Short communication: Chronology of different sexual behaviors and motion activity during estrus in dairy cows.

In studying the efficiency of a variety of methods for estrus detection in a large dairy herd, we suspected a definite sequence of estrus signs. Consequently, we observed a subset of animals continuously between 0400 and 2400 h, making a note of the precise timing and frequency of each sexual behavior. Sixteen Holstein-Friesian cows, >20 d postpartum, were equipped with motion activity-sensing neck collars and had milk progesterone profiles monitored simultaneously. The duration between the first and last observed estrus behavior was (mean ± SE) 14.0 ± 1.9 h, with a range 8.5 to 28.75 h. The duration of standing to be mounted (STBM) was 4.68 ± 1.49 h, with a range of 0.25 to 18.25 h. Sniffing the vulva of another cow occurred on average 5.5 ± 1.3 h (range = 0.25-18.25 h) before the first STBM. By ranking the first appearance of each behavior, we established that sniffing was followed by the active behaviors of mounting another cow and not accepting a mount, as well as the passive behaviors of being sniffed and STBM by another cow. Chin resting occurred before not accepting a mount and STBM. All these behaviors were observed in the reverse order after the last STBM. The mean profile of motion activity revealed an increase in motion activity with the onset of exploratory behaviors, and highest values occurred within the period of STBM. Such distinct behavioral sequences may be controlled by changes in peripheral progesterone and estradiol concentrations, as well as by subtle independent mechanisms via pheromones in differing concentrations or divergent composition.

Association of production diseases with motor activity-sensing devices and milk progesterone concentrations in dairy cows.

We have previously established that the efficiency of identifying oestrus with activity-sensing devices can be compromised by common production diseases; the present study was undertaken to determine how these diseases may affect device readings. A total of 67 Holstein-Friesian cows, >20 days postpartum, were equipped with activity-sensing neck collars and pedometers, and simultaneous milk progesterone profiles were also monitored twice a week. The influences of common production stressors on maximum activity and progesterone values were analysed. Approximately 30% potential oestrus events (low progesterone value between two high values) remained unrecognised by both activity methods, and progesterone values in these animals were higher on the potential day of oestrus when both activity methods did not detect an event (0.043 ±â€¯0.004 versus 0.029 ±â€¯0.004 ng/mL; P = 0.03). Data from a subset of 45 cows (two events each) were subjected to mixed models and multiple regression modelling to investigate associations with production diseases. Cow motor activity was lower in lame cows. Maximum progesterone concentrations prior to oestrus increased as time postpartum and body condition score (BCS) increased. There were also fewer days of low progesterone prior to oestrus associated with increases in BCS and maximum progesterone concentrations prior to oestrus. In conclusion, lameness was associated with lower activity values, but this suppression was insufficient to account for lowered oestrus detection efficiency of either device. However, associations were identified between production diseases and progesterone profiles.

KNDy neurone activation prior to the LH surge of the ewe is disrupted by LPS.

In the ewe, steroid hormones act on the hypothalamic arcuate nucleus (ARC) to initiate the GnRH/LH surge. Within the ARC, steroid signal transduction may be mediated by estrogen receptive dopamine-, ß-endorphin- or neuropeptide Y (NPY)-expressing cells, as well as those co-localising kisspeptin, neurokinin B (NKB) and dynorphin (termed KNDy). We investigated the time during the follicular phase when these cells become activated (i.e., co-localise c-Fos) relative to the timing of the LH surge onset and may therefore be involved in the surge generating mechanism. Furthermore, we aimed to elucidate whether these activation patterns are altered after lipopolysaccharide (LPS) administration, which is known to inhibit the LH surge. Follicular phases of ewes were synchronised by progesterone withdrawal and blood samples were collected every 2 h. Hypothalamic tissue was retrieved at various times during the follicular phase with or without the administration of LPS (100 ng/kg). The percentage of activated dopamine cells decreased before the onset of sexual behaviour, whereas activation of ß-endorphin decreased and NPY activation tended to increase during the LH surge. These patterns were not disturbed by LPS administration. Maximal co-expression of c-Fos in dynorphin immunoreactive neurons was observed earlier during the follicular phase, compared to kisspeptin and NKB, which were maximally activated during the surge. This indicates a distinct role for ARC dynorphin in the LH surge generation mechanism. Acute LPS decreased the percentage of activated dynorphin and kisspeptin immunoreactive cells. Thus, in the ovary-intact ewe, KNDy neurones are activated prior to the LH surge onset and this pattern is inhibited by the administration of LPS.

Kisspeptin, c-Fos and CRFR type 2 co-expression in the hypothalamus after insulin-induced hypoglycaemia.

Normal reproductive function is dependent upon availability of glucose and insulin-induced hypoglycaemia is a metabolic stressor known to disrupt the ovine oestrous cycle. We have recently shown that IIH has the ability to delay the LH surge of intact ewes. In the present study, we examined brain tissue to determine: (i) which hypothalamic regions are activated with respect to IIH and (ii) the effect of IIH on kisspeptin cell activation and CRFR type 2 immunoreactivity, all of which may be involved in disruptive mechanisms. Follicular phases were synchronized with progesterone vaginal pessaries and at 28 h after progesterone withdrawal (PW), animals received saline (n = 6) or insulin (4 IU/kg; n = 5) and were subsequently killed at 31 h after PW (i.e., 3 h after insulin administration). Peripheral hormone concentrations were evaluated, and hypothalamic sections were immunostained for either kisspeptin and c-Fos (a marker of neuronal activation) or CRFR type 2. Within 3 h of treatment, cortisol concentrations had increased whereas plasma oestradiol concentrations decreased in peripheral plasma (p < 0.05 for both). In the arcuate nucleus (ARC), insulin-treated ewes had an increased expression of c-Fos. Furthermore, the percentage of kisspeptin cells co-expressing c-Fos increased in the ARC (from 11 to 51%; p < 0.05), but there was no change in the medial pre-optic area (mPOA; 14 vs 19%). CRFR type 2 expression in the lower part of the ARC and the median eminence was not altered by insulin treatment. Thus, disruption of the LH surge after IIH in the follicular phase is not associated with decreased kisspeptin cell activation or an increase in CRFR type 2 in the ARC but may involve other cell types located in the ARC nucleus which are activated in response to IIH.

Effects of high somatic cell counts in milk on reproductive hormones and oestrus behaviour in dairy cows with special reference to those with concurrent lameness.

The present investigation aims to establish the reason(s) why dairy cows with high somatic cell counts (SCCs; >100,000 cells/ml) are less fertile than cows with low SCCs alone. The objective of Study One was to determine whether differences in steroid hormone profiles could explain the low incidence of ovulation in cows with combined High SCC and lameness. Between 30 and 80 days post-partum, animals were scored for SCC and lameness and three groups were formed: Healthy (n=22), High SCC alone (n=12) or High SCC + Lame (n=9). The ovarian follicular phases of all cows were synchronised by administering gonadotrophin releasing hormone (GnRH) followed seven days later by prostaglandin F2alpha (PG). Milk samples were collected daily throughout the entire study period; twice daily during the follicular phase, blood samples were taken and the ovaries were monitored using ultrasonography. Progesterone concentrations were similar in all three groups during each of five specific time periods, i.e. throughout the five days before PG injection, the peri-ovulatory period, on Day 5 and on Day 7, and during the mid luteal phase 12-17 days after ovulation (P>0.13). Mean plasma oestradiol concentrations monitored every 12h during the 36h period before ovulation were similar in all groups (Healthy, 2.80±0.30pg/ml; High SCC alone, 3.82±0.48pg/ml; High SCC+Lame 2.94±0.51pg/ml; P=0.175). The objective of Study Two was to establish whether cows with High SCC (scored and synchronised as above) display different behaviours, especially the intensity and timing of oestrus. Intervals from PG to the onset of oestrus or to the first stand-to-be-mounted (STBM) were longer for the High SCC cows than the Low SCC animals (n=8 and 20; P=0.011 and 0.002, respectively). Also, cows with High SCC tended to have a less intense oestrus and a lower maximum oestrus score per 30-min period than Low SCC cows (P=0.063 and 0.066, respectively). In conclusion, High SCC±lameness did not affect progesterone or twice daily oestradiol profiles but the onset of oestrus was delayed and oestrus tended to be less intense in cows with High SCC. These factors could explain low fertility associated with High SCC.

Kisspeptin, c-Fos and CRFR type 2 expression in the preoptic area and mediobasal hypothalamus during the follicular phase of intact ewes, and alteration after LPS.

Increasing estradiol concentrations during the late follicular phase stimulate sexual behavior and the GnRH/LH surge, and it is known that kisspeptin signaling is essential for the latter. Administration of LPS can block these events, but the mechanism involved is unclear. We examined brain tissue from intact ewes to determine: i) which regions are activated with respect to sexual behavior, the LH surge and LPS administration, ii) the location and activation pattern of kisspeptin cells in control and LPS treated animals, and iii) whether CRFR type 2 is involved in such disruptive mechanisms. Follicular phases were synchronized with progesterone vaginal pessaries and control animals were killed at 0 h, 16 h, 31 h or 40 h (n=4-6/group) after progesterone withdrawal (time zero). At 28 h, other animals received endotoxin (LPS; 100 ng/kg) and were subsequently killed at 31 h or 40 h (n=5/group). LH surges only occurred in control ewes, during which there was a marked increase in c-Fos expression within the ventromedial nucleus (VMN), arcuate nucleus (ARC), and medial preoptic area (mPOA), as well as an increase in the percentage of kisspeptin cells co-expressing c-Fos in the ARC and mPOA compared to animals sacrificed at all other times. Expression of c-Fos also increased in the bed nucleus of the stria terminalis (BNST) in animals just before the expected onset of sexual behavior. However, LPS treatment increased c-Fos expression within the VMN, ARC, mPOA and diagonal band of broca (dBb), along with CRFR type 2 immunoreactivity in the lower part of the ARC and median eminence (ME), compared to controls. Furthermore, the percentage of kisspeptin cells co-expressing c-Fos was lower in the ARC and mPOA. Thus, we hypothesize that in intact ewes, the BNST is involved in the initiation of sexual behavior while the VMN, ARC, and mPOA as well as kisspeptin cells located in the latter two areas are involved in estradiol positive feedback only during the LH surge. By contrast, disruption of sexual behavior and the LH surge after LPS involves cells located in the VMN, ARC, mPOA and dBb, as well as cells containing CRFR type 2 in the lower part of the ARC and ME, and is accompanied by inhibition of kisspeptin cell activation in both the ARC and mPOA.

Short-term individual housing induced social deficits in female Mongolian gerbils: attenuation by chronic but not acute imipramine.

Mongolian gerbils are highly sensitive to manipulations of their social environments. Housing females individually for short periods (in the order of 7-21 days) has been shown to produce robust and reliable impairments of their subsequent social behaviour. These effects are typified by a marked reduction in the social investigation of an unfamiliar male in a neutral arena and/or a marked increases in levels of freezing whilst and only whilst they are being socially investigated (Immobile in contact). These responses demonstrate housing induced impaired motivation to socially interact. These effects have also been shown to be sensitive to treatment with chronic (but not acute) administration of the selective serotonin reuptake inhibitor (SSRI) fluoxetine. It was therefore of interest to know if similar effects would be produced by treatment with the tricyclic antidepressant Imipramine. This mixed NA/5-HT reuptake inhibitor first developed in the 1950's is a commonly used standard in animal models of depression and remains in clinical use today. Female gerbils were individually housed for 7 days or maintained in single-sex groups of 4 for the same period. All animals were then randomly allocated to be administered with either 0, 10 or 20 mg/kg imipramine. Acute administration did not reverse the social impairments produced by the individual housing but did produce non-specific stimulant effects on locomotion in both housing conditions. These social impairments were however reduced after a further 14 days chronic treatment with 10 or 20 mg/kg imipramine and stimulant effects were no longer seen. Following chronic administration in group-housed animals locomotor stimulation was replaced with sedation, which resulted in a reduction in social behaviour. That is, opposite to the effect seen in Individual housed animals. It is therefore concluded that chronic treatment with imipramine serves to increase social behaviour but only in those animals with a pre-existing social deficit.

Comparison of oestrus detection methods in dairy cattle.

Sixty-seven Holstein-Friesian cows, from 20 days postpartum, were recruited into the study and fitted with both a pedometer (SAE Afikim) and a Heatime neck collar (SCR Engineers) and allocated a heat mount detector (either scratchcard [Dairymac] or KaMaR [KaMaR]) or left with none, relying only on farm staff observation. Common production stressors and other factors were assessed to determine their impact on the ability of each method to accurately detect oestrus and to investigate effects on the frequency of false-positive detections. Only 74 per cent of all potential oestrus periods (episodes of low progesterone) were identified by combining information from all methods. There was no difference between the methods in terms of sensitivity for detecting 'true oestrus events' (approximately 60 per cent), with the exception of scratchcards, which were less efficient (36 per cent). Pedometers and KaMaRs had higher numbers of false-positive identifications. No production stressors had any consequence on false-positives. The positive predictive values for neck collars or observation by farm staff were higher than those of other methods, and combining these two methods yielded the best results. Neck collars did not detect any of the nine oestrus events occurring in three cows with a body condition score (BCS) of less than 2, and the efficiency of correctly identifying oestrus was also reduced by high milk yield (odds ratio [OR]=0.34). Pedometer efficiency was reduced by lameness, low BCS or high milk yield (OR=0.42, 0.15 or 0.30, respectively).

Influence of lameness on follicular growth, ovulation, reproductive hormone concentrations and estrus behavior in dairy cows.

The objective of this study was to examine the effect of a chronic stressor, lameness, on reproductive parameters. Seventy cows 30-80 days post-partum were scored for lameness and follicular phases synchronized with GnRH followed seven days later by prostaglandin (PG). Fifteen Lame animals did not respond to GnRH ovarian stimulation. Milk progesterone for 5 days prior to PG was lower in the remaining Lame cows than Healthy herdmates. Fewer Lame cows ovulated (26/37 versus 17/18; P = 0.04) and the interval from PG to ovulation was shorter in Lame cows. In Subset 1 (20 animals), the LH pulse frequency was similar in ovulating animals (Lame and Healthy) but lower in Lame non-ovulators. An LH surge always preceded ovulation but lameness did not affect the interval from PG to LH surge onset or LH surge concentrations. Before the LH surge, estradiol was lower in non-ovulating cows compared to those that ovulated and estradiol concentrations were positively correlated with LH pulse frequency. In Subset 2 (45 cows), Lame ovulating cows had a less intense estrus than Healthy cows, although Lame cows began estrus and stood-to-be-mounted earlier than Healthy cows. In conclusion, we have identified several parameters to explain poor fertility in some chronically stressed animals. From 30 to 80 days post-partum, there was a graded effect that ranged from 29% Lame cows with absence of ovarian activity, whereas another 21% Lame cows failed to express estrus or ovulate a low estrogenic follicle; in 50% cows, many reproductive parameters were unaffected by lameness.

Retrospective audit of the investigation of patients with suspected acute subarachnoid haemorrhage.

INTRODUCTION: Recommended investigational care (RIC) of emergency department (ED) patients with suspected subarachnoid haemorrhage comprises lumbar puncture (LP) to detect xanthochromia if the preceding CT scan is negative. METHODS: Retrospective audit of the investigational care of 100 consecutive ED patients presenting with possible subarachnoid haemorrhage. RESULTS: Of the 100 patients, 91 had negative CT, and 36 (39.6%) of these patients had an LP performed to detect xanthochromia (i.e. RIC). Fifty-five of 91 (60.4%) patients did not receive RIC. Of the 55 patients who did not receive RIC, 25 (45.5%) had a documented senior clinical decision not to perform an LP; 15 (27.3%) had no documented reason; five (9.1%) refused consent; two (3.6%) had an LP but no xanthochromia requested, one patient did not have an LP because of technical issues, six patients underwent CT angiography (CTA), and one patient underwent magnetic resonance angiography (MRA), in the absence of a LP, following a negative CT. Two patients underwent CTA following a negative xanthochromia result. Patients admitted to the emergency extended care unit had 6.85 times the odds of receiving RIC (95% CI 2.20-21.4). CONCLUSIONS: Fifty-five (55) of 91 (60%) ED patients did not receive RIC. Fifteen of the 55 did not have any documented justification for not performing an LP with xanthochromia testing. Admission to an emergency extended care unit was a predictor of receiving RIC. Inappropriate use of CTA and MRA was identified. These findings have important implications for patient safety. Multifaceted strategies are required to close this evidence-practice gap.

The effect of a chronic stressor, lameness, on detailed sexual behaviour and hormonal profiles in milk and plasma of dairy cattle.

The objectives of the present study were to quantify the effects of a biological chronic stressor (lameness) on the duration and frequency of different oestrous behaviours in parallel with milk hormone profiles. Dairy cows 51.8 +/- 1.4 days postpartum (n = 59), including 18 non-lame control cows, were scored for lameness and closely observed for signs of oestrus having had their follicular phases synchronized by administration of gonadotrophin-releasing-hormone (GnRH) followed by prostaglandin F(2alpha) (PG) 7 days later. Lameness shortened the period when herd-mates attempted to mount the lame cows (1.83 +/- 0.69 h vs 5.20 +/- 1.53 h; p = 0.042) but did not affect the overall duration of total behaviours (lame 12.3 +/- 1.3 h vs non-lame 15.2 +/- 1.3 h). Lameness also lowered the intensity of oestrus [1417 +/- 206 points (n = 18) vs 2260 +/- 307 points (n = 15); p = 0.029]. Throughout the synchronized oestrous period, lame cows mounted the rear of herd-mates less frequently (p = 0.020) and tended to chin rest less (p = 0.075). Around the period of maximum oestrous intensity, lameness also diminished the proportion of cows mounting the rear of another cow and chin resting (p = 0.048, p = 0.037, respectively). Furthermore, lame cows had lower progesterone values during the 6 days before oestrous (p < or = 0.05). Fewer lame cows were observed in oestrus following PG (non-lame 83%, lame 53%; p = 0.030); however, if prior progesterone concentrations were elevated, lame cows were just as likely to be observed in oestrus. In conclusion, following endogenous progesterone exposure, lameness shortens the period when herd-mates attempt to mount lame cows but does not affect the incidence of oestrous. However, lame cows are mounted less frequently and express oestrus of lower intensity. This is associated with lower progesterone prior to oestrus but not with abnormal oestradiol or cortisol profiles in daily milk samples.

Further neurochemical and behavioural investigation of Brattleboro rats as a putative model of schizophrenia.

Brattleboro (BRAT) rats are a mutant variant of the Long-Evans (LE) strain deficient in the neurohormone vasopressin. BRAT rats show behavioural alterations relevant to schizophrenia. In particular, BRAT rats show deficits in prepulse inhibition (PPI) and alterations in various measures of cognition. The aim of this study was to replicate the reported PPI deficits in BRAT rats and its reversal by antipsychotic drugs and to investigate other behavioural and neurochemical characteristics. Acoustic startle reactivity, PPI, spontaneous and amphetamine-induced locomotor activity (LMA) and ex-vivo steady state neurochemistry were measured in male homozygous BRAT rats and LE rats. The effects of antipsychotics on PPI deficits were also determined. Relative to LE, BRAT rats showed enhanced startle reactivity, hyperactivity to a novel environment, PPI deficits and decreased levels of dopamine and DOPAC (dihydroxyphenylacetic acid) in the frontal cortex. BRAT and LE rats showed similar levels of hyperactivity following amphetamine (0.26 mg/kg s.c.). PPI deficits were attenuated by acute clozapine (5-10 mg/kg s.c.), risperidone (0.1-1 mg/kg i.p.), haloperidol (0.1-0.5 mg/kg p.o.) and less robustly by olanzapine (0.3-3 mg/kg s.c.). Chronic administration of clozapine (5 mg/kg s.c., once daily) attenuated baseline hyperactivity and elevated PPI of both strains. Clozapine concentrations were higher in BRAT brains compared with LE rats. These data confirm the reported PPI deficit in BRAT rats and its reversal by antipsychotic drugs, suggesting BRAT rats may represent a potential model for identifying novel antipsychotic drugs.

The Royal Australian and New Zealand College of Radiologists (RANZCR) relative value unit workload model, its limitations and the evolution to a safety, quality and performance framework.

The study reports on the evolution of the Australian radiologist relative value unit (RVU) model of measuring radiologist reporting workloads in teaching hospital departments, and aims to outline a way forward for the development of a broad national safety, quality and performance framework that enables value mapping, measurement and benchmarking. The Radiology International Benchmarking Project of Queensland Health provided a suitable high-level national forum where the existing Pitman-Jones RVU model was applied to contemporaneous data, and its shortcomings and potential avenues for future development were analysed. Application of the Pitman-Jones model to Queensland data and also a Victorian benchmark showed that the original recommendation of 40,000 crude RVU per full-time equivalent consultant radiologist (97-98 baseline level) has risen only moderately, to now lie around 45,000 crude RVU/full-time equivalent. Notwithstanding this, the model has a number of weaknesses and is becoming outdated, as it cannot capture newer time-consuming examinations particularly in CT. A significant re-evaluation of the value of medical imaging is required, and is now occurring. We must rethink how we measure, benchmark, display and continually improve medical imaging safety, quality and performance, throughout the imaging care cycle and beyond. It will be necessary to ensure alignment with patient needs, as well as clinical and organisational objectives. Clear recommendations for the development of an updated national reporting workload RVU system are available, and an opportunity now exists for developing a much broader national model. A more sophisticated and balanced multidimensional safety, quality and performance framework that enables measurement and benchmarking of all important elements of health-care service is needed.

Evaluation of the pro-cognitive effects of the AMPA receptor positive modulator, 5-(1-piperidinylcarbonyl)-2,1,3-benzoxadiazole (CX691), in the rat.

RATIONALE: Positive allosteric modulators of the glutamatergic alpha-amino-3-hydroxy-5-methyl-4-isoazolepropionic acid (AMPA) receptor do not stimulate AMPA receptors directly but delay deactivation of the receptor and/or slow its desensitisation. This results in increased synaptic responses and enhanced long-term potentiation. Thus, it has been suggested that such compounds may have utility for the treatment of cognitive impairment. OBJECTIVES: The objective of the study was to investigate the effect of an AMPA positive modulator, CX691, (1) in three rodent models of learning and memory, (2) on neurochemistry in the dorsal hippocampus and medial prefrontal cortex following acute administration, and (3) on brain-derived neurotrophic factor (BDNF) messenger RNA (mRNA) expression in the rat hippocampus following acute and sub-chronic administration. RESULTS: CX691 attenuated a scopolamine-induced impairment of cued fear conditioning following acute administration (0.1 mg/kg p.o.) and a temporally induced deficit in novel object recognition following both acute (0.1 and 1.0 mg/kg p.o.) and sub-chronic (bi-daily for 7 days) administration (0.01, 0.03, 0.1 mg/kg p.o.). It also improved attentional set-shifting following sub-chronic administration (0.3 mg/kg p.o.). Acute CX691 (0.1, 0.3 and 1.0 mg/kg, p.o.) increased extracellular levels of acetylcholine in the dorsal hippocampus and medial prefrontal cortex and dopamine in the medial prefrontal cortex. Sub-chronic administration of CX691 (0.1 mg/kg, p.o.) elevated BDNF mRNA expression in both the whole and CA(1) sub-region of the hippocampus (P < 0.05). CONCLUSIONS: Collectively, these data support the pro-cognitive activity reported for AMPA receptor positive modulators and suggest that these compounds may be of benefit in treating disorders characterised by cognitive deficits such as Alzheimer's disease and schizophrenia.

Influence of somatic cell count, body condition and lameness on follicular growth and ovulation in dairy cows.

The objective of this study was to investigate the effect of somatic cell count (SCC), body condition score (BCS) or lameness score on ovarian follicular growth and ovulation in dairy cows. Seventy four animals 30-80 days post-partum were monitored for all three conditions before synchronization of ovarian follicular phases by administration of gonadotrophin releasing hormone (GnRH) followed seven days later with prostaglandin F2alpha (PG). Ultrasonography of both ovaries twice daily throughout the follicular phase revealed that fewer animals with combined high SCC and lameness (4/9) ovulated compared to healthy animals (19/21; P=0.006) or animals with only high SCC (11/11; P=0.004) or only lameness (21/27; P=0.06). Overall, regardless of the presence of other concurrent conditions, fewer lame cows ovulated than Non Lame animals (30/42 and 30/32; P=0.015). Mean follicular growth and maximum follicular diameter were unaffected by any of the three conditions. However, dominant follicle growth and maximum diameter were greater in the 60 animals that ovulated compared to the 14 that did not; 1.83+/-0.16 versus 0.96+/-0.26mm/day (P=0.014) and 19.4+/-0.4 versus 16.4+/-1.2mm (P=0.003), respectively. In conclusion, lameness reduced the proportion of cows that ovulated and the synergistic effect of high SCC and lameness reduced that proportion further. However, follicular growth and maximum follicular diameter were unaffected by high SCC, low BCS or lameness.

Lameness, activity time-budgets, and estrus expression in dairy cattle.

The aim of the present study was to identify specific behavioral patterns that contribute to diminished estrus expression in lame cows. Behavioral scan and focal sampling were used to examine the effect of lameness on daily activity budgets, sexual behavior, feeding activities, and body condition score. A total of 59 milking cows (51.8 +/- 1.4 d postpartum) were monitored on a commercial dairy farm for 5 d following estrus synchronization. Overall, lame cows (n = 39) spent proportionately less time elevated on their feet and more time lying down compared with nonlame cows (n = 20). This included lame cows spending less time walking or standing. Overall, the total proportion of scans in which an estrous behavior was observed was very small but tended to be smaller for lame compared with nonlame cows. Throughout a day, lame cows displayed a lower proportion of estrous behaviors in the early morning. Lameness did not affect durations of drinking, grazing, or ruminating, or how these behavioral states fluctuated throughout the day. Similarly, rumination chewing rates were the same for lame and nonlame cows, and there was no association between lameness and dominance/displacement while feeding at a feed-fence. Lame cows did, however, have a slower bite rate at pasture and had a lower body condition score. Lame cows were also nearer the rear of the herd, both as they left the field and when entering the milking parlor. In conclusion, lame cows have longer lying times and spend less time standing, walking, and expressing an estrous behavior. Lame cows also have a lower bite rate at pasture and are more likely to be of lower body condition score.

Noradrenergic control of GnRH release from the ewe hypothalamus in vitro: sensitivity to oestradiol.

The present study investigates the influence of alpha(1)-adrenoreceptors in GnRH release in vitro and determines whether oestradiol modulates alpha(1)-adrenoreceptor-GnRH interaction. Within 10 min after ewe sacrifice, saggital midline hypothalamic slices were dissected, placed in oxygenated Minimum Essential Media-alpha (MEM-alpha) at 4 degrees C and within 2 h were singly perifused at 37 degrees C with oxygenated MEM-alpha (pH 7.4; flow rate 0.15 ml/min), either with or without oestradiol (24 pg/ml). After 4-h equilibration, 10-min fractions were collected for 4 h interposed with a 10-min exposure at 60 min to specific alpha(1)-adrenoreceptor agonist (methoxamine) or antagonist (thymoxamine) at various doses (0.1-10 mm). The alpha(1)-adrenoreceptor agonist (10 mm) increased (p < 0.05) GnRH release at 90 min both in presence and absence of oestradiol. However, in presence of oestradiol, alpha(1)-adrenoreceptor agonist (10 mm)-induced GnRH release remained elevated (p < 0.05) for at least 60 min. The bioactivity of the released GnRH was studied using a hypothalamus-pituitary sequential double-chamber perifusion. Only after exposure of hypothalamic slices to alpha(1)-adrenoreceptor agonist (10 mm), did the hypothalamic eluate stimulate LH release from pituitary fragments (n = 9, 7.8 +/- 12.3-36.2 +/- 21.6 ng/ml) confirming that the alpha(1)-adrenoreceptor agonist stimulated release of biologically active GnRH. In summary, GnRH release from the hypothalamus is under stimulatory noradrenergic control and this is potentiated in the presence of oestradiol.

GABA control of GnRH release from the ewe hypothalamus in vitro: sensitivity to oestradiol.

The present study examines the involvement of GABA(A or B) receptors in gonadotrophin-releasing hormone (GnRH) release in vitro and determines whether oestradiol modulates gamma-aminobutyric acid (GABA)-GnRH interaction. Within 10 min after ewe killing, hypothalamic slices were dissected and placed in oxygenated Minimum Essential Media (MEM)-alpha at 4 degrees C; within 2 h, slices were singly perifused at 37 degrees C with oxygenated MEM-alpha (0.15 ml/min), with or without oestradiol (24 pg/ml). After 4 h equilibration, fractions were collected for 4 h interposed with a 10 min exposure to specific GABA(A or B) receptor ligands (0.1-10 mM). The GABA(A or B) agonists (muscimol or baclofen) did not greatly influence GnRH release. However, GnRH increased (p < 0.05) after exposure to 10 mM GABA(A or B) antagonists (bicuculline or CGP52432, respectively). The GABA(A) antagonist stimulated greater sustained GnRH release (p < 0.05) in the absence of oestradiol than in its presence. The bioactivity of the released GnRH was studied using a hypothalamus-pituitary sequential double-chamber perifusion. Only after exposure of hypothalamic slices to the GABA(A) antagonist, did the hypothalamic eluate stimulate luteinizing hormone release from pituitary fragments (p < 0.05) confirming that the GABA(A) antagonist stimulated release of biologically active GnRH. In summary, GnRH release from the hypothalamus is predominantly under GABA(A) receptor inhibitory control and this is attenuated in the presence of oestradiol.

Chronic stress, hormone profiles and estrus intensity in dairy cattle.

The objectives of the present study were to determine if lameness, a model for a natural chronic stressor, affects hormone concentrations in milk prior to estrus and/or the subsequent expression of estrus in the postpartum period. Dairy cows>20 days postpartum were scored for lameness and observed for estrus intensity using a weighted scoring system (>100 points=estrus=Day 0). Increasing lameness score was not associated with daily profiles of milk progesterone (throughout Days -18 to 0), estradiol (Days -6 to 2) or cortisol (Days -18 to 2) around estrus, maximum estradiol values or estradiol concentrations on Day 0. However, post hoc pair wise comparisons revealed that prior to estrus, severely lame cows had lower maximum progesterone concentrations compared to nonlame cows (1.3+/-0.1, 1.2+/-0.2, 0.7+/-0.1 ng/ml milk; P=0.042). Furthermore, severely lame cows expressed behavioral estrus with lower intensity (284+/-128 points, n=9) compared to moderately lame (662+/-310 points, n=9) or nonlame animals (583+/-275 points, n=18; P=0.05 and P=0.02, respectively). Resting concentrations of cortisol (Days 20-80 postpartum) did not vary between days postpartum or lameness score. The incidence of behavioral estrus was not affected by increasing lameness score, as 54.2%, 56.2% and 50.0% periods with low progesterone were associated with spontaneous estrus expression, respectively. Concluding, in this biological model of chronic stress, lameness did not affect the incidence of behavioral estrus but did reduce estrus intensity once ovarian cyclicity had resumed after calving. This reduced intensity of estrus was associated with lower maximum progesterone values prior to estrus but not abnormal daily cortisol or estradiol values in milk.