Impacts of feeding intensity and breed on metabolism, negative energy balance and reproductive efficiency in dairy cows

Despite improved breeding, management, and nutritional strategies, decreased fertility in dairy cows is still widespread. Several studies have highlighted the unfavourable correlation between negative energy balance (NEB) and reproductive performance. However, there is a continuing need for more information regarding the effects of the interaction between different nutritional strategies with animals of different genetic background. This thesis evaluated the effect of high energy-diet (HE) and low energy-diet (LE) on Holstein and SRB dairy cows. The HE represents what is, in general, common practice among most high milk producing herds. The lower feeding intensity may be representative of e.g. organic dairy producing systems. In three studies, the metabolic status, milk yield, body condition score (BCS), and NEB were evaluated in cows fed either a HE or a LE diet. Associations between NEB, plasma adipokines, metabolism, and reproductive parameters were also investigated. Holstein cows had lower body condition score (BCS) than SRB cows within each energy-diet group. However, diet had no effect on BCS or subcutaneous adipose tissue thickness in Holstein cows irrespective of whether they received a HE or LE diet. The HE group tended to have a less severe energy deficit than the LE group. Holstein cows tended to be in a more severe energy deficit during the first 45 days after calving than SRB cows. Holstein had a lower nadir in energy balance than the SRB. In conclusion, our results indicate that nutritional strategies might have a stronger association to endocrine and traditional fertility traits than breed. However, breed had a stronger association to the energy balance variables than nutritional strategies. In addition, SRB cows prioritized energy differently when compared to Holstein cows in such a way that the SRB cows maintained homeostasis better than Holsteins who had a deeper energy deficit than SRB cows

Practical Method for Freezing Buck Semen

Although several protocols for cryopreserving buck semen are described in the literature, they differ widely in factors such as season and method of semen collection, extender and sperm concentration. Therefore, choosing a protocol that is suitable for a particular on-farm situation can be problematic. In the present study, semen was collected by artificial vagina from seven bucks on a farm located approximately 90 minutes’ drive away from the laboratory, about 6 weeks before the start of the goat breeding season. The semen was immediately extended in warm semen extender containing soy lecithin and was placed in an insulated box with a cold pack for up to 4 h, during semen collection from the remaining bucks and subsequent transport to the laboratory. Following centrifugation at 4 °C and resuspension in the soy lecithin extender to a sperm concentration of 800 × 106 spermatozoa/mL, 0.25 mL plastic straws were filled and frozen in racks 4 cm above the surface of liquid nitrogen. This simple protocol resulted in an acceptable post-thaw quality for all seven bucks, with a mean post-thaw motility of 55 ± 21% and mean fragmented chromatin of 3.27 ± 1.39%. Normal sperm morphology was >90% in all ejaculates. The semen was sent to a gamete bank for long-term storage

Milk fatty acids as indicators of delayed commencement of luteal activity in dairy cows in early lactation

Excessive mobilization of adipose reserves due to severe negative energy balance in early lactation may be detrimental to dairy cow fertility at individual and herd level. Reproductive efficiency is one of the main factors influencing herd profitability and a strategy for early detection and management of cows with delayed resumption of cyclicity will result higher conception rate, decreased proportion of cows with extended lactation, fewer inseminations per conception and lower culling rates due to reproductive disorders. Using two groups of dairy cows (Holstein n = 37, Swedish Red breed [SRB] n = 49), we investigated potential differences between cows with different commencement of luteal activity (CLA) and the feasibility of using milk fatty acids (MFAs) as predictors of delayed CLA. Milk samples for progesterone analysis were collected twice weekly during the first six weeks in milk. The concentrations of the MFAs C14:0, C16:0, C18:0 and C18:1 cis -9 in milk (g/100 g milk) and in milk fat (g/100 g fat) were analysed by Fourier transform infrared spectroscopy and individual MFA profiles were calculated by weeks in milk. Commencement of luteal activity was defined as the first day with milk progesterone concentrations > 3 ng/ml at two successive measurements. The study population was categorized as early (n = 42) or late (n = 44) CLA, using the median value of 21 DIM as the cut -off. Analysis of the data revealed that CLA was correlated with the proportion of some specific MFAs, where cows with delayed CLA had lower IGF-1 (92.9 +/- 7.9 vs. 114.1 +/- 7.9 ng/ml; p = .05) and C14:0 levels (10.4 +/- 0.2 vs. 11.5 +/- 0.2 g/100 fat; p < .01) and higher C18:0 (9.6 +/- 0.2 vs. 8.9 +/- 0.6 g/100 fat; p < .01) and C18:1 cis -9 levels (24.9 +/- 0.4 vs. 23.5 +/- 0.4 g/100 fat; p < .05). Delayed CLA (mean 34 days) was predictable for approximately 80% of cows based on C18:0 or C18:1 cis -9 concentrations in week 2 postpartum. Overall, MFAs (C18:0 and C18:1 cis -9) as biomarkers were better indicators than betahydroxybutyrate or non-esterified fatty acids in early detection of cows with delayed or normal CLA. The MFA concentrations in milk samples from cows in early lactation can thus be used as a non-invasive method to identify cows at risk of delayed CLA, acting as potential biomarkers for future reproductive performance

Deciphering sperm chromatin properties to predict stallion sperm fertility

Although previous studies have examined the relationship between the sperm DNA fragmentation index and fertility in stallions, other aspects of chromatin structure or packaging and fertility have not been explored. In the present study, relationships between fertility and DNA fragmentation index, protamine deficiency, total thiols, free thiols and disulfide bonds in stallion spermatozoa were investigated. Ejaculates (n = 36) were collected from 12 stallions and extended to prepare semen doses for insemination. One dose from each ejaculate was sent to the Swedish University of Agricultural Sciences. Aliquots of semen were stained for flow cytometry with acridine orange for the Sperm Chromatin Structure Assay (DNA fragmentation Index, %DFI), with chromomycin A3 (CMA) for protamine deficiency, and with monobromobimane (mBBr) for detection of total and free thiols and disulfide bonds. Per season pregnancy rates after insemination were obtained. Mixed linear models were used to analyze data. Negative correlations were found between pregnancy rate and %DFI (r = -0.35, P < 0.03) and pregnancy rate and free thiols (r = -0.60, P < 0.0001). Furthermore, there were positive correlations between total thiols and disulfide bonds (r = 0.95, P < 0.0001), and protamine and disulfide bonds (r = 0.4100, P < 0.01986). Since chromatin integrity, protamine deficiency and packaging were all associated with fertility, a combination of these factors could be used as a biomarker of fertility when assessing ejaculates

Variation among stallions in sperm quality after single layer centrifugation

Although single layer centrifugation (SLC) selects robust spermatozoa from stallion semen, the effect of individual variation has not been studied in detail. The objective of this study was to determine the variation among stallions in the effects of SLC on sperm quality during cooled storage for up to 48 hr. Semen samples from seven stallions (18 ejaculates) were split, with one portion being used for SLC and the other serving as a control (CON). Sperm quality (kinematics, reactive oxygen species (ROS) production, membrane integrity (MI) and chromatin integrity) were analysed at 0, 24 and 48 hr using computer-assisted sperm analysis and flow cytometry. Sperm quality was better in SLC than in CON at all timepoints, especially chromatin integrity and MI (p < .0001 for both), and some categories of ROS production (e.g. proportion of live hydrogen peroxide negative spermatozoa, p < .0001), but the degree of improvement varied among stallions and type of ROS (p p < .0001). Total and progressive motility were also better in SLC samples than in CON at 24 and 48 hr (p < .0001), although the effect on sperm kinematics varied. The interaction of treatment, time and stallion was not significant. In conclusion, sperm quality was better in SLC samples than in CON, although there was considerable individual variation among stallions. The improvement in sperm quality, particularly in chromatin integrity, was clearly beneficial, and therefore the use of this technique would be warranted for all stallion semen samples

Post-thaw semen quality in young bull ejaculates before being accepted for commercial semen doses

Background: Genomic selection enables bulls with desirable characteristics to be identified at a young age, but sperm quality can be poor in the ejaculates of young bulls. Few studies have been done on post-thaw sperm quality in bulls less than 10 months old. The objective of this study was to determine the age at which post-thaw sperm quality was acceptable for artificial insemination.Methods: Semen was collected by artificial vagina; samples containing 100-500 million spermatozoa/ml were frozen for this study. Post-thaw analyses of membrane integrity (MI), mitochondrial membrane potential (MMP), chromatin integrity, morphology, production of reactive oxygen species and sperm kinematics were made.Results: The age at which ejaculates exceeded the breeding company's thresholds of acceptance varied considerably among individuals, with 285 days being the earliest. Morphology (p < 0.003), MI (p = 0.0096), high MMP (p = 0.043) and superoxide production (p = 0.0084) increased between the first and last ejaculates but reached acceptable levels at different ages for individual bulls.Conclusions: It was possible to obtain acceptable post-thaw sperm quality from samples even though sperm concentration was lower than the breeding company's threshold. Therefore, it might be feasible to use ejaculates earlier than is currently considered possible, by modifying semen handling protocols

Sperm Quality in Young Bull Semen Can Be Improved by Single Layer Centrifugation

Simple Summary Genomic selection enables bulls with desirable genes to be identified early in life. Livestock producers need to use the semen from young bulls as early as possible for efficient milk and meat production with fewer greenhouse gas emissions. However, semen from young bulls is often of lower quality than needed for freezing for commercial artificial insemination. Colloid centrifugation selects spermatozoa with the desirable characteristics needed for fertilization from the rest of the ejaculate. In this study, split ejaculates from young bulls were prepared with or without colloid centrifugation. Using this technique, sperm doses of acceptable quality for artificial insemination could be produced from ejaculates that would otherwise be discarded. Thus, the semen from young bulls would be usable for artificial insemination sooner than is currently the case. Interest in using semen from young bulls is increasing due to identifying promising animals by genomic selection. However, sperm quality in these ejaculates may not reach currently accepted standards for the cattle breeding industry. The purpose of this study was to determine if centrifugation of semen from young bulls through the Bovicoll colloid could improve sperm quality sufficiently for the frozen semen to be acceptable for artificial insemination. Ejaculates from 19 young bulls were split and either processed by Single-Layer Centrifugation (SLC) or not (CON) before freezing. After thawing, sperm quality was evaluated by determination of membrane integrity, mitochondrial membrane potential, DNA integrity, production of reactive oxygen species, sperm morphology and motility. Approximately half of the CON samples reached acceptable post-thaw quality (membrane integrity >= 40%) despite being below the breeding company ' s desired sperm concentration threshold pre-freezing. In the remaining samples, sperm quality was improved by SLC such that 45% of them reached acceptable quality post-thaw. Almost 75% of the young bull sperm samples could have produced usable frozen semen doses by adjusting the breeding company ' s current processing protocols. Since lowering the generation interval has a direct effect on the genetic gain per year, SLC could aid genetic progress in cattle breeding

Stored Stallion Sperm Quality Depends on Sperm Preparation Method in INRA82 or INRA96

Removal of seminal plasma facilitates stallion sperm survival during storage, but washing may damage sperm chromatin. Therefore, sperm quality was compared in samples following single-layer centrifugation (SLC) or sperm washing and controls (extension only) in two extenders, INRA82 and INRA96. Ejaculates from six stallions were split among six treatments: SLC, sperm washing, and controls, in INRA82 and INRA96. Sperm motility and acrosome status were evaluated at 0, 24, 48, 72 and 96 hours; morphology at 0, 24, 48, 72 hours and chromatin integrity at 0 and 96 hours, with storage at 6 degrees C. Sperm samples in INRA96 had better motility, acrosome status, and normal morphology than samples in INRA82. The SLC samples had higher motility and fewer reacted acrosomes than controls, and lower fragmented chromatin than washed samples. Fewer spermatozoa with tail defects were observed after SLC than after sperm washing; spermatozoa washed in INRA82 had fewer tail defects than those washed in INRA96. In conclusion, sperm quality (except for morphology) was better in INRA96 than in INRA82 and was better in SLC samples than in washed samples or controls. The SLC method is a useful adjunct to stallion sperm preparation, especially for storage before artificial insemination. (C) 2020 The Author(s). Published by Elsevier Inc

Impact of the severity of negative energy balance on gene expression in the subcutaneous adipose tissue of periparturient primiparous Holstein dairy cows: Identification of potential novel metabolic signals for the reproductive system

The severity of negative energy balance (NEB) in high-producing dairy cows has a high incidence among health diseases. The cow's energy status during early lactation critically affects metabolic and reproductive parameters. The first objective of this study was to investigate by RNA-seq analysis and RT-qPCR the gene expression profile in white adipose tissue and by gene ontology and upstream regulation tools the relationships with energy metabolism and reproduction in two groups of primiparous dairy cows with extreme NEB statuses (NEB &lt; -9 Mcal/day vs. NEB > -9 Mcal/day) around parturition. The second objective was to determine the potential involvement of a new adipokine identified as a candidate for the regulation of ovarian function in our RNA-seq analysis by using bovine primary granulosa culture, thymidine incorporation to determine cell proliferation and ELISA assays to measure progesterone secretion. The RNA-seq analysis revealed that 514 genes were over-expressed and 695 were under-expressed in the adipose tissue of cows with severe NEB (SNEB) and cows with moderate NEB (MNEB) during the -4 and 16 wkpp period. In addition, 491 genes were over-expressed and 705 genes were under-expressed in the adipose tissue of SNEB cows compared to MNEB cows. Among these differently expressed genes (DEGs), 298 were related to metabolic functions and 264 to reproductive traits. A set of 19 DEGs were validated by RT-qPCR, including CCL21 (C-C motif chemokine ligand 21). Moreover, CCL21, a gene known to be secreted by adipose tissue, was chosen for further analysis in plasma and ovaries. The use of next-generation sequencing technologies allowed us to characterise the transcriptome of white adipose tissue from primiparous cows with different levels of NEB during lactation. This study highlighted the alteration of the expression of genes related to lipid metabolism, including CCL21, which is released in the bloodstream and associated with the in vitro regulation of ovarian functions