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Data on metabolic profiles of blood sampled at d 3, 6, 9, and 21 in lactation from 117 lactations (99 cows) were used for unsupervised k-means clustering. Blood metabolic parameters included β-hydroxybutyrate (BHB), nonesterified fatty acids, glucose, insulin-like growth factor-1 (IGF-1) and insulin. Clustering relied on the average and range of the 5 blood parameters of all 4 sampling days. The clusters were labeled as imbalanced (n = 42) and balanced (n = 72) metabolic status based on the values of the blood parameters. Various random forest models were built to predict the metabolic cluster of cows during early lactation from the milk composition. All the models were evaluated using a leave-group-out cross-validation, meaning data from a single cow were always present in either train or test data to avoid any data leakage. Features were either milk fatty acids (MFA) determined by gas chromatography (MFA [GC]) or features that could be determined during a routine dairy herd improvement (DHI) analysis, such as concentration of fat, protein, lactose, fat/protein ratio, urea, and somatic cell count (determined and reported routinely in DHI registrations), either or not in combination with MFA and BHB determined by mid-infrared (MIR), denoted as MFA [MIR] and BHB [MIR], respectively, which are routinely analyzed but not routinely reported in DHI registrations yet. Models solely based on fat, protein, lactose, fat/protein ratio, urea and somatic cell count (i.e., DHI model) were characterized by the lowest predictive performance [area under the receiver operating characteristic curve (AUCROC) = 0.69]. The combination of the features of the DHI model with BHB [MIR] and MFA [MIR] powerfully increased the predictive performance (AUCROC = 0.81). The model based on the detailed MFA profile determined by GC analysis did not outperform (AUCROC = 0.81) the model using the DHI-features in combination with BHB [MIR] and MFA [MIR]. Predictions solely based on samples at d 3 were characterized by lower performance (AUCROC DHI + BHB [MIR] + MFA [MIR] model at d 3: 0.75; AUCROC MFA [GC] model at d 3: 0.73). High predictive performance was found using samples from d 9 and 21. To conclude, overall, the DHI + BHB [MIR] + MFA [MIR] model allowed to predict metabolic status during early lactation. Accordingly, these parameters show potential for routine prediction of metabolic status.

Both ruminal microbial structure and functionality might play a role in inter-individual variation in susceptibility for subacute rumen acidosis (SARA) observed in dairy cows. The aims of this study were to determine whether differences between cows with distinct SARA susceptibility were reflected in distinct (1) ruminal microbial communities, (2) salivary bacterial communities, and (3) fermentative capacity of ruminal microbiota assessed in vitro.To test this hypothesis, rumen samples were collected via an esophageal tube on 21 d postpartum from 38 multiparous Holstein cows, which were classified into 4 groups differing in median and mean time of reticular pH below 6 as well as area under the curve of pH below 6.0. During the 21 d postpartum, all cows within a group fulfilled following criteria: susceptible (S, n = 10; mean or median ≥180 min/d), moderately susceptible (MS, n = 7; 60 min/d < mean time of pH below 6 < 180 min/d, and median time of pH below 6 Streptococcus,Sharpea,Prevotellaceae_YAB2003,Succinivibrionaceae_UCG-001,Ruminococcus, andRuminococcaceae_UCG-001were higher in S compared with U cows. In contrast,Lachnospiraceae_ND3007andOscillospiraceae_V9D2013were more abundant in U cows. Although pH-associated, inter-animal differences were also observed in the salivary bacteria, common differences in ruminal and salivary bacterial genera were limited. The functionality of the rumen microbiota was evaluated in vitro through exposure of the microbial inoculum of S and U cows to an anaerobic buffer at pH 5.8 and 6.8, in the presence of sterile supernatant of their own and of dry cows' rumen fluid (2 × 2 design). Generally, the S inoculum produced more volatile fatty acids, except at low pH with dry cows' supernatant, where volatile fatty acid production was completely impaired and lactate accumulation was highest. Compared with the microbes of U cows, microbes of S cows showed less fermentative activity in situations with 2 stress factors (low pH and an unfamiliar environment, i.e., rumen fluid supernatant of dry cows).

Both ruminal microbial structure and functionality might play a role in inter-individual variation in susceptibility for subacute rumen acidosis (SARA) observed in dairy cows. The aims of this study were to determine whether differences between cows with distinct SARA susceptibility were reflected in distinct (1) ruminal microbial communities, (2) salivary bacterial communities, and (3) fermentative capacity of ruminal microbiota assessed in vitro. To test this hypothesis, rumen samples were collected via an esophageal tube on 21 d postpartum from 38 multiparous Holstein cows, which were classified into 4 groups differing in median and mean time of reticular pH below 6 as well as area under the curve of pH below 6.0. During the 21 d postpartum, all cows within a group fulfilled following criteria: susceptible (S, n = 10; mean or median >= 180 min/d), moderately susceptible (MS, n = 7; 60 min/d < mean time of pH below 6 < 180 min/d, and median time of pH below 6

Funding Acknowledgements No disclosure Introduction. Pulmonary vein (PV) firing can trigger or act as a rapid driver to maintain atrial fibrillation (AF). Automated 1-minute measurement of fragmented and non-fragmented PV activity cycle length (CL) showed contradictory results to predict ablation outcome in persistent AF. Purpose. This study investigated the reproducibility of a novel non-automated simple method to measure non-fragmented fastest discrete consecutive AF signal cycle length and the value of this measurement in the PVs to predict long-term success after pulmonary vein isolation (PVI) only ablation in persistent AF. Methods. Consecutive 75 patients with persistent AF undergoing first-time PVI between 2015 and 2018 were included. The mean of 10 FAstest Repetitive Similar morphology discrete signal cycle lengths (FARS-AF CL) were measured twice with > 2 minutes between in the coronary sinus (CS), superior vena cava (SVC), left and right atrial appendage (LAA, RAA) and PVs. FARS discrete AF signals were defined as (I) signal duration ≤80 msec; (II) repetitive similar morphological characteristics; (III) fastest consecutive 10 intervals during 1-minute observation. The reproducibility of the FARS–AF CL measurement was compared to traditional 10 consecutive interval measurements of fragmented CS signals. The CL gradient between the PV and the LA was quantified by the computing the ratio of the PV and LAA or CS CL. Results. Good correlation was found between two FARS CL measurements in the CS, PVs, LAA and RAA (Correlation Kendall area: 0.882, 0.675-0.941, 0.859, 0.944, respectively). The correlation between two traditional CL measurements of fragmented CS signals was low (Correlation Kendall area:-0.006). After a mean follow-up of 20 months, freedom from atrial arrhythmias was achieved in 50 (66%) patients after the single PVI procedure with or without the use of AADs. Patient without recurrence were more likely to have FARS CL ≤140 msec (8 vs. 42%, p= 0.002), higher FARS PV CL/LAA CL and FARS PV CL/CS CL ratio (96 ± 13% vs. 86 ± 23 %, p= 0.04; 95 ± 13% vs. 82 ± 22%, p= 0.036). Patients with recurrence at follow-up had more dilated left atria (LAVI: 44 ± 12 vs. 38 ± 9 ml/m2, p= 0.02; LA diameter: 49 ± 6 vs. 45 ± 6 mm, p= 0.01), less AF termination during the procedure (16 vs. 37%, p= 0.049) and less first pass isolation (44 vs. 68%, p= 0.04). Multivariable Cox regression analysis showed that LAVI (p = 0.035) and FARS-PV CL (0.011) were significant predictors of arrhythmia free survival. After adjusting for LAVI, FARS-PV CL remained a significant predictor of AF recurrence (p = 0.028). Conclusions. Traditional non-automated AF CL measurement of fragmented CS signals is poorly reproducible. FARS-AF CL measurements in the PVs, RA and LA structures are highly reproducible. FARS-AF CL measurement in the PVs could predict the success of PVI-only procedure in persistent AF independent of left atrial size.

This study aimed to investigate the effect of pre and/or postnatal supplementation of a dry whole cell algae (DHA-Gold) to goat kids, on in vitro methane (CH(4)) production, animal growth, and rumen morphology at the age of 6 mo. Furthermore, the in vitro retreatment effect of DHA-Gold was evaluated. Twenty pregnant Saanen goats giving birth to 2 male kids were used. Half of these does were supplemented (D+) with 18.2 g/d of DHA-Gold in the last 3 wk of pregnancy, whereas the other half was not (D−). After kidding, one goat kid per doe in both groups was supplemented daily with 0.28 g/kg of body weight of DHA-Gold (k+) until 12 wk, whereas the other goat kids were untreated (k−). This resulted in 4 experimental groups D+k+, D+k−, D−k+, and D−k−. In vitro incubations were performed at the ages of 4 wk, 11 wk, and 6 mo. At the age of 6 mo, goat kids were euthanized and additional incubations were performed supplementing 4 doses of DHA-Gold (0, 0.4, 0.8, and 1.6 mg/mL). Additionally, rumen tissue of the atrium ruminis, ventral rumen, and dorsal blind sac were collected to assess rumen morphology. Rumen inocula of 4-wk-old goat kids supplemented D+ showed lower (P < 0.05) in vitro CH(4) production, however, this was mainly due to a reduction in the overall fermentation, while CH(4) expressed relatively to total volatile fatty acids (VFA) was higher when goat kids were treated D+ or k+. The detrimental D+ effect on VFA production diminished at 11 wk old but remained a tendency (0.05 < P < 0.1). As for 4 wk D+ as well as k+ supplementation of DHA-Gold stimulated rather than inhibited in vitro CH(4) production expressed relative to total VFA. Supplementation of DHA-Gold either D+ or k+ decreased density, width, and surface area of the ruminal papillae. However, no effect on animal growth was observed. Moreover, detrimental effects of D+ or k+ treatment on VFA production or stimulation of relative CH(4) production were no longer observed at 6 mo old. Nevertheless, direct exposure of DHA-Gold to 6-mo-old inoculum linearly (P < 0.05) decreased CH(4) and VFA production, which tended (P = 0.06) to be greater when using D-rumen inoculum. Accordingly, neither D+ nor k+ DHA-Gold supplementation showed potential for reduction of rumen methanogenesis. Furthermore, this early life intervention could represent some risk for impaired rumen papillae development, which, however, did not impair animal performance.

The aim of this study was to evaluate the dose response on in vitro methane (CH) production of PUFA to which the inoculum donor animals had been exposed early in life. Sixteen Holstein calves (160 ± 3 and 365 ± 2 kg BW) at 6 and 12 mo of age were used as inoculum donors. Half of the calves were given increasing amounts of extruded linseed from birth (22 g/d) until 4 mo of age (578 g/d) first mixed with milk and then included in their concentrate. Linseed oil (LSO) was supplemented in vitro at 5 different doses (0, 0.6, 1.2, 2.4, and 4.8 mg/mL). Supplementation of LSO in the rumen inocula at both ages linearly decreased (0.05) the in vitro CH production. Total in vitro VFA production was not affected by LSO supplementation. Inhibition of CH was smaller when using the rumen inoculum from calves that had received a similar treatment early in life (0.05). Differences in response to in vitro supplementation of a type of fatty acids similar to those applied during early life suggest some "changes" in the functioning of the rumen microbial community.

Within Annex 23 (`Multizone Air Flow Modelling') of the IEA (International Energy Agency), a large part of the effort went to the evaluation of the developed simulation tool COMIS [J.-M. Furbringer, C.-A. Roulet, R. Borchiellini, Evaluation of COMIS, IEA Annex 23: Multizone Air Flow Modelling, LESO-EPFL, Lausanne, 1996; J.-M. Furbringer, C.-A. Roulet, R. Borchiellini, Evaluation of COMIS—appendices, IEA Annex 23: Multizone Air Flow Modelling, LESO-EPFL, Lausanne, 1996]. The purpose of this work was to examine the reliability of the output of the program and also the impact of the input parameters on the simulation result. Therefore, the program was tested in different ways, for several cases with sometimes strongly different input parameters. Two cases were examined in detail by the BBRI (Belgian Building Research Institute): • Spread of contaminants in an apartment with natural ventilation: this case will be discussed in this text. • Single-sided ventilation in a PASSYS-test cell. Thanks to all these different simulations, one can have an increased confidence in the simulation tool COMIS. More exactly, it gives an idea of the possibilities and limitations of the tool.

The main purpose for human assisted reproductive technologies (ART) is clearly to bypass reproductive failures. This purpose is totally different from the aims in animal ART, being genetic improvement and more recently conservation of endangered species. Apoptosis or programmed cell death has been detected before implantation in in vivo as well as in vitro embryos and might contribute to lower developmental competence and embryonic losses of in vitro embryos. In vitro embryo quality is clearly jeopardized by quality of the gametes (oocytes and spermatozoa) and suboptimal culture condition, but there is no consensus on which factor is most important in the incidence of apoptosis during early embryo development. The objective of the present study was to unravel the contribution of both male and female gamete quality to the incidence of apoptosis of bovine embryos produced in vitro. In vivo bull fertility expressed a very low correlation with cleavage rate and blastocyst yield. In contrast, developmental kinetics and oocyte diameter are important markers of embryo developmental potential and embryo quality in terms of the appearance of apoptosis, indicating a significant maternal effect on embryo quality.

Immunofluorescent staining is often used to investigate the expression of specific proteins in pre-implantation embryos. The success of this method is determined by the specificity of the antibodies, but also by the protocol used for fixation and permeabilization of the samples. In this study, different fixatives are compared in combination with immunofluorescent staining of caudal-type homeobox 2 (CDX2), fibronectin 1 (FN1) and integrins (ITGs) on bovine blastocysts. For both CDX2 and the ITGs, the outcome of the staining was largely dependent on the fixation methods. Paraformaldehyde fixation was best for the intracellular CDX2 protein, whereas acetone fixation gave the best results for the transmembrane ITGs. No difference was observed for the FN1 staining between samples fixed with paraformaldehyde or acetone. These examples demonstrate that the choice of fixation and permeabilization agents is very important for the outcome of the experiment, and this choice is dictated by the (extra)cellular location of the protein under investigation. Inappropriate fixation and/or permeabilization methods can lead to erroneous conclusions regarding the site and amount of protein expression.

The CEC research project PASSYS (PASsive Solar Components and SYStems Testing, 1986–1992) is using high-grade outdoor test cells for testing the thermal and solar performances of building components as well as for model validation purposes. The test facilities are highly standardized: identical test cells, equipped with the same sensors, heating and cooling system and data acquisition hardware and software, are installed at 12 test sites throughout Europe, applying the same test method. A severe quality assurance policy is being developed in order to ensure reliable and comparable results. This paper describes the objectives, organization and main themes of the project. Two further papers report results.

In this work, we studied the incidence of DNA fragmentation, interpreted as apoptotic changes and assessed by the TUNEL assay, in cumulus cells and oocytes of immature Grade 1 cumulus-oocyte complexes (COCs) obtained from healthy bitches (n = 27) of three age groups: young (1-3 years; n = 13), adult (4-6 years; n = 8) and elderly (7-10 years; n = 6). Age affected (p 0.05) Grade 1 COCs recovery rates, with young animals yielding more (p 0.01) Grade 1 COCs than the other two age groups. Conversely, no differences were observed in the incidence of DNA fragmentation (TUNEL-positive) in cumulus cells or oocytes between the three age groups. Overall, more than 80% of Grade 1 COCs presented15% of TUNEL-positive cumulus cells and enclosed TUNEL-negative (intact DNA) oocytes. Despite a higher proportion of TUNEL-negative oocytes being found in the germinal vesicle stage, most of the oocytes with nuclear material compatible with meiosis resumption (MR) or with non-identifiable nuclear material (ND) did not present DNA fragmentation. No correlation was observed between DNA fragmentations in oocytes and in cumulus cells. We concluded that the morphological parameters used to classify canine Grade 1 COCs are reliable to select a homogeneous population of COCs with low incidence of DNA fragmentation. Furthermore, these results indicate that DNA fragmentation can only explain a minor proportion of the incidence of MR and degeneration in canine oocytes at collection.

This paper will briefly discuss the value of MRI in the detection and follow-up of bone marrow edema resulting from acute and chronic trauma in sports injuries. A meticulous pattern approach of the distribution of bone marrow changes around a joint can reveal in most instances the underlying mechanism of trauma. The use of fatsuppressed T2-weighted or STIR images is most appropriate to detect bone marrow edema.

The CT findings in a case of persistent sciatic artery complicated with an aneurysm in the gluteal area are reported. If on CT an aneurysm is found in the gluteal area, the possibility of an underlying persistent sciatic artery should be considered.