The effects of sire, lactation number, and time of the year on late embryo mortality in dairy cattle

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Published on: December 20, 2021

C. Bailey, J. Gibbons, P. Melendez

Texas Tech University, School of Veterinary Medicine, Amarillo, TX 79106

Introduction

In the dairy industry, there has been considerable drive to maximize birthing rates and profitability by using genetics from beef cattle in both lower performing and lower genetic quality cows. This has allowed cows that are of average quality or aged genetics to provide added value to the operation in the form of crossbred calves for the beef industry.

One measure of infertility in multiparous cows is late embryonic mortality. Although there is likely a higher percentage of embryonic loss during early pregnancy (prior to Day 30; [1]), late embryonic mortality (LEM; defined as pregnancy loss that occurs between 30 and 60 days of gestation) is also a contributor to low profitability. Other research has shown that roughly 12.8% of dairy cows will undergo LEM between 28-42 days post-breeding, averaging a 0.85% loss per day [1].  Some of the factors that cause LEM include genetics, infectious diseases, poor prophylactic management, sanitation, and season of the year.  However, genetic defects of the embryo may also account for up to 20% of early embryonic mortalities [2].

A common method of detecting pregnancy is by measuring Pregnancy Specific Protein B (PSPB) and plasma progesterone (P4) at specific times post insemination. An assay testing PSPB levels indicate a positive pregnancy if PSPB levels are a >10% above non-pregnant cow levels approximately 28 days post artificial insemination [3,4]. Elevated progesterone concentrations (>1 ng/ml) are also useful as a pregnancy detection aid, as they typically begin to taper off around 15 days post estrus in the non-pregnant cow but, are maintained in pregnant cows [5]. Both PSPB and P4 were evaluated and compared between groups in a subset of cows in this study.

The objective of this study was to compare the LEM of Holstein cows bred to Limousin bulls to Holstein cows bred to Holstein bulls. Optical density as a measure of PSPB and P4 concentrations were also evaluated from a blood sample collected at 30 days post breeding for comparison between a subset of 25 cows per group. Further, the LEM was evaluated based upon the time of year that the breeding occurred Summer (April 1 – September 30) and Non-Summer (October 1 – March 31).

Methods

The Limousin X Holstein and Holstein X Holstein breedings in this study were from a large dairy in the southeast US, which consisted of 12,847 Holstein cows. In this herd, 1,166 cows were dry with an average of 171 days open. The cows were milked 3 times a day with a rolling herd average of 14,600 kg of milk per year and were fed a total mixed ration based on corn silage, grass silage, and concentrates. The reproductive program consisted of a 60-day voluntary waiting period and timed artificial insemination using a variety of ovulation synchronization protocols. Cows were diagnosed for pregnancy status by trans-rectal ultrasound between 28 to 35 days post breeding. If the cow was diagnosed open, she was subjected to a resynchronization protocol. If the cow was diagnosed as pregnant, she was rechecked for pregnancy status between 50 to 57 days post breeding by rectal palpation. Blood was collected at random (n = 25 cows per group) at the time of ultrasound pregnancy diagnosis (28-35 days) and tested for PSPB levels expressed as optical densities and P4 concentrations measured by Radio-immuno Assay.

Results

Normal LEM for this herd is approximately 12%. Overall, for all cows bred in the summer the LEM was 15.2 ± 0.6% while those bred in the non-summer had an LEM of 9.9 ± 0.3%, (P<0.0001). Overall, for all Holstein X Holstein embryos the LEM was 15.2 ± 0.6% while the LEM for all Limousin X Holstein embryos was 9.8 ± 0.3%, (P<0.0001).  Differences (P<0.05) in LEM between Holstein x Holstein and Limousin X Holstein were observed during the non-summer months (15.1 ± 0.8% versus 8.3 ± 0.3%, respectively) but not for the summer months (15.1 ± 0.9% and 15.3 ± 0.9%, respectively). summer = 15.23 +/- 0.6%, non-summer = 9.88 +/- 0.3%    P<0.0001

In Figure 1, lactations within breed combination that do not share a common superscript are different (P<0.05). Further, there was a trend (P=0.08) for a difference between Lactation 2 and Lactation 3 for Limousin X Holstein embryos in the non-summer months and there was a breed combination difference (P<0.05) for all lactations during the non-summer months but not for any lactations during the summer months.

Figure 1.  Late embryo mortality (mean ± SEM) percentage over multiple lactation in Holstein cows either bred to Holstein or Limousin bulls that were confirmed pregnant at approximately 30 days post timed artificial insemination either during the summer or non-summer months (see text for more details).

Effects of Season on LEM 

Cows bred in the summer months (Figure 1), regardless of mating, had a higher LEM compared to cows bred in the non-summer months.  A substantial difference in fertility was noted among cows carrying crossbred embryos especially during the non-summer months. Taken together, these data suggest that heat stress during the summer months combined with the stress of milk production, may affect the uterine environment specifically and these stressors cannot be overcome by altering the breed of the sire.  Alternatively, in the non-summer months, the crossbred embryo apparently adapts more readily to the stress of milk production alone than do the Holstein X Holstein embryos.

Protein Specific Protein B and Progesterone

Optical densities of PSPB and P4 concentrations were compared between groups that were diagnosed pregnant at Day 30. The results showed a PSPB optical density of 4.44 in the Holstein X Holstein group and a 3.25 in the Limousin X Holstein group with a pooled standard error of ± 0.27 and a P-Value of 0.023.  Progesterone concentrations were not different (P>0.05) between groups and were 8.66 ng/ml in the Holstein X Holstein group and 8.96 ng/ml in the Limousin X Holstein group with a pooled standard error of ± 0.57.

Conclusion

This field study demonstrated that breeding Holstein cows with Limousin semen reduced LEM by approximately 4% overall and there were seasonal and lactational differences in LEM. During the summer months, the breed combination had a lesser effect on LEM perhaps as all cows exhibited some form of heat stress; however, during the non-summer months, purebred Holstein embryos had a higher LEM than the Limousin X Holstein crossbred embryos, indicating inherent embryo loss dynamics due to factors other than genetics or lactation. These results indicated that breeding beef bulls to lower quality genetic Holstein cows decreased LEM regardless of time of year. However, the lower LEM was especially evident in early lactation cows bred during the non-summer months which likely contains cows that can still contribute genetically to the herd.  The exact components of the “protective mechanism” associated with producing crossbred embryos may be due to hybrid vigor, but has not been fully elucidated; however, it seems to be mostly present during the non-summer months underscoring the ever-present stress of milk production on the females.  The heat induced stress associated with the summer months historically [6] leads to an increase in LEM rates in Holstein embryos and was lactation dependent in both groups in this study. Progesterone concentrations for both groups were similar indicating that corpus luteum function was likely not related to LEM.  Lower optical density which is reflective of the concentration of PSPB was seen in Holstein cows bred to Limousin bulls versus Holstein bulls. More research may be necessary to determine if PSPB plays a role in embryo viability during early pregnancy or if elevated optical density of PSPB is reflective of embryos in distress [7].

References

1)  Wiltbank MC, Baez GM, Garcia-Guerra A, Toledo MZ, Monteiro PLJ, Melo LF, Ochoa JC, Santos JEP, and Sartori R. (2016). Pivotal periods for pregnancy loss during the first trimester of gestation in lactating dairy cows. Theriogenology, 86(1), 239–253. https://doi.org/10.1016/j.theriogenology.2016.04.037 

2)  Alfieri AA, Leme RA, Agnol AMD, and Alfieri AF.  (2019). Sanitary program to reduce embryonic mortality associated with infectious diseases in cattle.  Animal reproduction vol. 16,3 386-393. 22 Oct. 2019, doi:10.21451/1984-3143-AR2019-0073 

3)  Humblot F, Camous S, Martal J, Charlery J, Jeanguyot N, ThibierM, and Sasser RG. (1988).  Pregnancy-specific protein B, progesterone concentrations and embryonic mortality during early pregnancy in dairy cows. J Reprod Fertil. 1988 May;83(1):215-23. doi: 10.1530/jrf.0.0830215. PMID: 3397939.

4)  Middleton EL, and Pursley JR. (2019).  Short communication: Blood samples before and after embryonic attachment accurately determine non-pregnant lactating dairy cows at 24 d post-artificial insemination using a commercially available assay for pregnancy-specific protein B. J Dairy Sci. 2019 Aug;102(8):7570-7575. doi: 10.3168/jds.2018-15961. Epub2019 Jun 6. PMID: 31178191.

5)  Thirapatsukun T, KW Entwistle, and Gartner, RJW.  (1978). Plasma Progesterone Levels as an Early Pregnancy Test in Beef Cattle.  Theriogenology, Vol. 9, no. 4, pp. 323-332.

https://doi.org/10.1016/0093-691x(78)90125-5.

6)  Morton, JM, Tranter, WP, Mayer, DG, and Jonsson, NN. (2007). Effects of environmental heat on conception rates in lactating dairy cows: Critical periods of exposure. Journal of Dairy Science, 90(5), 2271–2278. https://doi.org/10.3168/jds.2006-574 

7)  Thompson, IM, Tao, S, Branen, J, Ealy, AD, and Dahl, GE. (2013). Environmental regulation of pregnancy-specific protein B concentrations during late pregnancy in dairy cattle1. Journal of Animal Science, 91(1), 168–173. https://doi.org/10.2527/jas.2012-5730 

Dominant Follicle Removal Prior to Superovulation in Ewes

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Published on: December 20, 2021

 T. Mittleider a , C. Holcomba , D. Hobbs a, D. Davis c, L. Miller a , J. Gibbons a, b, d

a College of Veterinary Medicine, b DeBusk College of Osteopathic Medicine, Lincoln Memorial University, Harrogate, TN, 37752, c Laurel Highlands Animal Health, Somerset, PA, 15501, d Texas Tech University – School of Veterinary Medicine, Amarillo, TX, 79106

Link to poster: https://onedrive.live.com/view.aspx?resid=60555781396F53CD!1930&ithint=file%2cpptx&authkey=!AsTpH3b9DzphkGY

The Effects of Melatonin on Ovine Estrus Cyclicity

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Published on: December 20, 2021

C. Holcomb b , D. Hobbs b, J. Gibbons a, b, d, D. Davis c

a College of Veterinary Medicine, b DeBusk College of Osteopathic Medicine, Lincoln Memorial University, Harrogate, TN, 37752, c Laurel Highlands Animal Health, Somerset, PA, 15501, d Texas Tech University – School of Veterinary Medicine, Amarillo, TX, 79106

Link to poster: https://onedrive.live.com/view.aspx?resid=60555781396F53CD!2671&ithint=file%2cpptx&authkey=!AjrXPjYQPdeIIP8

Disappearance and uptake of [125I]FSH in the rat, rabbit, ewe and cow

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Published on: August 18, 2021

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D. B. LasterUSDA

Date of this Version

1972

Comments

Published in J. Reprod. Fert. (1972) 30, 407-415

Abstract

Follicle-stimulating hormone (NIH-FSH-S8) was labelled with 125I to determine its disappearance rate after a single intravenous injection and to determine the level of circulating [125I]fsh in the blood after a single intramuscular or subcutaneous injection in the rat, rabbit, ewe and cow. There was a difference in the disappearance and uptake rates among the four species, but the shape of the curve for rate of loss and uptake of labelled fsh was similar in all species. The disappearance of radioactivity occurred at two rates; the first from 1 to 8 min and the second from 16 to 96 min. The half-life, calculated from the total decay curve in each species was 94±21, 118±16, 334±41 and 301±23 min for the rats, rabbits, ewes and cows, respectively. Intramuscular injections resulted in an average of 56% higher [125I]fsh blood levels than subcutaneous injections for all species.

Embryo Transfer in Cattle

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Published on: August 18, 2021

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Reproductive Physiology Review

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Published on: August 18, 2021

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In Vitro Fertilization in Cattle

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Published on: August 18, 2021

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Articles of Interest

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Published on: August 18, 2021

https://www.animal-reproduction.org/article/5b5a604bf7783717068b46a0

https://www.journalofdairyscience.org/article/S0022-0302(19)30635-6/fulltext

https://www.animal-reproduction.org/article/doi/10.21451/1984-3143-AR1002

https://www.semanticscholar.org/paper/Artificial-Insemination-and-Embryo-Transfer-in-Farin/da20551c1a0fad2bafd1dd931de027691a6bebe0

https://www.sciencedirect.com/science/article/pii/S0022030201746905

https://www.mdpi.com/2076-2615/11/6/1666/htm

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7830735/

https://www.sciencedirect.com/science/article/pii/S0022030208712153

https://rep.bioscientifica.com/view/journals/rep/154/6/REP-17-0357.xml

Equine chorionic gonadotropin increases estradiol levels in the bovine oviduct and drives the transcription of genes related to fertilization in superstimulated cows

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Published on: February 18, 2021

Patricia K Fontes 1Eduardo M Razza 1Antônio G R Pupulim 2Ciro M Barros 1Anthony C de Souza Castilho 3

1Departament of Pharmacology, Institute of Biosciences, Universidade Estadual Paulista (UNESP), Botucatu, São Paulo, Brazil.

2Centro Universitário Cesumar (UNICESUMAR), Maringá, Paraná, Brazil.

3Universidade do Oeste Paulista (UNOESTE), Presidente Prudente, São Paulo, Brazil.

https://pubmed.ncbi.nlm.nih.gov/31353672/

Mol Reprod Dev. 2019 Nov;86(11):1582-1591. doi: 10.1002/mrd.23243. Epub 2019 Jul 29.

Abstract

In the bovine oviduct, estradiol (E2) stimulates secretion and cell proliferation, whereas progesterone (P4) suppresses them. In this study, we have evaluated the effect of two superstimulatory protocols (follicle-stimulating hormone [FSH] or FSH combined with equine chorionic gonadotropin [eCG]) on the oviductal levels of E2 and P4 and its outcome on oviductal cells. Compared with the control group (a single pre-ovulatory follicle), we have observed that the cows submitted to FSH/eCG treatment showed a higher concentration of E2 in the oviduct tissue, together with a higher abundance of messenger RNA encoding steroid receptors (ESR1 and progesterone receptor), and genes linked to gamete interactions and regulation of polyspermy (oviduct-specific glycoprotein 1, heat-shock protein family A member 5, α-l-fucosidase 1 [FUCA1], and FUCA2) in the infundibulum and ampulla segments of the oviduct. However, we did not observe any modulation of gene expression in the isthmus segment. Even though the FSH protocol upregulated some of the genes analyzed, we may infer that the steady effect of FSH combined with eCG on oviduct regulation might benefit fertilization and may potentially increase pregnancy rates.

Keywords: cattle; female reproductive tract; gametes; gene expression; steroids; superovulation.

Effect of superstimulation on the expression of microRNAs and genes involved in steroidogenesis and ovulation in Nelore cows

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Published on: February 18, 2021

P H Santos 1R A Satrapa 2P K Fontes 1F F Franchi 1E M Razza 1F Mani 3M F G Nogueira 4C M Barros 1A C S Castilho 5

1Universidade Estadual Paulista (UNESP), Institute of Biosciences, Campus Botucatu, Department of Pharmacology, Botucatu, São Paulo, Brazil.

2Universidade Federal do Acre (UFAC), Center of Biological and Natural Sciences, Rio Branco, Acre, Brazil.

3Universidade Estadual Paulista (UNESP), Institute of Biosciences, Campus Botucatu, Department of Chemistry and Biochemistry, Botucatu, São Paulo, Brazil.

4Universidade Estadual Paulista (UNESP), School of Sciences and Languages, Campus Assis, Department of Biological Sciences, Assis, São Paulo, Brazil.

5Universidade Estadual Paulista (UNESP), School of Sciences and Languages, Campus Assis, Department of Biological Sciences, Assis, São Paulo, Brazil. Electronic address: anthony@unoeste.br.

https://pubmed.ncbi.nlm.nih.gov/29407901/

Theriogenology 2018 Apr 1;110:192-200. doi: 10.1016/j.theriogenology.2017.12.045. Epub 2018 Jan 11.

Abstract

To better understand the impact of ovarian superstimulation on bovine follicular microenvironment, Nelore cows (Bos taurus indicus) were subjected to ovarian superstimulation with follicle stimulating hormone (FSH, n = 10; P-36 protocol) or FSH combined with eCG (n = 10; P-36/eCG protocol). Follicular fluid was analyzed for cholesterol concentration. Granulosa cells were analyzed by RT-qPCR to assess the expression of genes involved in steroidogenic and ovulatory and expression of microRNAs involved in final follicular development and luteinizing hormone/choriogonadotropin receptor (LHCGR) expression. Plasma concentration of estradiol was also measured. Follicular fluid from the P-36 group showed higher concentration of cholesterol than that of control (non-superstimulated) cows. Plasma concentration of estradiol was higher in the P-36/eCG group. Abundance of STAR and FSHR mRNAs were lower in granulosa cells from the P-36/eCG group. In contrast, LHCGR mRNA abundance was higher in superstimulated granulosa cells from the P-36 group and showed a pattern opposite to that of miR-222 expression. Ovarian superstimulation did not affect the expression of other markers (mmu-miR-202-5p, has-miR-873, has-miR-144, and their target genes, CREB, TGFBR2, and ATG7) of antral follicle development. However, the mRNA expression of VEGF pathway components was modulated by P-36 treatment. Taken together, these results demonstrate that superstimulatory protocols modify steroidogenic capacity, increase plasma estradiol, and regulate the abundance of VEGF system, LHCGR mRNA and suppress the expression of miR-222 in bovine granulosa cells.

The incompletely fulfilled promise of embryo transfer in cattle—why aren’t pregnancy rates greater and what can we do about it?

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Published on: February 18, 2021

Peter J Hansen

Department of Animal Sciences, D.H. Barron Reproductive and Perinatal Biology Research Program, and Genetics Institute, University of Florida, Gainesville, FL

https://academic.oup.com/jas/article/98/11/skaa288/5954183

Journal of Animal Science, Volume 98, Issue 11, November 2020, skaa288 https://doi.org/10.1093/jas/skaa288

Abstract

Typically, bovine embryos are transferred into recipient females about day 7 after estrus or anticipated ovulation, when the embryo has reached the blastocyst stage of development. All the biological and technical causes for failure of a female to produce a blastocyst 7 d after natural or artificial insemination (AI) are avoided when a blastocyst-stage embryo is transferred into the female. It is reasonable to expect, therefore, that pregnancy success would be higher for embryo transfer (ET) recipients than for inseminated females. This expectation is not usually met unless the recipient is exposed to heat stress or is classified as a repeat-breeder female. Rather, pregnancy success is generally similar for ET and AI. The implication is that either one or more of the technical aspects of ET have not yet been optimized or that underlying female fertility that causes an embryo to die before day 7 also causes it to die later in pregnancy. Improvements in pregnancy success after ET will depend upon making a better embryo, improving uterine receptivity, and forging new tools for production and transfer of embryos. Key to accelerating progress in improving pregnancy rates will be the identification of phenotypes or phenomes that allow the prediction of embryo competence for survival and maternal capacity to support embryonic development.

‘There is only one thing that is truly important in an IVF laboratory: everything’ Cairo Consensus Guidelines on IVF Culture Conditions

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Published on: February 18, 2021

Cairo Consensus Group

https://www.sciencedirect.com/science/article/pii/S1472648319307540#!

Reproductive BioMedicine Online Volume 40, Issue 1, January 2020, Pages 33-60

Abstract

This proceedings report presents the outcomes from an international expert meeting to establish consensus guidelines on IVF culture conditions. Topics reviewed and discussed were: embryo culture – basic principles and interactions; temperature in the IVF laboratory; humidity in culture; carbon dioxide control and medium pH; oxygen tension for embryo culture; workstations – design and engineering; incubators – maintaining the culture environment; micromanipulation – maintaining a steady physcochemical environment; handling practices; assessment practices; culture media – buffering and pH, general composition and protein supplementation, sequential or single-step media for human embryo culture; use and management – cold chain and storage; test equipment – calibration and certification; and laboratory equipment and real-time monitoring. More than 50 consensus guideline points were established under these general headings.

Keywords: Culture conditions, Incubators, IVFMaintenance, Medium, Quality control

Direct transfer of frozen-thawed bovine embryos and its application in cattle reproduction management

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Published on: February 18, 2021

Osamu DOCHI1

1)Rakuno Gakuen University, Hokkaido 069-0851, Japan

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6815740/

J Reprod Dev. 2019 Oct; 65(5): 389–396.Published online 2019 Jun 13. doi: 10.1262/jrd.2019-025

Abstract

Embryo transfer entails many procedures and techniques, of which embryo freezing is an important component in bovine embryo transfer. Embryo freezing techniques have been developed over the last 40 years, allowing practical availability, and have become essential for cattle reproduction management under field conditions. The direct transfer methods of frozen-thawed, in vivo-derived, and in vitro-produced (IVF) bovine embryos using 1.5 M ethylene glycol (EG) with or without sucrose (SUC) are used widely under on-farm conditions, not only in Japan but also globally. The direct transfer method using 1.5 M glycerol (GLY) and 0.25 M SUC (GLY-SUC) is used mainly in Japan. The pregnancy rate with direct transfer of frozen-thawed bovine embryos in either EG or GLY-SUC has been found to not differ from conventional freezing with GLY and traditional dilution techniques. Pregnancy rates following direct transfer of frozen-thawed bovine embryos were affected by the developmental stage of the embryos and the parity of the recipients. The use of ultrasound-guided on-farm ovum pickup is ushering in a new revolution for the commercial application of IVF embryos. Globally, for the first time more IVF bovine embryos were transferred in 2017 than produced in vivo. More than 60% of IVF embryos were transferred fresh due to a low pregnancy rate of frozen-thawed IVF embryos. Many factors seemed to be involved in improving the survival rate of frozen-thawed IVF embryos. Therefore, further research is needed to improve the freezing tolerance of IVF embryos to develop efficient direct transfer methods analogous to those used for in vivo embryos.

Keywords: Bovine embryo, Direct transfer, Ethylene glycol, Freezing, Pregnancy rate

Articles of Interest

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Published on: February 18, 2021

Ano-genital distance relates to fertility

Effect of dose and timing of prostaglandin F2α treatments during a 7-d Ovsynch protocol on progesterone concentration at the end of the protocol and pregnancy outcomes in lactating Holstein cows

Estradiol cypionate administered at the end of a progesterone-based protocol for FTAI induces ovulation and improves postovulatory luteal function and uterine environment in anestrous beef cows

Association between heat stress during intrauterine development and the calving-to-conception and calving-to-first-service intervals in Holstein cows

Impact of ethanol and heat stress–dependent effect of ultra-diluted Arnica montana 6 cH on in vitro embryo production in cattle

Analysis of bovine blastocysts indicates ovarian stimulation does not induce chromosome errors, nor discordance between inner-cell mass and trophectoderm lineages

Developmental kinetics and viability of bovine embryos produced in vitro with sex-sorted semen

Heat stress and reproduction – A foreword

Growing cattle embryos beyond Day 8 – An investigation of media components

Melatonin enhances in vitro developmental competence of cumulus-oocyte complexes collected by ovum pick-up in prepubertal and adult dairy cattle

Effect of expression of estrus and treatment with GnRH on pregnancies per AI in beef cattle synchronized with an estradiol/progesterone-based protocol

The incompletely fulfilled promise of embryo transfer in cattle—why aren’t pregnancy rates greater and what can we do about it?

2021 IETS proceedings

Effects of epidermal growth factor and progesterone on oocyte meiotic resumption and the expression of maturation-related transcripts during prematuration of oocytes from small and medium-sized bovine antral follicles              

Effect of Supplemental Trace Minerals on Standard and Novel Measures of Bull Fertility

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Published on: November 2, 2020

T. W. Geary – a2, R. C. Waterman – a, M. L. Van Emon – b, C. R. Ratzburg – c, S. Lake – c, B. A. Eik – a, D. R. Armstrong – a, A. L. Zezeski – a, and J. S. Heldt – d

aUSDA-ARS, Fort Keogh Livestock and Range Research Laboratory, Miles City, MT 59301; bDepartment of Animal and Range Sciences, Montana State University, Bozeman, MT 59717; cDepartment of Animal Sciences, University of Wyoming, Laramie, WY; dMicronutrients USA LLC, 2601 Fortune Circle Drive E. Suite 200C, Indianapolis, IN 46241

Abstract

Two studies were conducted to evaluate the effects of trace mineral supplementation on traditional and novel measures of bull fertility. In experiment 1, 37 mature bulls received one of three dietary supplements daily for 71 d: 1) Supplement without Cu, Zn, and Mn (CON); 2) Supplement with Cu, Zn, and Mn sulfate (SULF); and 3) Supplement with basic Cu chloride, and Zn and Mn hydroxychloride (CHLR). In experiment 2, 128 Angus or Angus-Hereford calves were maintained on a growing diet for 75 d (year 1) or 119 d (year 2) in Calan gate equipped pens without mineral supplementation. Bulls (n = 32 head/treatment) received one of four trace mineral supplements daily for 84 d: 1) Zn with no Cu (ZN), 2) Cu with no Zn (CU), 3) Cu and Zn (ZNCU), or 4) no Cu or Zn (CON). Fertility measures included a breeding soundness examination (BSE) and novel fertility measures conducted using flow cytometry. In mature bulls, final liver Zn concentration was positively correlated (P = 0.02) with sperm concentration (r = 0.31) and tended (P = 0.06) to be negatively correlated with acrosome damage (r = -0.39). Peripubertal bulls receiving ZNCU had greater ADG than CU bulls (P = 0.05). Each BSE and novel fertility component improved from d 0 to 84 in peripubertal bulls and were not affected (P > 0.10) by mineral supplementation. Bulls that received no supplement (CON) had greater (P < 0.01) percentage of sperm with distal midpiece reflex and Dag defect in their ejaculates. Sperm viability after 30 min of incubation were not affected by trace mineral supplementation, but after 3 h incubation, sperm viability tended to differ (P = 0.06) between treatments and tended to be less for CON bulls compared to ZNCU bulls. Among contrast comparisons, trace mineral supplemented bulls had greater (P < 0.05) percentage of viable sperm at 3 h post collection and reactive oxygen resistant sperm than CON bulls. Addition of Zn to trace mineral containing Cu (ZNCU) improved (P < 0.05) percentage of sperm in the ejaculate with high mitochondrial energy potential and viable sperm with intact acrosome membrane. In summary, it appears the homeostasis mechanisms for bull trace mineral maintenance are extremely efficient and mineral supplementation of mature and peripubertal bulls did not have major improvements in any laboratory or chute-side measures of bull fertility, however bulls exposed to breeding or in environments with diet antagonists might respond differently.

Out of Season Artificial Insemination and Embryo Transfer Results in Ewes: A Field Trial

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Published on: November 2, 2020

B Price, T Mittleider, S Collins, P Gibbons, and J Gibbons

College of Veterinary Medicine, Lincoln Memorial University, Harrogate, TN

Introduction: 

Ewes are seasonally polyestrous short-day breeders with an estrous cycle of approximately 16 – 17 days.  In the northern hemisphere ewes have active estrous cycles and are naturally receptive to rams from late September to late December when there is less than 12 hours of daylength.  However, progressive sheep breeders often prefer to breed sheep earlier in the year, during periods where there is more than twelve hours of daylength, (July and August) in order to have lambs that are appropriate to target specific show markets.  In order to facilitate this out of season breeding and accelerate genetic gain, producers rely on Assisted Reproductive Techniques such as Laparoscopic Artificial Insemination (LAI), ovarian hyper-stimulation, embryo collection from valuable embryo donors, and embryo transfer (ET) into synchronized recipients (1,2,3.)  This field trial was conducted during late July through early August in southwest Virginia (latitude 36-38’12” N), during a daylight period of about 14 hours. Pregnancy rates of ewes bred by means of AI were compared to those that underwent ET.

Hatching Blastocysts

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Published on: November 2, 2020

Day 7 in vitro produced bovine hatching blastocysts.

By Dr Daniela Demetrio – Ruann Genetics – Riverdale, CA

7 & 7 Synch: An Estrus Synchronization Protocol for Postpartum Beef Cows

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Published on: November 2, 2020

Overview


Researchers at the University of Missouri recently
evaluated a new protocol for synchronization of estrus
among postpartum beef cows. This protocol was found
to be highly effective both for cows receiving embryo
transfer (ET) and cows receiving fixed-time artificial
insemination (AI). Extensive field trials with 7 & 7
Synch observed improvements in the proportion of
cows expressing estrus and in the proportion of cows
becoming pregnant to embryo transfer or to AI.

How can we improve embryo production and pregnancy outcomes of Holstein embryos produced in vitro? (12 years of practical results at a California dairy farm)

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Published on: November 2, 2020

Daniela Garcia Borges Demetrio, 1 ,* Eduardo Benedetti, 2 Clarice Garcia Borges Demetrio, 3 Julio Fonseca, 1 Mayara Oliveira, 1 Alvaro Magalhaes, 1 and Ricarda Maria dos Santos 4

1RuAnn Genetics, Riverdale, CA, United States
2Arizona Dairy Co, Mesa, AZ, United States
3Escola Superior de Agricultura “Luiz de Queiroz”, Universidade do Estado de São Paulo, Piracicaba, SP, Brasil
4Faculdade de Medicina Veterinária, Universidade Federal de Uberlândia, Uberlândia, MG, Brasil

Abstract


Genomic evaluations have revolutionized dairy cattle breeding, and the demand for embryos produced from very young heifers with high genetic merit has increased over time. The combination of low oocyte recovery, young age of donors, and milk production status can make the in vitro embryo production (IVP) of Holstein cattle incredibly challenging. Several factors need to be coordinated to obtain a live calf from an IVP embryo, but the quality of the oocyte at the start of the process is one of the key factors. Aspects related to oocyte quality, laboratory quality control, embryo quality and recipient selection are addressed here, based on the measures that the RuAnn Genetics Laboratory (Riverdale, California, USA) adopted in the last 12 years, with the goal of improving production of live, healthy calves from Holstein embryos. Follicular wave synchronization and stimulation with follicular stimulating hormone (FSH) is necessary to improve oocyte quality and consequently embryo production. Laboratory quality control and the use of high-quality supplies are essential to reduce variability in production and facilitate identification of other factors that might interfere with embryo production. High pregnancy rates can be achieved with good quality embryos selected at optimal time and stage of development, transferred by an experienced embryo transfer (ET) technician, to well managed recipients 7 or 8 days after estrus. Attention to detail at every step of the process is crucial to success.
Keywords: embryo, Holstein, in vitro production, pregnancy, recipient.

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One-step automated bioprinting-based method for cumulus-oocyte complex microencapsulation for 3D in vitro maturation

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Published on: November 2, 2020

Antonella Mastrorocco1¤a, Ludovica Cacopardo2, Nicola Antonio Martino1¤b, Diana Fanelli3, Francesco Camillo3, Elena Ciani1, Bernard A. J. Roelen4, Arti Ahluwalia2,5☯, Maria Elena Dell’Aquila1☯

1 Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari Aldo Moro, Bari, Italy; 2 Research Centre E. Piaggio, University of Pisa, Pisa, Italy; 3 Department of Veterinary Sciences, University of Pisa, Pisa, Italy; 4 Department of Clinical Sciences, Embryology, Anatomy and Physiology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands; 5 Department of Information Engineering, University of Pisa, Pisa, Italy

☯ These authors contributed equally to this work.
¤a Current address: Faculty of Veterinary Medicine, University of Teramo, Teramo, Italy
¤b Current address: Department of Veterinary Sciences, University of Torino, Torino, Italy

Abstract

Three-dimensional in vitro maturation (3D IVM) is a promising approach to improve IVM efficiency as it could prevent cumulus-oocyte complex (COC) flattening and preserve its structural and functional integrity. Methods reported to date have low reproducibility and validation studies are limited. In this study, a bioprinting based production process for generating microbeads containing a COC (COC-microbeads) was optimized and its validity tested in a large animal model (sheep). Alginate microbeads were produced and characterized for size, shape and stability under culture conditions. COC encapsulation had high efficiency and reproducibility and cumulus integrity was preserved. COC-microbeads underwent IVM, with COCs cultured in standard 2D IVM as controls. After IVM, oocytes were analyzed for nuclear chromatin configuration, bioenergetic/oxidative status and transcriptional activity of genes biomarker of mitochondrial activity (TFAMATP6ATP8) and oocyte developmental competence (KHDC3NLRP5OOEP and TLE6). The 3D system supported oocyte nuclear maturation more efficiently than the 2D control (P<0.05). Ooplasmic mitochondrial activity and reactive oxygen species (ROS) generation ability were increased (P<0.05). Up-regulation of TFAMATP6 and ATP8 and down-regulation of KHDC3NLRP5 expression were observed in 3D IVM. In conclusion, the new bioprinting method for producing COC-microbeads has high reproducibility and efficiency. Moreover, 3D IVM improves oocyte nuclear maturation and relevant parameters of oocyte cytoplasmic maturation and could be used for clinical and toxicological applications.

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