Heart rate, cardiac vagal tone, respiratory rate, and rectal temperature in dairy calves exposed to heat stress in a continental region.

Changes in non-invasive stress biomarkers were studied in shaded (n = 8) and non-shaded (n = 8) Holstein bull calves exposed to extreme heat load conditions in a continental region. Ambient temperature and humidity data were recorded for the S and NS hutch and exercise pen environments. Temperature-humidity-index (THI) was also calculated. Respiratory rate, rectal temperature, heart rate, and the root mean square of successive differences between R-R intervals (RMSSD) were recorded as animal-based indicators during three periods: (1) day 1, control day, during which all calves were shaded for 24 h (shade removal from non-shaded calves at 2400 h); (2) day 2, heat stress day, with shade over shaded calves; and (3) days 3-5, post-stress period, with shade over shaded calves. On the heat stress day, the maximum temperatures were 44.3 and 46.7 °C for the non-shaded hutch and pen environments, respectively. The temperatures were with 6.2 and 6.9 °C (P = 0.015 and P = 0.008) and the THIs were with 5.9 and 4.2 units higher (P = 0.020 and P = 0.032) in the non-shaded than in the shaded environment for the hutch and exercise pen, respectively. Shaded calves had with 42.3 ± 3.2 breaths/min higher respiratory rate than non-shaded ones on the heat stress day at 1200 h (P = 0.001), which was moderated to 20.1 ± 2.4 breaths/min at 1600 h (P = 0.023). Significant differences in respiratory rate occurred earlier than in any other animal-based parameter between shaded and non-shaded calves on day 2. The only significant group difference in rectal temperature was found at 1200 h on day 2 when THI exceeded 91 units, with 0.59 °C higher values for non-shaded calves (P = 0.045). The heart rate on days 2 and 3 was higher for non-shaded calves than for shaded ones. Group differences were also significant at 0800 h (18.2 ± 1.2 beats/min, P = 0.008), 1200 h (22.3 ± 1.4 beats/min, P = 0.003), 1600 h (15.3 ± 0.8 beats/min, P = 0.012), and 2000 h (19.0 ± 1.1 beats/min, P = 0.010) on day 2. Following a rapid daytime reduction, RMSSD showed a nighttime overcompensation in non-shaded calves on day 3 (between 0000 and 0600 h) and day 4 (between 0000 and 0800 h), exceeding the levels recorded on day 1 (control) and the levels for shaded calves, thus suggesting a recovery of the autonomic nervous system from heat stress. Based on our results, shading effectively reduced heat stress as evidenced by heart rate and RMSSD in addition to traditional measures of heat stress. Respiratory rate and heart rates exhibited by non-shaded calves support that the well-being of pre-weaned calves can be impaired in continental regions during following a heat stress day without providing shade.

Heart rate, heart rate variability, faecal glucocorticoid metabolites and avoidance response of dairy cows before and after changeover to an automatic milking system.

The heart rate variability (HRV) parameters of dairy cows were monitored during parlour (PARL) and the later installed automatic (AMS) milking on a small-scale commercial dairy farm in Hungary. The aim of the study was to assess stress in relation to the type of milking and the frequency of human interaction. Parlour milking involved regular moving and crowding of the animals with frequent human interaction, which were much less frequent in automatic milking. The first phase of the study was conducted prior to the changeover [n = 27] and the second two months afterwards [n = 19 (of the cows from the first phase)]. Heart rate (HR) was recorded by the Polar RS800 CX recording system. HRV parameters indicative of sympathovagal balance were calculated for periods of lying and standing in the barn, waiting before milking and milking, respectively. Morning and evening faecal glucocorticoid concentrations were also measured. Fear of humans was tested by an avoidance distance test. Baseline HRV parameters showed no difference (P > 0.05) between the two systems. In the periods before, during and after milking a higher sympathetic tone was detected in cows in the PARL phase. Mean faecal glucocorticoid concentrations were higher at the time of parlour milking. The avoidance distance did not differ between the two phases. The results suggest that automatic milking might be less stressful for cows than parlour milking, possibly due to the shorter duration of restraint after milking and the less frequent human interaction.

Salivary cortisol as a non-invasive approach to assess stress in dystocic dairy calves.

The intensity and the magnitude of saliva cortisol responses were investigated during the first 48 h following birth in newborn dairy calves which underwent normal (eutocic, EUT, n = 88) and difficult (dystocic, DYS, n = 70) calvings. The effects of parity and body condition of the dam, the duration of parturition, the time spent licking the calf, the sex and birth weight of the calf were also analyzed. Neonatal salivary cortisol concentrations were influenced neither by factors related to the dam (parity, body condition) nor the calf (sex, birth weight). The duration of parturition and the time spent licking the calf also had no effect on salivary cortisol levels. Salivary cortisol concentrations increased rapidly after delivery in both groups to reach their peak levels at 45 and 60 min after delivery in EUT and DYS calves, respectively supporting that the birth process means considerable stress for calves and the immediate postnatal period also appears to be stressful for newborn calves. DYS calves exhibited higher salivary cortisol concentrations compared to EUT ones for 0 (P = 0.022), 15 (P = 0.016), 30 (P = 0.007), 45 (P = 0.003), 60 (P = 0.001) and 120 min (P = 0.001), and for 24 h (P = 0.040), respectively. Peak levels of salivary cortisol and the cortisol release into saliva calculated as AUC were higher in DYS than in EUT calves for the 48-h of the sampling period (P = 0.009 and P = 0.003, respectively). The greater magnitude of saliva cortisol levels in DYS calves compared to EUT ones suggest that difficult parturition means severe stress for bovine neonates and salivary cortisol could be an opportunity for non-invasive assessment of stress during the early neonatal period in cattle.

Assessment of heat stress in 7-week old dairy calves with non-invasive physiological parameters in different thermal environments.

We estimated thermal stress in 7-week old Holstein bull calves during a warm episode in summer to study acute physiological responses of calves to heat stress. Data were collected over a 5-day period: day 1 (control), day 2 (heat stress), and a 3-day post-stress period in shaded (n = 8) and unshaded (n = 8) thermal environments. On the control day, both groups were shaded. Thermal environment was characterized by relative humidity, ambient temperature, and the temperature-humidity index (THI). Physiological variables included respiratory rate, rectal temperature, ear skin temperature and heart rate. Correlations between animal-based and meteorological indices were calculated, and ambient temperature correlated slightly better with physiological measures than THI. Rectal temperature was the only animal-based parameter that showed stronger correlations with the thermal indices when calculated for the shaded than for the unshaded environment [r = 0.42 vs. r = 0.47, P = 0.032 (ambient temperature), r = -0.39 vs. r = -0.45, P = 0.012 P = 0.015 (relative humidity), r = 0.41 vs. r = 0.46, P = 0.022 (THI)]. No differences were found between groups during the control day for any of the physiological parameters. During days 2 and 3, average and maximal values of respiratory and heart rates were higher in unshaded calves than in shaded ones. Maximal respiratory rates were in average by 25.9, 17.8 and 10.1 breaths/min lower in shaded calves than in unshaded calves for days 2, 3 and 4, respectively (P < 0.001, P < 0.001 and P = 0.024). Maximal heart rate was 127.4 ± 8.5 vs. 99.2 ± 6.3 beats/min on the heat stress day (P < 0.001), and 121.0 ± 6.9 vs. 103.4 ± 7.7 beats/min on day 3 (P = 0.006) in unshaded and shaded calves, respectively. Maximal body temperatures were higher measured either in the rectum or on the ear skin in unshaded calves than in shaded ones (with 0.5 and 1.6°C, P = 0.040 and P = 0.018, respectively), but only on the heat stress day. Based on our results, shading of young calves may be adequate for alleviating acute heat stress in continental regions. Ambient temperature is appropriate to estimate acute heat stress in dairy calves.

Correction: Assessment of heat stress in 7-week old dairy calves with non-invasive physiological parameters in different thermal environments.

[This corrects the article DOI: 10.1371/journal.pone.0200622.].

Timing of obstetrical assistance affects peripartal cardiac autonomic function and early maternal behavior of dairy cows.

Peripartal autonomic nervous system function and early maternal behavior were investigated in 79 multiparous Holstein-Friesian cows. Animals were allocated into four groups based on the technology of calving management: 1) unassisted calving in a group pen (UCG; N=19), 2) unassisted calving in an individual pen (UCI; N=21), 3) assisted calving with appropriately timed obstetrical assistance (ACA; N=20), and 4) assisted calving with premature obstetrical assistance (ACP; N=19). Heart rate, the high frequency (HF) component of heart rate variability (HRV) as a measure of vagal activity and the ratio between the low frequency (LF) and HF components (LF/HF ratio) as a parameter of sympathetic nervous system activity were calculated. Heart rate and HRV parameters were presented as areas under the curves (AUC) for the following periods: 1) prepartum period (between 96h before the onset of calving restlessness and the onset of restlessness), 2) parturition (between the onset calving restlessness and delivery), and 3) postpartum period (during a 48-h period after delivery). Pain-related behaviors were recorded during parturition (i.e., the occurrence of vocalization and stretching the neck towards the abdomen) and during a 2-h observation period after calving (i.e., the occurrence of vocalization, stretching the neck towards the abdomen and the duration of standing with an arched back). Early maternal behavior was observed during the first 2h following calving as follows: 1) latency and duration of sniffing calf's head/body, and 2) latency and duration of licking calf's head/body. No difference was found across groups in autonomic function before the onset of calving restlessness. Area under the heart rate curve was higher in ACP cows during parturition (39.6±2.5beats/min×h) compared to UCG, UCI and ACA animals (AUC=13.1±0.9beats/min×h, AUC=22.3±1.4beats/min×h and AUC=25.0±2.1beats/min×h, respectively). Area under the heart rate curve did not differ across the UCG, UCI and ACA groups during the postpartum period (AUC=65.2±16.7beats/min×h, AUC=58.0±14.2beats/min×h and AUC=62.9±12.1beats/min×h, respectively) but it was higher in ACP cows compared to the former groups (AUC=269.1±36.3beats/min×h). During parturition, area under the HF curve reflected a lower vagal tone (AUC=-30.5±1.6n.u.×h) in cows with premature obstetrical assistance than in animals that calved individually without farmer assistance (AUC=2.7±0.4n.u.×h) or with appropriately timed assistance (AUC=3.2±1.2n.u.×h). During parturition, LF/HF ratio showed greater sympathetic activity in ACP cows than in animals from any other group. Area under the HF curve was similar across UCG, UCI and ACA cows (AUC=-232.1±42.0n.u.×h, AUC=-163.4±35.6n.u.×h and AUC=-331.4±56.2n.u.×h, respectively) during the postpartum period and was the lowest in ACP cows (AUC=-1025.6±44.2n.u.×h) reflecting a long-term stress load in the latter group. During parturition, both vocalization and stretching the neck towards the abdomen occurred more often in UCG cows than in cows from any other groups, and the incidence of both behaviors was statistically higher in ACP cows than in UCI and ACA animals. There were no significant differences across groups in these behaviors during the 2-h postpartum observation. UCG cows had a shorter latency and a longer duration of maternal grooming during the first 2h following delivery compared to any other groups. UCI and ACA dams spent more time with licking the calf within the 2-h period after calving and had a shorter latency to sniff and lick the offspring compared to cows that received premature assistance. Group calving is less stressful for cows than calving in an individual pen either with or without obstetrical assistance. Calving in a group or with appropriately timed farmer assistance supports the expression of early maternal behavior and lead to a rapid postpartum recovery of the autonomic nervous system. Premature obstetrical assistance means stress for cows during parturition, leads to a prolonged postpartum recovery of the autonomic nervous system and inhibits the expression of early maternal behavior.

Cardiac autonomic activity has a circadian rhythm in summer but not in winter in non-lactating pregnant dairy cows.

This investigation was conducted to examine circadian and seasonal rhythms of heart rate and heart rate variability (HRV) by means of hour-by-hour recordings over 24h in a large population of non-lactating Holstein-Friesian pregnant cows [N=56, summer (June-July); N=61, winter (November-December)]. Data were collected during a 5-day period from each animal. Besides parameters of cardiac autonomic function [the high-frequency (HF) component of HRV and the ratio between the low-frequency (LF) and the HF components (LF/HF ratio)], the RR triangular index and Lmax were calculated. A clear circadian profile was observed for every parameter in summer. Heart rate elevated gradually with the course of the day from 7:00 to 17:00 o'clock and then slightly decreased from 18:00 to 6:00. Sympathovagal balance shifted towards sympathetic dominance during the daytime (increased LF/HF ratio), whereas parasympathetic activity was predominant during the night (increased HF). Lmax reflected a chaotic behavior of heart rate fluctuations during the afternoon in summer. Decreased values of RR triangular index indicated a sensitive period for cows between 14:00 and 16:00 o'clock in summer. During winter, except for the RR triangular (RRtri) index reflecting a high overall variability in R-R intervals between 12:00 and 23:00 o'clock, heart rate and HRV showed no periodicity over the 24-h period. The results suggest an impaired cardiac autonomic function during daytime in summer. HF, Lmax and RRtri index showed seasonal differences for both daytime and nighttime. Heart rate was higher in summer than in winter during the daytime, whereas the LF/HF ratio was higher in winter during the nighttime. Circadian and seasonal rhythms of cardiovascular function are presumably related to the differing temperature, and animal activity associated with summer and winter. As all of the investigated parameters are commonly used in bovine HRV research, these findings have practical implications for behavioral, physiological and welfare studies on dairy cattle.

Diverse associations in the ternary systems of ß-cyclodextrin, simple carbohydrates and phenyl derivatives of inorganic oxoacids.

Complex formation reactions of phenylboronic, phenylphosphonic, phenylarsonic and 4-aminophenyl arsonic acids with ß-cyclodextrin (cycloheptaamylose, ß-CD) and some simple carbohydrates (mannitol, sorbitol, glucose) have been studied using spectrophotometric, potentiometric methods and solubility measurements, supplemented with HPLC and IR analyses of the solid samples. Equilibrium constants have been determined at ionic strength of 0.2M (NaCl) and 25°C. ß-CD forms the most stable complexes with the neutral, undissociated forms of the acids, the stability constants are as follows: phenylboronic acid: 320 ± 36, phenylphosphonic acid: 108 ± 25, phenylarsonic acid: 97 ± 4 and 4-aminophenyl arsonic acid: 107 ± 10. The stability constants for the ß-CD-complexes of the ionic forms are much lower. Ternary complexes of low stability could be detected in the case of phenylphosphonic acid and sorbitol with the undissociated form and with glucose and the dianion. In more concentrated solutions phenylboronic acid forms insoluble complexes with mannitol, sorbitol and ß-CD. The solid phases obtained in the ternary systems are predominantly mixtures of ester type 3:1 complexes with the carbohydrate and 1:1 inclusion complex with the ß-CD. No significant interaction has been found with glucose. The phenomena can be explained by the differences in the structures of the components and by the changes in the H-bonding network of ß-CD on the complex formation.

Effect of substituents on activation parameters in aliphatic S(N)2 reactions. A DFT study.

The activation parameters and optimized structures of the reactants and transition states in the S(N)2 reactions of substituted pyridines and N,N-dimethylanilines with methyl iodide were computed at the DFT level in different solvents. The measured and calculated deltaG/deltaH/deltaS versus sigma plots proved to be linear, and their slopes, the deltadeltaG, deltadeltaH, and deltadeltaS reaction constants, were determined. The least solvent-dependent deltadeltaG reaction constants can be computed with acceptable accuracy. The calculated deltadeltaS data decrease only very slightly with the jointly increasing electron-withdrawing effect of the substituents and tightness of the transition states. The measured deltadeltaS values are influenced mainly by the change of solvation in the reactions, and deltadeltaH is also influenced by the reorganization of the solvent. Consequently, the experimental and calculated deltadeltaS and deltadeltaH reaction constants may deviate considerably from each other. In dipolar aprotic solvents the measured deltadeltaS was less than zero, and in protic solvents it was greater than zero. The ordering of the solvent molecules around the transition state with increasing charge is increased in the former but decreased in the latter media, as compared to the bulk of the solvents. The calculated deltaG(o), deltaH(o), and deltaS(o) parameters of the unsubstituted compounds agree relatively well with the experimental data for reactions of neutral molecules in dipolar aprotic solvents (e.g., XC6H4N(CH3)2 + CH3I). On the other hand, the measured and calculated activation parameters may show considerable deviations for reactions of ions (e.g., XC5H4NCH3+ + I-) and for any reaction in protic solvents.

The effect of peptide length on the cleavage kinetics of 2-chlorotrityl resin-bound ethers.

Different characteristics of cleavage kinetics of resin-bound amino alcohols and their peptide derivatives were observed in acid containing protic and aprotic solvent mixtures. The hydrolysis reactions are hindered by steric crowding around the cleaving C--O bond and accelerated by the special solvation effect of CF(3)CH(2)OH on the peptide chain as well as the increase of the strength and concentration of the acid. In trifluoroacetic acid containing mixtures, trifluoroacetylation of the peptide alcohols was detected. The appearance of O-trifluoroacetyl serine and threonine derivatives is detected in cleavage mixtures containing trifluoroacetic acid in anhydrous solvent.