Plasma cell-free DNA concentration increases during luteolysis in beef cows.

During cell death, DNA is fragmented and reaches the bloodstream in the form of cell-free DNA (cfDNA). Luteal cells must undergo an apoptotic process during structural luteolysis to begin a new oestrous cycle. We hypothesized that cfDNA concentrations would increase when inducing luteolysis by applying prostaglandin F2α (PGF2α) analog to the cycling cow. Multiparous non-pregnant and non-lactating Angus cows (Bos taurus; n = 15) were synchronized using the 7-day CoSynch + CIDR protocol. Ten days after oestrus was detected, two treatments were applied (PGF2α, n = 10; or Con, n = 5). Twice a day, grey mode and colour Doppler ultrasonography were used to calculate area (CL-A) and luteal blood perfusion (LBP%). Additionally, we collected one blood sample for plasma progesterone (P4) and cfDNA concentrations for four consecutive days. Data analysis was performed using the GLM procedure of SAS. The luteolysis induction was demonstrated by a decrease in P4 concentrations (p ≤ .01) and CL-A (p ≤ .01) in the PGF2α group after 12 h of the PGF2α injection. Reduction of LBP% (p ≤ .01) in the PGF2α group after 36 h of the injection. The concentration of cfDNA showed a significant increase (p = .05) after 48 h of the PGF2α application in the PGF2α group. In conclusion, cfDNA showed a significantly increased concentration after luteolysis induction, which can imply that cfDNA could be used as a luteolysis biomarker in plasma.

Actions of DKK1 on the preimplantation bovine embryo to affect pregnancy establishment, placental function, and postnatal phenotype†.

One mechanism by which the maternal environment regulates the early embryo is by secretion of cell-signaling molecules. One of these is dickkopf WNT signaling pathway inhibitor 1. Objectives were to (A) resolve discrepancies in the literature regarding effects of dickkopf WNT signaling pathway inhibitor 1 in the bovine embryo on development of trophectoderm and competence to establish pregnancy after embryo transfer and (B) determine whether there are long-term consequences of dickkopf WNT signaling pathway inhibitor 1 on placental function and postnatal phenotype. Embryos produced in vitro were cultured with vehicle or 100 ng/mL recombinant human dickkopf WNT signaling pathway inhibitor 1 from Days 5 to 7.5 of development (i.e., the morula and blastocyst stages of development). dickkopf WNT signaling pathway inhibitor 1 increased the number of cells positive for the trophectoderm marker CDX2 at Day 7.5 of development while having no effect on number of cells positive for the inner cell mass marker SOX2. There was no effect of dickkopf WNT signaling pathway inhibitor 1 on pregnancy or calving rate after transfer of blastocysts produced with Y-sorted semen to either lactating dairy cows or suckling beef cows. Treatment with dickkopf WNT signaling pathway inhibitor 1 at the morula-to-blastocyst stages programmed placental function, as measured by an effect of dickkopf WNT signaling pathway inhibitor 1 on plasma concentrations of pregnancy associated glycoproteins and placental lactogen at Day 160 of gestation (although not on other days examined). dickkopf WNT signaling pathway inhibitor 1 treatment also resulted in calves that were heavier at birth as compared to calves derived from control embryos. After birth, dickkopf WNT signaling pathway inhibitor 1 calves grew slower than controls. Results confirm that dickkopf WNT signaling pathway inhibitor 1 alters the developmental program of the bovine embryo to affect both prenatal and postnatal phenotypes.

Impact of Supplementing a Backgrounding Diet with Nonprotein Nitrogen on In Vitro Methane Production, Nutrient Digestibility, and Steer Performance.

Two experiments were conducted to evaluate the effect of nonprotein nitrogen (NPN) supplementation on in vitro fermentation and animal performance using a backgrounding diet. In experiment 1, incubations were conducted on three separate days (replicates). Treatments were control (CTL, without NPN), urea (U), urea-biuret (UB), and urea-biuret-nitrate (UBN) mixtures. Except for control, treatments were isonitrogenous using 1% U inclusion as a reference. Ruminal fluid was collected from two Angus-crossbred steers fed a backgrounding diet plus 100 g of a UBN mixture for at least 35 d. The concentration of volatile fatty acids (VFA) and ammonia nitrogen (NH3-N), in vitro organic matter digestibility (IVOMD), and total gas and methane (CH4) production were determined at 24 h of incubation. In experiment 2, 72 Angus-crossbred yearling steers (303 ±â€…29 kg of body weight [BW]) were stratified by BW and randomly allocated in nine pens (eight animals/pen and three pens/treatment). Steers consumed a backgrounding diet formulated to match the diet used in the in vitro fermentation experiment. Treatments were U, UB, and UBN and were isonitrogenous using 1% U inclusion as a reference. Steers were adapted to the NPN supplementation for 17 d. Then, digestibility evaluation was performed after 13 d of full NPN supplementation for 4 d using 36 steers (12 steers/treatment). After that, steer performance was evaluated for 56 d (24 steers/treatment). In experiment 1, NPN supplementation increased the concentration of NH3-N and VFA (P < 0.01) without affecting the IVOMD (P = 0.48), total gas (P = 0.51), and CH4 production (P = 0.57). Additionally, in vitro fermentation parameters did not differ (P > 0.05) among NPN sources. In experiment 2, NPN supplementation did not change dry matter and nutrient intake (P > 0.05). However, UB and UBN showed lower (P < 0.05) nutrient digestibility than U, except for starch (P = 0.20). Dry matter intake (P = 0.28), average daily gain (P = 0.88), and gain:feed (P = 0.63) did not differ among steers receiving NPN mixtures. In conclusion, tested NPN mixtures have the potential to be included in the backgrounding diets without any apparent negative effects on animal performance and warrant further studies to evaluate other variables to fully assess the response of feeding these novel NPN mixtures.

Nonprotein nitrogen (NPN) supplements can be used as a nitrogen source for ruminants fed low-protein diets. The most common NPN source is urea, included typically at a range between 0.5% and 1% of the diet dry matter in growing beef cattle. Although other NPN sources and mixtures are available, there is scarce information regarding their use in ruminant production. Two experiments were conducted to evaluate the effect of NPN sources on in vitro fermentation and animal performance using a backgrounding diet. In experiment 1, three different incubations were performed for 24 h. Treatments were control (without NPN), urea (U), urea­biuret (UB), and urea­biuret­nitrate (UBN) mixtures. In experiment 2, 72 crossbred yearling steers were randomly assigned to one of the following treatments: U, UB, and UBN mixtures. Diets were formulated to contain the same nitrogen concentration in both experiments. In experiment 1, supplementation of NPN increased the in vitro fermentation, but there were no differences among NPN sources. In experiment 2, steers performed similarly among NPN sources. These findings suggest that NPN mixtures have the potential to be included in the backgrounding diets without detrimental effects. Further studies should evaluate other variables (e.g., fermentation dynamic and microbial protein supply) when using these novel mixtures.

Extracellular vesicle-microRNAs mediated response of bovine ovaries to seasonal environmental changes.

BACKGROUND: Among the various seasonal environmental changes, elevated ambient temperature during the summer season is a main cause of stress in dairy and beef cows, leading to impaired reproductive function and fertility. Follicular fluid extracellular vesicles (FF-EVs) play an important role in intrafollicular cellular communication by, in part, mediating the deleterious effects of heat stress (HS). Here we aimed to investigate the changes in FF-EV miRNA cargoes in beef cows in response to seasonal changes: summer (SUM) compared to the winter (WIN) season using high throughput sequencing of FF-EV-coupled miRNAs. In addition to their biological relevance, the potential mechanisms involved in the packaging and release of those miRNAs as a response to environmental HS were elucidated. RESULTS: Sequencing analysis revealed that an average of 6.6% of the EV-RNA mapped reads were annotated to bovine miRNAs. Interestingly, miR-148a, miR-99a-5p, miR-10b, and miR-143 were the top four miRNAs in both groups accounting for approximately 52 and 62% of the total miRNA sequence reads in the SUM and WIN groups, respectively. A group of 16 miRNAs was up-regulated and 8 miRNAs were down-regulated in the SUM compared to the WIN group. Five DE-miRNAs (miR-10a, miR-10b, miR-26a, let-7f, and miR-1246) were among the top 20 expressed miRNA lists. Sequence motif analysis revealed the appearance of two specific motifs in 13 out of the 16 upregulated miRNAs under HS conditions. Both motifs were found to be potentially bonded by specific RNA binding proteins including Y-box binding proteins (YBX1 and YBX2) and RBM42. CONCLUSION: Our findings indicate that FF EV-coupled miRNA profile varies under seasonal changes. These miRNAs could be a good indicator of the cellular mechanism in mediating HS response and the potential interplay between miRNA motifs and RNA binding proteins can be one of the mechanisms governing the packaging and release of miRNAs via EVs to facilitate cellular survival.

Effects of Aspergillus oryzae prebiotic on animal performance, nutrients digestibility, and feeding behavior of backgrounding beef heifers fed with either a sorghum silage- or a byproducts-based diet.

Eighty-four Angus crossbred heifers (13 ± 1 mo of age, 329.5 ± 61.92 kg of body weight [BW]) were used in a generalized randomized block design with a 2 × 2 factorial arrangement of treatments. The factors evaluated were: 1) diet type (whole plant sorghum silage [SS] vs. byproducts-based [BP]), and 2) feed additive: Aspergillus oryzae prebiotic (AOP; 2 g/d) vs. Negative control (CTL; 0 g/d), resulting in four treatments: sorghum silage-control (SC), sorghum silage-AOP (SA), byproducts-control (BC), and byproducts-AOP (BA). Heifers were stratified by body weight (BW), randomly assigned to treatments (21 heifers per treatment) and housed in 12 pens equipped with two GrowSafe feed bunks each to measure individual dry matter intake (DMI). After a 14-d adaptation, BW was measured every 14 d for 56 d. Chewing activity was monitored through collar-mounted HR-Tags (heat-related tags). Following the performance period, apparent total tract digestibility was measured in 40 heifers, using indigestible neutral detergent fiber as a marker. Heifers fed with the BP diets had greater DMI (2.92% vs. 2.59% of BW, P < 0.01) and average daily gain (ADG; 1.16 vs. 0.68 kg, P ≤ 0.01) than heifers fed with SS diets. Compared with BP-fed animals, heifers consuming the SS diets had 23 more visits/d to the feed bunks (P ≤ 0.01), consumed 53% less dry matter on each visit (P ≤ 0.01), and spent 39% more min chewing/d and 63% more min chewing/kg of DMI (P ≤ 0.01). However, chewing measured in min/kg of neutral detergent fiber intake was not affected by treatment (average 111.3 min/kg of NDF intake). Feeding AOP improved gain:feed (GF) by 15% in BP-fed heifers (0.120 vs. 0.104 kg/kg; P < 0.05). Inclusion of AOP increased organic matter digestibility (OMD) in SS diets (55.88% vs. 49.83%; P < 0.01), whereas it decreased OMD in BP diets (61.67% vs. 65.77%; P < 0.05). In conclusion, ADG and GF of BP-fed heifers was greater than SS-fed heifers, and GF was greater with AOP supplementation in BP-fed heifers. Improvement in GF in BP-fed heifers was likely not related to differences in nutrient digestibility as AOP inclusion did not enhance digestibility in the BP diet. Additionally, the effects of the AOP inclusion appear to be diet-dependent, where the 15% improvement in GF by AOP occurred in heifers fed with the more fermentable diet. Therefore, further research should explore the mechanisms responsible for the observed improvements in growth performance when feeding AOP to BP-fed heifers.

This experiment evaluated the effects of the dietary inclusion or not of Aspergillus oryzae prebiotic (AOP; 2 g/d) in two contrasting diets: sorghum silage-based (SS) vs. byproducts-based (BP), on growth performance, nutrient digestibility, and feeding behavior of growing heifers. A total of 84 Angus crossbred heifers were used in the study. Heifers fed with the BP diets had greater feed intake, average daily gain, and final body weight. In addition, heifers fed with the BP diets had reduced number of visits to the feed bunk but consumed more in each visit than heifers fed with the SS diets. Additionally, heifers fed with the BP diets had lesser chewing activity measured in total min/d and in min/kg of dry matter intake; however, chewing activity measured in min/kg of neutral detergent fiber was not influenced by treatments. The inclusion of AOP increased the gain:feed ratio by 15% in heifers fed with the BP diet but did not influence this variable in the SS diet. The inclusion of AOP increased nutrient digestibility in heifers fed with the SS diet and decreased nutrient digestibility in heifers fed with the BP diet. These results show that feeding AOP can enhance growth performance in beef heifers in a diet-dependent manner.

Evaluation of carinata meal or cottonseed meal as protein sources in silage-based diets on behavior, nutrient digestibility, and performance in backgrounding beef heifers.

Changing climatic conditions are imposing risks and diminishing yields in agriculture. Sorghum (Sorghum bicolor) silage is a feasible option for backgrounding beef cattle in terms of economic risk management and animal productivity when compared with corn (Zea mays) silage, due to its drought adaptability. Similarly, Brassica carinata meal has proven to be a viable alternative as a protein supplement in forage-based beef cattle systems, when included at 10% of the diet dry matter (DM). However, research is scarce regarding its inclusion in silage-based diets for backgrounding animals. The objective of this trial was to compare a processor-chopped sorghum silage (SS) against a typical corn silage (CS) in a digestibility and performance trial while supplementing two protein sources; one traditionally used like cottonseed meal (CSM) and one novel like B. carinata meal (BCM). A total of 84 Angus crossbred heifers (307 ± 33 kg BW) were evaluated in a randomized block design with a 2 × 2 factorial treatment arrangement with type of silage and protein source as factors. Diets were fed ad libitum, consisting of 89% silage source plus 10% protein source, and 1% mineral inclusion on DM basis. The experimental period consisted of 14 d of adaptation followed by 5 d of apparent total tract digestibility measurements and 56 d of animal performance and intake behavior measurements. Heifers fed SS showed greater number of daily meals but decreased meal sizes (P ≤ 0.05), not differing in meal length (P > 0.10) when compared with CS. Dry matter and organic matter (OM) digestibility showed a silage type × protein source interaction (P ≤ 0.01), where in CS diets, OM tended to be more digestible with CSM vs. BCM, and it did not differ between protein sources in SS based diets. There was an effect of protein (P ≤ 0.01) on ADF digestibility, where CSM was greater than BCM. No effect of treatment was observed (P ≥ 0.10) on DM intake. Average daily gain (ADG) and gain-to-feed ratio were greater for CS than SS (P ≤ 0.01) regardless of protein source. Although heifers fed CS had greater feed efficiency and digestibility, SS can still be considered a viable option for backgrounding beef heifers, obtaining adequate ADG rates of 0.945 kg/d. Lastly, BCM did not differ from CSM in terms of feed efficiency and animal performance, proving to be a viable alternative protein source in silage-based diets.

Increased atmospheric CO2, rising temperatures, and altered patterns of precipitation can limit the production of certain crops commonly used in agriculture, increasing risk, cost, and availability of feedstuffs. The search for alternative plants that could thrive in these changing scenarios is necessary to provide producers with a broader array of options to feed cattle. In this study, sorghum silage was compared with corn silage as the main dietary ingredient, with either Brassica carinata (carinata) or cottonseed meal as the protein source for growing beef heifers. Variables assessed included intake behavior, digestibility, and performance of beef heifers. Heifers fed sorghum silage gained less than heifers fed corn silage, though they grew at an adequate rate for a replacement heifer. Carinata meal showed similar performance results compared with cottonseed meal, despite some of its fiber components being less digestible in the total tract. Therefore, sorghum silage has potential to be a viable feedstuff for growing beef heifers although it may result in decreased performance compared with corn silage. Alternatively, carinata meal can be a practical alternative to cottonseed meal as a protein source in terms of animal performance. This could translate in an increase in the planted area of both sorghum and carinata in Southern United States, as they are adapted to drought and high temperatures, enhancing the resilience of beef production systems in a context of increased climate variability.