scholarly journals Sex-sorted sperm in farm animals: scale of use, the effectiveness of insemination, risk factors

2017 ◽  
Vol 73 (5) ◽  
pp. 272-279
Author(s):  
Bartłomiej M. Jaśkowski ◽  
Magdalena Woźna ◽  
Marek Gehrke
Keyword(s):  
Risk Factors ◽  
Embryo Transfer ◽  
Conception Rate ◽  
Farm Animals ◽  
Lower Percentage ◽  
Technical Parameters ◽  
Cattle Herds ◽  
The One

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.

164 EFFECTS OF KINETICS AND MORPHOLOGY ON EARLY EMBRYONIC DEVELOPMENT IN BOVINE OPU-IVF EMBRYOS

2016 ◽  
Vol 28 (2) ◽  
pp. 212
Author(s):  
M. Takayama ◽  
O. Dochi ◽  
K. Imai
Keyword(s):  
Embryo Transfer ◽  
Calf Serum ◽  
Conception Rate ◽  
Culture Dish ◽  
Fresh Embryo Transfer ◽  
Cell Stage ◽  
Vitro Fertilization ◽  
Observation Method

In recent years, the use of ovum pick up (OPU) and IVF for embryo production has increased worldwide; however, the conception rate of embryo transfer is lower for OPU-IVF embryos than for in vivo-derived embryos. This study aimed to determine the efficacy of embryo selection by a 3-step observation method by using a micro-well culture dish (Dai Nippon Printing, Tokyo, Japan). In this study, 9 Holstein and 15 Japanese Black cows were used, and the OPU-IVF was conducted from October 2014 to May 2015. The collected cumulus-oocyte complexes (COC) were cultured for 22 h in 25 mM HEPES-buffered TCM-199 supplemented with 5% calf serum (CS) and 0.02 AU mL–1 of FSH. Sperm (at a final concentration of 5 × 106 spermatozoa mL–1) were incubated with COC for 6 h. After insemination, presumptive zygotes were separated from cumulus cells and sperm by pipetting. Then, the presumptive zygotes were cultured for 9 days in CR1aa supplemented with 5% CS by using a micro-well culture dish. Kinetics and morphology were observed at 27, 31, and 55 h post-insemination (hpi). The presumptive zygotes were divided to 3 groups (more than 2 cells, 2 cells, and no cleavage) at 27 and 31 hpi. Then, embryos at the 2-cell stage at 31 hpi were divided into 2 groups: 2-cell with normal cleavage and 2-cell embryos with abnormal cleavage (unequal cleavage, 2-cell with fragments, and 2-cell with protrusion). Subsequently, embryos were classified as 8-cell and more than 8 cell, or less than 8 cell at 55 hpi. The blastocyst rate (BL%) was analysed at 7, 8, and 9 days post IVF. Embryos selected by the 3-step observation method were used for fresh embryo transfer. The data were analysed by chi-squared test. In total, 856 oocytes were collected by OPU and 633 oocytes were cultured, of which 39.7% (263/663) developed to the blastocyst stage. The BL% of 2-cell embryos (72.5%, 116/160) was significantly higher (P < 0.01) than that of no cleavage (47.0%, 117/249) at 27 hpi. The BL% of 2-cell (65.4%, 206/315) and more than 2-cell (53.0%, 35/66) was significantly higher (P < 0.01 and P < 0.05) than that of embryos divided as no cleavage (25.9%, 22/85) at 31 hpi. The BL% was not significantly different between 2-cell with normal cleavage (68.5%, 172/251) and abnormal cleavage (53.1%, 34/64). The BL% of 8-cell and more than 8-cell stage (72.8%, 182/250) was significantly higher (P < 0.01) than that of embryos with less than 8 cells (38.8%, 81/209) at 55 hpi. Overall, 2-cell embryos at 27 hpi, 2-cell embryos with normal cleavage at 31 hpi, and 8-cell and more than 8 cell at 55 hpi showed the highest BL% (82.1%, 78/91). The conception rate was higher for following the selected fresh embryo transfer that was 70.6% (12/17) than average of in vitro fertilization embryos transfer that was 40.0%. These results demonstrate that the 3-step observation method used in this study can be effectively applied for the selection of IVF embryos that have a strong ability to develop into blastocysts and high competence for conception.

47 CRYOPRESERVATION OF BOVINE GERM CELL USING ANTIFREEZE POLYAMINO-ACID (CARBOXYLATED POLY-L-LYSINE)

2017 ◽  
Vol 29 (1) ◽  
pp. 131
Author(s):  
T. Fujikawa ◽  
C. Kubota ◽  
T. Ando ◽  
S. Imamura ◽  
M. Tokumaru ◽  
...  
Keyword(s):  
Survival Rate ◽  
Germ Cell ◽  
Embryo Transfer ◽  
Conception Rate ◽  
Control Group ◽  
Vitro Fertilization ◽  
Polyamino Acid ◽  
Significant Difference

Carboxylated poly-l-lysine (CPLL) is an ampholytic polymer compound, and it is obtained by converting 65% amino groups to carboxyl groups after synthesising ε-poly-l-lysine aqueous solution and succinic anhydride. CPLL has cryoprotective property similar to antifreeze protein, and addition of CPLL into cryopreservation medium improves the post-thaw survival rate of cells and embryos. In this research, we examined the effectiveness of CPLL as a bovine germ cell cryoprotective material. In experiment 1 (in sperm), the conventional cryopreservation medium used for control group was consisted of 6.5% (vol/vol) glycerin, and the cryopreservation medium used for CPLL group was consisted of 3.25% (vol/vol) glycerin and 0.5% CPLL (wt/vol). The post-thaw survival and motility were assessed by using Sperm Motility Analysis System (DITECT Corp., Tokyo, Japan). There was no significant difference for post-thaw survival rate and motility (control v. CPLL; 98.8% v. 96.6% and 69.7% v. 62.2%, respectively). Artificial insemination was carried out in 65 cows (control v. CPLL; 34 v. 31), and the conception rate of the CPLL group was higher than that of the control group (80.6% v. 67.6%; P = 0.23). In experiment 2 (embryos), the conventional cryopreservation medium used for control group was consisted of 5% (vol/vol) ethylene glycol and 6% (vol/vol) propylene glycol in PBS. In the CPLL group, 7% (wt/vol) CPLL was added to the conventional medium. In vitro fertilization embryos were cryopreserved at Day 7 and Day 8. There was no significant difference in survival rate at 0, 24, and 48 h and hatched rate until 72 h after thawing (control v. CPLL: 93.6% v. 93.2%, 69.0% v. 64.7%, 56.1% v. 56.3%, 12.9% v. 10.2%, respectively). Embryos obtained by superovulation treatment and in vivo fertilization at Day 7 were cryopreserved using above 2 media, and transferred non-surgically into synchronized recipient cows (1 embryo per animal). Embryo transfer (ET) was carried out in 81 cows (control v. CPLL: 31 v. 50), and recipients were diagnosed for pregnancy ultrasonically 50 days after embryo transfer. Conception rate of CPLL group was higher than control group (50.0% v. 29.0%; P = 0.063). In both experiments, the significant differences between control group and CPLL group were determined by chi-squared test. The effectiveness of CPLL in cells and embryos has been reported; however, there is no report using CPLL in bovine germ cells. In this research, CPLL improved the conception rate of AI and ET, probably due to its low toxicity and protection of the cell membrane. These results suggest that CPLL is available as a new cryoprotective material for bovine sperm and embryo in slow freezing methods.

136 IMPACT OF SELECTION SYSTEM BY KINETICS ON THE EARLY EMBRYONIC DEVELOPMENT IN BOVINE OVUM PICKUP-IVF EMBRYOS

2017 ◽  
Vol 29 (1) ◽  
pp. 176
Author(s):  
M. Takayama ◽  
M. Moriyoshi ◽  
O. Dochi ◽  
K. Imai
Keyword(s):  
Embryo Transfer ◽  
Calf Serum ◽  
Cumulus Cells ◽  
Conception Rate ◽  
Culture Dish ◽  
Fresh Embryo Transfer ◽  
Chi Square ◽  
Predicting Factors

Recently, in vitro-produced (IVP) embryos have been increasingly produced using ovum pickup (OPU) and IVF in cows worldwide. However, the conception rate of IVP embryos is lower than that of in vivo-derived embryos. This study was conducted to determine the proportion of embryos that led to a high conception rate when the embryos were selected according to the 4 predicting factors. A total of 30 Holstein and 20 Japanese Black cows were used, and 81 OPU-IVF sessions were performed from October 2014 to May 2016. The collected cumulus-oocyte complexes (COC) were cultured for 22 h. Capacitated sperm (at a final concentration of 5 × 106 spermatozoa/mL) were incubated with COC for 6 h. After insemination, presumptive zygotes were separated from cumulus cells and sperm by pipetting. Then, the presumptive zygotes were cultured for 9 days in CR1aa supplemented with 5% calf serum by using a micro-well culture dish (Dai Nippon Printing, Tokyo, Japan). The kinetics of embryo development was observed at 27, 31, and 55 h post-insemination (hpi). The 4 factors used to select embryos were as follows: (1) time at which first cleavage occurred (less than 27 hpi, or less than 31 hpi, in case any of the zygotes did not cleave at 27 hpi in each culture dish); (2) 2 blastomeres after first cleavage at 31 hpi; (3) absence of fragments after first cleavage at 31 hpi; and (4) 8 or more blastomeres at 55 hpi. The number of blastocysts was analysed at 7, 8, and 9 days post IVF. Additionally, the number of produced embryos that could be used for embryo transfer (ET) was determined. The data were analysed using the chi-square test. The total numbers of blastocysts and produced embryos were 615 and 503, respectively. The numbers of blastocysts and produced embryos selected using the combination of factors 1 to 4 were 200 (32.5%) and 169 (27.5%), respectively. The numbers of blastocysts and produced embryos selected using factor 1 were 397 (64.6%) and 340 (67.6%), using factor 2 were 445 (71.3%) and 378 (75.1%), using factor 3 were 364 (81.8%) and 307 (81.2%), and using factor 4 were 374 (60.8%) and 308 (61.2%), respectively. The numbers of blastocysts and produced embryos that were rejected using a combination of factors 1 to 4 were 123 (27.5%) and 90 (17.9%), respectively. The conception rate of fresh embryo transfer was 46.6% (n = 73). We found that the conception rate of the embryos selected using factors 1 to 4 was significantly (P < 0.05) higher than that of embryos without one factor or more [60.0% (n = 35) v. 29.4% (n = 34)]. These results show the applicability and efficiency of the 4 factors for producing embryos with a high competence for conception.

scholarly journals Effectiveness of embryo transfer in cows - risk factors including in vivo derived and in vitro produced embryos

2021 ◽  
Vol 9 (3) ◽  
pp. 123-131
Author(s):  
Maria Wieczorkiewicz ◽  
Jędrzej M. Jaśkowski ◽  
Agnieszka Wichtowska ◽  
Monika Olszewska-Tomczyk ◽  
Bartłomiej M. Jaśkowski
Keyword(s):  
Risk Factors ◽  
Embryo Transfer ◽  
Body Condition Score ◽  
Production Method ◽  
Development Stage ◽  
Individual Characteristics ◽  
Multiple Ovulation ◽  
Condition Score

Abstract Multiple Ovulation Embryo Transfer is a biotech method with more than 50 years of history and an established position in cattle breeding. This procedure is beneficial in many ways, but it also carries a risk of failure. The study presents the overview of the most important risk factors that may affect conception rates in the course of embryo transfer, including the factors associated with the embryo sourcing (embryo production method, embryo quality, development stage and breed, embryo storage method), embryo transfer procedure (synchrony/asynchrony, embryo transfer difficulty, the time of the transcervical insemination gun passage, depth of embryo deposition, localization and structure of the corpus luteum relative to the follicle and both individual characteristics of donors and recipients (level of concentration of progesterone, the state of health of the udder, lactation level, body condition score and age) and some environmental factors.

ECONOMIC PERSPECTIVES OF BIOTECHNOLOGIES USING IN ANIMAL FARMING

1970 ◽  
pp. 57-60
Author(s):  
I. F. Gorlov ◽  
A. A. Mosolov ◽  
G. V. Komlatskiy ◽  
M. A. Nesterenko ◽  
K. D. Nimbona ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Economic Effect ◽  
Dairy Herd ◽  
State Level ◽  
Modern Level ◽  
Economic Perspectives ◽  
The Cost ◽  
Short Time

The article presents materials on the study of the possibility of reproduction and increase in the herd of highly productive cows through the use of embryo transplantation technology. The classical (in vivo) and more modern, developing (in vitro) methods of embryotransfer, their positive and negative sides are considered in detail. The possibility of accelerating the breeding process by using the method of transplantation, in which from one cow can be obtained from 10 to 100 calves, which will allow for 4-5 years, almost any herd (of any size and breed) with the help of biotechnology to turn into a cattle-breeding enterprise of the most modern level. At the same time, heifers obtained from unproductive cows can be used as "surrogate" mothers who are transplanted with the best donor embryos, which allows to obtain a full-fledged offspring adapted to local environmental conditions. A detailed scheme of obtaining, evaluation, storage, as well as the cost and economic effect of embryo transplantation was calculated, the market was evaluated, the required annual volume of transplants and the number of donor cows for large livestock farms were determined. As a positive example of "Scientific-production enterprise "Centre of biotechnology and embryo transfer" in 2014, implemented a project for accelerated replacement and genetic improvement of the dairy herd, engraftment averaged 57-69%, and the economic effect of the enterprise from getting a single animal by the method of embryo transfer, compared with imports of similar close in quality, ranged from 60 to 100 thousand rubles on his head. It is shown that it is necessary to organize at the state level a developed service for embryo transplantation to reduce the cost of embryo transfer and the possibility of creating in a short time in the country's own highly productive breeding nucleus of dairy and beef cattle, which will reduce, and in the future completely eliminate, import dependence on cattle products.

Effects of rate of development of in vitro-produced ovine embryos on sex ratio and in vivo survival after embryo transfer

1997 ◽  
Vol 48 (8) ◽  
pp. 1369-1378 ◽  
Author(s):  
S.L. Catt ◽  
J.K. O'Brien ◽  
W.M.C. Maxwell ◽  
G. Evans
Keyword(s):  
Sex Ratio ◽  
Embryo Transfer ◽  
Rate Of Development

scholarly journals Mammary gland 3D cell culture systems in farm animals

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Laurence Finot ◽  
Eric Chanat ◽  
Frederic Dessauge
Keyword(s):  
Cell Culture ◽  
Mammary Gland ◽  
Bacterial Infections ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Farm Animals ◽  
Culture Systems ◽  
Cell Culture Systems

AbstractIn vivo study of tissue or organ biology in mammals is very complex and progress is slowed by poor accessibility of samples and ethical concerns. Fortunately, however, advances in stem cell identification and culture have made it possible to derive in vitro 3D “tissues” called organoids, these three-dimensional structures partly or fully mimicking the in vivo functioning of organs. The mammary gland produces milk, the source of nutrition for newborn mammals. Milk is synthesized and secreted by the differentiated polarized mammary epithelial cells of the gland. Reconstructing in vitro a mammary-like structure mimicking the functional tissue represents a major challenge in mammary gland biology, especially for farm animals for which specific agronomic questions arise. This would greatly facilitate the study of mammary gland development, milk secretion processes and pathological effects of viral or bacterial infections at the cellular level, all with the objective of improving milk production at the animal level. With this aim, various 3D cell culture models have been developed such as mammospheres and, more recently, efforts to develop organoids in vitro have been considerable. Researchers are now starting to draw inspiration from other fields, such as bioengineering, to generate organoids that would be more physiologically relevant. In this chapter, we will discuss 3D cell culture systems as organoids and their relevance for agronomic research.

Effect of microbial isolates on microbial yield estimation in RUSITEC system

1998 ◽  
Vol 22 ◽  
pp. 306-308
Author(s):  
M. D. Carro ◽  
E. L. Miller
Keyword(s):  
Protein Synthesis ◽  
Liquid Phase ◽  
Solid Phase ◽  
Liquid Fraction ◽  
Microbial Protein ◽  
Microbial Protein Synthesis ◽  
Continuous Culture System ◽  
The One

The estimation of rumen microbial protein synthesis is one of the main points in the nitrogen (N)-rationing systems for ruminants, as microbial protein provides proportionately 0.4 to 0.9 of amino acids entering the small intestine in ruminants receiving conventional diets (Russell et al., 1992). Methods of estimating microbial protein synthesis rely on marker techniques in which a particular microbial constituent is related to the microbial N content. Marker : N values have generally been established in mixed bacteria isolated from the liquid fraction of rumen digesta and it has been assumed that the same relationship holds in the total population leaving the rumen (Merry and McAllan, 1983). However, several studies have demonstrated differences in composition between solid-associated (SAB) and fluid-associated bacteria in vivo (Legay-Carmier and Bauchart, 1989) and in vitro (Molina Alcaide et al, 1996), as well in marker : N values (Pérez et al., 1996). This problem could be more pronounced in the in vitro semi-continuous culture system RUSITEC, in which there are three well defined components (a free liquid phase, a liquid phase associated with the solid phase and a solid phase), each one having associated microbial populations.The objective of this experiment was to investigate the effect of using different bacterial isolates (BI) on the estimation of microbial production of four different diets in RUSITEC (Czerkawski and Breckenridge, 1977), using (15NH4)2 SO4 as microbial marker, and to assess what effects any differences would have on the comparison of microbial protein synthesis between diets.This study was conducted in conjunction with an in vitro experiment described by Carro and Miller (1997). Two 14-day incubation trials were carried out with the rumen simulation technique RUSITEC (Czerkawski and Breckenridge, 1977). The general incubation procedure was the one described by Czerkawski and Breckenridge (1977) and more details about the procedures of this experiment are given elsewhere (Carro and Miller, 1997).

scholarly journals Biofilm Formation by and Antifungal Susceptibility of Candida Isolates from Urine

2007 ◽  
Vol 73 (6) ◽  
pp. 1697-1703 ◽  
Author(s):  
N. Jain ◽  
R. Kohli ◽  
E. Cook ◽  
P. Gialanella ◽  
T. Chang ◽  
...  
Keyword(s):  
Risk Factors ◽  
Growth Medium ◽  
Biofilm Formation ◽  
Chart Review ◽  
Antifungal Susceptibility ◽  
Retrospective Chart Review ◽  
Urine Samples ◽  
Rpmi Medium

ABSTRACT Biofilm formation (BF) in the setting of candiduria has not been well studied. We determined BF and MIC to antifungals in Candida spp. isolates grown from urine samples of patients and performed a retrospective chart review to examine the correlation with risk factors. A total of 67 Candida spp. isolates were grown from urine samples from 55 patients. The species distribution was C. albicans (54%), C. glabrata (36%), and C. tropicalis (10%). BF varied greatly among individual Candida isolates but was stable in sequential isolates during chronic infection. BF also depended on the growth medium and especially in C. albicans was significantly enhanced in artificial urine (AU) compared to RPMI medium. In nine of the C. albicans strains BF was 4- to 10-fold higher in AU, whereas in three of the C. albicans strains and two of the C. glabrata strains higher BF was measured in RPMI medium than in AU. Determination of the MICs showed that planktonic cells of all strains were susceptible to amphotericin B (AMB) and caspofungin (CASPO) and that three of the C. glabrata strains and two of the C. albicans strains were resistant to fluconazole (FLU). In contrast, all biofilm-associated adherent cells were resistant to CASPO and FLU. The biofilms of 14 strains (28%) were sensitive to AMB (MIC50 of <1 μg/ml). Correlation between degree of BF and MIC of AMB was not seen in RPMI grown biofilms but was present when grown in AU. A retrospective chart review demonstrated no correlation of known risk factors of candiduria with BF in AU or RPMI. We conclude that BF is a stable characteristic of Candida strains that varies greatly among clinical strains and is dependent on the growth medium. Resistance to AMB is associated with higher BF in AU, which may represent the more physiologic medium to test BF. Future studies should address whether in vitro BF can predict treatment failure in vivo.

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