Effect of spironolactone on diuresis and urine sodium and potassium excretion in healthy dogs

ObjectivesTo document the diuretic effect of different oral doses of spironolactone (SP) in healthy dogs.BackgroundSP is currently mentioned as a diuretic agent in the dog. However, the recommended doses were empirically defined and their corresponding diuretic effect has never been documented in dogs.Animals, materials and methodsEight adult Beagle dogs were used for two separate 2 * 2 cross-over designs. In the first cross-over, 4 dogs received SP orally for 8 days at 1 and 2 mg/kg per day. In the second cross-over the 4 other dogs received SP similarly, but at 4 and 8 mg/kg per day. Dogs were weighed on the first and last day of each period. Plasma SP and canrenone (the main active metabolite of SP) were assayed by high performance liquid chromatography (HPLC). Daily water consumption, urine weight, urine specific gravity, and urine excretion of sodium and potassium were measured during the SP treatment.ResultsTwo hours after SP administration, SP was metabolized into canrenone. A significant 14 and 22% decrease in urine potassium excretion was observed at 1 and 2 mg/kg, respectively, but not at the two other dose levels. Daily water consumption, urine weight, urine specific gravity, and urine excretion of sodium were not significantly altered by the SP treatment regardless of dose.ConclusionsRepeated oral administration of SP at 1, 2, 4 or 8 mg/kg for 8 days had no effect on water and sodium diuresis in healthy dogs.     

Influence of combined angiotensin-converting enzyme inhibitors and spironolactone on serum K, Mg, and Na concentrations in small dogs with degenerative mitral valve disease

ObjectiveTo determine if the administration of an angiotensin-converting enzyme inhibitor (ACEI) plus spironolactone caused hyperkalemia, hypermagnesemia, or hyponatremia in elderly small dogs with degenerative mitral valve disease (MVD).BackgroundACEIs and spironolactone can increase serum potassium and magnesium concentrations and lower serum sodium concentrations. It has been recommended to either not combine these drugs or to do so with caution.Animals, material and methodsFifty client-owned dogs with MVD, left atrial dilation, and without congestive heart failure or azotemia were evaluated retrospectively. Baseline data sets, followed by 1–9 (mean = 2.66) data sets, comprised of serum urea nitrogen (SUN), creatinine, sodium, potassium, and magnesium concentrations, were tabulated. Each dog received an ACEI plus spironolactone for a mean of 23.8 ± 26.6 weeks (range: 2–150) and a median of 15 weeks. No cardiac drugs other than an ACEI and spironolactone were administered during the study period.ResultsThere were no significant differences between baseline and follow-up serum sodium or potassium concentrations. Serum magnesium concentrations increased significantly (p = 0.02) with time >20 weeks compared to baseline.ConclusionsThe combination of an ACEI and spironolactone results in no significant difference between baseline and follow-up serum sodium or potassium concentrations. Although serum magnesium concentrations may increase significantly with time >20 weeks compared to baseline concentrations, hypermagnesemia appears to be rare, mild, and unlikely to be of clinical importance. The combination of an ACEI and spironolactone is safe in elderly small dogs with MVD with normal SUN and creatinine concentrations.     

Quadricuspid aortic valve and associated abnormalities in the dog: Report of six cases

ObjectivesTo describe the clinical and echocardiographic findings in dogs with quadricuspid aortic valve (QAV).BackgroundQAV is a rare canine congenital heart disease which has been reported only three times in the young dog.Animals, materials and methodsSix dogs (0.3- to 13-year-old) with QAV diagnosed by two-dimensional echocardiography were retrospectively evaluated. Medical records, echocardiograms, and follow-ups were reviewed.ResultsAccording to aortic cusp morphology, QAV was classified as type A (n = 1), type B (n = 4) or type C (n = 1). QAV was associated with at least one other heart disease in all of the dogs including, ventricular septal defect (n = 1), enlarged left coronary ostium (n = 4), degenerative mitral valve disease (MVD, n = 1) and patent ductus arteriosus (PDA, n = 3). Mild to moderate aortic regurgitation was also detected in all dogs by continuous-wave and color-flow Doppler echocardiography. QAV was diagnosed in four asymptomatic dogs referred for evaluation of a heart murmur. The remaining two dogs had QAV and PDA with evidence of mild exercise intolerance and moderately retarded growth. The PDA was surgically corrected in both dogs and at the time of writing, 1–2.5 years after the initial diagnosis, none of the six animals shows evidence of clinical signs.ConclusionQAV is a cause of aortic insufficiency. It may incidentally be found by two-dimensional echocardiography in dogs of various ages in association with other congenital or acquired cardiac abnormalities.     

Weekly variability of plasma and serum NT-proBNP measurements in normal dogs

ObjectivesTo determine the weekly variability of serum and plasma N-terminal pro-B-type natriuretic peptide (NT-proBNP) concentrations in healthy dogs.Animals, materials and methodsFifty-three normal dogs were examined prospectively. Serum (n = 25) or plasma (n = 28) samples were obtained for NT-proBNP assay at one week interval for 3 consecutive weeks.ResultsMedian serum or plasma NT-proBNP concentration did not change over 3 consecutive weeks. Twenty-two of 53 dogs (42%) had at least one NT-proBNP value >500 pmol/L, including 14 dogs with at least one serum NT-proBNP concentration >500 pmol/L and 8 dogs with at least one plasma NT-proBNP concentration >500 pmol/L during the 3-week sampling period. The difference between the maximum and minimum NT-proBNP value obtained over the 3-week sampling period was <100 pmol/L in 40% of dogs, between 100 and 200 pmol/L in 40% of dogs, and >200 pmol/L in 20% of dogs. Of the 19 dogs with a value >500 pmol/L on either week 1 or 2, 11 dogs (58%) had a subsequent NT-proBNP value <500 pmol/L on either week 2 or 3.ConclusionsThere is a high degree of variability in weekly serum and plasma NT-proBNP values in healthy dogs. Individual variability should be considered when interpreting NT-proBNP results in dogs.     

Effect of azotemia on serum N-terminal proBNP concentration in dogs with normal cardiac function: A pilot study

ObjectivesTo evaluate amino-terminal pro-B type natriuretic peptide (NT-proBNP) concentration in dogs with renal dysfunction and normal cardiac structure and function.AnimalsEight dogs with renal disease, 23 healthy control dogs.MethodsSerum NT-proBNP concentration was measured in healthy dogs and dogs with renal disease using an ELISA validated for use in dogs. Affected dogs were eligible for inclusion if renal dysfunction was diagnosed based on urinalysis and serum chemistry, and if they were free of cardiovascular disease based on physical exam, systolic blood pressure, and echocardiography.ResultsThe geometric mean serum NT-proBNP concentration was significantly higher in dogs with renal disease (617 pmol/L; 95% CI, 260–1467 pmol/L) than in healthy control dogs (261 pmol/L; 95% CI, 225–303 pmol/L; P = 0.0014). There was a modest positive correlation between NT-proBNP and BUN and creatinine. Median NT-proBNP concentration was not significantly different between groups when indexed to BUN (median NT-proBNP:BUN ratio; renal, 14.2, IQR, 3.93–17.7 vs. control, 16.3, IQR, 9.94–21.2; P = 0.29) or creatinine (median NT-proBNP:creatinine ratio; renal, 204, IQR, 72.6–448 vs. control, 227, IQR, 179–308; P = 0.67).ConclusionDogs with renal disease had significantly higher mean serum concentration of NT-proBNP than control dogs. Renal function should be considered when interpreting NT-proBNP results as concentrations may be falsely elevated in dogs with renal dysfunction and normal cardiac function. The effect of renal disease was lessened by indexing NT-proBNP to BUN or creatinine. Future studies in dogs with both renal and heart disease are warranted.     

The effect of maropitant on intraoperative isoflurane requirements and postoperative nausea and vomiting in dogs: a randomized clinical trial

ObjectiveTo establish if preoperative maropitant significantly reduced intraoperative isoflurane requirements and reduced clinical signs associated with postoperative nausea and vomiting (PONV) in dogs.Study designRandomized clinical trial.AnimalsTwenty-four healthy, client-owned dogs undergoing routine ovariohysterectomy.MethodsPremedication involved acepromazine (0.03 mg kg⁻¹) combined with methadone (0.3 mg kg⁻¹) intramuscularly 45 minutes before anaesthetic induction with intravenous (IV) propofol, dosed to effect. Meloxicam (0.2 mg kg⁻¹) was administered intravenously. Dogs were randomly assigned to administration of saline (group S; 0.1 mL kg⁻¹, n = 12) or maropitant (group M; 1 mg kg⁻¹, n = 12) subcutaneously at time of premedication. Methadone (0.1 mg kg⁻¹ IV) was repeated 4 hours later. Anaesthesia was maintained with isoflurane in oxygen, dosed to effect by an observer unaware of group allocation. The dogs were assessed hourly, starting 1 hour postoperatively, using the short form of the Glasgow Composite Pain Score (GCPS), and for ptyalism and signs attributable to PONV [score from 0 (none) to 3 (severe)] by blinded observers. Owners completed a questionnaire at the postoperative recheck.ResultsOverall mean ± standard deviation end-tidal isoflurane percentage was lower in group M (1.19 ± 0.26%) than group S (1.44 ± 0.23%) (p = 0.022), but was not significantly different between groups at specific noxious events (skin incision, ovarian pedicle clamp application, cervical clamp application, wound closure). Cardiorespiratory variables and postoperative GCPS were not significantly different between groups. Overall, 50% of dogs displayed signs attributable to PONV, with no difference in PONV scores between groups (p = 0.198). No difference in anaesthetic recovery was noted by owners between groups.ConclusionsMaropitant reduced overall intraoperative isoflurane requirements but did not affect the incidence of PONV.Clinical relevanceMaropitant provided no significant benefits to dogs undergoing ovariohysterectomy with this anaesthetic and analgesic protocol, although clinically significant reductions in isoflurane requirements were noted.     

Noninvasive assessment of systolic left ventricular torsion by 2-dimensional speckle tracking imaging in the awake dog: repeatability, reproducibility, and comparison with tissue Doppler imaging variables

BACKGROUND: Left ventricular (LV) torsional deformation plays an important role in myocardial function. However, it has never been assessed in the awake dog, because magnetic resonance imaging and sonomicrometry have been the only methods available so far. HYPOTHESIS: Two dimensional speckle tracking echocardiography (STE), a new ultrasound imaging technique, provides a repeatable and reproducible noninvasive assessment of systolic LV wringing motion in the awake dog. ANIMALS: Six healthy dogs were used to determine the repeatability and reproducibility of STE variables (study 1). These variables also were prospectively assessed in a population of 35 healthy dogs (study 2). METHODS: Peak LV basal and apical systolic rotations were measured by STE from right parasternal short-axis views using automatic frame-to-frame tracking of gray-scale speckle patterns. Systolic LV torsion (LVtor, degrees ) was defined as apical rotation relative to the base. RESULTS: All within-day and between-day coefficients of variation were <20% (6.8-18.0%). Amplitude of apical systolic rotation was significantly higher (P < .001) than the basal value (5.4 +/- 3.2 degrees and -3.1 +/- 1.3 degrees , respectively). Global LVtor was significantly correlated with systolic longitudinal LV myocardial velocity gradient assessed by tissue Doppler (P < .05), but not with either systolic radial LV myocardial velocity gradient or the ratio of early mitral inflow velocity to early mitral annular velocity (Em/Ea). CONCLUSIONS AND CLINICAL IMPORTANCE: Speckle tracking echocardiography is a repeatable and reproducible method for assessing systolic LV torsional deformation. The combination of these new STE indices with tissue Doppler variables could provide a new approach for quantifying canine LV systolic function.     

Arrhythmogenic right ventricular cardiomyopathy in Boxer dogs is associated with calstabin2 deficiency

ObjectiveTo examine the presence and effect of calstabin2-deficiency in Boxer dogs with arrhythmogenic right ventricular cardiomyopathy (ARVC).AnimalsThirteen Boxer dogs with ARVC.Materials and methodsTissue samples were collected for histopathology, oligonucleotide microarray, PCR, immunoelectrophoresis, ryanodine channel immunoprecipitation and single-channel recordings, and calstabin2 DNA sequencing.ResultsIn cardiomyopathic Boxer dogs, myocardial calstabin2 mRNA and protein were significantly decreased as compared to healthy control dogs (calstabin2 protein normalized to tetrameric cardiac ryanodine receptor (RyR2) complex: affected, 0.51 ± 0.04; control, 3.81 ± 0.22; P < 0.0001). Calstabin2 deficiency in diseased dog hearts was associated with a significantly increased open probability of single RyR2 channels indicating intracellular Ca²⁺ leak. PCR-based sequencing of the promoter, exonic and splice site regions of the canine calstabin2 gene did not identify any causative mutations.ConclusionsCalstabin2 deficiency is a potential mechanism of Ca²⁺ leak-induced ventricular arrhythmias and heart disease in Boxer dogs with ARVC.     

Methods and normal values for echocardiography in adult dairy cattle

ObjectiveThe objective of the study was to report normal ultrasonographic appearance and intra-cardiac dimensions in two dairy breeds and to calculate cardiac output (CO) using echocardiography.BackgroundIntra-cardiac dimensions, time indices and CO estimation have not previously been reported in adult cattle.Animals, materials and methodsEchocardiograms were obtained from healthy adult dairy cows (10 Jersey (J) and 12 Holstein Friesians (HF)) in the body weight range of 400 to 700 kg. Standard echocardiographic images were obtained from the left and right hemithoraces. Velocity time integrals were obtained in order to calculate CO using pulsed wave Doppler of aortic flow in the J cows. Measurements obtained included pulmonary artery and aortic diameters, left and right ventricular diameters (and calculated fractional shortening and left ventricular ejection fraction), left atrial size and time indices assessing valve function.ResultsHF cows had significantly (p < 0.05) larger pulmonary artery and aortic diameters, larger left atrial diameters and left ventricular internal diameters during diastole, but these were not different when corrected for body weight. Left and right ventricular dimensions, adjusted for body weight, were significantly larger (p = 0.02 and p = 0.035 respectively) in J cows when compared to HF cows. No differences were noted in the time indices between the two groups. No significant differences were noted in intra-operator variability and the only significant difference in inter-operator variability was in measurement of the pulmonary artery (p = 0.03; ICC = 0.63).ConclusionsIt is possible to obtain repeatable, reliable echocardiograms in order that meaningful intra-cardiac dimensions can be obtained in adult dairy cattle.     

Plasma atrial natriuretic peptide concentration in warmblood horses with heart valve regurgitations

ObjectivesThis study measured plasma atrial natriuretic peptide (ANP) concentration in horses with heart valve regurgitations (HVR) with and without atrial and ventricular dilatation.BackgroundIn humans and small animals, plasma ANP concentration is increased in heart disease and correlates with the severity of clinical signs and heart enlargement.Animals, materials and methodsTen healthy horses (control) and 36 horses with HVR were evaluated by auscultation, electrocardiography, echocardiography, and determination of plasma ANP.ResultsControl horses demonstrated mean plasma ANP concentration of 21 ± 5.4 pg/mL. Of the 36 horses with HVR, 17 horses possessed normal echocardiographic heart size (group 1), 10 horses had a left atrial dilatation (group 2) and 9 horses had both left atrial and ventricular dilatation (group 3). Mean plasma ANP concentration of groups 1, 2 and 3 was 20.1 ± 5.6 pg/mL, 22.9 ± 11.0 pg/mL and 27.6 ± 17.4 pg/mL, respectively. The plasma ANP concentrations of HVR and control horses were not significantly different. The highest ANP concentrations were observed in horses with atrial and ventricular dilatation. No correlation between left atrial or ventricular size, weight, or sex and the plasma ANP concentration was found.ConclusionsNo significant differences in plasma ANP concentration was observed between groups. Further study, especially in horses with clinical signs of heart failure is needed.     

Evaluation of transformation growth factor beta1, interleukin-10, and interferon-gamma in male symptomatic and asymptomatic dogs naturally infected by Leishmania (Leishmania) chagasi

The aims of this study were to evaluate the immunomodulatory role of TGF-β₁, IL-10, and INF-γ in spleen and liver extracts and supernatant cultures of white spleen cells from male symptomatic and asymptomatic dogs, naturally infected by Leishmania (Leishmania) chagasi. Thirty dogs from Araçatuba, São Paulo, Brazil, an endemic leishmaniosis area, were selected by positive ELISA serological reaction for Leishmania sp. and divided into two groups: asymptomatic (n = 15) and symptomatic (n = 15) consisting of animals with at least three characteristic signs (fever, dermatitis, lymphoadenopathy, onychogryphosis, weight loss, cachexia, locomotion problems, conjunctivitis, epistaxis, hepatosplenomegaly, edema, and apathy). After euthanasia, spleen and liver fragments were collected for ex vivo quantification of TGF-β₁, IL-10, and INF-γ. Naturally active in vitro produced TGF-β₁ was also evaluated in spleen cell culture supernatant. Spleen and liver extract of asymptomatic dogs had higher mean TGF-β₁ levels than symptomatic dogs. High concentrations of IL-10 were found in spleen, and mainly in liver extract of both groups. Higher INF-γ concentrations were found in spleen extracts of symptomatic dogs, and in liver extracts of asymptomatic dogs. Extract of this cytokine was lower in spleen extract. Although INF-γ is being produced in canine infection, mean levels of TGF-β₁ and IL-10 from spleen and liver extracts were quantitatively much higher; suggesting that immune response in both asymptomatic and symptomatic dogs was predominantly type Th2.     

Canine heartworm (Dirofilaria immitis) in Fresno and Madera Counties, California: prevalence differences between foothill and valley habitats

The aim of this study was to assess the prevalence of heartworm in domestic dogs in Madera and Fresno Counties, California, dependent on habitat and other host and environmental factors. Dogs were screened for presence of heartworm antigen using the PetChek^® ELISA on blood samples (N = 519) collected at seven sites during April-July 2009. Eighteen dogs were heartworm antigen positive. Pearson Chi-square analyses were conducted testing the presence of heartworm antigen against the following independent variables: elevation range, percentage of time spent outdoors during the day, percentage of time spent outdoors during the night, pet coat length, weight class, prevention status, and sex. Dogs that spent at least 50% of their time outdoors during the day were significantly more likely to have heartworm that those who spent less time outside (N = 519, df = 1, p = 0.031). Overall prevalence (3.47%) was lower than expected, with Madera County having 3.8% positive samples and Fresno County 3.5%; this prevalence is lower than in many previous studies. The effect of time spent outdoors on heartworm prevalence was similar to previous studies. The impact of elevation on infection, though not significant, requires further investigation, as does the prevalence and viability of larval stages in mosquitoes.     

Expression of trefoil factor genes in the duodenum and colon of dogs with inflammatory bowel disease and healthy dogs

Trefoil factors (TFF) are small peptides produced by goblet cells, which are crucial for epithelial restitution. In humans with inflammatory bowel disease (IBD), TFF expression is up-regulated as part of an unspecific repair mechanism. The goal of this study was to assess TFF gene expression in the gastrointestinal tract from dogs with IBD compared to healthy controls. Preliminary assessment by PCR revealed TFF1 and 3 expression in the small and large intestine, whereas TFF2 was amplified only in the stomach. Subsequent RT-qPCR (with relative quantification against 3 reference genes) on endoscopic duodenal (IBD n = 22, healthy controls n = 18) and colonic (IBD n = 12, controls n = 11) biopsies revealed that TFF1 expression was significantly up-regulated in the duodenum from IBD dogs (Mann–Whitney p = 0.001), whereas TFF3 expression was significantly lower in IBD colon compared to controls (t-test p = 0.018).This study demonstrates evidence for dysregulation of TFF gene expression in canine IBD. Up-regulation of TFF1 could signify ectopic expression as a compensatory repair-mechanism, whereas down-regulation of TFF3 could contribute to defective epithelial barrier function, respectively. Whether this is a cause or consequence of IBD could not be established.     

Resveratrol decreases oxidative burst capacity and alters stimulated leukocyte cytokine production in vitro

IntroductionResveratrol, a naturally-occurring phytophenol, has been shown to bolster immune surveillance and reverse immunosenescence in a dose dependent manner in rodents and humans. Although safety and pharmacokinetic studies have been completed in dogs, the immunomodulatory effects of resveratrol in dogs has not yet been investigated. The objective of this study was to determine the effect of resveratrol on canine innate immune system function in vitro. The hypothesis was that similar to other species, low concentrations of resveratrol would stimulate while high concentrations would depress innate immune system function.MethodsWhole blood was collected from six healthy, adult, client-owned dogs and was incubated with resveratrol at final concentrations of 6000 ng ml⁻¹, 3000 ng ml⁻¹, 1000 ng ml⁻¹, or control solution for 4 h. Following incubation, phagocytosis and oxidative burst were evaluated using flow cytometry, and LPS-, lipoteichoic acid (LTA) – and peptidoglycan (PG)-stimulated leukocyte production of TNF, IL-6, and IL-10 were measured using a canine specific multiplex assay.ResultsPhagocytosis was not altered by resveratrol at any concentration compared to control. However, while the number of PMNs capable of performing oxidative burst did not change, the robustness of the reaction following stimulation with Escherichia coli and PMA was reduced in a dose dependent manner. In addition, LPS-, LTA-, PG, and PBS-stimulated TNF production was increased following incubation with all concentrations of resveratrol compared to control, and this effect was dose dependent. LTA-stimulated IL-6 was increased with resveratrol compared to control. Furthermore, LTA-stimulated IL-10 was decreased with 6000 ng ml⁻¹ and 3000 ng ml⁻¹ concentrations of resveratrol and PG-stimulated IL-10 production was decreased with all concentrations of resveratrol compared to control. The LPS-, LTA-, and PG-stimulated TNF:IL-10 ratio was increased with 6000 ng ml⁻¹ of resveratrol compared to control and lower resveratrol concentrations.ConclusionWhile resveratrol was sparing to PMN phagocytosis, it reduced the robustness of PMN oxidative burst. Resveratrol also increased pro-inflammatory and decreased anti-inflammatory leukocyte cytokine production capacity in vitro. These data suggest that resveratrol supplementation may depress oxidative burst reactions while promoting pro-inflammatory leukocyte cytokine production and decreasing anti-inflammatory cytokine production. Based on these findings, further in vivo study regarding the effects of resveratrol on PMN oxidative burst capability and leukocyte cytokine production capacity are indicated prior to routine supplementation.     

Immunohistochemical detection of IgM and IgG in lung tissue of dogs with leptospiral pulmonary haemorrhage syndrome (LPHS)

Leptospiral pulmonary haemorrhage syndrome (LPHS) is a severe form of leptospirosis. Pathogenic mechanisms are poorly understood. Lung tissues from 26 dogs with LPHS, 5 dogs with pulmonary haemorrhage due to other causes and 6 healthy lungs were labelled for IgG (n = 26), IgM (n = 25) and leptospiral antigens (n = 26). Three general staining patterns for IgG/IgM were observed in lungs of dogs with LPHS with most tissues showing more than one staining pattern: (1) alveolar septal wall staining, (2) staining favouring alveolar surfaces and (3) staining of intra-alveolar fluid. Healthy control lung showed no staining, whereas haemorrhagic lung from dogs not infected with Leptospira showed staining of intra-alveolar fluid and occasionally alveolar septa. Leptospiral antigens were not detected. We conclude that deposition of IgG/IgM is demonstrable in the majority of canine lungs with naturally occurring LPHS, similar to what has been described in other species. Our findings suggest involvement of the host humoral immunity in the pathogenesis of LPHS and provide further evidence to support the dog as a natural disease model for human LPHS.     

Azotemia in cats with feline hypertrophic cardiomyopathy: Prevalence and relationships with echocardiographic variables

ObjectivesThe prevalence of renal azotemia in cats with acquired heart disease is not well documented. The aims of this study were therefore (1) to determine the prevalence of azotemia within a hospital population of cats with hypertrophic cardiomyopathy (HCM), and (2) to evaluate the relationship between echocardiographic variables and plasma urea and creatinine.Animals, materials and methods134 client-owned cats were retrospectively studied including 102 cats with HCM and 32 control cats. A complete physical examination, electrocardiography, systolic arterial blood pressure measurement, thoracic radiographs, and echocardiography were performed. Plasma creatinine and urea were determined in all cats. The animal was considered azotemic if plasma creatinine was >1.8 mg/dL and/or urea >65 mg/dL (i.e. BUN> 30 mg/dL).ResultsThe prevalence of azotemia was lower in control cats (25.0%) than in cats with HCM (58.8%) (P = 0.003). No significant differences in plasma urea and creatinine were observed between the HCM and control cats. There was no effect of plasma creatinine and urea on conventional echocardiographic variables in cats with HCM.ConclusionsAzotemia is a frequent finding in cats with HCM but is not dependent on echocardiographic variables.     

IL-6 and TNF-alpha production during active canine visceral leishmaniasis

We investigated the production of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) during canine visceral leishmaniasis (VL) to gain a better understanding of the role of such multi-functional cytokines in parasite resistance. IL-6 and TNF-α levels were measured by capture ELISA in sera from 8 healthy dogs from a non-endemic area (control group) and in sera from 16 dogs from Araçatuba, SP, Brazil, an area endemic for leishmaniosis. The dogs from the endemic area were selected by positive ELISA serology against total Leishmania chagasi antigen, positive spleen imprints for Leishmania, and the presence of at least three clinical signs associated with active visceral leishmaniasis (fever, dermatitis, lymphoadenopathy, onychogryphosis, weight loss, cachexia, locomotory difficulty, conjunctivitis, epistaxis, hepatosplenomegaly, edema, and apathy).Enhanced systemic IL-6 production was found in sera from dogs with the active disease compared to healthy dogs (t-test, P < 0.05). In contrast, TNF-α did not differ between the two groups studied. There was no correlation between IL-6 production and anti-leishmanial antibody titers in the sera. Our findings suggest that IL-6 is a good marker of active disease during leishmaniasis, and that other cytokines may be involved in the hypergammaglobulinemia characteristic of canine visceral leishmaniasis.     

Identification and in vitro screening of avian yeasts for use as probiotic

The objective of this study was to isolate and identify yeast strains from broilers excreta and to evaluate in vitro their potential for probiotic use in animal production.Methods and resultsNine yeast strains were isolated and presumptively pre-identified by biochemical assays. These isolates were grouped in six different molecular profiles using PCR-fingerprinting technique. Each profile was identified by sequencing of the D1/D2 domains of the large subunit of the 26S rRNA gene. These yeasts were identified as: Trichosporon sp. (LV-2), Wickerhamomyces anomalus (LV-6), Pichia kudriavzevii (LV-8), Kodamaea ohmeri (LV-9) and Trichosporon asahii (LV-10). A pre-screening of the strains for probiotic use was based on their ability to agglutinate pathogenic micro-organisms. These yeast strains were characterized for specific growth rate, duplication time, their cell surface hydrophobicity, medium acidification, resistance to low pH (2.0, 2.5 and 3.0) and concentrations of bile salts (0.3% and 0.6%). The isolate of W. anomalus (LV-6) had the highest agglutinating and adherence capacity, a growth rate of 2.07 × 10⁸ cfu/mL in 24 h at 30 °C, decreasing the medium pH from 6.5 to 5.23, a 25% hydrophobicity and an elevated capacity to grow under stress conditions.ConclusionsW. anomalus strain LV-6 showed the best characteristics for use as a probiotic candidate.Significance and impact of the Study:The data from this study helped in choosing a probiotic candidate from yeast to use in broiler production.     

Contrast echocardiography in Boxer dogs with and without aortic stenosis

ObjectivesThe aim of this study was to investigate whether contrast echocardiography could enhance the subcostal Doppler signal for aortic flow measurements and achieve myocardial opacification, in Boxer dogs with and without AS.BackgroundIn evaluating dogs for aortic stenosis (AS) subcostal Doppler echocardiography was used for measurement of the aortic flow velocity, a measurement that can sometimes be difficult to perform in Boxer dogs.Animals, materials, and methodsCardiac auscultation, phonocardiographic and echocardiographic examinations, including a contrast study with Optison, were performed on 29 Boxer dogs selected based on previous examinations.ResultsThe initial subcostal Doppler signal was weak in 66% of the dogs and a marked improvement was seen in all dogs after contrast injection. The peak aortic flow velocity increased 5% from 2.58 ± 1.42 m/s before contrast to 2.71 ± 1.54 m/s after contrast (p = 0.003). This corresponds to a 2.8 mmHg increase in the pressure gradient from 26.6 mmHg before to 29.4 mmHg after contrast. A dose of 0.05–0.1 mL of Optison administered intravenously resulted in approximately 4 min of Doppler signal enhancement. With the present technique contrast echocardiography did not achieve myocardial opacification.ConclusionsSingle use of the contrast agent Optison can be recommended for enhancement of the subcostal Doppler signal in dogs, in which plain Doppler signals are difficult to obtain. Albeit statistically significant, the mild increase in peak aortic flow velocity after contrast was not considered biologically or clinically significant.     

The effect of a pressure wrap (ThunderShirt®) on heart rate and behavior in canines diagnosed with anxiety disorder

The effect of moderate-to-deep pressure on soft tissue has been studied in both humans and nonhuman animals with positive results shown in reducing anxiety and promoting relaxation. Canine anxiety disorders are often treated with medications and behavior modification programming. Pressure wraps are sometimes used to treat canine anxiety, but the effects of this investigation have not been rigorously investigated. The aims of this study were to investigate the use of a pressure wrap (ThunderShirt®; ThunderWorks, Durham, NC) on heart rate and behavior in dogs diagnosed with an anxiety disorder. Subjects were 90 dogs (39 males and 51 females) that were diagnosed with Separation Anxiety or Generalized Anxiety Disorder. Dogs were randomly assigned to 1 of the 3 groups, namely Experimental Group 1 (dogs wore the ThunderShirt® per manufacturer's recommendations), Experimental Group 2 (dogs wore the ThunderShirt® loosely without pressure), and Control Group (dogs had no specific treatment). Average and maximum heart rates were measured at baseline and after 15 minutes alone in the kennel. Dogs were video-recorded and analysis completed for 12 behaviors, namely pacing, panting, yawning, tongue-flicking, drooling, elimination, barking, stress whining, howling, licking, door orientation, and calmness. The results were as follows: Dogs in Experimental Group 1 showed significantly less increase from baseline in average heart rate than the Control Group dogs (P < 0.001) and dogs in Experimental Group 2 (P < 0.001). Dogs in Experimental Group 1did not differ significantly from Control Group dogs in maximum heart rate when all dogs were considered, but did differ significantly from the Control Group (P = 0.04) when only those dogs not currently on anxiety medication were considered. There were no statistically significant differences among groups on behavioral outcomes, except that the dogs in the Control Group were significantly more likely to orient toward the door than the dogs in both Experimental Groups 1 (OR = 7.46, P = 0.01) and 2 (OR = 4.42, P = 0.04). Dogs in the Experimental Group 1 presented a trend of less tongue-flicking (P = 0.06) and yawning (P = 0.07) stress reduction behaviors when compared with the other 2 groups. More research should be conducted with the ThunderShirt® on behavioral outcomes. The ThunderShirt® can be used with behavior modification programming and medication as an adjunct treatment option to assist dogs diagnosed with anxiety disorder to reduce heart rate.