The relationship between feed efficiency and behaviour differs between lactating Holstein and Jersey cows.

In dairy production, high feed efficiency (FE) is important to reduce feed costs and negative impacts of milk production on the climate and environment, yet little is known about the relationship between FE, eating behaviour and activity. This research communication describes how cows differing in FE, expressed as daily energy corrected milk production per unit of feed intake, differed in eating behaviour and activity. We used data from a study of 253 lactations obtained from 97 Holstein and 91 Jersey cows milked in an automatic milking system. Automated feed troughs recorded feed intake behaviour and cows wore a sensor that recorded activity from 5 to 200 d in milk (DIM). We used a mixed linear model to estimate random solutions for individual cows for traits of steps, lying and eating behaviour and calculated their correlation with FE during four periods (5-35, 36-75, 76-120 and 121-200 DIM). Separate analyses were performed for each breed and period. We found that individual level correlations between FE and behaviour traits were stronger in Jersey than in Holstein cows. Eating rate correlated weakly negatively to FE in Holstein cows and more strongly so in Jersey cows, such that efficient Jerseys were slower eaters. The physical activity of Jersey cows was weakly and negatively correlated to FE, but this was not the case in Holstein cows. We conclude that eating rate was consistently negatively associated with FE throughout lactation for Jersey cows, but not for Holstein cows.

Metabolome and proteome changes in skeletal muscle and blood of pre-weaning calves fed leucine and threonine supplemented diets.

In pre-weaning calves, both leucine and threonine play important roles in growth and muscle metabolism. In this study, metabolomics, proteomics and clinical chemistry were used to assess the effects of leucine and threonine supplementation added to milk replacer on 14 newborn Holstein male calves: 7 were fed a control diet (Ctrl) and 7 were fed the Ctrl diet supplemented with 0.3% leucine and 0.3% threonine (LT) from 5.6 days of age to 53.6 days. At this time, blood and semitendinosus muscle biopsies were collected for analysis. Integrated metabolomics and proteomics showed that branched-chain amino acids (BCAA) degradation and mitochondrial oxidative metabolism (citrate cycle and respiratory chain) were the main activated pathways in muscle because of the supplementation. BCAA derivatives and metabolites related to lipid mobilization showed the major changes. The deleterious effects of activated oxidative phosphorylation were balanced by the upregulation of antioxidant proteins. An increase in protein synthesis was indicated by elevated aminoacyl-tRNA biosynthesis and increased S6 ribosomal protein phosphorylation in skeletal muscle. In conclusion, LT group showed greater BCAA availability and mitochondrial oxidative activity; as the muscle cells undergo greater aerobic metabolism, antioxidant defenses were activated to compensate for possible cell damage. Data are available via ProteomeXchange (PXD016098). SIGNIFICANCE: Leucine and threonine are essential amino acids for the pre-weaning calf, being of high importance for growth. In this study, we found that leucine and threonine supplementation of milk replacer to feed pre-weaning calves led to differences in the proteome, metabolome and clinical chemistry analytes in skeletal muscle and plasma, albeit no differences in productive performance were recorded. This study extends our understanding on the metabolism in dairy calves and helps optimizing their nutritional status.

Symposium review: Decomposing efficiency of milk production and maximizing profit.

The dairy industry has focused on maximizing milk yield, as it is believed that this maximizes profit mainly through dilution of maintenance costs. Efficiency of milk production has received, until recently, considerably less attention. The most common method to determine biological efficiency of milk production is feed efficiency (FE), which is defined as the amount of milk produced relative to the amount of nutrients consumed. Economic efficiency is best measured as income over feed cost or gross margin obtained from feed investments. Feed efficiency is affected by a myriad of factors, but overall they could be clustered as follows: (1) physiological status of the cow (e.g., age, state of lactation, health, level of production, environmental conditions), (2) digestive function (e.g., feeding behavior, passage rate, rumen fermentation, rumen and hindgut microbiome), (3) metabolic partitioning (e.g., homeorhesis, insulin sensitivity, hormonal profile), (4) genetics (ultimately dictating the 2 previous aspects), and (5) nutrition (e.g., ration formulation, nutrient balance). Over the years, energy requirements for maintenance seem to have progressively increased, but efficiency of overall nutrient use for milk production has also increased due to dilution of nutrient requirements for maintenance. However, empirical evidence from the literature suggests that marginal increases in milk require progressively greater marginal increases in nutrient supply. Thus, the dilution of maintenance requirements associated with increases in production is partially overcome by a progressive diminishing marginal biological response to incremental energy and protein supplies. Because FE follows the law of diminishing returns, and because marginal feed costs increase progressively with milk production, profits associated with improving milk yield might, in some cases, be considerably lower than expected.

Feeding Pasteurized Waste Milk to Preweaned Dairy Calves Changes Fecal and Upper Respiratory Tract Microbiota.

In the present study bacterial communities from both, the gastrointestinal and respiratory tract of pre-weaned dairy calves fed two different milk-feeding programs were characterized using 16S rRNA gene sequencing. Twenty female Holstein calves (38.8 ± 1.40 kg of BW) were fed pasteurized waste milk (pWM) containing residues of various antimicrobials. Twenty additional calves (38.1 ± 1.19 kg of BW) were fed milk replacer (MR) with similar nutrient composition (27.5% crude protein, 32.1% fat) compared to waste milk (28.6% crude protein, 30.0% fat) from day 1 to weaning at day 49 of study. Fecal samples and nasal swabs were collected on day 42 only from calves that were not treated with therapeutic antibiotics throughout the study, which were 8 MR and 10 pWM calves. To assess the impact of the two feeding regimes on the fecal and nasal microbiota, α and ß-diversity measures were calculated, and the relative abundance of operational taxonomic units (OTUs) at different taxonomic levels was determined for each sample. In general, Chao1, PD Whole Tree, and Shannon diversity indices were similar for the fecal and nasal bacterial communities of calves regardless of the feeding regime. However, principal coordinate analysis based on unweighted Unifrac distances indicated differences in the structure of bacterial communities of calves fed milk replacer compared with those from calves fed pasteurized waste milk. The relative abundance of the Streptococcaceae family and the genus Histophilus was greater (P < 0.05) in the nasal microbiota of calves fed milk replacer than in those fed pasteurized waste milk. However, the genus Prevotella tended (P = 0.06) to be more relatively abundant in the respiratory tract of calves fed pasteurized waste milk than in those fed milk replacer. Differences in relative abundances of bacterial taxa in gut microbiota were only observed at the phylum level, suggesting that antimicrobial residues present in waste milk have a non-specific influence at a lower taxonomical level.

Age-Related Serum Biochemical Reference Intervals Established for Unweaned Calves and Piglets in the Post-weaning Period.

The purpose of the present study is to establish the influence of age on serum biochemistry reference intervals (RIs) for unweaned calves and recently-weaned piglets using large number of animals sampled at different ages from populations under different season trials. Specifically, milk replacer (MR)-fed calves from April-July 2017 (n = 60); from December 2016-March 2017 (n = 76) and from April-August 2018 (n = 57) and one group of healthy weaned piglets (n = 72) were subjected to the study. Serum enzymes and metabolites of calves at age of 24 h (24 h after colostrum intake), 2, 5, and 7 weeks from merged trials and piglets at 0, 7, and 14 days post-weaning (at 21, 28, and 35 days of age) were studied. The main variable is age whereas no major trial- or sex-biased differences were noticed. In calves, ALT, AST, GGT, GPx, SOD, NEFAs, triglycerides, glucose, creatinine, total protein, and urea were greatly elevated (p < 0.001) at 24 h compared with other ages; glucose, creatinine, total protein, and urea constantly decreased through the age; cholesterol's lowest level (p < 0.001) was found in 24 h compared with other ages and the levels of haptoglobin remained unchanged (p > 0.1) during the study. In comparison with the adult RIs, creatinine from 24 h, NEFAs from 2 w, GGT from 5 w, and urea from 7 w are fully comparable with RIs or lie within RIs determined for adult. In piglets, no changes were noticed on glucose (p > 0.1) and haptoglobin (p > 0.1) and there were no major changes on hepatic enzymes (ALT, AST, and GGT), total protein, creatinine and urea even though several statistical differences were noticed on 7 days post-weaning. Cholesterol, triglycerides, NEFAs, cortisol and PigMAP were found increased (p < 0.05) while TNF-alpha was found less concentrated (p < 0.001) at 0 days post-weaning compared with other times. Moreover, the RIs of creatinine and GGT are fully comparable with RIs or lie within RIs determined for adult. In conclusion, clinical biochemistry analytes RIs were established for unweaned calves and recently-weaned piglets and among them some can vary at different ages.

Escalating Threat Levels of Bacterial Infection Can Be Discriminated by Distinct MAPK and NF-κB Signaling Dynamics in Single Host Cells.

During an infection, immune cells must identify the specific level of threat posed by a given bacterial input in order to generate an appropriate response. Given that they use a general non-self-recognition system, known as Toll-like receptors (TLRs), to detect bacteria, it remains unclear how they transmit information about a particular threat. To determine whether host cells can use signaling dynamics to transmit contextual information about a bacterial stimulus, we use live-cell imaging to make simultaneous quantitative measurements of host MAPK and NF-κB signaling, two key pathways downstream of TLRs, and bacterial infection and load. This combined, single-cell approach reveals that NF-κB and MAPK signaling dynamics are sufficient to discriminate between (1) pathogen-associated molecular patterns (PAMPs) versus bacteria, (2) extracellular versus intracellular bacteria, (3) pathogenic versus non-pathogenic bacteria, and (4) the presence or absence of features indicating an active intracellular bacterial infection, such as replication and effector secretion.

Western diet regulates immune status and the response to LPS-driven sepsis independent of diet-associated microbiome.

Sepsis is a deleterious immune response to infection that leads to organ failure and is the 11th most common cause of death worldwide. Despite plaguing humanity for thousands of years, the host factors that regulate this immunological response and subsequent sepsis severity and outcome are not fully understood. Here we describe how the Western diet (WD), a diet high in fat and sucrose and low in fiber, found rampant in industrialized countries, leads to worse disease and poorer outcomes in an LPS-driven sepsis model in WD-fed mice compared with mice fed standard fiber-rich chow (SC). We find that WD-fed mice have higher baseline inflammation (metaflammation) and signs of sepsis-associated immunoparalysis compared with SC-fed mice. WD mice also have an increased frequency of neutrophils, some with an "aged" phenotype, in the blood during sepsis compared with SC mice. Importantly, we found that the WD-dependent increase in sepsis severity and higher mortality is independent of the microbiome, suggesting that the diet may be directly regulating the innate immune system through an unknown mechanism. Strikingly, we could predict LPS-driven sepsis outcome by tracking specific WD-dependent disease factors (e.g., hypothermia and frequency of neutrophils in the blood) during disease progression and recovery. We conclude that the WD is reprogramming the basal immune status and acute response to LPS-driven sepsis and that this correlates with alternative disease paths that lead to more severe disease and poorer outcomes.

Skeletal muscle metabolomics and blood biochemistry analysis reveal metabolic changes associated with dietary amino acid supplementation in dairy calves.

The effects of different amino acid (AA) supplementations of milk protein-based milk replacers in pre-ruminant calves from 3 days to 7 weeks of age were studied. Animals were divided into 4 groups: Ctrl) Control group fed with milk protein-based milk replacer without supplementation; GP) supplementation with 0.1% glycine and 0.3% proline; FY) supplementation with 0.2% phenylalanine and 0.2% tyrosine; MKT) supplementation with 0.62% lysine, 0.22% methionine and 0.61% threonine. For statistical analysis, t-test was used to compare AA-supplemented animals to the Ctrl group. At week 7, body weight and average daily gain (ADG) were measured and blood samples and skeletal muscle biopsies were taken. Blood biochemistry analytes related to energy metabolism were determined and it was shown that MKT group had higher serum creatinine and higher plasma concentration of three supplemented AAs as well as arginine compared with the Ctrl group. GP group had similar glycine/proline plasma concentration compared with the other groups while in FY group only plasma phenylalanine concentration was higher compared with Control. Although the AA supplementations in the GP and FY groups did not affect average daily gain and metabolic health profile from serum, the metabolome analysis from skeletal muscle biopsy revealed several differences between the GP-FY groups and the Ctrl-MKT groups, suggesting a metabolic adaptation especially in GP and FY groups.

Surveying selected European feed and livestock production chains for features enabling the case-specific post-market monitoring of livestock for intake and potential health impacts of animal feeds derived from genetically modified crops.

This review, which has been prepared within the frame of the European Union (EU)-funded project MARLON, surveys the organisation and characteristics of specific livestock and feed production chains (conventional, organic, GM-free) within the EU, with an emphasis on controls, regulations, traceability, and common production practices. Furthermore, an overview of the origin of animal feed used in the EU as well as an examination of the use of genetically modified organisms (GMOs) in feed is provided. From the data, it shows that livestock is traceable at the herd or individual level, depending on the species. Husbandry practices can vary widely according to geography and animal species, whilst controls and checks are in place for notifiable diseases and general health symptoms (such as mortality, disease, productive performance). For feeds, it would be possible only to make coarse estimates, at best, for the amount of GM feed ingredients that an animal is exposed to. Labeling requirements are apparently correctly followed. Provided that confounding factors are taken into account, practices such as organic agriculture that explicitly involve the use of non-GM feeds could be used for comparison to those involving the use of GM feed.

Influence of milk processing temperature on growth performance, nitrogen retention, and hindgut's inflammatory status and bacterial populations in a calf model.

This research communication describes a study aimed at evaluating the effects of heat treatment of milk on growth performance, N retention, and hindgut's inflammatory status and bacterial populations using young dairy calves as a model. Twenty-one Holstein calves were randomly allocated to one of three treatments: raw milk (RM), pasteurised milk (PAST), or UHT milk (UHT). Calves were submitted to a N balance study, and a biopsy from the distal colon and a faecal sample were obtained from 5 animals per treatment to determine expression of several genes and potential changes in the hindgut's bacterial population. Milk furosine content was 33-fold greater in UHT than in RM and PAST milks. Calves receiving RM grew more than those fed UHT, and urinary N excretion was greatest in calves fed UHT. Quantification of Lactobacillus was lower in calves consuming PAST or UHT, and Gram negative bacteria were greater in UHT than in PAST calves. The expression of IL-8 in the hindgut's mucosa was lowest and that of IL-10 tended to be lowest in RM calves, and expression of claudin-4 tended to be greatest in UHT calves. In conclusion, the nutritional value of UHT-treated milk may be hampered because it compromises growth and increases N excretion in young calves and may have deleterious effects on the gut's bacterial population and inflammation status.

EMPOWERING MULTI-COHORT GENE EXPRESSION ANALYSIS TO INCREASE REPRODUCIBILITY.

A major contributor to the scientific reproducibility crisis has been that the results from homogeneous, single-center studies do not generalize to heterogeneous, real world populations. Multi-cohort gene expression analysis has helped to increase reproducibility by aggregating data from diverse populations into a single analysis. To make the multi-cohort analysis process more feasible, we have assembled an analysis pipeline which implements rigorously studied meta-analysis best practices. We have compiled and made publicly available the results of our own multi-cohort gene expression analysis of 103 diseases, spanning 615 studies and 36,915 samples, through a novel and interactive web application. As a result, we have made both the process of and the results from multi-cohort gene expression analysis more approachable for non-technical users.

Integrated, Multi-cohort Analysis Identifies Conserved Transcriptional Signatures across Multiple Respiratory Viruses.

Respiratory viral infections are a significant burden to healthcare worldwide. Many whole genome expression profiles have identified different respiratory viral infection signatures, but these have not translated to clinical practice. Here, we performed two integrated, multi-cohort analyses of publicly available transcriptional data of viral infections. First, we identified a common host signature across different respiratory viral infections that could distinguish (1) individuals with viral infections from healthy controls and from those with bacterial infections, and (2) symptomatic from asymptomatic subjects prior to symptom onset in challenge studies. Second, we identified an influenza-specific host response signature that (1) could distinguish influenza-infected samples from those with bacterial and other respiratory viral infections, (2) was a diagnostic and prognostic marker in influenza-pneumonia patients and influenza challenge studies, and (3) was predictive of response to influenza vaccine. Our results have applications in the diagnosis, prognosis, and identification of drug targets in viral infections.

Long-term effects on heifer performance of an enhanced-growth feeding programme applied during the preweaning period.

Sixty female dairy calves (body weight, BW, 43.2+/-0.58 kg and age 9.8+/-0.61 d) were arranged in two groups to compare the short-term and long-term effects of an enhanced-growth feeding programme (EF) with those of a conventional-growth feeding programme (CF). After 1 week of adaptation to a milk replacer (MR), CF calves were fed 4 l/d of MR (25% crude protein, CP; 19.2% ether extract) at 12% dry matter (DM) from days 1 to 27 and 2 l/d at 12% DM from days 28 to 34, and the EF calves were offered the same MR at 18% DM: 4 l/d from days 1 to 6, 6 l/d from days 7 to 13, 7 l/d from days 14 to 20, 6 l/d from days 21 to 27 and 3 l/d from day 28 to weaning at day 34 of the study (50 d of age). Individual calf starter (20.7% CP) intake was recorded daily from the beginning until day 41 of study (57 d of age). Then, calves were placed in groups of six and they received a total mixed ration (TMR) containing 18.5% CP until day 56 d of study (72 d of age). Then, heifers were moved to larger pens and were fed the same TMR in both treatments at each subsequent stage of growth throughout the study. Calves were weighed weekly until day 56 of study and before every pen change (days 94, 149, 200, 387 of study). When heifers were 400 d old and weighed >380 kg, they were moved to a breeding pen where oestruses were checked three times a day. Heifers were inseminated 12 h after the detection of oestrus. One month before calving, heifers were returned to their original farm and milk yield at 305 days in milk was recorded from 28 cows. Starter intake was greater (P<0.001) in CF than in EF calves (0.79 v. 0.29+/-0.043 kg/d, respectively) during the preweaning period, but TMR consumption was similar in both treatments from days 42 to 56 of study. BW of EF calves was greater (P<0.01) than that of CF calves at weaning (76.4 v. 71.6+/-1.10 kg, respectively), but BW was not different at day 387 of study (405 d of age) (406.3 v. 401.3+/-4.05 kg, respectively). There were numerical differences in age at first breeding, fertility at first artificial insemination, age at pregnancy, and milk yield but some of these differences might have reached statistical significance with more replication.

Performance and behaviour of calves reared in groups or individually following an enhanced-growth feeding programme.

Nineteen calves were reared in individual pens (IP), and twenty calves were grouped in four pens of five calves each (GP) to study the effects of grouping on behaviour and performance. All calves were on an enhanced-growth feeding programme and had unlimited access to starter. Total dry matter intake (DMI) body weight (BW), serum urea, non-esterified fatty acid (NEFA), cortisol, insulin, and glucose concentrations, and immune response to vaccination were measured. Behaviour patterns categorized as: non-nutritive oral behaviour, cross-sucking, inter-sucking, and self-grooming, were monitored 20 min following the morning and afternoon milk replacer consumption, and at the same times after weaning. There were no differences in final BW and starter DMI between treatments. The decrease of average daily gain (ADG) during the week following weaning was more pronounced in GP than in IP calves. Serum NEFA concentration increased at the beginning of the study in IP calves but serum urea concentrations were greater in GP than in IP calves the week after weaning. Self-grooming increased in both treatments throughout the study but GP calves increased non-nutritive oral behaviour and decreased cross-sucking and inter-sucking behaviours throughout the study. Rearing calves in groups following an enhanced-growth feeding programme did not increase starter intake. Furthermore, cross-sucking and inter-sucking behaviours in calves reared in groups did not seem to cause increased metabolic or immunological distress.