and Technology R&D Program (No. 0630006-5B, 07109006), Guangxi Science Foundation (No. 0832012) and Guangxi University Key Program (No. 2005ZD05).

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Cited by (27)

Review: Development, adoption, and impact of assisted reproduction in domestic buffaloes
2023, Animal
The domestic buffalo (Bubalus bubalis), also known as water buffalo, comprises two sub-species the River buffalo (B. bubalis ssp. bubalis; 50 chromosomes) and the Swamp buffalo (ssp. carabanensis; 48 chromosomes). Domestic buffaloes are a globally significant livestock species. In South Asia, the River buffalo is a primary source of milk and meat and has a very important role in food security. The River buffalo also supports high-value, differentiated food production in Europe and the Americas. The Swamp buffalo is an important draft animal and a source of food in Southeast Asia and East Asia. The growing importance of buffaloes requires that they undergo an accelerated rate of genetic gain for efficiency of production, product quality, and sustainability. This will involve the increased use of assisted reproduction. The initial application of reproductive technology in buffaloes had variable success as it relied on the adoption of procedures developed for cattle. This included artificial insemination (AI), sperm cryopreservation, and embryo technologies such as cloning and in vitro embryo production (IVEP). Reproductive technology has been progressively refined in buffaloes, and today, the success of AI and IVEP is comparable to cattle. Ovarian follicular superstimulation (superovulation) combined with in vivo embryo production results in low embryo recovery in buffaloes and has limited practical application. The contribution of elite female buffaloes to future genetic improvement will therefore rely mainly on oocyte pickup and IVEP. This will include IVEP from females before puberty to reduce generation intervals. This review provides for the first time a clear chronology on the development, adoption, and impact, of assisted reproduction in domestic buffaloes.
Reproductive management in buffalo by artificial insemination
2020, Theriogenology
Citation Excerpt :
Deposition in the body of the uterus makes the use of sexed semen more practical and will facilitate adoption for genetic improvement. More recently, sexed semen of River buffalo gave good pregnancy rates in Swanp buffalo and F1 River x Swamp buffalo inseminated at spontaneous estrus [82,83]. This demonstrated the potential for accelerating genetic improvement in Swamp buffalo with sexed semen of highly selected River buffalo.
Artificial insemination (AI) is important for genetic improvement and to control the period of breeding in buffalo and has increased significantly over the past 20 years. AI is more difficult in buffalo compared with cattle due to variable estrous cycles, reduced estrous behavior, and reproductive seasonality. The latter is associated with a higher incidence of anestrus and increased embryonic mortality during the nonbreeding season. Protocols to control the stage of the estrous cycle have undergone recent development in buffalo. These protocols are based on the control of both the luteal phase of the cycle, mainly by prostaglandins and progesterone, and follicle development and ovulation, by prostaglandins, progesterone, GnRH, hCG, eCG and estradiol. Protocols that synchronize the time of ovulation enable fixed timed AI, avoiding estrous detection. Factors to consider when selecting an AI protocol include animal category (heifers, primiparous or pluriparous), reproductive status (cyclic or anestrus), and season. This review looks at the current status of estrus synchronization and AI in buffalo and provides some practical suggestions for application of AI in the field.
Reproductive technologies in the buffalo (Bubalus bubalis)
2020, Reproductive Technologies in Animals
The domestic buffalo (Bubalus bubalis) is characterized by a worldwide distribution and its production is expressed within both intensive systems and marginal areas. Although a domesticated species since long time, over the years, the buffalo has scarcely benefitted of the strategies held in place for genetic improvement, as opposed to the cattle counterpart. Only at the beginning of the 1990s, the problems related to the enhancement of buffalo productivity through the implementation of reproductive technologies, namely the use of AI, have begun to be tackled. In this chapter, a report on the use of reproductive technologies in buffaloes will be presented, highlighting the early attempts at such and the remarkable more recent achievements in different areas, ranging from in vivo and in vitro embryo production to transgenesis, cloning, and stem cell technologies. Finally, an outlook to more pragmatic aspects will be given, such as the cost-to-benefit ratio of some of the reproductive strategies discussed.
A field study on artificial insemination of swamp and crossbred buffaloes with sexed semen from river buffaloes
2015, Theriogenology
Citation Excerpt :
In the present study, we applied the sexing technology to buffalo breeding under field trail level and obtained a pregnancy of 48.5%. This is ∼73% of the pregnancy rate obtained in AI using unsexed semen in a previous study (66.5%, 1545 of 2325) [8] and agrees with a previous report for Holstein cows where the pregnancy rate of AI using sexed semen was about 70% to 80% of that achieved by unsexed sperm [10]. The difference in pregnancy rate might be due to the lower number of sexed sperm used in insemination, which was only one-tenth of unsexed semen.
Sex preselection by flow sorting of X- and Y-sperm has been proven to be an efficient and economically feasible strategy for use in Holstein dairy cow breeding, and previous reports have demonstrated the feasibility of altering the sex ratio in buffalo species by using sexed semen in either artificial insemination or IVF. However, because buffalo reproductive physiology and farm management are different from Holsteins, factors involved in artificial insemination by sexed semen need to be further addressed before being applied in buffalo breeding at village-level husbandry. In this study, a total of 4521 swamp or crossbred (F1 or F2) buffaloes with natural estrus were inseminated with X-sorted sperm from river buffaloes, resulting in a 48.5% (2194 of 4521) pregnancy rate and 87.6% (1895 of 2163) sex accuracy in the derived calves. The pregnancy rate obtained with sexed semen from Murrah bulls was higher than that of Nili-Ravi, 52.5% (895 of 1706) versus 46.1% (1299 of 2815; P < 0.01), respectively. Also, significant variations were seen in pregnancy rates from inseminations performed in different seasons (P < 0.01) and by different technicians (P < 0.01). In contrast to Holsteins, no difference was seen in the pregnancy rate between heifers and parous buffalo cows, and buffalo cows with different genetic backgrounds (swamp type, crossbred F1 and F2) showed similar fertility after insemination with sexed semen. The findings in the present study under field conditions pave the way for application of sexing technology to buffalo breeding under village-level husbandry and diverse genetic backgrounds.
Minimum number of spermatozoa per dose in Mediterranean Italian buffalo (Bubalus bubalis) using sexed frozen semen and conventional artificial insemination
2013, Theriogenology
Citation Excerpt :
Controversial outcomes reported from different authors are the result of varied conditions in which the conclusions are drawn, especially with regard to the buffalo species which exhibits a marked seasonality. Lu et al. [31], using one Murrah and one Nili-Ravi bull in southern China, did not find differences between heifers and parous cows, and speculated that the relatively high fertility rate (nearly 70%) in AI with sexed buffalo sperm obtained in their work was largely because of a seasonal effect. In eastern Thailand, other authors [32] evaluated fertility in swamp buffalo after the Ovsynch protocol combined with fixed-time artificial insemination, and reported a marked effect of the heifer with respect to cows, with a reduction in pregnancy rates of the first compared with the second.
In buffaloes, AI with sexed semen is not fully optimized, and the procedure has only been performed using the approach currently in use for cattle. The objective of the present work was to compare the pregnancy rates in Mediterranean Italian buffalo cows inseminated with sexed frozen–thawed semen at 2, 4, 6, and 8 million sperm per dose, using the Ovsynch protocol and conventional AI at a fixed time. Fresh ejaculates from three buffalo bulls were processed according to Beltsville sperm sorting technology, and packaged in 0.25-mL straws with two total concentrations of 2 and 4 million live sorted sperm per straw. After thawing, semen was evaluated for total motility, forward motility, average path velocity, membrane and DNA integrity, and membrane fluidity. Sorting efficiency was estimated using a real time polymerase chain reaction method developed and validated in our laboratory. The artificial inseminations were conducted during the breeding season on 849 Italian Mediterranean buffalo heifers and cows distributed in 13 farms in northern and central Italy. No significant difference in quality parameters was reported between nonsexed and sexed straws produced with 2 and 4 million sperm. Lower pregnancy rate (P < 0.001) was reported when inseminating doses of sexed semen at 2 million were used (53/170; 31.2%), with respect to conventional nonsexed (78/142; 54.9%), and sexed doses at 4, 6, and 8 million spermatozoa (102/205, 49.8%; 84/175, 48.0%; and 74/157, 47.1%, respectively). No differences were evident using conventional doses and sexed semen with sperm numbers equal or higher than 4 million per dose. Pregnancies were not affected by the sire; 39/82 (47.6%), 120/270 (44.4%), and 151/355 (42.5%), respectively, for the three bulls. Variability in pregnancy rates observed in different herds was not significant. Furthermore, no significant difference was reported between pregnancies obtained with sexed semen in heifers and multiparous, respectively, 179/407 (44.0%) and 131/300 (43.7%). The results of the present work indicate that in Mediterranean Italian buffalo the dose of 4 million represents an optimal compromise when using sexed semen with conventional technologies of insemination, together with estrus synchronization, and the minimum number of spermatozoa per dose. In addition, the real time polymerase chain reaction method was optimized and is now available for estimating sorting efficiency in buffalo.
Pregnancy rates following AI with sexed semen in Mediterranean Italian buffalo heifers (Bubalus bubalis)
2011, Theriogenology
Citation Excerpt :
In this study, a straw containing total 2 million live sorted spermatozoa has been found sufficient in obtaining similar rates of pregnancy when compared with straw content of 20 million nonsexed spermatozoa. Also, in contrast to the report of Lu et al. [51], the season of the year of increasing daylight did not undermine the final pregnancy outcome in our study. Additional important information derived from this study comes from the feasibility of achieving pregnancy rates with sexed semen similar to what is usually obtained with conventional semen, without the use of special precautions or inserting devices and particular skills [50].
The use of sexed semen in farm animal production and genetic improvement has been shown to be feasible with variable degree of efficiency in a number of species, and proved to be economically viable in cattle. In the last two decades, various newly developed reproductive technologies applicable in buffaloes have mushroomed. Recently, following the birth of the first buffalo calves using AI with sexed semen, commercial interest to exploit sexing of semen in this species too is aroused. In order to verify the successful adoption of this technology in the buffalo, the present study on the use of sexed semen for AI was carried out and compared with conventional artificial insemination using nonsexed semen. A total of 379 buffalo heifers were used for synchronization of ovulation using the Presynch protocol in the South of Italy. Selected animals at the time of AI were randomly allocated to three different experiment groups: (1) 102 animals subjected to AI in the body of the uterus with sexed semen (SS body); (2) 104 animals subjected to AI in the horn of the uterus with sexed semen (SS horn); and (3) 106 animals subjected to AI in the body of the uterus with conventional nonsexed semen (NSS body). Semen of three buffalo bulls was sexed by a collaborating company and commercially distributed in 0.25 mL straws with a total of 2 million sexed spermatozoa. Pregnancy rates were first assessed at Day 28 following AI, and rechecked at Day 45 by ultrasound. Pregnancy rates were nonsignificantly different between animals inseminated with sexed or nonsexed semen: 80/206 (38.8%) and 40/106 (37.7%), respectively (P = 0.85). However, site of insemination of sexed semen affected pregnancy rate significantly as higher pregnancy rates were obtained when sexed semen was deposited into the body rather than the horn of the uterus: 46/101 (45.5%) and 34/105 (32.3%), respectively (P = 0.05). In conclusion, the use of sexed semen in buffalo heifers gave satisfactory and similar pregnancy rates when compared with conventional nonsexed semen. Deposition of sexed semen into the body of the uterus, however, increased pregnancy rates significantly.

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Sex-preselected buffalo (Bubalus bubalis) calves derived from artificial insemination with sexed sperm

Abstract

Introduction

Section snippets

Animals

Sperm sorting

Conception rates in AI with sexed and non-sexed buffalo sperm

Acknowledgements

Theriogenology

Theriogenology

J. Dairy Sci.

Theriogenology

Anim. Reprod. Sci.

J. Dairy Sci.

Theriogenology

Theriogenology

J. Dairy Sci.

Anim. Reprod. Sci.

Anim. Reprod. Sci.