Comparison of kinematic variables in defining lameness caused by naturally occurring rupture of the cranial cruciate ligament in dogs.

OBJECTIVE: To (1) discover kinematic variables that differ between dogs with cranial cruciate ligament rupture (CCLR) and control dogs and (2) to identify suitable outcome variables for longitudinal studies on the effects of surgical interventions for CCLR in dogs. STUDY DESIGN: In vivo case-control comparison. ANIMALS: Dogs with unilateral CCLR (n=16), and no other detectable neurologic or orthopedic disease and 10 dogs without CCLR, neurologic, or orthopedic disease. METHODS: Kinematic data was collected from dogs as they walked at constant speed on a treadmill, using an infrared camera system and adhesive reflective markers applied to the pelvic limbs. Data on 5 selected variables was extracted and comparisons made between paired pelvic limbs in both CCLR and control dogs and between control and CCLR dogs. RESULTS: Comparisons in CCLR dogs revealed significant differences between affected and unaffected limbs for many examined variables; the largest differences were in paw velocity and stifle angular velocity. There were highly significant differences between CCLR and control dogs when examining paired limb ratios for 4 of the variables, the largest differences were in stride length and paw velocity. CONCLUSIONS: Kinematic analysis provides straightforward and objective methods for defining the lameness associated with CCLR in dogs. Paw velocity and stride length were most notably reliable and not susceptible to systematic alterations in stifle joint dynamics that might be associated with specific surgical procedures. CLINICAL RELEVANCE: This study identifies several variables that can provide the objective measurements essential to evaluate the efficacy of surgical interventions for CCLR in dogs.

Use of behavioural and physiological responses for scoring sound sensitivity in dogs.

Sound sensitive dogs have exaggerated responses to sound stimuli that can negatively impact the welfare of the dog. Behavioural reactions combined with the response to sound involve a marked autonomic imbalance towards sympathetic predominance and release of cortisol. The purpose of the present study was to evaluate, in the laboratory, the cardiac autonomic modulation using heart rate variability (HRV) analysis, serum cortisol levels and behavioural parameters in response to sounds of fireworks in dogs with a history of sensitivity to fireworks. Based on these data, and combining qualitative measures and categorical measures, we propose one short and one full index of sound sensitivity in dogs. Six privately owned dogs with no history and another twelve dogs with a history of sound sensitivity to fireworks were used. The sound stimulus consisted of a standardised recording of fireworks (180-seconds long) with a peak intensity of 103-104 dB. The cardiac intervals were recorded using a frequency meter (Polar® RS800CX model) to evaluate the HRV, and the acquired data were processed using CardioSeries 2.4.1 software. Twenty-one behavioural parameters were analysed quantitatively by time, frequency or categorically by scores and were grouped in behavioural categories of arousal, fear, relaxation and "other". Sound sensitive dogs had exacerbated autonomic responses to the sound stimulus in the laboratory compared to non-sensitive dogs, with higher LF/HF ratios suggesting autonomic imbalance towards sympathetic predominance, but the cortisol levels were similar between the sensitive and non-sensitive dogs. Sound sensitive dogs showed pronounced responses for the parameters: alert and attention, search sound, startle, trembling, hiding, run away and less intense responses for the parameters rest and wink/sleep. Furthermore, the behavioural categories of arousal, fear, relaxation (lack of) and LF/HF were correlated to the caregiver's perception of the sound sensitivity of the dogs. Not only the short index for sound sensitivity (behavioural categories arousal, fear and relaxation, and LF/HF ratio) but also the full index for sound sensitivity (all behavioural categories, LF/HF and cortisol levels) was highly correlated to sound fear response at home. These indexes can contribute to the development of strategies to treat sound sensitive dogs.

Stress-induced c-Fos expression is differentially modulated by dexamethasone, diazepam and imipramine.

Immobilization stress upregulates c-Fos expression in several CNS areas. Repeated stress or the use of drugs can modulate stress-induced c-Fos expression. Here, we investigated in 40 different areas of the rat brain the effects of dexamethasone (SDX, a synthetic glucocorticoid), diazepam (SBDZ, a benzodiazepine), and imipramine (IMI, an antidepressant) on the c-Fos expression induced by restraint stress. Wistar rats were divided into four groups and submitted to 20 days of daily injection of saline (three first groups) or imipramine, 15 mg/kg, i.p. On day 21, animals were submitted to injections of saline (somatosensory, SS), SDX (1 mg/kg, i.p.), SBDZ (5 mg/kg, i.p.), or IMI (15 mg/kg, i.p.) before being submitted to restraint. Immediately after stress, the animals were perfused and their brains processed with immunohistochemistry for c-Fos (Ab-5 Oncogene Science). Dexamethasone reduced stress-induced c-Fos expression in SS cortex, hippocampus, paraventricular nucleus of the hypothalamus (PVH), and locus coeruleus (LC), whereas diazepam reduced c-Fos staining in the SS cortex, hippocampus, bed nucleus of stria terminalis, septal area, and hypothalamus (preoptic area and supramammillary nucleus). Chronic administration of imipramine decreased staining in the hippocampus, PVH, and LC, while increasing it in the nucleus raphe pallidus. We conclude that dexamethasone, diazepam and imipramine differentially modulate stress-induced Fos expression. The present study provides an important comparative background that may help in the further understanding of the effects of these compounds and on the brain activation as well as on the behavioral, neuroendocrine, and autonomic responses to stress.

Dipsogenic stimulation in ibotenic DRN-lesioned rats induces concomitant sodium appetite.

The main purpose of this study was to investigate whether dipsogenic stimuli influences the sodium appetite of rats with ibotenic acid lesion of the dorsal raphe nucleus (IBO-DRN). Compared to control, rats microinjected with phosphate buffer (PB-DRN), the ingestion of 0.3M NaCl was enhanced in IBO-DRN at 21 and 35 days after DRN lesion under a protocol of fluids and food deprivation. Despite of similar dipsogenic response observed both in IBO-DRN and PB-DRN treated with isoproterenol (ISO, 300 microg/kg, sc), the 0.3M NaCl intake was again significantly enhanced in IBO-DRN at 21 and 35 days post-lesion. Finally, treatment with polyethylene glycol (PEG, MW=20,000, 20%, w/v, 16.7 ml/kg, sc) induced higher dipsogenic response in IBO-DRN than PB-DRN at 21 day after lesion. In addition, IBO-DRN also expressed higher sodium appetite than PB-DRN, concomitantly with a drinking response. These results suggest that ibotenic lesion of DRN promote an increase of the brain angiotensinergic response, possibly settled within the subfornical organ, through paradigms which increase circulating ANG II levels. The current paper supports the hypothesis that the ibotenic lesion of DRN suppresses a serotonergic component implicated on the modulation of the sodium appetite and, therefore, furthering homeostatic restoration of extracellular fluid volume.

Acupuncture Affects Autonomic and Endocrine but Not Behavioural Responses Induced by Startle in Horses.

Startle is a fast response elicited by sudden acoustic, tactile, or visual stimuli in a variety of animals and in humans. As the magnitude of startle response can be modulated by external and internal variables, it can be a useful tool to study reaction to stress. Our study evaluated whether acupuncture can change cardiac autonomic modulation (heart rate variability); and behavioural (reactivity) and endocrine (cortisol levels) parameters in response to startle. Brazilian Sport horses (n = 6) were subjected to a model of startle in which an umbrella was abruptly opened near the horse. Before startle, the horses were subjected to a 20-minute session of acupuncture in acupoints GV1, HT7, GV20, and BL52 (ACUP) and in nonpoints (NP) or left undisturbed (CTL). For analysis of the heart rate variability, ultrashort-term (64 s) heart rate series were interpolated (4 Hz) and divided into 256-point segments and the spectra integrated into low (LF; 0.01-0.07 Hz; index of sympathetic modulation) and high (HF; 0.07-0.50 Hz; index of parasympathetic modulation) frequency bands. Acupuncture (ACUP) changed the sympathovagal balance with a shift towards parasympathetic modulation, reducing the prompt startle-induced increase in LF/HF and reducing cortisol levels 30 min after startle. However, acupuncture elicited no changes in behavioural parameters.

Analgesia and c-Fos expression in the periaqueductal gray induced by electroacupuncture at the Zusanli point in rats.

The need to use anaesthetised or restrained animals in acupuncture research in laboratory animals may represent a confounding variable, since both anaesthesia and stress alter the pain threshold and the activity of pain-related brain areas. In the current study we assessed the participation of the periaqueductal gray (PAG) in electroacupuncture's (EA) analgesic effects applied to the Zusanli point (36S) under carefully controlled stress conditions. Repeated immobilisation protocols (6 days, 1 h/day and 13 days, 2 h/day) were used to diminish the influence of acute immobilisation stress on c-Fos expression and analgesia (tail-flick test) induced by electroacupuncture on the 36S point (EA36S). Animals submitted to immobilisation alone (IMMO) or to electroacupuncture (100 Hz, 2-4 V, faradic wave) on a non-point region (EANP) were compared with animals submitted to electroacupuncture on the 36S point. In animals not previously submitted to repeated immobilisation, electroacupuncture on the 36S point induced analgesia and c-Fos expression in the PAG was not different from that induced by electroacupuncture at a non-acupuncture point. In animals submitted to repeated immobilisation (repeated immobilisation for 6 days or repeated immobilisation for 13 days), however, electroacupuncture on point 36S led to higher levels of analgesia and c-Fos expression, specifically in the ventrolateral PAG (vlPAG), as compared with animal groups subjected only to immobilisation or to electroacupuncture on a non-point. Our findings endorse previous results, and point to a specific part of the PAG involved in the effects of electroacupuncture at the Zusanli point.

Sertraline, a selective serotonin reuptake inhibitor, affects thirst, salt appetite and plasma levels of oxytocin and vasopressin in rats.

We investigated the effects of chronic administration of sertraline (SERT; approximately 20 mg kg(-1) day(-1) in drinking water), a selective serotonin reuptake inhibitor, on water and sodium intake and on plasma levels of oxytocin (OT) and vasopressin (AVP) in basal and stimulated conditions. Basal water intake was reduced in SERT-treated rats. After 24 h of water deprivation, rats treated with SERT for 21 days ingested less water than the control rats (9.7 +/- 0.5 versus 20.0 +/- 0.9 ml, respectively, at 300 min after water presentation, P < 0.0001). Subcutaneous injection of 2 m NaCl or isoproterenol evoked a lower dipsogenic response in rats treated with SERT for 21 days. Fluid and food deprivation also induced a weaker dipsogenic response in SERT-treated rats (1.6 +/- 0.5 versus 10.2 +/- 1.2 ml, at 300 min, P < 0.0001) but had no effect on saline intake. Sodium depletion induced a higher natriorexigenic response in the SERT group (5.6 +/- 1.3 versus 1.2 +/- 0.3 ml, at 300 min, P < 0.0002). Higher urinary density and lower plasma sodium levels were observed after SERT treatment. Sertraline also increased plasma levels of vasopressin and oxytocin (AVP, 2.65 +/- 0.36 versus 1.31 +/- 0.16 pg ml(-1), P < 0.005; OT, 17.16 +/- 1.06 versus 11.3 +/- 1.03 pg ml(-1), P < 0.0009, at the third week post-treatment). These data constitute the first evidence that chronic SERT treatment affects water and sodium intake in rats. These effects seem to be related to the hyponatraemia caused by the higher plasma levels of AVP and OT.