Prediction of key milk biomarkers in dairy cows through milk mid-infrared spectra and international collaborations.

At the individual cow level, suboptimum fertility, mastitis, negative energy balance, and ketosis are major issues in dairy farming. These problems are widespread on dairy farms and have an important economic impact. The objectives of this study were (1) to assess the potential of milk mid-infrared (MIR) spectra to predict key biomarkers of energy deficit (citrate, isocitrate, glucose-6 phosphate [glucose-6P], free glucose), ketosis (ß-hydroxybutyrate [BHB] and acetone), mastitis (N-acetyl-ß-d-glucosaminidase activity [NAGase] and lactate dehydrogenase), and fertility (progesterone); (2) to test alternative methodologies to partial least squares (PLS) regression to better account for the specific asymmetric distribution of the biomarkers; and (3) to create robust models by merging large datasets from 5 international or national projects. Benefiting from this international collaboration, the dataset comprised a total of 9,143 milk samples from 3,758 cows located in 589 herds across 10 countries and represented 7 breeds. The samples were analyzed by reference chemistry for biomarker contents, whereas the MIR analyses were performed on 30 instruments from different models and brands, with spectra harmonized into a common format. Four quantitative methodologies were evaluated to address the strongly skewed distribution of some biomarkers. Partial least squares regression was used as the reference basis, and compared with a random modification of distribution associated with PLS (random-downsampling-PLS), an optimized modification of distribution associated with PLS (KennardStone-downsampling-PLS), and support vector machine (SVM). When the ability of MIR to predict biomarkers was too low for quantification, different qualitative methodologies were tested to discriminate low versus high values of biomarkers. For each biomarker, 20% of the herds were randomly removed within all countries to be used as the validation dataset. The remaining 80% of herds were used as the calibration dataset. In calibration, the 3 alternative methodologies outperform the PLS performances for the majority of biomarkers. However, in the external herd validation, PLS provided the best results for isocitrate, glucose-6P, free glucose, and lactate dehydrogenase (coefficient of determination in external herd validation [R2v] = 0.48, 0.58, 0.28, and 0.24, respectively). For other molecules, PLS-random-downsampling and PLS-KennardStone-downsampling outperformed PLS in the majority of cases, but the best results were provided by SVM for citrate, BHB, acetone, NAGase, and progesterone (R2v = 0.94, 0.58, 0.76, 0.68, and 0.15, respectively). Hence, PLS and SVM based on the entire dataset provided the best results for normal and skewed distributions, respectively. Complementary to the quantitative methods, the qualitative discriminant models enabled the discrimination of high and low values for BHB, acetone, and NAGase with a global accuracy around 90%, and glucose-6P with an accuracy of 83%. In conclusion, MIR spectra of milk can enable quantitative screening of citrate as a biomarker of energy deficit and discrimination of low and high values of BHB, acetone, and NAGase, as biomarkers of ketosis and mastitis. Finally, progesterone could not be predicted with sufficient accuracy from milk MIR spectra to be further considered. Consequently, MIR spectrometry can bring valuable information regarding the occurrence of energy deficit, ketosis, and mastitis in dairy cows, which in turn have major influences on their fertility and survival.

Multivariate analysis of milk metabolite measures shows potential for deriving new resilience phenotypes.

In a context of growing interest in breeding more resilient animals, a noninvasive indicator of resilience would be very valuable. We hypothesized that the time-course of concentrations of several milk metabolites through a short-term underfeeding challenge could reflect the variation of resilience mechanisms to such a challenge. We submitted 138 one-year-old primiparous goats, selected for extreme functional longevity (i.e., productive longevity corrected for milk yield [60 low longevity line goats and 78 high longevity line goats]), to a 2-d underfeeding challenge during early lactation. We measured the concentration of 13 milk metabolites and the activity of 1 enzyme during prechallenge, challenge, and recovery periods. Functional principal component analysis summarized the trends of milk metabolite concentration over time efficiently without preliminary assumptions concerning the shapes of the curves. We first ran a supervised prediction of the longevity line of the goats based on the milk metabolite curves. The partial least square analysis could not predict the longevity line accurately. We thus decided to explore the large overall variability of milk metabolite curves with an unsupervised clustering. The large year × facility effect on the metabolite concentrations was precorrected for. This resulted in 3 clusters of goats defined by different metabolic responses to underfeeding. The cluster that showed higher ß-hydroxybutyrate, cholesterol, and triacylglycerols increase during the underfeeding challenge was associated with poorer survival compared with the other 2 clusters. These results suggest that multivariate analysis of noninvasive milk measures show potential for deriving new resilience phenotypes.

Milk metabolites and fatty acids as noninvasive biomarkers of metabolic status and energy balance in early-lactation cows.

The objective was to study the effects of week of lactation (WOL) and experimental nutrient restriction on concentrations of selected milk metabolites and fatty acids (FA), and assess their potential as biomarkers of energy status in early-lactation cows. To study WOL effects, 17 multiparous Holstein cows were phenotyped from calving until 7 WOL while allowed ad libitum intake of a lactation diet. Further, to study the effects of nutrient restriction, 8 of these cows received a diet containing 48% straw (high-straw) for 4 d starting at 24 ± 3 days in milk (mean ± SD), and 8 cows maintained on the lactation diet were sampled to serve as controls. Blood and milk samples were collected weekly for the WOL data set, and daily from d -1 to 3 of nutrient restriction (or control) for the nutritional challenge data set. Milk ß-hydroxybutyrate (BHB), isocitrate, glucose, glucose-6-phosphate (glucose-6P), galactose, glutamate, creatinine, uric acid, and N-acetyl-ß-d-glucosaminidase activity (NAGase) were analyzed in p.m. and a.m. samples, and milk FA were analyzed in pooled p.m. and a.m. samples. Average energy balance (EB) per day ranged from -27 MJ/d to neutral when cows received the lactation total mixed ration, and from -109 to -87 ± 7 MJ/d for high-straw (least squares means ± standard error of the mean). Plasma nonesterified FA concentration was 1.67 ± 0.13 mM and BHB was 2.96 ± 0.39 mM on the d 3 of high-straw (least squares means ± standard error of the mean). Milk concentrations of BHB, glucose, glucose-6P, glutamate, and uric acid differed significantly between p.m. and a.m. milkings. Milk isocitrate, glucose-6P, creatinine, and NAGase decreased, whereas milk glucose and galactose increased with WOL. Changes in milk BHB, isocitrate, glucose, glucose-6P, and creatinine were concordant during early lactation and in response to nutrient restriction. Milk galactose and NAGase were modulated by WOL only, whereas glutamate and uric acid concentrations responded to nutrient restriction only. The high-straw increased milk concentrations of FA potentially mobilized from adipose tissue (e.g., C18:0 and cis-9 C18:1 and sum of odd- and branched-chain FA (OBCFA) with carbon chain greater than 16; ∑ OBCFA >C16), and decreased concentrations of FA synthesized de novo by the mammary gland (e.g., sum of FA with 6 to 15 carbons; ∑ C6:0 to C15:0). Similar observations were made during early lactation. Plasma nonesterified FA concentrations had the best single linear regression with EB (R2 = 0.62). Milk isocitrate, Σ C6:0 to C15:0. and cis-9 C18:1 had the best single linear regressions with EB (R2 ≥ 0.44). Milk BHB, isocitrate, galactose, glutamate, and creatinine explained up to 64% of the EB variation observed in the current study using multiple linear regression. Milk concentrations of ∑ C6:0 to C15:0, C18:0, cis-9 C18:1, and ∑ OBCFA >C16 presented some of the best correlations and regressions with other indicators of metabolic status, lipomobilization, and EB, and their responses were concordant during early lactation and during experimental nutrient restriction. Metabolites and FA secreted in milk may serve as noninvasive indicators of metabolic status and EB of early-lactation cows.

Elements in Mental Health Interventions Associated with Effects on Emotion Regulation in Adolescents: A Meta-Analysis.

PURPOSE: Adolescence is a sensitive period for developing mental health problems. Interventions targeting emotion regulation have shown promising transdiagnostic effects for this group, but optimization efforts are called for. In the current study, we used an element-based approach to identify potentially active ingredients in interventions measuring emotion regulation, to guide further optimization. METHODS: We coded practice elements in 30 studies based on a systematic review of mental health interventions measuring emotion regulation in adolescents (N = 2,389 participants, mean age 13-17 years). Using a three-level modeling approach, we then investigated the difference in effect on emotion regulation between studies of interventions with and without these practice elements. RESULTS: We identified 75 practice elements and 15 element categories used in the included interventions. Results showed significantly stronger effects on emotion regulation when interventions included the practice elements Setting goals for treatment (difference in d = 0.40, 95% CI [0.09, 0.70], p = .012) and Psychoeducation about acceptance (difference in d = 0.58, 95% CI [0.09, 1.07], p = .021). Furthermore, a total of 11 elements and four overall categories were identified as potentially active ingredients, based on an effect size difference of > 0.20 between interventions with and without the elements. CONCLUSION: The results can direct experimental research into the selection of practices that are most likely key to mechanisms of change in interventions addressing emotion regulation for adolescents. The challenge of measuring emotion regulation is discussed.

Developmental stage and morphology of the competent blastocyst are associated with sex of the child but not with other obstetric outcomes: a multicenter cohort study.

STUDY QUESTION: Are transfer day, developmental stage and morphology of the competent blastocyst in pregnancies leading to live birth associated with preterm birth, birthweight, length at birth and sex of the child? SUMMARY ANSWER: A high score in blastocyst developmental stage and in trophectoderm (TE) showed a significant association with the sex of the child, while no other associations with obstetric outcomes were observed. WHAT IS KNOWN ALREADY: The association between blastocyst assessment scores and obstetric outcomes have been reported in small single-center studies and the results are conflicting. STUDY DESIGN, SIZE, DURATION: Multicenter historical cohort study based on exposure data (transfer day (blastocyst developmental stage reached by Day 5 or Day 6)) blastocyst developmental stage (1-6) and morphology (TE and inner cell mass (ICM): A, B, C)) and outcome data (preterm birth, birthweight, length at birth, and sex of the child) from women undergoing single blastocyst transfer resulting in a singleton pregnancy and live birth. PARTICIPANTS/MATERIALS, SETTING, METHODS: Data from 16 private and university-based facilities for clinical services and research were used. A total of 7246 women, who in 2014-2018 underwent fresh-embryo transfer with a single blastocyst or frozen-thawed embryo transfer (FET) with a single blastocyst resulting in a singleton pregnancy were identified. Linking to the Danish Medical Birth Registry resulted in a total of 4842 women with a live birth being included. Cycles with pre-implantation genetic testing and donated gametes were excluded. The analyses were adjusted for female age (n = 4842), female BMI (n = 4302), female smoking (n = 4290), parity (n = 4365), infertility diagnosis (n = 4765), type of treatment (n = 4842) and center (n = 4842); some analyses additionally included gestational age (n = 4368) and sex of the child (n = 4833). MAIN RESULTS AND THE ROLE OF CHANCE: No statistically significant associations between blastocyst assessment scores (transfer day, developmental stage, TE, ICM) and preterm birth (8.3%) or birthweight (mean 3461.7 g) were found. The adjusted association between blastocysts with a TE score of C and a TE score of A and length at birth (mean 51.6 cm) were statistically significant (adjusted mean difference 0.4 cm (95% CI: 0.02; 0.77)). Blastocysts transferred with developmental stage score 5 compared to blastocysts transferred with score 3 had a 34% increased probability of being a boy (odds ratio (OR) 1.34 (95% CI: 1.09; 1.64). Further, TE score B blastocysts compared to TE score A blastocysts had a 31% reduced probability of being a boy (OR 0.69 (95% CI: 0.60; 0.80)). LIMITATIONS, REASONS FOR CAUTION: It is possible that some residual confounding remains. WIDER IMPLICATIONS OF THE FINDINGS: Blastocyst selection during ART does not appear to introduce any negative effects on obstetric outcome. Therefore, clinicians and patients can be reassured that the assessment scores of the selected blastocyst will not in themselves pose a risk of preterm birth or affect birthweight and the length at birth. STUDY FUNDING/COMPETING INTEREST(S): Unrestricted grant from Gedeon Richter Nordics AB, Sweden. None of the authors have any competing interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Andreev Modes from Phase Winding in a Full-Shell Nanowire-Based Transmon.

We investigate transmon qubits made from semiconductor nanowires with a fully surrounding superconducting shell. In the regime of reentrant superconductivity associated with the destructive Little-Parks effect, numerous coherent transitions are observed in the first reentrant lobe, where the shell carries 2π winding of superconducting phase, and are absent in the zeroth lobe. As junction density was increased by gate voltage, qubit coherence was suppressed then lost in the first lobe. These observations and numerical simulations highlight the role of winding-induced Andreev states in the junction.

Fungal quinones: diversity, producers, and applications of quinones from Aspergillus, Penicillium, Talaromyces, Fusarium, and Arthrinium.

Quinones represent an important group of highly structurally diverse, mainly polyketide-derived secondary metabolites widely distributed among filamentous fungi. Many quinones have been reported to have important biological functions such as inhibition of bacteria or repression of the immune response in insects. Other quinones, such as ubiquinones are known to be essential molecules in cellular respiration, and many quinones are known to protect their producing organisms from exposure to sunlight. Most recently, quinones have also attracted a lot of industrial interest since their electron-donating and -accepting properties make them good candidates as electrolytes in redox flow batteries, like their often highly conjugated double bond systems make them attractive as pigments. On an industrial level, quinones are mainly synthesized from raw components in coal tar. However, the possibility of producing quinones by fungal cultivation has great prospects since fungi can often be grown in industrially scaled bioreactors, producing valuable metabolites on cheap substrates. In order to give a better overview of the secondary metabolite quinones produced by and shared between various fungi, mainly belonging to the genera Aspergillus, Penicillium, Talaromyces, Fusarium, and Arthrinium, this review categorizes quinones into families such as emodins, fumigatins, sorbicillinoids, yanuthones, and xanthomegnins, depending on structural similarities and information about the biosynthetic pathway from which they are derived, whenever applicable. The production of these quinone families is compared between the different genera, based on recently revised taxonomy. KEY POINTS: • Quinones represent an important group of secondary metabolites widely distributed in important fungal genera such as Aspergillus, Penicillium, Talaromyces, Fusarium, and Arthrinium. • Quinones are of industrial interest and can be used in pharmacology, as colorants and pigments, and as electrolytes in redox flow batteries. • Quinones are grouped into families and compared between genera according to the revised taxonomy.

Fava beans can substitute soybean meal and rapeseed meal as protein source in diets for lactating dairy cows.

The effect of replacing mixtures of wheat and soybean meal and wheat and rapeseed meal by toasted fava beans, and the effect of toasting fava beans on feed intake, milk yield, and composition of milk and feces were investigated using 40 Holstein cows in each of two 4 × 4 Latin square design trials conducted simultaneously. In trial 1, the 4 treatment concentrates were untreated fava beans, toasted fava beans, 42% soybean meal + 58% rolled wheat, and a 21, 29, and 50% mix of soybean meal, rolled wheat, and toasted fava beans, respectively [on dry matter (DM) basis]. In trial 2, the 4 experimental treatments were untreated fava beans, toasted fava beans, 64% rapeseed meal + 36% rolled wheat, and a 32, 18, and 50% mix of rapeseed meal, rolled wheat, and toasted fava beans, respectively (on DM basis). In each trial, 16 primiparous and 24 multiparous cows were fed the treatment concentrates as part of a partial mixed ration, of which the forage consisted of 50% corn silage and 50% grass-clover silage. Substitution of soybean meal and wheat or rapeseed meal and wheat with toasted fava beans did not affect total DM intake, and no linear effects were observed on milk yield or energy-corrected milk (ECM) yield. However, in trial 2, a quadratic effect was observed on milk yield when substituting rapeseed meal and wheat with toasted fava beans. In both trials, substitution of soybean meal and wheat or rapeseed meal and wheat with toasted fava beans increased milk lactose concentration and decreased milk protein yield and concentration of protein in milk. In both trials, fecal concentration of starch increased linearly when substituting soybean meal and wheat or rapeseed meal and wheat with toasted fava beans. In trial 2, fecal concentration of P decreased when substituting rapeseed meal and wheat with toasted fava beans. In situ investigations showed increased rumen undegradable protein concentration and thereby increased estimated metabolizable protein supply when toasting fava beans. However, in both trials, milk protein yield and concentration decreased when cows were fed toasted compared with untreated fava beans. Furthermore, when cows were fed toasted compared with untreated fava beans in trial 1, milk yield, ECM yield, and nitrogen efficiency decreased. We conclude that toasted fava beans could substitute soybean meal and wheat or rapeseed meal and wheat with regard to ECM yield. However, milk protein yield decreased when substituting soybean meal and wheat or rapeseed meal and wheat with toasted fava beans. Compared with untreated fava beans, toasting had no positive effect on milk production and nitrogen efficiency.

Suppressed Charge Dispersion via Resonant Tunneling in a Single-Channel Transmon.

We demonstrate strong suppression of charge dispersion in a semiconductor-based transmon qubit across Josephson resonances associated with a quantum dot in the junction. On resonance, dispersion is drastically reduced compared to conventional transmons with corresponding Josephson and charging energies. We develop a model of qubit dispersion for a single-channel resonance, which is in quantitative agreement with experimental data.

Parity-Protected Superconductor-Semiconductor Qubit.

Coherence of superconducting qubits can be improved by implementing designs that protect the parity of Cooper pairs on superconducting islands. Here, we introduce a parity-protected qubit based on voltage-controlled semiconductor nanowire Josephson junctions, taking advantage of the higher harmonic content in the energy-phase relation of few-channel junctions. A symmetric interferometer formed by two such junctions, gate-tuned into balance and frustrated by a half-quantum of applied flux, yields a cos(2φ) Josephson element, reflecting coherent transport of pairs of Cooper pairs. We demonstrate that relaxation of the qubit can be suppressed tenfold by tuning into the protected regime.

Controlled dc Monitoring of a Superconducting Qubit.

Creating a transmon qubit using semiconductor-superconductor hybrid materials not only provides electrostatic control of the qubit frequency, it also allows parts of the circuit to be electrically connected and disconnected in situ by operating a semiconductor region of the device as a field-effect transistor. Here, we exploit this feature to compare in the same device characteristics of the qubit, such as frequency and relaxation time, with related transport properties such as critical supercurrent and normal-state resistance. Gradually opening the field-effect transistor to the monitoring circuit allows the influence of weak-to-strong dc monitoring of a "live" qubit to be measured. A model of this influence yields excellent agreement with experiment, demonstrating a relaxation rate mediated by a gate-controlled environmental coupling.

The association of dietary animal and plant protein with putative risk markers of colorectal cancer in overweight pre-diabetic individuals during a weight-reducing programme: a PREVIEW sub-study.

PURPOSE: Diets with increased protein content are popular strategies for body weight regulation, but the effect of such diets for the colonic luminal environment is unclear. We aimed to investigate the associations between putative colorectal cancer-related markers and total protein intake, plant and animal proteins, and protein from red and processed meat in pre-diabetic adults (> 25 years). METHODS: Analyses were based on clinical and dietary assessments at baseline and after 1 year of intervention. Protein intake was assessed from 4-day dietary records. Putative colorectal cancer-related markers identified from 24-h faecal samples collected over three consecutive days were: concentration of short-chain fatty acids, phenols, ammonia, and pH. RESULTS: In total, 79 participants were included in the analyses. We found a positive association between change in total protein intake (slope: 74.72 ± 28.84 µmol per g faeces/E%, p = 0.01), including animal protein intake (slope: 87.63 ± 32.04 µmol per g faeces/E%, p = 0.009), and change in faecal ammonia concentration. For change in ammonia, there was a dose-response trend from the most negative (lowest tertile) to the most positive (highest tertile) association (p = 0.01): in the high tertile, a change in intake of red meat was positively associated with an increase in ammonia excretion (slope: 2.0 ± 0.5 µmol per g faeces/g/day, p < 0.001), whereas no such association was found in the low and medium tertile groups. CONCLUSION: Increases in total and animal protein intakes were associated with higher excretion of ammonia in faeces after 1 year in overweight pre-diabetic adults undertaking a weight-loss intervention. An increase in total or relative protein intake, or in the ratio of animal to plant protein, was not associated with an increase in faeces of any of the other putative colorectal cancer risk markers. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT01777893.

Borderline anaemia and postoperative outcome in women undergoing major abdominal surgery: a retrospective cohort study.

Pre-operative anaemia is typically diagnosed with a haemoglobin concentration < 120 g.l-1 for women and < 130 g.l-1 for men on the basis of limited evidence. This retrospective cohort study stratified women undergoing elective, major abdominal surgery based on pre-operative haemoglobin concentration: anaemic (< 120 g.l-1 ); borderline anaemic (120-129 g.l-1 ); and non-anaemic (> 130 g.l-1 ). Data from 1554 women were analysed. Women with borderline anaemia had a greater incidence of postoperative complications (55 (16%) vs. 110 (11%); p = 0.026), longer duration of hospital stay (median (IQR [range]) 3 (1-6 [0-69]) days vs. 2 (1-5 [0-80]) days; p = 0.017) and fewer days alive and out of hospital at postoperative day 30 (median (IQR [range]) 27 (23-29 [0-30]) vs. 28 (25-29 [0-30]) days; p = 0.017) compared with non-anaemic women. However, after matched cohort analysis, these outcome differences no longer remained statistically significant. After multivariable adjustment for procedure, Charlson comorbidity index and patient age, a negative relationship between logarithmic pre-operative haemoglobin concentration and duration of stay was found (parameter estimate (standard error) -0.006 (0.003) vs. 0.003 (0.003) for a haemoglobin concentration < 130 g.l-1 vs. > 130 g.l-1 , respectively; p = 0.03); the difference in duration of stay was approximately 50% greater for women with a haemoglobin concentration of 120 g.l-1 compared with those with a haemoglobin concentration of 130 g.l-1 . Although the contribution of borderline anaemia to the incidence of postoperative complications is uncertain, the current diagnostic criteria should be re-assessed.

Short communication: Diets supplemented with starch and corn oil, marine algae, or hydrogenated palm oil differently affect selected metabolite concentrations in cow and goat milk.

The objective was to investigate the effects of species (cow vs. goat) and of various dietary lipid supplements, known to modulate milk fat content, on selected metabolites and enzymes in milk and to explore their correlations with performance traits. Twelve Holstein cows and 12 Alpine goats, all multiparous and nonpregnant, and at 86 ± 24.9 and 61 ± 1.8 DIM, respectively, were fed a basal diet (45% forage + 55% concentrate) not supplemented (CTL) or supplemented with corn oil plus wheat starch [COS, 5% of diet dry matter (DM)], marine algae powder (MAP, 1.5% of diet DM), or hydrogenated palm oil (HPO, 3% of diet DM) in a replicated 4 × 4 Latin square design with 28-d experimental periods. Intake, milk production and composition, milk fatty acid profile, and plasma metabolite concentrations were previously reported. Concentrations of 9 milk metabolites [ß-hydroxybutyrate (BHB), glucose, glucose-6-phosphate, isocitrate, choline, glutamate, urea, cholesterol, and free amino groups] and 2 milk enzyme activities (alkaline phosphatase and lactate dehydrogenase) were measured on d 24 of each experimental period. Dairy performance data showed marked species and diet effects on milk fat content. Irrespective of diet, cow milk was richer in alkaline phosphatase and glucose compared with goat milk (16 and 3 times more, respectively), whereas goat milk had greater urea and glucose-6-phosphate concentrations compared with cow milk (1.9 and 5.3 times more, respectively). In cows, COS decreased milk BHB and choline (-25 and -43%, respectively) compared with CTL, whereas no effects were observed in goats. The COS and MAP diets increased milk isocitrate compared with CTL in cows, but COS decreased isocitrate concentrations in goat milk. Milk choline was correlated with milk fat content in cows (Spearman r, rS = +0.73) and goats (rs = +0.58), and lactate dehydrogenase activity was correlated with milk somatic cell count (rs = +0.66) in cows but not in goats. We provide evidence of different milk metabolite responses according to species and diets. Metabolites and enzymes secreted in milk may be indicators of specificities of lipid metabolism among ruminant species and may contribute to a better understanding of mechanisms regulating milk fat secretion. Changes in the concentrations of some metabolites considered minor components of milk may be valuable diagnostic tools of mammary gland and animal metabolism as well as of milk processing characteristics.

Oral administration of lipopolysaccharides from Escherichia coli (serotype O111:B4) does not induce an effective systemic immune response in milk-fed Holstein calves.

It is well established that intravenous administration of lipopolysaccharides (LPS)-cell wall components from gram-negative bacteria-induce acute inflammatory responses in dairy calves, but the effect of oral administration of LPS to dairy calves is currently unknown. To evaluate the effects of oral administration of LPS derived from Escherichia coli (serotype O111:B4) on innate immune responses in milk-fed Holstein calves, 20 visually healthy calves (34 ± 1 d) received 4 L of milk with LPS (12 µg/kg body weight; n = 10; LPS) or without LPS (n = 10; control) at the morning feeding. Samples were collected at 0.5 h before the morning feeding and at 3, 6, 24, 48, 72, and 168 h after the morning feeding to measure rectal temperature and heart rate, as well as plasma-negative and plasma-positive acute phase proteins (i.e., haptoglobin, serum amyloid A, albumin, total protein, and fibrinogen) and immunoglobulin concentrations (IgG, IgM, and IgA). None of these measurements was affected by the oral administration of LPS. Oral administration of LPS at 12 µg/kg of body weight did not induce an acute inflammatory response in visually healthy milk-fed Holstein calves when administered in milk.

Inclusion of lemon leaves and rice straw into compound feed and its effect on nutrient balance, milk yield, and methane emissions in dairy goats.

The objective of this experiment was to study the effects of incorporating lemon leaves and rice straw into the compound feed of diets for dairy goats. Ten Murciano-Granadina dairy goats (n = 5 per group) in mid-lactation were used in a crossover design experiment (2 treatments across 2 periods). Goats were fed a mixed ration with barley grain (control, CON) or CON plus lemon leaves [189 g/kg of dry matter (DM)] and rice straw (120 g/kg of DM) in place of barley grain (LRS). Soybean oil (19 g/kg of DM) was added to the LRS diet to make it isoenergetic (17 MJ of gross energy/kg of DM) relative to CON. After 14 d on their respective treatments, goats were allocated to individual metabolism cages for another 7 d. Subsequently, feed intake, total fecal and urine output, and milk yield were recorded daily over the first 5 d. During the last 2 d, ruminal fluid and blood samples were collected, along with individual gas exchange measurements recorded by a mobile open-circuit indirect calorimetry system using a head box. No differences in DM intake were detected, and ME intake in LRS was lower than in CON (1,095 vs. 1,180 kJ/kg of metabolic body weight). No differences were observed in milk production, but milk fat content was greater in LRS (6.4%) than in CON (5.6%). Greater concentrations of monounsaturated (14.94 vs. 11.96 g/100 g of milk fat) and polyunsaturated fatty acids (4.53 vs. 4.03 g/100 g of milk fat) were detected in the milk of goats fed LRS compared with CON. Atherogenicity (2.68 vs.1.91) and thrombogenic (4.58 vs. 2.81) indices were lower with LRS compared with CON. Enteric CH4 emission was lower in LRS (24.3 g/d) compared with CON (31.1 g/d), probably due to the greater lipid content and unsaturated fatty acid profile of lemon leaves and the soybean oil added in the LRS diet. Overall, data suggest that incorporating lemon leaves and rice straw into lactating goat diets is effective in reducing CH4 emissions while allowing improvements in milk fat production and milk thrombogenic index without affecting production performance. Thus, their inclusion in compound feeds fed to small ruminants appears warranted and would have multiple positive effects, as on efficiency of nutrient use, human health, and the environment.

Milk metabolites as noninvasive indicators of nutritional status of mid-lactation Holstein and Montbéliarde cows.

The objective was to investigate the effects of feed restriction on concentrations of selected milk metabolites in mid-lactation Holstein and Montbéliarde cows and to explore their correlations with energy balance and classic plasma and milk indicators of nutritional status. Eight Holstein and 10 Montbéliarde cows (165 ± 21 d in milk) underwent 6 d of feed restriction during which feed allowance was reduced to meet 50% of their net energy for lactation (NEL) requirements. The experiment was divided in 4 periods: control (CON; d -3 to -1), restriction (RES; d 1 to 6), wk 1 (W1; d 7 to 13), and wk 2 (W2; d 14 to 18) after refeeding at ad libitum intake. Intake, milk production, energy balance and plasma metabolites were used to validate the feed restriction model. Concentrations of 7 milk metabolites: ß-hydroxybutyrate (BHB), glucose, glucose-6-phosphate, isocitrate, glutamate, uric acid, and free amino groups were measured in morning milk samples, and fatty acids were measured in pooled p.m. and a.m. samples. Feed restriction induced a negative energy balance (-42.5 ± 4.4 MJ/d), increased plasma nonesterified fatty acids and BHB, and decreased plasma glucose concentrations. Feed restriction increased milk glucose-6-phosphate and isocitrate (+38% and +39%, respectively) and decreased milk BHB, glucose, glutamate, uric acid and free amino group concentrations (-20%, -57%, -65%, -42%, and -14%, respectively), compared with pre- restriction. Milk concentrations of medium-chain fatty acids (e.g., sum of C10 to C15) decreased and those of long chain (e.g., 18:0, cis-9 18:1) increased during restriction. Breed differences were not detected for the majority of variables. All studied milk metabolites were significantly correlated with energy balance (Spearman correlation = 0.48, 0.63, -0.31, -0.45, and 0.61 for BHB, glucose, glucose-6-phosphate, isocitrate, and glutamate, respectively). Milk glucose and glutamate showed the strongest correlations with plasma metabolites and milk FA associated with lipomobilization. These results suggest that milk metabolites may be used as noninvasive indicators of negative energy balance and metabolic status of dairy cows.

Only few benefits from propylene glycol drench in early lactation for cows identified as physiologically imbalanced based on milk spectra analyses.

The main objective of this study was to test the efficiency of a management system combining metabolic clustering of cows based on Fourier-transform mid-infrared (FT-MIR) spectra of milk and targeted treatment of metabolically imbalanced cows with propylene glycol drench. We hypothesized that cows identified in a metabolically imbalanced status during early lactation were associated with subsequent impaired health, reproduction, and production, and that treatment with propylene glycol treatment would improve health, reproduction, and production relatively more in these cows than in control cows. We completed a prospective, randomized controlled trial with 356 early-lactation cows in 2 private dairy herds in Denmark from December 2017 to April 2018. Milk samples of cows were collected before treatment, from 4 to 9 d in milk, and after treatment, from 22 to 27 d in milk. Milk samples were analyzed using FT-MIR spectroscopy. We also measured 4 milk metabolites (ß-hydroxybutyrate, isocitrate, malate, and glutamate) and fat and protein contents. Based on FT-MIR spectra and cluster analyses, cows were clustered into groups of metabolically imbalanced and healthy cows. Within each group, cows were allocated randomly to treatment with propylene glycol (500 mL for 5 d) or no treatment. We analyzed the effect of the treatment on cow-level variables: metabolic cluster, milk metabolites, fat and protein contents, and fat-to-protein ratio at a milk sampling after the treatment. Furthermore, we analyzed daily milk yield, calving to first service interval, and disease occurrence. Results showed only a few effects of propylene glycol treatment and few interactions between treatment and metabolic clusters. We found no significant main effects of propylene glycol treatment in any of these analyses. A negative effect of the imbalanced metabolic cluster was found for the outcome of calving to first service interval for multiparous cows. In conclusion, we found a longer calving to first service interval in metabolically imbalanced cows, but we were not able to demonstrate overall benefits from the applied detection of cows in imbalanced metabolic status in early lactation and follow-up by treatment with propylene glycol.

Taxonomy of Aspergillus section Flavi and their production of aflatoxins, ochratoxins and other mycotoxins.

Aflatoxins and ochratoxins are among the most important mycotoxins of all and producers of both types of mycotoxins are present in Aspergillus section Flavi, albeit never in the same species. Some of the most efficient producers of aflatoxins and ochratoxins have not been described yet. Using a polyphasic approach combining phenotype, physiology, sequence and extrolite data, we describe here eight new species in section Flavi. Phylogenetically, section Flavi is split in eight clades and the section currently contains 33 species. Two species only produce aflatoxin B1 and B2 (A. pseudotamarii and A. togoensis), and 14 species are able to produce aflatoxin B1, B2, G1 and G2: three newly described species A. aflatoxiformans, A. austwickii and A. cerealis in addition to A. arachidicola, A. minisclerotigenes, A. mottae, A. luteovirescens (formerly A. bombycis), A. nomius, A. novoparasiticus, A. parasiticus, A. pseudocaelatus, A. pseudonomius, A. sergii and A. transmontanensis. It is generally accepted that A. flavus is unable to produce type G aflatoxins, but here we report on Korean strains that also produce aflatoxin G1 and G2. One strain of A. bertholletius can produce the immediate aflatoxin precursor 3-O-methylsterigmatocystin, and one strain of Aspergillus sojae and two strains of Aspergillus alliaceus produced versicolorins. Strains of the domesticated forms of A. flavus and A. parasiticus, A. oryzae and A. sojae, respectively, lost their ability to produce aflatoxins, and from the remaining phylogenetically closely related species (belonging to the A. flavus-, A. tamarii-, A. bertholletius- and A. nomius-clades), only A. caelatus, A. subflavus and A. tamarii are unable to produce aflatoxins. With exception of A. togoensis in the A. coremiiformis-clade, all species in the phylogenetically more distant clades (A. alliaceus-, A. coremiiformis-, A. leporis- and A. avenaceus-clade) are unable to produce aflatoxins. Three out of the four species in the A. alliaceus-clade can produce the mycotoxin ochratoxin A: A. alliaceus s. str. and two new species described here as A. neoalliaceus and A. vandermerwei. Eight species produced the mycotoxin tenuazonic acid: A. bertholletius, A. caelatus, A. luteovirescens, A. nomius, A. pseudocaelatus, A. pseudonomius, A. pseudotamarii and A. tamarii while the related mycotoxin cyclopiazonic acid was produced by 13 species: A. aflatoxiformans, A. austwickii, A. bertholletius, A. cerealis, A. flavus, A. minisclerotigenes, A. mottae, A. oryzae, A. pipericola, A. pseudocaelatus, A. pseudotamarii, A. sergii and A. tamarii. Furthermore, A. hancockii produced speradine A, a compound related to cyclopiazonic acid. Selected A. aflatoxiformans, A. austwickii, A. cerealis, A. flavus, A. minisclerotigenes, A. pipericola and A. sergii strains produced small sclerotia containing the mycotoxin aflatrem. Kojic acid has been found in all species in section Flavi, except A. avenaceus and A. coremiiformis. Only six species in the section did not produce any known mycotoxins: A. aspearensis, A. coremiiformis, A. lanosus, A. leporis, A. sojae and A. subflavus. An overview of other small molecule extrolites produced in Aspergillus section Flavi is given.

Pre-treatment microbial Prevotella-to-Bacteroides ratio, determines body fat loss success during a 6-month randomized controlled diet intervention.

On the basis of the abundance of specific bacterial genera, the human gut microbiota can be divided into two relatively stable groups that might have a role in personalized nutrition. We studied these simplified enterotypes as prognostic markers for successful body fat loss on two different diets. A total of 62 participants with increased waist circumference were randomly assigned to receive an ad libitum New Nordic Diet (NND) high in fiber/whole grain or an Average Danish Diet for 26 weeks. Participants were grouped into two discrete enterotypes by their relative abundance of Prevotella spp. divided by Bacteroides spp. (P/B ratio) obtained by quantitative PCR analysis. Modifications of dietary effects of pre-treatment P/B group were examined by linear mixed models. Among individuals with high P/B the NND resulted in a 3.15 kg (95% confidence interval (CI): 1.55; 4.76, P<0.001) larger body fat loss compared with ADD, whereas no differences was observed among individuals with low P/B (0.88 kg (95% CI: -0.61; 2.37, P=0.25)). Consequently, a 2.27 kg (95% CI: 0.09; 4.45, P=0.041) difference in responsiveness to the diets were found between the two groups. In summary, subjects with high P/B ratio appeared more susceptible to lose body fat on diets high in fiber and whole grain than subjects with a low P/B ratio.