Steven W. Eicker

Analysis of correlated continuous repeated observations: modelling the effect of ketosis on milk yield in dairy cows

This study used mixed models analysis to demonstrate the advantages of a repeated measures technique for a continuous variable over a single measure technique. As an illustration, the loss of milk yield due to ketosis was studied in 2604 multiparous New York State Holstein cows belonging to eight herds, calving between 1991 and 1993. Two methods of analysis were presented: The first treated milk yield as a continuous, summary measure (projected 305-day milk yield); the second treated milk yield as repeated measurements (test-day milk yields). In the first example, with 305-day milk yield as the outcome, ketosis was treated as a binary covariate. Ketosis had no effect on the 305-day milk yield. In the second example, with monthly test-day milk yields as the outcome, four different covariance structures (simple, compound symmetry, autoregressive, and unstructured) to model the association among the repeated measurements were compared. With this approach, ketotic cows yielded significantly less milk per day both before and immediately after diagnosis than did non-ketotic cows. Based on the goodness-of-fit statistics, it was unclear whether an autoregressive or unstructured covariance structure was best. However, an autoregressive structure, in which the previous and current test-day milk yields are assumed to be correlated, was considered more suitable in this study; it is a simpler and more appropriate covariance structure for this particular problem than is an unstructured covariance structure. Nevertheless, with the test-day approach, any of these correlation structures could be used to estimate milk loss after disease. Based on these findings, it is recommended that a repeated measures approach, rather than a single measure approach, be used to study the short-term effect of disease on milk yield.

The effect of strenuous exercise on mRNA concentrations of interleukin-12, interferon-gamma and interleukin-4 in equine pulmonary and peripheral blood mononuclear cells.

The effect of strenuous exercise on the mRNA concentrations of interleukin-12p35 subunit (IL-12p35), interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) in equine pulmonary and peripheral blood mononuclear cells (PBMCs) was investigated. We hypothesized that strenuous exercise would suppress the expression of IL-12p35, IFN-gamma and augment the expression of IL-4. Eleven horses were randomly divided into two groups, a stall-confined control group (n=5) and an exercise-conditioned treatment group (n=6). Bronchoalveolar and PBMCs were obtained from horses in the treatment group prior to the commencement of a 9-week conditioning program and 24h after the completion of a maximum exercise test conducted in week 12. Samples were obtained simultaneously from control horses. Differential counts were performed on the bronchoalveolar lavage cells. Real-time PCR was performed on the pulmonary and PBMCs to quantitate cytokine expression using equine-specific primers and Taqman probes. Target gene expression was normalized to 18s rRNA expression. With the exception of IL-4 in the BALF cells, mRNA for the three cytokines was detected in the mononuclear cells from all horses at both sampling times. There were no significant differences in the cytokine mRNA concentrations between the two groups of horses at either of the sampling times. These findings demonstrate that strenuous treadmill exercise does not exert a deleterious effect on gene expression for IL-12p35, IFN-gamma or IL-4 when assessed in horses 24h following the intense physical activity.

Pulmonary-locomotory interactions in exercising dogs and horses

In exercising quadrupeds, limb movement is often coupled with breathing frequency. This finding has lead some investigators to conclude that locomotory forces, associated with foot plant, abdominal visceral displacements or lumbo-sacral flexion, are the primary determinants of airflow generation. Analysis of respiratory muscle electrical activation (EMG) and contraction profiles in chronically instrumented dogs and horses, along with measurements of esophageal pressure (Pes) changes and limb movements, provide evidence that each breath during the exercise hyperpnea is determined by respiratory neuromuscular events. Specifically: (1) Phasic diaphragmatic EMG and tidal shortening are always synchronous with decreases in Pes; (2) decrements in Pes are always associated with inspiratory flow generation; and (3) strict phase coupling between breathing and stride frequency is not obligatory. Thus, although locomotory-associated forces may minimally assist with flow generation, they are not the primary determinants of breathing during exercise.

The effect of exercise on diaphragmatic activation in horses

Horses chronically-instrumented with costal diaphragmatic electromyographic electrodes were studied during exercise while unencumbered by a breathing mask. Exercise-associated changes in esophageal (Pes), gastric (Pga) and transdiaphragmatic (Pdi) pressures were measured and related to diaphragmatic electromyographic activity (CS EMG) and to left forelimb impact. In all breaths examined, CS EMG always coincided with decrements in Pes. For all exercise trials, linear increases in CS EMG, Pga and Pdi and linear decreases in Pes, as a function of exercise intensity, always occurred. During all gaits, breathing frequency (fR) was entrained with stride frequency (fS) one for one. However, a constant phase-coupling relationship between fR and fS, observed when horses cantered and galloped, was absent when horses walked or trotted. We conclude that biomechanical forces contribute minimally to ventilation in exercising horses, that the diaphragm is always phasically active during each breath and its total electrical activity and mechanical output are proportional to the exercise hyperpnea.