Development of an in Vivo Lipopolysaccharide Inflammation Model to Study the Pharmacodynamics of COX-2 Inhibitors Celecoxib, Mavacoxib, and Meloxicam in Cockatiels (Nymphicus hollandicus).

Nonsteroidal anti-inflammatory drugs (NSAIDs) are used frequently in avian medicine for their antipyretic, analgesic, and anti-inflammatory properties during surgery and for diseases that cause tissue damage and inflammation. NSAIDs inhibit cyclooxygenase (COX) enzymes, which are responsible for the induction of pyresis, pain, and inflammation. In our study, a lipopolysaccharide-induced (LPS) pyresis model was optimized using cockatiels (Nymphicus hollandicus) as subject birds (four males/three females) and validated in two females and one male, characterized by an intravenous bolus injection of LPS (7.5 mg/kg) administered at T0 and T24 (24 hours following the first LPS injection). To demonstrate the feasibility of the model to assess pharmacodynamic (PD) parameters of different NSAIDs, mavacoxib 4 mg/kg (four males/four females), celecoxib 10 mg/kg (four males/four females) and meloxicam 1 mg/kg (four males/four females) were evaluated in the model at dosages used frequently in practice. The PD parameters (body temperature, mentation, posture, preference of location in the cage, and prostaglandin E2 [PGE2] plasma concentrations) were determined for 10 hours following the second LPS injection. At the doses evaluated, mavacoxib and celecoxib significantly reduced LPS-induced hypothermia, but had no clear effects on other clinical signs of illness. In contrast, no effect on hypothermia or clinical appearance was observed in the LPS-challenged cockatiels treated with meloxicam. All three NSAIDs were able to inhibit the increase in LPS-induced PGE2 plasma concentrations, yet the effect was most pronounced in the birds treated with meloxicam. Consequently, the presented model opens perspectives for future dose-effect PD studies to optimize analgesic protocols in cockatiels.

Biomechanical properties of celiotomy wounds closed with tape and cyanoacrylate versus intradermal sutures.

OBJECTIVE: To compare cutaneous wound closure with mesh and 2-octyl cyanoacrylate (MOC) vs an intradermal suture pattern (ISP) in terms of time of application and biomechanical properties. SAMPLE POPULATION: Experimental study with 12 female beagle dogs. METHODS: A skin incision was created from the xiphoid to the pubis as part of an ovariohysterectomy; the linea alba and subcutaneous tissue were closed routinely. Half of the skin incision was closed with MOC, and the other half was closed with an ISP. Tissue samples were collected from both sections at days 14 and 28 and tested for ultimate strength and stiffness. RESULTS: Closure with MOC (72.8 ± 14.0 s) was faster than with an ISP (398.4 ± 36.4 s; P = .001). The ultimate load and stiffness increased with time for MOC (P = .005 and P = .005, respectively) and ISP (P < .001 and P < .001, respectively). On day 14, ultimate load and stiffness were greater in wounds closed with MOC compared with ISP (P = .014 and P = .02, respectively). No difference between groups was detected at day 28. CONCLUSION: Cutaneous wound closure with MOC was faster and resulted in superior strength at 14 days compared with closure with an ISP in this healthy population. CLINICAL SIGNIFICANCE: Mesh and 2-octyl cyanoacrylate offers an attractive alternative to ISP for skin closure after celiotomy in dogs, especially if surgical/anesthesia time is a concern.

Detection of innocent systolic murmurs by auscultation and their relation to hematologic and echocardiographic findings in clinically normal Whippets.

OBJECTIVE: To determine murmur prevalence by auscultation of 105 apparently healthy Whippets without signs of cardiac disease, to determine the origin of these murmurs, and to evaluate the influence of sex, type of pedigree (ie, bred for showing or racing), and training on these murmurs. DESIGN: Cross-sectional study. ANIMALS: 105 client-owned Whippets. PROCEDURES: All dogs were auscultated by the first author and underwent a complete physical and cardiological examination, together with a hematologic assessment. Several RBC variables and echocardiographic variables were compared between dogs with or without a murmur at the level of the aortic valve. RESULTS: 44 of 105 (41.9%) dogs had no murmur. A soft systolic murmur was present with point of maximal intensity at the level of the aortic valve in 50 (47.6%) dogs, at the level of the pulmonic valve in 8 (7.6%) dogs, and at the level of the mitral valve in 3 (2.9%) dogs. No significant differences were found in heart rate, rhythm, murmur presence, point of maximal intensity, and murmur grade between males and females, between dogs with race- and show-type pedigrees, or between dogs in training and not in training. Dogs with a murmur at the level of the aortic valve had a significantly higher aortic and pulmonic blood flow velocity and cardiac output, compared with dogs without a murmur. CONCLUSIONS AND CLINICAL RELEVANCE: Whippets have a high prevalence of soft systolic murmurs in the absence of any structural abnormalities, which fit the description of innocent murmurs. No influence of sex, pedigree type, or training was found on the occurrence of these murmurs in Whippets.

Immunomodulatory properties of gamithromycin and ketoprofen in lipopolysaccharide-challenged calves with emphasis on the acute-phase response.

Macrolide antibiotics and non-steroidal anti-inflammatory drugs (NSAIDs) have been reported to be modulators of the innate immune response, irrespectively of their antimicrobial and anti-inflammatory actions. Therefore, it was our objective to evaluate whether the macrolide gamithromycin (GAM) and the NSAID ketoprofen (KETO) attenuate the acute-phase response in calves, and whether their combined administration is beneficial due to synergistic and/or additive effects. To this end, both drugs, as well as their combination, were studied in a previously developed inflammation model, i.e., the induction of an acute-phase response by an intravenous lipopolysaccharide (LPS) challenge (0.5 µg/kg body weight). Sixteen 4-week-old Holstein-Friesian calves were randomized into 4 groups: a positive control (+CONTR) group, receiving LPS but no pharmacological treatment (n=4) and a GAM (n=4), a KETO (n=4) and a GAM-KETO (n=4) group, receiving the respective drugs 1h prior to LPS administration. Clinical scoring and blood collection were performed at regular time points until 72 h post LPS challenge. Plasma concentrations of the selected cytokines (tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6)), acute-phase protein (serum amyloid A (SAA)) and prostaglandin E2 (PGE2) were subsequently quantified. Pre-treatment with GAM had no effect in the inflammation model compared to the +CONTR group. KETO, on the other hand, completely inhibited depression, anorexia and fever. This remarkable influence was associated with a significant reduction of PGE2 synthesis by KETO, while the effect on TNF-α, IL-6 and SAA was not straightforward. The combined administration of GAM and KETO provided no synergistic or additive effects in this model, neither clinically nor regarding the studied inflammatory mediators. In conclusion, KETO entirely inhibited PGE2 synthesis, fever development and depression, while GAM did not exert any effect in this model. These results promote the concomitant use of an antimicrobial drug and a NSAID in the treatment of calf diseases associated with LPS, both to enhance clinical recovery and to improve animal welfare.

Neuregulin-1 induces a negative inotropic effect in cardiac muscle: role of nitric oxide synthase.

BACKGROUND: Deficient cardiac neuregulin/ErbB signaling increases susceptibility to heart failure. In this study, we examined the effects of neuregulin-1 (NRG-1) on myocardial contractility. METHODS AND RESULTS: NRG-1 (alpha and beta isoforms) induced a negative inotropic effect in isolated rabbit papillary muscles and a rightward shift of the dose-response curve to isoproterenol. Both effects were attenuated by L-NMMA, which suggests a role for NO synthase. In cultured rat cardiomyocytes, NRG-1beta enhanced nitrite production and resulted in phosphorylation of endothelial NO synthase and the serine/threonine kinase Akt. CONCLUSIONS: NRG-1 has negative inotropic effects that are preserved during beta-adrenergic stimulation and activates endothelial NO synthase in cardiomyocytes.

Serologic screening for 13 infectious agents in roe deer (Capreolus capreolus) in Flanders.

INTRODUCTION: In order to investigate the role of roe deer in the maintenance and transmission of infectious animal and human diseases in Flanders, we conducted a serologic screening in 12 hunting areas. MATERIALS AND METHODS: Roe deer sera collected between 2008 and 2013 (n=190) were examined for antibodies against 13 infectious agents, using indirect enzyme-linked immunosorbent assay, virus neutralisation, immunofluorescence, or microagglutination test, depending on the agent. RESULTS AND DISCUSSION: High numbers of seropositives were found for Anaplasma phagocytophilum (45.8%), Toxoplasma gondii (43.2%) and Schmallenberg virus (27.9%), the latter with a distinct temporal distribution pattern following the outbreak in domestic ruminants. Lower antibody prevalence was found for Chlamydia abortus (6.7%), tick-borne encephalitis virus (5.1%), Neospora caninum (4.8%), and Mycobacterium avium subsp paratuberculosis (4.1%). The lowest prevalences were found for Leptospira (1.7%), bovine viral diarrhoea virus 1 (1.3%), and Coxiella burnetii (1.2%). No antibodies were found against Brucella sp., bovine herpesvirus 1, and bluetongue virus. A significant difference in seroprevalence between ages (higher in adults >1 year) was found for N. caninum. Four doubtful reacting sera accounted for a significant difference in seroprevalence between sexes for C. abortus (higher in females). CONCLUSIONS: Despite the more intensive landscape use in Flanders, the results are consistent with other European studies. Apart from maintaining C. abortus and MAP, roe deer do not seem to play an important role in the epidemiology of the examined zoonotic and domestic animal pathogens. Nevertheless, their meaning as sentinels should not be neglected in the absence of other wild cervid species.

The influence of morphine on cerebral 5-HT2A availability in dogs: a SPECT study.

UNLABELLED: The opioid and serotonergic systems are closely involved in pain processing and mood disorders. The aim of this study was to assess the influence of systemic morphine on cerebral serotonin 2A receptor (5-HT(2A)) binding in dogs using SPECT with the 5-HT(2A) radioligand (123)I-5I-R91150. METHODS: 5-HT(2A) binding was estimated with and without morphine pretreatment in 8 dogs. The 5-HT(2A) binding indices in the frontal, parietal, temporal, and occipital cortex and in the subcortical region were obtained by semiquantification. RESULTS: A significantly decreased 5-HT(2A) binding index was found in the morphine group for the right (morphine, 1.41 ± 0.06; control, 1.52 ± 0.10) and left (morphine, 1.44 ± 0.08; control, 1.55 ± 0.11) frontal cortices, with P = 0.012 and P = 0.040, respectively. No significant differences were noted for the other regions. CONCLUSION: Morphine decreased the frontocortical 5-HT(2A) availability, confirming an interaction between the 5-HTergic and the opioid systems. Whether this interaction is caused by decreased receptor density due to direct internalization or is the result of indirect actions, such as increased endogenous serotonin release, remains to be elucidated.

Stromal cell-derived factor-1 retention and cardioprotection for ischemic myocardium.

BACKGROUND: Stromal cell-derived factor-1 (SDF-1) is a chemoattractant of stem/progenitor cells, and several studies have shown that SDF-1 may improve ventricular function after infarction. SDF-1 is cleaved by proteases including matrix metalloproteinase-2 (MMP-2) and CD26/dipeptidylpeptidase-4 (DPP-4), which are activated in injured tissues. METHODS AND RESULTS: We investigated the biodistribution and functional roles of SDF-1 in experimental ischemia/reperfusion injury in rats. Radiolabeled SDF-1 given by intracoronary injection was selectively concentrated in ischemic myocardium. The enhanced uptake of SDF-1 in ischemic myocardium was not mediated by its receptor, CXCR4. Mass spectrometry and Western analyses showed that SDF-1 was cleaved by DPP-4 in plasma and myocardium, whereas a bioengineered MMP-2/DPP-4-resistant form of SDF-1, SSDF-1(S4V), was highly stable. A single dose of SSDF-1(S4V) exhibited greater potency for cardioprotection than wild-type SDF-1. SSDF-1(S4V) improved cardiac function in rats even after a 3-hour ischemic period. CONCLUSIONS: These results show that a single dose of protease-resistant SSDF-1(S4V) after myocardial infarction leads to dramatic improvement in angiogenesis and ventricular function even 3 hours after the onset of ischemia, revealing a simple, clinically feasible approach to prevention of heart failure.

Electrocardiographic reference values in whippets.

The aim of this study was to determine the electrocardiographic characteristics of whippets and to compare the results with published reference values for a general dog population. Electrocardiographic parameters from 105 healthy whippets were used to establish reference values for the breed. The most important differences compared to published reference values were the higher median R-wave amplitudes in leads II, CV(6)LL and CV(6)LU. For some parameters (P-wave amplitude, ST-segment deflection and T-wave amplitude in lead II; R-wave amplitude in CV(5)RL), a marked percentage of the whippet values were above the published maximum reference data. The results confirmed that whippets have electrocardiographic characteristics similar to those reported in athletic heart syndrome in humans. Some of these characteristics could be erroneously taken as evidence of cardiac disease and clinicians should be aware of these factors to prevent unnecessary investigations in healthy dogs.

Enteral feeding reduces endothelial nitric oxide synthase in the caudal intestinal microvasculature of preterm piglets.

The initiation of enteral feeding represents a challenge to the neonatal intestinal microcirculation, especially in preterms where it predisposes to necrotizing enterocolitis (NEC). We hypothesized that a structural microvascular deficiency may occur when enteral feeding is initiated in preterm piglets susceptible to NEC. Stereologic volume densities of a pan-endothelial marker (vWF), and the main vasodilator endothelial nitric oxide synthase (eNOS), were determined along the small intestine of 1) unfed preterm piglets, 2) piglets receiving total parenteral nutrition (TPN) for 2-3 d, and 3) piglets fed 2 d sow's colostrum (TPN+SOW) or milk formula (TPN+FOR) following TPN. In the mucosa, vWF-density decreased in a cranio-caudal direction. A corresponding mucosal eNOS gradient appeared only after initiating enteral feeding. In TPN+SOW, eNOS induction may lag behind the mucosal growth of the caudal region. In TPN+FOR, formula-related factors (i.e. bacteria, cytokines) may suppress mucosal eNOS, indicated by increased stress-sensitive nuclear HIF1alpha staining. The low mucosal endothelial eNOS density was related to the presence of NEC lesions, maybe via increased hypoxia-sensitivity, especially in the caudal region as indicated by nuclear HIF1alpha-staining. Our results suggest an insufficient structural adaptation of the microvasculature to enteral feeding, especially of mucosal eNOS, which may lead to NEC.

Echocardiographic reference values in whippets.

The aim of the study was to establish reference echocardiographic values for whippets, to compare these values with previously published reference values for the general dog population, and to determine whether there is an influence of gender and breeding lines on echocardiographic measurements. Echocardiographic parameters from 105 apparently healthy whippets without cardiac symptoms were used to establish reference values for the breed and to compare these values with two previously reported reference ranges. The coefficients of the allometric equation Y= aM(b), useful to reconstruct normal M-mode and two-dimensional average values for whippets of varying weights, were calculated, as well as the lower and upper limits of the 95% prediction interval. First, we found that whippets have a significantly larger left ventricular diameter, increased left ventricular wall, and interventricular septum thickness than expected, in diastole as well as in systole. Fractional shortening was significantly lower than the reference value. Second, comparing males and females, taking body weight differences into account, females had a significantly larger left ventricular diameter in diastole and systole. Minor differences were found between racing and show pedigree dogs. In conclusion, the results of this study confirm that breed-specific reference values are needed in echocardiography. In whippets, the values found in this study can be used as references in order to avoid overinterpretation of cardiac dilation, hypertrophy, and/or decreased contractility in these dogs.

Inhibition of heme oxygenase-1 impairs cardiac muscle sensitivity to beta-adrenergic stimulation.

UNLABELLED: Heme oxygenase-1 (HO-1) is the inducible isoform of heme oxygenase and plays a role in defense against cellular stress. The effects of HO-1 on cardiac muscle contractility, however, are unknown. METHODS: HO-1 was induced by intraperitoneal injection of hemin in rabbits 24 and 48 h before isolating right ventricular papillary muscles for mechanical in vitro analysis at baseline and during stimulation with isoprenalin. Western blotting and activity measurement con.rmed upregulation of HO-1 in ventricular tissue, and immunohistochemical stainings showed localization in the cardiac endothelium. RESULTS: Baseline mechanical performance of papillary muscles and maximal inotropic response to ISO was not significantly affected by HO-1 induction. Also, the log(EC50) of the ISO concentration-response curve was not affected by HO-1 induction. Inhibition of heme oxygenase with stanneous mesoporphyrin or chromium mesoporphyrin in muscles with induced HO-1, however, shifted the log(EC50) of the ISO concentration-response curve from -6.9 +/- 0.2 to -6.0 +/- 0.2 (p = 0.008). CONCLUSION: These results indicate that induction of cardiac HO-1 has no direct effect on baseline contractility. Pharmacological inhibition of HO-1 upon induction, however, diminishes cardiac muscle sensitivity to beta-adrenergic stimulation. These results caution against pharmacologically targeting HO-1 when an activated adrenergic system is important for hemodynamic stability.

Vertebral heart size ranges specific for whippets.

To assess the influence of breed, breeding lines, and training on heart size, the vertebral heart size (VHS) was evaluated on left-to-right lateral, right-to-left lateral, dorsoventral, and ventrodorsal thoracic radiographs from 44 whippets free from cardiac and pulmonary disease. In lateral views, the VHS was 11.0 +/- 0.5 vertebrae (mean +/- SD) on right-to-left lateral and 11.3 +/- 0.5 vertebrae on left-to-right lateral radiographs, being larger than the 9.7 +/- 0.5 vertebrae proposed by Buchanan (P<0.0001). The VHS on left-to-right lateral views was larger than on right-to-left lateral views (P<0.0001). The VHS was 10.5 +/- 0.6 vertebrae on dorsoventral radiographs and 11.1 +/- 0.6 vertebrae on ventrodorsal radiographs. Both values were larger than the 10.2 +/- 1.5 vertebrae (dorsoventral) (P<0.0082) or 10.2 +/- 0.8 vertebrae (ventrodorsal) (P<0.0001) proposed by Buchanan. In addition, the VHS on ventrodorsal views was larger than on dorsoventral views (P<0.0001). Dogs out of racing pedigree lines had a significantly larger VHS than those out of show pedigree lines, and trained dogs had a significantly larger VHS than nontrained dogs. Because most trained dogs came out of racing pedigree lines, and all nontrained dogs came out of show pedigree lines, however, it is difficult to determine whether the higher VHS for trained dogs is due to genetic influences or training, or both. In conclusion, it is important to take into account the breed and the radiographic view when evaluating heart size in thoracic radiographs of whippets to avoid overinterpretation of cardiac enlargement in these dogs.

Coronary perfusion and muscle lengthening increase cardiac contraction: different stretch-triggered mechanisms.

An increase in coronary perfusion, transversal stretch of the myocardium, increases developed force (F(dev)) (Gregg effect) through activation of stretch-activated ion channels (SACs). Lengthening of the muscle, longitudinal stretch of the myocardium, causes an immediate increase in F(dev) followed by a slow F(dev) increase (Anrep effect). In isometrically contracting perfused papillary muscles of Wistar rats, we investigated whether both effects were based on similar stretch-induced mechanisms by measuring F(dev) and intracellular Ca(2+) concentration ([Ca(2+)](i)) after a muscle length increase from 85% to 95% L(max) (length at which maximal isometric force develops) at low and high coronary perfusion before and after inhibition of SACs with gadolinium (10 micromol/l Gd(3+)). The increase of F(dev) and peak [Ca(2+)](i) by the Gregg effect was of similar magnitude as the Anrep effect (from 3.5 +/- 0.8 to 3.9 +/- 1.2 mN/mm(2) and from 3.0 +/- 0.7% to 3.8 +/- 0.9% normalized [Ca(2+)](i), means +/- SE). SAC blockade completely blunted the increase of F(dev) and peak [Ca(2+)](i) by the Gregg effect; however, it did not affect the Anrep effect. The slow force response, but not the calcium response, was augmented by an increase in coronary perfusion. Therefore, increased coronary perfusion, transversal stretch of the myocardium, and muscle lengthening, longitudinal stretch of the myocardium, increase myocardial contraction in the rat through different stretch-triggered mechanisms.