Viktor Jurkovich

Welfare implication of measuring heart rate and heart rate variability in dairy cattle: literature review and conclusions for future research

Heart rate (HR) measurements have been used to determine stress in livestock species since the beginning of the 1970s. However, according to the latest studies in veterinary and behaviour-physiological sciences, heart rate variability (HRV) proved to be more precise for studying the activity of the autonomic nervous system. In dairy cattle, HR and HRV indices have been used to detect stress caused by routine management practices, pain or milking. This review provides the significance of HR and HRV measurements in dairy cattle by summarising current knowledge and research results in this area. First, the biological background and the interrelation of the autonomic regulation of cardiovascular function, stress, HR and HRV are discussed. Equipment and methodological approaches developed to measure interbeat intervals and estimate HRV in dairy cattle are described. The methods of HRV analysis in time, frequency and non-linear domains are also explained in detail emphasising their physiological background. Finally, the most important scientific results and potential possibilities for future research are presented.

Prevalence of Coxiella burnetii in Hungary: screening of dairy cows, sheep, commercial milk samples, and ticks.

Q fever is an important zoonotic disease caused by Coxiella burnetii. There are few reliable data about C. burnetii infection available. The aim of this study was to assess the importance and potential infectious sources of Q fever in Hungary. A total of 215 milk samples (10 individual samples from each herd and 1 bulk tank milk sample from each cattle herd), and 400 serum samples (20 from each herd) were tested from 15 dairy cattle herds and 5 sheep flocks located in different parts of Hungary. The study found 19.3% (58/300) and 38.0% (57/150) seropositivity in cattle, and 0% (0/100) and 6.0% (3/50) seropositivity in sheep, by complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA), respectively. C. burnetii DNA was detected by IS1111 element-based TaqMan real-time polymerase chain reaction (PCR) in 8.7% (13/150) of individual dairy cow milk samples, 4.0% (2/50) of individual sheep milk samples, and 66.7% (10/15) of dairy bulk tank milk samples. Samples taken from nine different commercially-available pasteurized cow milk products from different Hungarian producers were also tested for the presence of C. burnetii DNA, and eight of these samples were found to be positive (88.9%). The real-time PCR examination of 5402 ixodid ticks collected from different parts of the country yielded negative results. Knowledge of the true prevalence of Q fever is crucial for policymakers involved in evidence-based decision making.

Heart Rate Variability as an Indicator of Chronic Stress Caused by Lameness in Dairy Cows

Most experimental studies on animal stress physiology have focused on acute stress, while chronic stress, which is also encountered in intensive dairy cattle farming–e.g. in case of lameness–, has received little attention. We investigated heart rate (HR) and heart rate variability (HRV) as indicators of the autonomic nervous system activity and fecal glucocorticoid concentrations as the indicator of the hypothalamic–pituitary–adrenal axis activity in lame (with locomotion scores 4 and 5; n = 51) and non-lame (with locomotion scores 1 and 2; n = 52) Holstein-Friesian cows. Data recorded during the periods of undisturbed lying–representing baseline cardiac activity–were involved in the analysis. Besides linear analysis methods of the cardiac inter-beat interval (time-domain geometric, frequency domain and Poincaré analyses) non-linear HRV parameters were also evaluated. With the exception of standard deviation 1 (SD1), all HRV indices were affected by lameness. Heart rate was lower in lame cows than in non-lame ones. Vagal tone parameters were higher in lame cows than in non-lame animals, while indices of the sympathovagal balance reflected on a decreased sympathetic activity in lame cows. All geometric and non-linear HRV measures were lower in lame cows compared to non-lame ones suggesting that chronic stress influenced linear and non-linear characteristics of cardiac function. Lameness had no effect on fecal glucocorticoid concentrations. Our results demonstrate that HRV analysis is a reliable method in the assessment of chronic stress, however, it requires further studies to fully understand the elevated parasympathetic and decreased sympathetic tone in lame animals.

Risk assessment of postpartum uterine disease and consequences of puerperal metritis for subsequent metabolic status, reproduction and milk yield in dairy cows

The objective of this study was to determine some metabolic and other factors predicting the risk of postpartum uterine disease (PUD), and the effects of puerperal metritis (PM) on metabolic status, reproduction and milk yield were analysed. A total of 105 Holstein-Friesian cows were included, and sampled on day < -14 prepartum and days 4, 10-14, 28-35 and 56-63 postpartum for metabolic tests. From day 4 the development of PUD, and from days 28-35 the ovarian activity was monitored. When grade > or = 1 + ketonuria was present on day 4 postpartum, this indicated a higher probability of PUD [odds ratio (OR) 2.64; P < 0.05] including PM occurring on days 10-14 (OR: 2.65; P < 0.05). Plasma nonesterified fatty acid (NEFA) concentrations > 0.200 mmol/l on days < -14 prepartum indicated a higher risk of uterine diseases (OR: 3.44; P < 0.05). The odds of PUD increased, depending on whether a body condition score (BCS) loss of > or = 1.0 occurred between days < -14 and 28-35 (OR: 2.82; P < 0.05), between days < -14 and 10-14 (OR: 4.79; P < 0.01) or between days 10-14 and 28-35 (OR: 10.81; P < 0.01). PM was more probable (OR: 27.3; P < 0.001) in cows with retained placenta. The risk of uterine diseases was lower in multiparous than in primiparous cows (OR: 0.29; P < 0.01). PM increased the risk of ovarian inactivity between days 28 and 35 (OR: 2.83; P < 0.05). Cows affected with PM (PM+ cows) showed lower milk production on day 4 (kg; P < 0.05) and lower milk production (P < 0.05), milk fat and milk protein production (kg; P < 0.01; P < 0.01) in the first 100 days of lactation than did PM-cows.

Short communication: Changes in heart rate variability of dairy cows during conventional milking with nonvoluntary exit

Heart rate variability (HRV), as a physiological measure of animal welfare, was investigated in 36 cows milked in a parallel milking parlor with nonvoluntary exit. Heart rate variability parameters measured during the morning resting (baseline period) were compared with those measured during different stages of the entire milking process. No differences were found in HRV parameters between the baseline period, preparation, and main milking. A considerable reduction in vagal activity was detected during the movement of the cows to the milking parlor (driving) and while cows were in the holding area. The parasympathetic measures of HRV decreased whereas the sympatho-vagal balance increased compared with baseline. The same pattern was observed regarding the stage between removing the teat cups and leaving the milking parlor (waiting). No differences in any sympathetic measures were observed between the baseline period and any of the milking stages. These findings indicate that the milking process itself (preparation and main milking) is not stressful for cows. Decreased parasympathetic activity during driving might be the result of the physical activity of the cows, whereas waiting in the holding area and in the milking stall after milking caused stress for animals.

Characterisation and effects of a xylanase enzyme preparation extracted from Thermomyces lanuginosus cultures

This paper describes the production of an enzyme preparation from the fungus Thermomyces lanuginosus. Thermal resistance, pH stability and lignocellulolytic activity of the enzyme preparation high in xylanase were studied on a variety of grains and forages. The enzyme preparation preserved more than 70% of its original xylanase activity for 4 and 1 h at 60 and 70 degrees C, respectively. The xylanase activity remained over 80% when the preparation was incubated for 30 min at pH 4.5. In vitro digestibility studies indicated that the enzyme digested 7.5, 8.5 and 8.0% of the dry matter (DM) of barley meal, wheat bran and oat meal samples, respectively. When applying 60-min incubation, 7.5, 7.3 and 8.4% of DM of the oat straw, alfalfa hay and triticale straw was digested, respectively. When the time of digestion was increased to 360 min, the sunflower hull showed 15.8% DM digestibility.

Associations between Heart Rate Variability Parameters and Housing- and Individual-Related Variables in Dairy Cows Using Canonical Correspondence Analysis.

We investigated the associations between heart rate variability (HRV) parameters and some housing- and individual-related variables using the canonical correspondence analysis (CCOA) method in lactating Holstein-Friesian dairy cows. We collected a total of 5200 5-min interbeat interval (IBI) samples from 260 animals on five commercial dairy farms [smaller-scale farms with 70 (Farm 1, n = 50) and 80 cows per farm (Farm 2, n = 40), and larger-scale farms with 850 (Farm 3, n = 66), 1900 (Farm 4, n = 60) and 1200 (Farm 5, n = 45) cows. Dependent variables included HRV parameters, which reflect the activity of the autonomic nervous system: heart rate (HR), the root mean square of successive differences (RMSSD) in IBIs, the standard deviation 1 (SD1), the high frequency (HF) component of HRV and the ratio between the low frequency (LF) and the HF parameter (LF/HF). Explanatory variables were group size, space allowance, milking frequency, parity, daily milk yield, body condition score, locomotion score, farm, season and physical activity (lying, lying and rumination, standing, standing and rumination and feeding). Physical activity involved in standing, feeding and in rumination was associated with HRV parameters, indicating a decreasing sympathetic and an increasing vagal tone in the following order: feeding, standing, standing and rumination, lying and rumination, lying. Objects representing summer positioned close to HR and LF and far from SD1, RMSSD and HF indicate a higher sympathetic and a lower vagal activity. Objects representing autumn, spring and winter associated with increasing vagal activity, in this order. Time-domain measures of HRV were associated with most of the housing- and individual-related explanatory variables. Higher HR and lower RMSSD and SD1 were associated with higher group size, milking frequency, parity and milk yield, and low space allowance. Higher parity and milk yield were associated with higher sympathetic activity as well (higher LF/HF), while individuals with lower locomotion scores (lower degree of lameness) were characterized with a higher sympathetic and a lower vagal tone (higher HR and LF/HF and lower RMSSD and SD1). Our findings indicate that the CCOA method is useful in demonstrating associations between HRV and selected explanatory variables. We consider physical activity, space allowance, group size, milking frequency, parity, daily milk yield, locomotion score and season to be the most important variables in further HRV studies on dairy cows.

The effects of Saccharomyces cerevisiae strains on the rumen fermentation in sheep fed with diets of different forage to concentrate ratios

The objective of this study was to compare the effects of two different Saccharomyces cerevisiae yeast strains on the rumen fermentation of sheep fed with two different total mixed rations. Three rumen-cannulated Merino wethers were used in a pilot study in which a phase of feeding a higher proportion of concentrate in the diet (HC; forage to concentrate ratio 40:60, as fed) was followed by a phase of lower proportion of concentrate in the diet (LC; forage to concentrate ratio 49:51, as fed). Both phases consisted of three 3-week periods as follows: no yeast supplementation, trehalose non-producing and trehalose-producing S. cerevisiae supplementations, respectively. Rumen fluid samples taken in the last days of each period were measured for pH, short chain fatty acid (SCFA) concentrations and ammonia content. The degradability of neutral detergent fibre (NDF) and starch was estimated by the in sacco method. Supplementation with the trehalose non-producing strain did not alter any variables tested in the HC phase. In the LC phase, though degradability was not altered, total SCFA concentration increased, resulting in a decrease of rumen fluid pH. The trehalose-producing strain increased total SCFA content and effectively prevented the consequent decrease in pH in both phases. These effects were more pronounced in the high-concentrate phase, as shown by higher molar proportion of propionate, increased level of ammonia content, and higher ruminal degradability of NDF and starch compared to control and the trehalose non-producing yeast strain.

Hypothalamic-pituitary-adrenal and cardiac autonomic responses to transrectal examination differ with behavioral reactivity in dairy cows.

Behavior, hypothalamic-pituitary-adrenal axis, and cardiac autonomic nervous system (ANS) activity were evaluated in response to transrectal examination in nonlactating Holstein-Friesian cows with different behavioral reactivity. According to behavioral reactions shown to the procedure of fixing the heart rate (HR) monitors, the 20 cows with the highest and the 20 cows with the lowest behavioral reactivity were involved in the study (high responder, n=20; and low responder, n=20, respectively). Activity of the ANS was assessed by HR and HR variability parameters. Blood and saliva were collected at 5 min before (baseline) and 0, 5 10, 15, 20, 30, 40, 60, and 120 min after the examination to determine cortisol concentrations. The examination lasted for 5 min. Cardiac parameters included HR, the root mean square of successive differences between the consecutive interbeat intervals, the high frequency (HF) component of heart rate variability, and the ratio between the low frequency (LF) and HF parameter (LF/HF). Following the examination, peak plasma and saliva cortisol levels and the amplitude of the plasma and saliva cortisol response were higher in high responder cows than in low responders. Areas under the plasma and saliva cortisol response curves were greater in high responder cows. Plasma and salivary cortisol levels correlated significantly at baseline (r=0.91), right after examination (r=0.98), and at peak levels (r=0.96). Area under the HR response curve was higher in low responder cows; however, maximum HR and the amplitude of the HR response showed no differences between groups. Minimum values of both parameters calculated for the examination were higher in high responders. Following the examination, response parameters of root mean square of successive differences and HF did not differ between groups. The maximum and the amplitude of LF/HF response and area under the LF/HF response curve were lower in low responder cows, suggesting a lower sympathetic activation of the ANS. Although changes in behaviors indicated that the procedure was painful for the animals, no differences were observed either in vocalization or in attendant behavior between groups during the examination. Our results demonstrate that behaviorally more reactive animals exhibit increased plasma and salivary cortisol concentrations and higher cardiac autonomic responsiveness to transrectal examination than less reactive cows. Salivary cortisol may substitute for plasma cortisol when assessing response of cattle to stress.

Detection of 70 kDa heat shock protein in the saliva of dairy cows.

This Research Communication describes, for the first time, the detection of HSP70 in saliva of dairy cows. Thermal stress is a major environmental stress that limits animal growth, metabolism, and productivity. The cellular response to heat stress involves the synthesis of heat shock proteins (HSPs), presumably to protect the functional stability of cells at increasing temperatures. HSP70 has been found to be present in cattle blood serum and may also be present in other secretory fluids, such as saliva, as already observed in humans. The aim of this study was to detect heat shock protein HSP70 in bovine saliva. Saliva samples were taken from higher- (n = 5) and lower milk producing (n = 5) Holstein-Friesian cows in summer and in winter for the detection of HSP70. HSP70 concentrations were assayed using the ELISA technique. Salivary HSP70 concentrations ranged from 0·524 to 12·174 ng/ml in cows. Higher salivary HSP70 concentrations were significantly associated with higher milk production and higher environmental temperature, but not with rectal temperature.