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Flow cytometric detection and procoagulant activity of circulating canine platelet-derived microparticles
2013
Helmond, Sarah E. | Catalfamo, James L. | Brooks, Marjory B.
Objective: To measure platelet membrane–derived microparticle (PMP) content and thrombin-generating capacity of canine plasma subjected to specific processing and storage conditions. Animals: 31 clinically normal dogs (19 males and 12 females). Procedures: Citrate-anticoagulated blood samples obtained from each dog were centrifuged at 2,500 × g to isolate platelet-poor plasma (PPP), then PPP was centrifuged at 21,000 × g to isolate microparticle-free plasma (MPF) and microparticle-enriched plasma (MPEP). Whole blood and paired samples of fresh and frozen-thawed PPP, MPF, and MPEP were dual labeled for flow cytometric detection of membrane CD61 (constitutive platelet antigen) and annexin V (indicating phosphatidylserine externalization). Platelets and PMPs were enumerated with fluorescent, size-calibrated beads. Thrombin generation in fresh and frozen-thawed PPP, MPF, and MPEP was measured via kinetic fluorometric assays configured with low tissue factor and low phospholipid concentrations. Results: Initial centrifugation yielded PPP with < 0.5% the platelets of whole blood, with median counts of 413 PMPs/μL for males and 711 PMPs/μL for females. Sequential centrifugation resulted in a 10-fold concentration of PMPs in MPEP and virtually depleted PMPs from MPF. Thrombin generation depended on PMP content, with median endogenous thrombin potential of 0, 893, and 3,650 nmol•min for MPF, PPP, and MPEP, respectively. Freeze-thaw cycling caused significant increases in PMP counts and phosphatidylserine externalization. Conclusions and Clinical Relevance: Canine PMPs were major determinants of thrombin-generating capacity; preanalytic variables influenced plasma PMP content. Processing conditions described here may provide a basis for characterization of PMPs in clinical studies of thrombosis in dogs.
Mostrar más [+] Menos [-]Effects of interstitial cystitis on the acoustic startle reflex in cats
2013
Hague, Devon W. | Stella, Judi L. | Buffington, C.A Tony
Objective: To compare acoustic startle reflexes (ASRs) of healthy cats and cats with interstitial cystitis (IC). Animals: 28 healthy cats (11 males and 17 females) and 20 cats with IC (13 males and 7 females). Procedures: To evaluate the effect of neutering on ASRs, ASRs in neutered and unneutered healthy cats were measured. To evaluate the effect of housing facility acclimation on ASRs in cats with IC, ASRs were measured in cats with IC within 1 month after arrival at the housing facility and again 2 to 3 months after arrival. To evaluate the effect of the environment on ASRs, ASRs were evaluated in all cats with and without IC after acclimation but before and then after environmental enrichment. Results: Neutering led to a significant decrease in overall ASR in the healthy cats. Habituation to the housing facility resulted in a significant decrease in overall ASR of female but not male cats with IC. Environmental enrichment led to a significant decrease in ASR in cats with IC but not in healthy cats. Conclusions and Clinical Relevance: The magnitude of the ASR appeared to be sensitive to environmental conditions and affected by sex, both in healthy cats and cats with IC. It was also higher in cats with IC versus healthy cats, except when cats were housed in a highly enriched environment. Impact for Human Medicine: Treatment approaches that include reduction of a patient's perception of environmental unpredictability may benefit humans with IC.
Mostrar más [+] Menos [-]Plasma concentrations and therapeutic effects of budesonide in dogs with inflammatory bowel disease
2013
Pietra, Marco | Fracassi, Federico | Diana, Alessia | Gazzotti, Teresa | Bettini, Giuliano | Peli, Angelo | Morini, Maria | Pagliuca, Giampiero | Roncada, Paola
Objective: To evaluate the pharmacokinetics and clinical efficacy of budesonide in dogs with inflammatory bowel disease (IBD). Animals: 11 dogs (mean ± SD age, 5.7 ± 3.9 years; various breeds and body weights) with moderate or severe IBD. Procedures: Each dog received a controlled-release formulation of budesonide (3 mg/m2, PO, q 24 h) for 30 days (first day of administration was day 1). The concentration of budesonide and its metabolite (16-α-hydroxyprednisolone) was measured via liquid chromatography–tandem mass spectrometry in plasma and urine samples obtained on days 1 and 8 of treatment. On those days, plasma samples were obtained before the daily budesonide administration and 0.5, 1, 2, 4, and 7 hours after drug administration, whereas urine samples were obtained after collection of the last blood sample. A clinical evaluation was performed on the dogs before onset of drug administration and on days 20 and 30 after start of drug administration. Results: The highest plasma concentration of budesonide and 16-α-hydroxyprednisolone on day 1 was detected at 1 hour and at 2 hours after drug administration, respectively. After standardization on the basis of specific gravity, the ratio between urinary concentrations of budesonide and 16-α-hydroxyprednisolone was 0.006 and 0.012 on days 1 and 8, respectively. The clinical response was adequate in 8 of 11 dogs. Conclusions and Clinical Relevance: Budesonide was rapidly absorbed and metabolized in dogs with IBD. The drug gradually accumulated, and there was an adequate therapeutic response and no adverse effects.
Mostrar más [+] Menos [-]Effects of chemical restraint on electroretinograms recorded sequentially in awake, sedated, and anesthetized dogs
2013
Freeman, Kate S. | Good, Kathryn L. | Kass, Philip H. | Park, Shin Ae | Nestorowicz, Natalia | Ofri, Ron
Objective-To quantitatively and qualitatively compare electroretinography (ERG) recordings in awake, sedated, and anesthetized dogs. Animals-Six 6-month-old Beagles. Procedures-A brief ERG protocol for dogs was used. Following 1-minute and subsequent 5-minute dark adaptation, mixed rod-cone responses were recorded bilaterally with a handheld multispecies ERG device with dogs in each of 3 states of consciousness: awake, sedated (dexmedetomidine and butorphanol), and anesthetized (atropine and hydromorphone, followed by propofol and midazolam and anesthetic maintenance with isoflurane). Low- and high-frequency noise levels were quantified via Fourier analysis, and the effect of consciousness state on signal amplitude, implicit time, and noise was analyzed via repeated-measures ANOVA. In addition, 13 veterinary ophthalmologists who were unaware of the dogs' consciousness states subjectively graded the ERG recording quality, and scores for each tracing were compared. Results-ERG amplitudes were highest in awake dogs and lowest in anesthetized dogs. Implicit times were shortest in awake dogs and longest in anesthetized dogs. Differences in b-wave amplitudes and a-wave implicit times were significant. Neither low- nor high-frequency noise levels differed significantly among consciousness states. Furthermore, no significant differences were identified among observers' scores assigned to ERG tracings. Conclusions and Clinical Relevance-Anesthesia and sedation resulted in significant attenuation and delay of ERG responses in dogs. Chemical restraint of dogs had no consistently significant effect on low- or high-frequency noise levels or on observer perception of signal quality.
Mostrar más [+] Menos [-]A temporal study of Salmonella serovars from environmental samples from poultry breeder flocks in Ontario between 1998 and 2008
2013
Sivaramalingam, Theva | McEwen, Scott A. | Pearl, David L. | Ojkic, Davor | Guerin, Michele T.
A temporal study was carried out to determine Salmonella prevalence, trends, major serovars, and their clusters from environmental samples, in poultry breeder flocks in Ontario between January 1998 and December 2008. Surveillance data were obtained from the Ontario Hatchery and Supply Flock Policy. Logistic regression with a random effect for flock was used to identify factors [poultry type, year (trend) and season] associated with the prevalence of Salmonella. A cluster detection test was used to identify clusters of common serovars. The period prevalence of Salmonella was 47.4% in broiler-breeder, 25.7% in layer-breeder, and 19.6% in turkey-breeder flocks. The overall trend in the prevalence of Salmonella was decreasing for all breeder types, due primarily to decreasing trends of Salmonella Heidelberg. The seasonal effects varied by year with the highest probability of Salmonella occurring in different seasons. The 4 most common serovars identified were Salmonella Heidelberg, Kentucky, Hadar, and Typhimurium in broiler-breeders; Salmonella Heidelberg, Brandenburg, Thompson, and Typhimurium in layer-breeders; and Salmonella Heidelberg, Saintpaul, Brandenburg, and Muenster in turkey-breeders. Salmonella Enteritidis was infrequently isolated in all poultry breeder types. Temporal clusters of different serovars were identified in all poultry breeder types. Clusters of Salmonella Heidelberg, Typhimurium, and Hadar from environmental samples from breeder flocks were detected during a similar period to clusters from hatchery fluff samples from the same population. Therefore, interventions at the breeder flock-level might help to reduce transmission of Salmonella from breeder flocks to hatcheries and possibly, to lower levels of the poultry production chain.
Mostrar más [+] Menos [-]Snatch-farrowed, porcine-colostrum-deprived (SF-pCD) pigs as a model for swine infectious disease research
2013
Huang, Yanyun | Haines, Deborah M. | Harding, John C.S.
The current study tested the benefit of commercially available spray-dried bovine colostrum (The Saskatoon Colostrum Company, Saskatoon, Saskatchewan) in raising snatch-farrowed, porcine-colostrum-deprived (SF-pCD) pigs. In experiment 1, 12 SF-pCD pigs received a liquid diet composed mainly of bovine colostrum from birth to day 10; 6 remained on the same liquid diet (COL), and the other 6 were fed a diet composed mainly of milk replacer (RPL) until weaning. In experiment 2, 12 SF-pCD pigs were fed mainly bovine colostrum before weaning; after weaning, 6 were fed a starter diet containing 20% (w/w) bovine colostrum powder (STARTER-COL), and the other 6 were fed a starter diet without any bovine colostrum (STARTER-CTRL) until termination (day 42 or day 49). In experiment 1 the COL pigs had significantly fewer fever-days than did the RPL pigs. In experiment 2 diarrhea, typhlocolitis, and pancreatic degeneration developed in 4 of the STARTER-COL pigs after weaning. In both experiments all the pigs fed mainly bovine colostrum before weaning survived until termination. All pigs tested free of swine influenza virus H1N1 and H3N2, Porcine reproductive and respiratory syndrome virus, and Porcine parvovirus. In experiment 2 all the pigs tested free of Porcine circovirus type 2 (PCV2), but some in both groups tested positive for Torque teno virus genogroups 1 and 2. In conclusion, with the use of snatch-farrowing and bovine colostrum, pigs can be raised in the absence of porcine maternal antibodies with 100% survival and freedom from most porcine pathogens of biologic relevance. This model is potentially suitable for animal disease research.
Mostrar más [+] Menos [-]Prediction of serum IgG concentration by indirect techniques with adjustment for age and clinical and laboratory covariates in critically ill newborn calves
2013
Fecteau, Gilles | Arsenault, Julie | Paré, Julie | Van Metre, David C. | Holmberg, Charles A. | Smith, Bradford P.
The objective of this study was to develop prediction models for the serum IgG concentration in critically ill calves based on indirect assays and to assess if the predictive ability of the models could be improved by inclusion of age, clinical covariates, and/or laboratory covariates. Seventy-eight critically ill calves between 1 and 13 days old were selected from 1 farm. Statistical models to predict IgG concentration from the results of the radial immunodiffusion test, the gold standard, were built as a function of indirect assays of serum and plasma protein concentrations, zinc sulfate (ZnSO4) turbidity and transmittance, and serum γ-glutamyl transferase (GGT) activity. For each assay 4 models were built: without covariates, with age, with age and clinical covariates (infection and dehydration status), and with age and laboratory covariates (fibrinogen concentration and packed cell volume). For the protein models, dehydration status (clinical model) and fibrinogen concentration (laboratory model) were selected for inclusion owing to their statistical significance. These variables increased the coefficient of determination (R2) of the models by ≥ 7% but did not significantly improve the sensitivity or specificity of the models to predict passive transfer with a cutoff IgG concentration of 1000 mg/dL. For the GGT assay, including age as a covariate increased the R2 of the model by 3%. For the ZnSO4 turbidity test, none of the covariates were statistically significant. Overall, the R2 of the models ranged from 34% to 62%. This study has provided insight into the importance of adjusting for covariates when using indirect assays to predict IgG concentration in critically ill calves. Results also indicate that ZnSO4 transmittance and turbidity assays could be used advantageously in a field setting.
Mostrar más [+] Menos [-]Effects of reduction of inspired oxygen fraction or application of positive end-expiratory pressure after an alveolar recruitment maneuver on respiratory mechanics, gas exchange, and lung aeration in dogs during anesthesia and neuromuscular blockade
2013
De Monte, Valentina | Grasso, Salvatore | De Marzo, Carmelinda | Crovace, Antonio | Staffieri, Francesco
Objective: To evaluate the effectiveness of reduction of inspired oxygen fraction (Fio2) or application of positive end-expiratory pressure (PEEP) after an alveolar recruitment maneuver (ARM) in minimizing anesthesia-induced atelectasis in dogs. Animals: 30 healthy female dogs. Procedures: During anesthesia and neuromuscular blockade, dogs were mechanically ventilated under baseline conditions (tidal volume, 12 mL/kg; inspiratory-to-expiratory ratio, 1:2; Fio2, 1; and zero end-expiratory pressure [ZEEP]). After 40 minutes, lungs were inflated (airway pressure, 40 cm H2O) for 20 seconds. Dogs were then exposed to baseline conditions (ZEEP100 group), baseline conditions with Fio2 reduced to 0.4 (ZEEP40 group), or baseline conditions with PEEP at 5 cm H2O (PEEP100 group; 10 dogs/group). For each dog, arterial blood gas variables and respiratory system mechanics were evaluated and CT scans of the thorax were obtained before and at 5 (T5) and 30 (T30) minutes after the ARM. Results: Compared with pre-ARM findings, atelectasis decreased and Pao2:Fio2 ratio increased at T5 in all groups. At T30, atelectasis and oxygenation returned to pre-ARM findings in the ZEEP100 group but remained similar to T5 findings in the other groups. At T5 and T30, lung static compliance in the PEEP100 group was higher than values in the other groups. Conclusions and Clinical Relevance: Application of airway pressure of 40 cm H2O for 20 seconds followed by Fio2 reduction to 0.4 or ventilation with PEEP (5 cm H2O) was effective in diminishing anesthesia-induced atelectasis and maintaining lung function in dogs, compared with the effects of mechanical ventilation providing an Fio2 of 1.
Mostrar más [+] Menos [-]Effect of transportation during periods of high ambient temperature on physiologic and behavioral indices of beef heifers
2013
Theurer, Miles E. | White, Brad J. | Anderson, David E. | Miesner, Matt D. | Mosier, Derek A. | Coetzee, Johann F. | Amrine, David E.
Objective: To determine the effect of transportation during periods of high ambient temperature on physiologic and behavioral indices of beef heifers. Animals: 20 heifers (mean body weight, 217.8 kg). Procedures: Ten heifers were transported 518 km when the maximum ambient temperature was ≥ 32.2°C while the other 10 heifers served as untransported controls. Blood samples were collected from transported heifers at predetermined intervals during the transportation period. For all heifers, body weights, nasal and rectal temperatures, and behavioral indices were measured at predetermined intervals for 3 days after transportation. A week later, the entire process was repeated such that each group was transported twice and served as the control twice. Results: Transported heifers spent more time near the hay feeder on the day of transportation, had lower nasal and rectal temperatures for 24 hours after transportation, and spent more time lying down for 2 days after transportation, compared with those indices for control heifers. Eight hours after transportation, the weight of transported heifers decreased 6%, whereas that of control heifers increased 0.6%. At 48 hours after initiation of transportation, weight, rectal temperature, and time spent at various pen locations did not differ between transported and control heifers. Cortisol concentrations were higher 4 hours after initiation of transportation, compared with those determined just prior to transportation. Conclusions and Clinical Relevance: Results indicated transportation during periods of high ambient temperatures caused transient changes in physiologic and behavioral indices of beef heifers.
Mostrar más [+] Menos [-]Contrast-enhanced ultrasonographic evaluation of adrenal glands in dogs with pituitary-dependent hyperadrenocorticism
2013
Pey, Pascalline | Daminet, Sylvie | Smets, Pascale Marcel Yvonne | Duchateau, Luc | Fornel-Thibaud, Pauline de | Rosenberg, Dan | Saunders, Jimmy C.H.
Objective: To assess vascular changes induced by hyperadrenocorticism of hyperplastic adrenal glands in dogs via contrast-enhanced ultrasonography. Animals: 12 dogs with pituitary-dependent hyperadrenocorticism (PDH) and 7 healthy control dogs ≥ 7 years old. Procedures: Dogs were assigned to the PDH and control groups and to small-breed (n = 6), medium-breed (4), and large-breed (9) subgroups. Contrast-enhanced ultrasonography of both adrenal glands in each dog was performed with IV injections of contrast agent. Time-intensity curves for the adrenal cortex, adrenal medulla, and ipsilateral renal artery of both adrenal glands were generated. Perfusion variables (time to peak [TTP], upslope of wash-in phase, and downslope of washout phase) were calculated. Results: Contrast-enhanced ultrasonography revealed no qualitative difference between PDH and control groups. Quantitatively, TTPs were longer in the adrenal cortex and adrenal medulla of the PDH group, compared with values for the control group, particularly in the adrenal cortex and adrenal medulla of the small-breed subgroup. Washout downslopes were lower for the renal artery, adrenal cortex, and adrenal medulla of the small-breed subgroup between the PDH and control groups. No other perfusion variables differed between groups. Conclusions and Clinical Relevance: Contrast-enhanced ultrasonography of the adrenal glands in dogs with PDH revealed a delayed TTP in the adrenal cortex and adrenal medulla, compared with values for control dogs. Contrast-enhanced ultrasonography was able to detect vascular changes induced by hyperadrenocorticism. Further studies are needed to evaluate whether reference ranges for clinically normal dogs and dogs with PDH can be determined and applied in clinical settings.
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