Invited review: Large-scale indirect measurements for enteric methane emissions in dairy cattle: A review of proxies and their potential for use in management and breeding decisions

Efforts to reduce the carbon footprint of milk production through selection and management of low-emitting cows require accurate and large-scale measurements of methane (CH₄) emissions from individual cows. Several techniques have been developed to measure CH₄ in a research setting but most are not suitable for large-scale recording on farm. Several groups have explored proxies (i.e., indicators or indirect traits) for CH₄; ideally these should be accurate, inexpensive, and amenable to being recorded individually on a large scale. This review (1) systematically describes the biological basis of current potential CH₄ proxies for dairy cattle; (2) assesses the accuracy and predictive power of single proxies and determines the added value of combining proxies; (3) provides a critical evaluation of the relative merit of the main proxies in terms of their simplicity, cost, accuracy, invasiveness, and throughput; and (4) discusses their suitability as selection traits. The proxies range from simple and low-cost measurements such as body weight and high-throughput milk mid-infrared spectroscopy (MIR) to more challenging measures such as rumen morphology, rumen metabolites, or microbiome profiling. Proxies based on rumen samples are generally poor to moderately accurate predictors of CH₄, and are costly and difficult to measure routinely on-farm. Proxies related to body weight or milk yield and composition, on the other hand, are relatively simple, inexpensive, and high throughput, and are easier to implement in practice. In particular, milk MIR, along with covariates such as lactation stage, are a promising option for prediction of CH₄ emission in dairy cows. No single proxy was found to accurately predict CH₄, and combinations of 2 or more proxies are likely to be a better solution. Combining proxies can increase the accuracy of predictions by 15 to 35%, mainly because different proxies describe independent sources of variation in CH₄ and one proxy can correct for shortcomings in the other(s). The most important applications of CH₄ proxies are in dairy cattle management and breeding for lower environmental impact. When breeding for traits of lower environmental impact, single or multiple proxies can be used as indirect criteria for the breeding objective, but care should be taken to avoid unfavorable correlated responses. Finally, although combinations of proxies appear to provide the most accurate estimates of CH₄, the greatest limitation today is the lack of robustness in their general applicability. Future efforts should therefore be directed toward developing combinations of proxies that are robust and applicable across diverse production systems and environments.     

Dispersive Liquid–Liquid Microextraction Followed by Capillary High-Performance Liquid Chromatography for the Determination of Six Sulfonylurea Herbicides in Fruit Juices

In this study a simple, rapid, and efficient method has been developed for the determination of six sulfonylurea herbicides (SUHs): triasulfuron, metsulfuron-methyl, chlorsulfuron, flazasulfuron, chlorimuron-ethyl, and primisulfuron-methyl in commercial grape and apple juice samples, using dispersive liquid–liquid microextraction coupled with capillary high-performance liquid chromatography with diode array detection. Various parameters that influence the extraction efficiency, such as the type and volume of extraction and disperser solvents, sample pH, and salt addition, were investigated and optimized. Under the optimum conditions, limits of detection and quantification of the method were in the ranges of 2–9 and 8–29 μg L^?1, respectively, lower than the maximum residue limits set by the European Union for the raw fruits, such as grape and apple. The intra- and inter-day relative standard deviations varied from 1.0 to 8.2 and 1.8 to 9.8 %, respectively, with recoveries between 72.0 and 109.5 % for commercial grape (both white and red) and apple juice samples, showing satisfactory accuracy for the determination of SUHs in fruit juices.     

Response to recombinant human erythropoietin in patients with myelodysplastic syndromes

OBJECTIVES: The impact of a positive surgical margin in otherwise confined prostate cancer after radical prostatectomy remains unclear. We analyzed the outcome of a large number of patients with organ-confined prostate cancer according to the presence and anatomic site of margin positivity. METHODS: We evaluated 2712 prostatectomy patients with Stage pT2N0 cancer (ie, no evidence of extra-prostatic disease, seminal vesicle or regional node involvement) and no prior therapy who were treated by radical prostatectomy between 1987 and 1995 at Mayo Clinic. A total of 697 patients (26%) had positive margins. To assess the effect of margin status in the absence of treatment, 378 patients with postoperative adjuvant therapy were not considered for the study group: the final group consisted of 2334 patients. RESULTS: Overall, 253 (58%) tumors were positive at the apex and/or urethra, 85 (19%) at the prostate base, 11 (2.5%) at the anterior prostate, and 174 (40%) at the posterior prostate; 89 (20%) had at least two margins involved and 21 (8.3%) had more than two involved. The apex/urethra was the only positive anatomic site in 183 (42%). Five-year survival free of clinical recurrence or prostate-specific antigen (PSA) biochemical failure (postoperative serum PSA of 0.2 ng/mL or more) for patients with a single positive margin was 79% for apex or urethra, 78% for anterior/posterior, and 56% for prostate base. Five-year survival free of clinical recurrence or PSA (biochemical) failure was slightly higher for those with one versus two margin-positive regions (77% versus 68%, respectively). Multivariate analysis revealed that positive surgical margins were a significant predictor of clinical recurrence and PSA (biochemical) failure (relative risk [95% confidence interval]: 1.65 [1.24, 2.18]) after controlling for Gleason grade, preoperative PSA, and deoxyribonucleic acid (DNA) ploidy. The effect of margin positivity on recurrence at a specific anatomic site (versus negative margins or positive at a different anatomic site) revealed the prostate base to be the only significant anatomic site when adjusted for grade, PSA, and ploidy. Five-year survival free of the combined clinical or PSA failure end point for those with versus those without positive margins at the prostate base was 56% versus 85%, respectively (P < 0.0001). CONCLUSIONS: Positive surgical margins are a significant predictor of recurrence in Stage pT2N0 cancer, which is independent of grade, PSA, and DNA ploidy. The impact of positive margin status on recurrence-free survival appears to be anatomic and site-specific, with prostate base positivity significantly associated with poor outcome. The benefit of adjuvant therapy based on anatomic site-specific margin positivity remains to be tested in order to optimize recurrence-free survival.     

Genetic association of clinical mastitis with test-day somatic cell score and milk yield during first lactation of Finnish Ayrshire cows

In this study the genetic association during lactation of 2 clinical mastitis (CM) traits: CM1 (7 d before to 30 d after calving) and CM2 (31 to 300 d after calving) with test-day somatic cell score (SCS) and milk yield (MY) was assessed using multitrait random regression sire models. The data analyzed were from 27,557 first-lactation Finnish Ayrshire cows. Random regressions on second- and third-order Legendre polynomials were used to model the daily genetic and permanent environmental variances of test-day SCS and MY, respectively, while only the intercept term was fitted for CM. Results showed that genetic correlations between CM and the test-day traits varied during lactation. Genetic correlations between CM1 and CM2 and test-day SCS during lactation varied from 0.41 to 0.77 and from 0.34 to 0.71, respectively. Genetic correlations of test-day MY with CM1 and CM2 ranged from 0.13 to 0.51 and from 0.49 to 0.66, respectively. Correlations between CM1 and SCS were strongest during early lactation, whereas correlations between CM2 and SCS were strongest in late lactation. Genetic correlations lower than unity indicate that CM and SCS measure different aspects of the trait mastitis. Milk yield in early lactation was more strongly correlated with both CM1 and CM2 than milk yield in later lactation. This suggests that selection for higher lactation MY through selection on increased milk yield in early lactation will have a more deleterious effect on genetic resistance to mastitis than selection for higher yield in late lactation. The approach used in this study for the estimation of the genetic associations between test-day and CM traits could be used to combine information from traits with different data structures, such as test-day SCS and CM traits in a multitrait random regression model for the genetic evaluation of udder health.     

Reliability of breeding values for feed intake and feed efficiency traits in dairy cattle: When dry matter intake recordings are sparse under different scenarios

Currently, routine recordings of dry matter intake (DMI) in commercial herds are practically nonexistent. Recording DMI from commercial herds is a prerequisite for the inclusion of feed efficiency (FE) traits in dairy cattle breeding goals. To develop future on-farm phenotyping strategies, recording strategies that are low cost and less demanding logistically and that give relatively accurate estimates of the animal's genetic merit are therefore needed. The objectives of this study were (1) to estimate genetic parameters for daily DMI and FE traits and use the estimated parameters to simulate daily DMI phenotypes under different DMI recording scenarios (SCN) and (2) to use the simulated data to estimate for different scenarios the associated reliability of estimated breeding value and accuracies of genomic prediction for varying sizes of reference populations. Five on-farm daily DMI recording scenarios were simulated: once weekly (SCN1), once monthly (SCN2), every 2 mo (SCN3), every 3 mo (SCN4), and every 4 mo (SCN5). To estimate reliability of estimated breeding values, DMI and FE observations and true breeding values were simulated based on variance components estimated for daily observations of Nordic Red cows. To emulate realistic on-farm recording, 5 data set replicates, each with 36,037 DMI and FE records, were simulated for real pedigree and data structure of 789 Holstein cows. Observations for the 5 DMI recording scenarios were generated by discarding data in a step-wise manner from the full simulated data per the scenario's definitions. For each of these scenarios, reliabilities were calculated as correlation between the true and estimated breeding values. Variance components and genetic parameters were estimated for daily DMI, residual feed intake (RFI), and energy conversion efficiency (ECE) fitting the random regression model. Data for variance components were from 227 primiparous Nordic Red dairy cows covering 8 to 280 d in milk. Lactation-wise heritability for DMI, RFI, and ECE was 0.33, 0.12, and 0.32, respectively, and daily heritability estimates during lactation ranged from 0.18 to 0.45, 0.08 to 0.32, and 0.08 to 0.45 for DMI, RFI, and ECE, respectively. Genetic correlations for DMI between different stages of lactation ranged from ?0.50 to 0.99. The comparison of different on-farm DMI recording scenarios indicated that adopting a less-frequent recording scenario (SCN3) gave a similar level of accuracy as SCN1 when 17 more daughters are recorded per sire over the 46 needed for SCN1. Such a strategy is less demanding logistically and is low cost because fewer observations need to be collected per animal. The accuracy of genomic predictions associated with the 5 recording scenarios indicated that setting up a relatively larger reference population and adopting a less-frequent DMI sampling scenario (e.g., SCN3) is promising. When the same reference population size was considered, the genomic prediction accuracy of SCN3 was only 5.0 to 7.0 percentage points lower than that for the most expensive DMI recording strategy (SCN1). We concluded that DMI recording strategies that are sparse in terms of records per cow but with slightly more cows recorded per sire are advantageous both in genomic selection and in traditional progeny testing schemes when accuracy, logistics, and cost implications are considered.     

Chemical composition of naturally fermented buttermilk

Naturally fermented buttermilk, prepared from soured cream or milk, was collected during two seasons from sixteen farms in northern Ethiopia, to study chemical composition and flavour compounds. Protein, fat, organic acids, carbohydrates and volatile compounds were quantified using Kjeldahl, Gerber, high‐pressure liquid chromatography (HPLC) and headspace GC methods, respectively. Widely differing concentrations of organic acids and volatile compounds among samples indicated variable fermentation in the products. This indicates the need for the introduction of the standardisation of the process to supply the market with homogenous buttermilk products.     

Genetic and phenotypic relationships among milk yield and somatic cell count before and after clinical mastitis

This paper studies whether cows with originally lower somatic cell count (SCC) are more susceptible to clinical mastitis (CM) than cows with higher somatic cell count, and evaluates the correlations between CM, SCC, and milk yield. Data were extracted from the Finnish national milk-recording database and from the health recording system. First and second lactation records of 87,861 Ayrshire cows calving between January 1998 and December 2000 were included. Traits studied were incidence of CM, test-day SCC, and test-day milk yield before and following CM. Genetic parameters were estimated using multitrait REML with a sire model. Results did not indicate that cows with genetically low SCC would be more susceptible to CM. The genetic correlation between CM in the first and second lactation was reasonably high (0.73), suggesting that susceptibility to mastitis remains similar across lactations. The genetic correlation between CM and milk yield traits was positive (from 0.38 to 0.56), confirming the genetic antagonism between production and udder health traits. The genetic correlation between SCC and milk was positive in the first lactation, but negative, or near zero in the second lactation. This indicates that breeding for lower SCC might not affect milk production in later lactations. The results of this study support the use of SCC as an indicator of mastitis and a tool for selection for mastitis resistance.     

Genetic background in partitioning of metabolizable energy efficiency in dairy cows

The main objective of this study was to assess the genetic differences in metabolizable energy efficiency and efficiency in partitioning metabolizable energy in different pathways: maintenance, milk production, and growth in primiparous dairy cows. Repeatability models for residual energy intake (REI) and metabolizable energy intake (MEI) were compared and the genetic and permanent environmental variations in MEI were partitioned into its energy sinks using random regression models. We proposed 2 new feed efficiency traits: metabolizable energy efficiency (MEE), which is formed by modeling MEI fitting regressions on energy sinks [metabolic body weight (BW^0.75), energy-corrected milk, body weight gain, and body weight loss] directly; and partial MEE (pMEE), where the model for MEE is extended with regressions on energy sinks nested within additive genetic and permanent environmental effects. The data used were collected from Luke's experimental farms Rehtijärvi and Minkiö between 1998 and 2014. There were altogether 12,350 weekly MEI records on 495 primiparous Nordic Red dairy cows from wk 2 to 40 of lactation. Heritability estimates for REI and MEE were moderate, 0.33 and 0.26, respectively. The estimate of the residual variance was smaller for MEE than for REI, indicating that analyzing weekly MEI observations simultaneously with energy sinks is preferable. Model validation based on Akaike's information criterion showed that pMEE models fitted the data even better and also resulted in smaller residual variance estimates. However, models that included random regression on BW^0.75 converged slowly. The resulting genetic standard deviation estimate from the pMEE coefficient for milk production was 0.75 MJ of MEI/kg of energy-corrected milk. The derived partial heritabilities for energy efficiency in maintenance, milk production, and growth were 0.02, 0.06, and 0.04, respectively, indicating that some genetic variation may exist in the efficiency of using metabolizable energy for different pathways in dairy cows.     

Effect of grain tef [Eragrostis tef (Zucc.) Trotter] flour substitution with flaxseed on quality and functionality of injera

Injera from tef substituted with two flaxseed forms at 3%, 6% and 9% and control injera showed substitution had a significant effect on injera proximate, energy, titratable acidity (TA), total phenolics (TP) contents and sensory acceptability. Flaxseed forms had influence on moisture, fibre, total carbohydrate (TC) and TA. With 9% flour and whole flaxseed substitution, percentage energy, moisture, ash, crude protein, fibre and TA increase were 3.5, 27.3, 25.9, 20.4, 114.3 and 10.1, respectively. TC and pH were high for control. In all injera samples, condensed tannins and free fatty acid were insignificant. With an increase in the flaxseed substitution, most sensory acceptance increased, whereas injera eyes and colour decreased and appeared superior for control (100% tef injera). The 9% flaxseed‐substituted injera showed good proximate nutritional and energy contents of functional potential of high in dietary fibre, alpha linolenic acid, lignans, proteins and TP of anti‐oxidant nature.     

A new selective liquid membrane extraction method for the determination of basic herbicides in agro-processed fruit juices and Ethiopian honey wine (Tej) samples

Supported liquid membrane (SLM) extraction was optimised for trace extraction and enrichment of selected triazine herbicides from a variety of agro-processed fruit juices and Ethiopian honey wine (Tej) samples. In the extraction process, a 1:1 mixture of n-undecane and di-n-hexylether was immobilised in a thin porous PTFE membrane that forms a barrier between two aqueous phases (the donor and acceptor phases) in a flow system. The extracts constitute the selectively enriched analytes collected from the acceptor phase and were analysed by transferring to a HPLC-UV system using a diode array detector at 235?nm. High enrichment factors were obtained with very good repeatability of results, and the detection limit was lower than 3.00?μg l?1 for ametryn in apple juice. The optimised method showed very good linearity of over 50-500?μg l?1 with a correlation coefficient of >0.990 or better for triplicate analysis. All chromatograms gave well resolved peaks with no interfering peaks at the retention times of the selected triazines, showing high selectivity of the SLM extraction method in combination with HPLC-UV for the selected matrices. The optimised method can be used as an alternative solventless extraction method for microgram-level extraction of other triazine herbicides and a variety of pesticides from liquid and semi-liquid environmental, biological and food matrices.