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Reduced oestrus behaviour expression or its absence (silent oestrus) results in subfertility in high-producing dairy cows. Insight into the genomic regulation of oestrus behaviour is likely to help alleviate reproduction problems. Here, gene expression was recorded in the ventral tegmental area (VTA) of high milk production dairy cows differing in the degree of showing oestrus behaviour (H - highly expressing versus L - lowly expressing), which was then analysed. Genes regulating cell morphology and adhesion or coding for immunoglobulin G (IgG) chains were differentially expressed in VTA between cows around day 0 and 12 of the oestrus cycle, but only in cows that earlier in life tended to show high levels of oestrus behaviour (H0 versus H12). The comparisons between H and L groups of cows also revealed differential expression of several genes (e.g. those of the IgG family or encoding for pro-melanin-concentrating hormone). However, any significant changes in VTA genes expression were detected in the comparison of L0 versus L12 cows. Altogether, the genes expression profile in VTA of cows highly expressing oestrus behaviour changes together with phases of the oestrus cycle, while in case of cows expressing oestrus behaviour lowly it remains stable. This supports the existence of genomic regulation by centrally expressed genes on the expression of oestrus behaviour in dairy cows.

Milk progesterone profiles were determined from samples obtained twice weekly for 100 d postpartum in 100 Holstein primiparous cows at a Dutch experimental farm. Three treatments were applied in a 2 x 2 x 2 factorial arrangement with high-low genetic merit for overall production, high-low caloric density diet, and 2-3 times milking/day as factors. Milk progesterone profiles were characterized by start of first ovarian cyclical activity (commencement of luteal activity, C-LA), length and peak milk progesterone concentration of first ovarian cycle, and number of ovarian cycles in first 100 d postpartum, as well as classified into normal, delayed, prolonged, and interrupted ovarian cyclical activity. Cows with a greater milk production had lower peak progesterone concentrations, especially if the high milk production was caused by milking 3 times a day. A more negative energy and protein balance was associated with later C-LA and less ovarian cycles within 100 d postpartum. Relationships between protein balance and C-LA differed between cows with a high genetic merit and a low genetic merit. Cows with a high genetic merit for production showed delayed C-LA with more negative protein balances, whereas this association was not observed among cows with a low genetic merit. Cows in negative energy balance had greater risk for prolonged ovarian cycles when there was no delay in C-LA than when C-LA was delayed.

Stress parameters that can be measured noninvasively may help to identify poor welfare in dogs that live in private homes and institutions. Behavioural parameters are potentially useful to identify stress, but require further investigation to establish which behaviours are appropriate. In the present study, behaviours were recorded and analysed for signs of acute stress in dogs. Simultaneously, saliva cortisol and heart rate were measured to support the interpretation of the behavioural data with regard to stress. Ten dogs of either sex, different ages and various breeds were each subjected to six different stimuli: sound blasts, short electric shocks, a falling bag, an opening umbrella and two forms of restraint. Each type of stimulus had been selected for its assumed aversive properties and was administered intermittently for 1 min. The stimuli that could not be anticipated by the dogs, sound blasts, shocks and a falling bag, tended to induce saliva cortisol responses and a very low posture. The remainder of the stimuli, which were administered by the experimenter visibly to the dog, did not change the cortisol levels but did induce restlessness, a moderate lowering of the posture, body shaking, oral behaviours, and to a lesser extent, yawning and open mouth. Pronounced increases in the heart rate were nonspecifically induced by each type of stimulus. Heart rate levels normalized within 8 min after stressor administration had stopped. Saliva cortisol levels decreased to normal within the hour. Correlations between behavioural and physiological stress parameters were not significant. From the present results, we conclude that in dogs a very low posture may indicate intense acute stress since dogs show a very low posture concomitant with saliva cortisol responses. Dogs may typically show increased restlessness, oral behaviours, yawning, open mouth and a moderate lowering of the posture when they experienced moderate stress in a social setting. The nonspecific character of canine heart rate responses complicates its interpretation with regard to acute stress.

A problem in assessing animal welfare is that collecting data in itself may be stressful to the animals. Therefore, noninvasive methods for collecting data have to be devised and tested. A first step in investigating saliva cortisol, urinary cortisol, and urinary catecholamine as noninvasive indicators of canine well-being is the validation of these hormonal measures as alternatives for those in plasma. Using a model of insulin (0.2 U/kg)-induced hypoglycemia, we report on stress-induced responses in saliva cortisol, urinary cortisol, and urinary catacholamines relative to cortisol and catecholamine responses in plasma. Hypoglycemia in six dogs induced significant (P0.05) increases in plasma cortisol and adrenaline but not noradrenaline. Saliva cortisol responses expressed as net area under the response curve correlated significantly with plasma cortisol responses (r0.92). Saliva cortisol levels measured 7 to 12% of plasma cortisol concentrations. Cortisol/creatinine rations in urine were significantly higher when voided after insulin administeration, compared to when voided after saline treatment. Insulin-induced increments in cortisol/ creatinine ratios were nonsignificant when urine samples were assayed after dichloromethane extraction. Although urinary adrenaline/creatinine (A/C) ratios were significantly correlated with maximum plasma adrenaline values after insulin administration, A/C ratios did not differ significantly between insulin and saline treatment. The present experiment provides strong support for using saliva sampling and urine collection as noninvasive methods to establish stress-induced cortisol responses. For measuring acute plasma adrenaline responses, measuring A/C ratios may not be a valid alternative.

Selection in dairy cattle for a higher milk yield has coincided with declined fertility. One of the factors is reduced expression of estrous behavior. Changes in systems that regulate the estrous behavior could be manifested by altered gene expression. This literature review describes the current knowledge on mechanisms and genes involved in the regulation of estrous behavior. The endocrinological regulation of the estrous cycle in dairy cows is well described. Estradiol (E2) is assumed to be the key regulator that synchronizes endocrine and behavioral events. Other pivotal hormones are, for example, progesterone, gonadotropin releasing hormone and insulin-like growth factor-1. Interactions between the latter and E2 may play a role in the unfavorable effects of milk yield-related metabolic stress on fertility in high milk-producing dairy cows. However, a clear understanding of how endocrine mechanisms are tied to estrous behavior in cows is only starting to emerge. Recent studies on gene expression and signaling pathways in rodents and other animals contribute to our understanding of genes and mechanisms involved in estrous behavior. Studies in rodents, for example, show that estrogen-induced gene expression in specific brain areas such as the hypothalamus play an important role. Through these estrogen-induced gene expressions, E2 alters the functioning of neuronal networks that underlie estrous behavior, by affecting dendritic connections between cells, receptor populations and neurotransmitter releases. To improve the understanding of complex biological networks, like estrus regulation, and to deal with the increasing amount of genomic information that becomes available, mathematical models can be helpful. Systems biology combines physiological and genomic data with mathematical modeling. Possible applications of systems biology approaches in the field of female fertility and estrous behavior are discussed.

Background The decline noticed in several fertility traits of dairy cattle over the past few decades is of major concern. Understanding of the genomic factors underlying fertility, which could have potential applications to improve fertility, is very limited. Here, we aimed to identify and study those genes that associated with a key fertility trait namely estrous behavior, among genes expressed in four bovine brain areas (hippocampus, amygdala, dorsal hypothalamus and ventral hypothalamus), either at the start of estrous cycle, or at mid cycle, or regardless of the phase of cycle. Results An average heat score was calculated for each of 28 primiparous cows in which estrous behavior was recorded for at least two consecutive estrous cycles starting from 30 days post-partum. Gene expression was then measured in brain tissue samples collected from these cows, 14 of which were sacrificed at the start of estrus and 14 around mid cycle. For each brain area, gene expression was modeled as a function of the orthogonally transformed average heat score values using a Bayesian hierarchical mixed model. Genes whose expression patterns showed significant linear or quadratic relationships with heat scores were identified. These included genes expected to be related to estrous behavior as they influence states like socio-sexual behavior, anxiety, stress and feeding motivation (OXT, AVP, POMC, MCHR1), but also genes whose association with estrous behavior is novel and warrants further investigation. Conclusions Several genes were identified whose expression levels in the bovine brain associated with the level of expression of estrous behavior. The genes OXT and AVP play major roles in regulating estrous behavior in dairy cows. Genes related to neurotransmission and neuronal plasticity are also involved in estrous regulation, with several genes and processes expressed in mid-cycle probably contributing to proper expression of estrous behavior in the next estrus. Studying these genes and the processes they control improves our understanding of the genomic regulation of estrous behavior expression.

Dog diets may differ in their effectiveness of maintaining satiety after a meal. Consequently, sensations of hunger, feeding motivation, physical activity, and sensitivity to environmental stressors may be increased. Dietary fibre may be effective in prolonging postprandial satiety depending on type and inclusion level. This study evaluated the effect of fibre fermentability on behaviour in dogs. Sixteen healthy adult dogs were housed individually and fed a low-fermentable fibre (LFF) diet containing 8.5% cellulose or a high-fermentable fibre (HFF) diet containing 8.5% sugar beet pulp and 2% inulin. Dogs were fed two equal portions at 8:30 and 18:30 according to energy requirements. Behaviour of dogs in their home-cage was recorded and analyzed by instantaneous scan sampling (2 × 24 h with 15 min intervals) and focal sampling continuous recordings (10 min per animal per hour, from 9:00 until 18:00). Dogs were subjected to a behaviour test composed of the subtests open-field, sudden-silence, novel-object, and acoustic-startle. The behavioural responses of each dog were recorded. Scores for the scan and focal samples were expressed per clock hour and DIET × TIME effects were tested statistically using Residual Maximum Likelihood (REML). Data from the tests were examined using principal component analysis resulting in the compilation of two components. Data were tested statistically for DIET and DIET × SUBTEST effects using REML. Variables specific for the open-field and novel-object test were analyzed using analysis of variance. For the scans, a significant DIET × TIME effect was found for resting. At night and in the morning, HFF dogs rested more compared to LFF dogs, but they rested less between 14:00 and 17:00. For the continuous recordings, the main findings were a tendency for DIET × TIME effect for time spent resting with a pattern consistent with that for the scans. The interaction was significant for inactive-alert (lie with head up or sitting) with HFF-fed dogs having lower values around 10:00–11:00 and higher values hereafter. Finally, time spent tail wagging was significantly higher for LFF-fed dogs just before the evening meal that may indicate higher level of arousal. For the behaviour tests, no significant DIET or DIET × SUBTEST effects were detected. It is concluded that compared to the LFF diet, the HFF diet increased inactivity in kennelled beagle dogs likely through the prolongation of postprandial satiety. This effect did not change the reaction to stressful events in kennelled laboratory dogs. Enhanced susceptibility to environmental stressors at times of hunger in sensitive companion dogs may occur but requires further study.

This study was part of a larger study that addressed whether milk production levels affect health risks in dairy cows as influenced by genetic merit for milk yield and management factors. Plasma samples were collected from 80 Holstein Friesian heifers at 2 weeks pre-partum and at 4 and 8 weeks post-partum in a 2 × 2 × 2 factorial arrangement design with the factors breeding value for milk production (high or low), milking frequency (2 or 3 times a day) and feed energy density (high or low). The following parameters indicative of the oxidative status were measured in plasma: ferric reducing ability of plasma (FRAP), α-tocopherol level, glutathione peroxidase activity (GSH-Px), superoxide dismutase activity (SOD), thiobarbituric acid reactive substances (TBARS) concentration and paraoxonase activity (PON). There was a significant effect of time for FRAP, α-tocopherol, GSH-Px and SOD, indicating changes in the plasma oxidative status around parturition. A high merit for milk yield, three-times-daily milking and especially feeding the high energy feed all stimulated milk production level. However, the influence on oxidative status parameters was minimal. Only the FRAP value was significantly lower in the high compared to the low energy group. Cows seem to undergo some oxidative stress around parturition, which can make them more susceptible to disorders. However, there were no clear indications that breeding value, milk frequency or feed energy level jeopardizes heifers in a way oxidative stress became critical.

International audience; Measuring HPA axis activity is the standard approach to the study of stress and welfare in farm animals. Although the reference technique is the use of blood plasma to measure glucocorticoid hormones (cortisol or corticosterone), several alternative methods such as the measurement of corticosteroids in saliva, urine or faeces have been developed to overcome the stress induced by blood sampling itself. In chronic stress situations, as is frequently the case in studies about farm animal welfare, hormonal secretions are usually unchanged but dynamic testing allows the demonstration of functional changes at several levels of the system, including the sensitization of the adrenal cortex to ACTH and the resistance of the axis to feedback inhibition by corticosteroids (dexamethasone suppression test). Beyond these procedural aspects, the main pitfall in the use of HPA axis activity is in the interpretation of experimental data. The large variability of the system has to be taken into consideration, since corticosteroid hormone secretion is usually pulsatile, follows diurnal and seasonal rhythms, is influenced by feed intake and environmental factors such as temperature and humidity, age and physiological state, just to cite the main sources of variation. The corresponding changes reflect the important role of glucocorticoid hormones in a number of basic physiological processes such as energy metabolism and central nervous system functioning. Furthermore, large differences have been found across species, breeds and individuals, which reflect the contribution of genetic factors and environmental influences, especially during development, in HPA axis functioning. Usually, these results will be integrated with data from behavioral observation, production and pathology records in a comprehensive approach of farm animal welfare.

Each year, millions of dogs worldwide are abandoned by their owners, relinquished to animal shelters, and euthanised because of behaviour problems. Nutrition is rarely considered as one of the possible contributing factors of problem behaviour. This contribution presents an overview of current knowledge on the influence of nutrition on canine behaviour and explores the underlying mechanisms by which diet may affect behaviour in animals. Behaviour is regulated by neurotransmitters and hormones, and changes in the availability of their precursors may influence behaviour. Tryptophan, the precursor of serotonin, may affect the incidence of aggression, self-mutilation and stress resistance. The latter may also be influenced by dietary tyrosine, a precursor to catecholamines. As diet composition, nutrient availability and nutrient interactions affect the availability of these precursors in the brain, behaviour or stress resistance may be affected. PUFA, especially DHA, have an important role as structural constituents in brain development, and dietary supply ofn-3 andn-6 PUFA could modify aspects of the dopaminergic and serotonergic system and, consequently, cognitive performance and behaviour. Finally, persistent feeding motivation between meals can increase stereotyped behaviour and aggression and decrease resting time. This feeding motivation may be altered by dietary fibre content and source. At present, few studies have been conducted to evaluate the role of nutrition in canine (problem) behaviour through the above mentioned mechanisms. Studies that explore this relationship may help to improve the welfare of dogs and their owners.

Increasing milk yields in modern dairy cows cause concern that high yield may impair the cows' health and welfare, for example, via negative effects on metabolic status and hypothalamo-pituitary-adrenocortical (HPA) function. This study aims to investigate whether a high level of milk production, and the associated metabolic status, affects HPA function in dairy cows and changes their adaptive capacity. Additionally, it aims to establish whether possible effects of milk production level only show under challenging conditions. Holstein-Friesian cows, which produced on average 11,443 and 7727 kg of fat and protein-corrected milk (FPCM)/305 d in their previous lactation, were compared. During the dry period, the cows were fed to requirements or overfed. High milk yield and the concomitant large energy deficit were associated with 1) increased pituitary (re)activity, i.e., increased ACTH baseline concentrations and higher ACTH concentrations after corticotropin-releasing hormone (CRH) administration, and 2) decreased adrenocortical reactivity, i.e., lower cortisol responses after ACTH administration. Although significant, the effects of milk production level on HPA function were relatively small. Animals showed seemingly normal hormonal responses to CRH and ACTH administration. Also, cortisol baseline concentrations were unaffected. It seems, therefore, unlikely that the adaptive capacity of the high-producing cows was significantly impaired compared with their low-producing herdmates.

Selection for a higher milk yield increases metabolic load via a higher yield per se and/or via physiological processes that facilitate milk yield, and it is difficult to differentiate between these two. Here, we aim to identify important pathways that contribute to the reduction in fertility following selection for higher yield. The associations between milk yield and fertility may run via pleiotropic effects, i.e. via functional pathways (for example related to intake), or linkage of genes and may involve changes in levels of hormones and metabolites. A number of studies have investigated the effects of genetic merit for milk yield on fertility, feed intake, energy balance and levels of metabolic and fertility hormones or metabolites. Differences in genetic merit were associated with differences in: (1) feed intake; (2) energy balance; and (3) plasma levels especially of GH, IGF-I, prolactin, progesterone, insulin, glucose, NEFAs and ketones. In the discussion we focus on the possible roles that energy balance, the growth hormone axis, and glucose together with insulin may have in the reduced fertility that is associated with high yield. The overall conclusion is that many minor pathways probably contribute, but that reduced metabolic fuel availability, rather than direct effects of hormone concentrations, is an important cause of poorer fertility with increasing genetic merit. (C) 2003 Elsevier B.V. All rights reserved.