Identification of hepatic biomarkers for physiological imbalance of dairy cows in early and mid lactation using proteomic technology.

Identification of biomarkers for degree of physiological imbalance (PI), a situation in which physiological parameters deviate from normal, is needed to reduce disease risk and improve production and reproduction performance of cows. The objective was to describe the liver proteome in early and mid lactation for cows with different degrees of PI with a special focus on biomarkers and pathways involved in periparturient disease complexes. Twenty-nine cows in early [49 ± 22d in milk (DIM); n=14] and mid (159 ± 39 DIM; n=15) lactation were nutrient restricted for 4d to increase PI by supplementing the ration with 60% wheat straw. Liver biopsies were collected -1 and 3d relative to restriction. Before restriction, an index for PI was calculated based on plasma nonesterified fatty acids, ß-hydroxybutyrate, and glucose concentrations. Within E and M cows, a subsets of 6 cow was classified as having either the greatest (PI) or least (normal; N) degree of PI and were used for isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative profiling in liver using liquid chromatography-tandem mass spectrometry. We identified pyruvate carboxylase and isocitrate dehydrogenase as potential hepatic biomarkers for PI for cows during early lactation and alcohol dehydrogenase-4 and methylmalonate-semialdehyde dehydrogenase for cows in mid lactation. This preliminary study identified new biomarkers in liver for PI and provided a better understanding of the differences in coping strategies used for cows in PI. Despite the small sample size (n=3/group), the results lay a foundation for future research focused on the usefulness of the hepatic biomarkers for predicting PI and thereby cows at risk for disease during lactation.

Changes in protein abundance between tender and tough meat from bovine longissimus thoracis muscle assessed by isobaric Tag for Relative and Absolute Quantitation (iTRAQ) and 2-dimensional gel electrophoresis analysis.

The aim of this study was to find potential biomarkers for meat tenderness in bovine Longissimus thoracis muscle and to compare results from isobaric Tag for Relative and Absolute Quantitation (iTRAQ) and 2-dimensional gel electrophoresis (2-DE) analysis. The experiment included 4 tender and 4 tough samples, based on shear force measurements at 7 d postmortem, from young Norwegian red (NRF) bulls, taken at 1 h postmortem. A number of the proteins which have previously been related to tenderness were found to change in abundance between tender and tough samples, both in iTRAQ (P < 0.1) and 2-DE analysis (P < 0.05). Furthermore, 3 proteins that have not previously been related to tenderness were found to change significantly in abundance between tender and tough meat samples in the present study. These include proteins related to control of flux through the tricarboxylate cycle [2-oxoglutarate dehydrogenase complex component E2 (OGDC-E2)], apoptosis (galectin-1) and regulatory role in the release of Ca(2+) from intracellular stores (annexin A6). Even though the overlap in significantly changing proteins was relatively low between iTRAQ and 2-DE analysis, certain proteins predicted to have the same function were found in both analyses and showed similar changes between the groups, such as structural proteins and proteins related to apoptosis and energy metabolism.

Differential smoothing of time-series measurements to identify disturbances in performance and quantify animal response characteristics: An example using milk yield profiles in dairy cows.

Recent advances in on-farm technology now provide us with multiple time-series of reliably measured indicators of animal performance and status at the level of the individual. This paper presents a smoothing approach for extracting biologically meaningful features from such time series using bovine milk yield data as an example. The main goal of this study was to illustrate how the method can be used to detect production deviations, extract quantifiable features of the deviation profiles, and thus provide means to examine hypotheses concerning the nature of the deviations. The effectiveness of the method was assessed with complete lactation curves from 47 Holstein cows. Within their lactations, the cows were each subjected to 1 nutritional challenge for a period of 4 d (their standard diet: a maize silage-based total mixed ration was diluted with 60% wheat straw), which provoked a decline in the milk yield in all cows. The challenge was imposed between the same calendar days for all cows. Thus, the cows were at different stages of lactation: early (n = 14), mid (n = 15), and late (n = 18). Each milk-yield curve was decomposed into components that capture the short-term deviations of the cow such as the response to the nutritional challenge and describe the phenotypic potential yield function of that cow throughout its lactation. The difference between the 2 components gives a measure of the milk loss. In all, 480 deviations were detected from the complete lactations of 47 cows. The milk loss provoked by the feeding challenge (n = 47) was significantly related to the milk yield immediately before the challenge (r = 0.86, P < 0.01). The correlation between the rate of recovery and milk loss was (r = 0.94, P < 0.01). Further, there was no significant slope (P > 0.1) to the relationship between the ratio (rate of recovery/milk loss) and days from calving, indicating that the force of recovery was unaffected by stage of lactation. These results suggest that differential smoothing can be a useful tool for quantifying biological disturbances in animal performance and for extracting features that relate to the potential and robustness of an animal.

Proteomic characterization of a temperature-sensitive conditional lethal in Drosophila melanogaster.

Genetic variation that is expressed only under specific environmental conditions can contribute to additional adverse effects of inbreeding if environmental conditions change. We present a proteomic characterization of a conditional lethal found in an inbred line of Drosophila melanogaster. The lethal effect is apparent as a large increase in early mortality at the restrictive temperature (29 degrees C) as opposed to normal survival at the permissive temperature (20 degrees C). The increased mortality in response to the restrictive temperature is probably caused by a single recessive major locus. A quantitative trait locus (QTL) region segregating variation affecting the lethal effect has been identified, allowing for a separation of primary/causal effects and secondary consequences in the proteome expression patterns observed. In this study, the proteomic response to the restrictive temperature in the lethal-line (L-line) was compared with the response in an inbred-control-line (IC-line) and an outbred-control-line (OC-line). Quantitative protein changes were detected using isobaric tags for relative and absolute quantitation (iTRAQ) two-dimensional liquid chromatography-tandem mass spectrometry. In all, 45 proteins were found to be significantly differently regulated in response to the restrictive temperature in the L-line as compared with the IC-line. No proteins were significantly differently regulated between the IC-line and the OC-line, verifying that differential protein regulation was specific to a genetic defect in the L-line. Proteins associated with oxidative phosphorylation and mitochondria were significantly overrepresented within the list of differentially expressed proteins. Proteins related to muscle contraction were also found to be differentially expressed in the L-line in response to the restrictive temperature, supporting phenotypic observations of moribund muscle hyper-contraction.