Comparación de los valores de glucosa en caninos obtenidos con glucómetros portátiles y la prueba estándar de laboratorio / Comparação dos valores de glicose em caninos obtidos com glicômetros portáteis e o teste laboratorial padrão / Comparison of glucose values in dogs obtained with portable glucometers and the standard laboratory test

La medición de glucosa en caninos es un procedimiento habitual en la clínica diaria, actualmente este valor se puede obtener mediante dispositivos portátiles y pruebas laboratoriales. Se realizó esta investigación con el fin de aportar mayor conocimiento sobre la importancia de la medición de glucosa, ya que en los últimos años ha perdido valor entre las pruebas hematológicas a considerar debido a que solo se relaciona con determinadas patologías como la diabetes u otras enfermedades metabólicas. El presente trabajo tiene como objetivo comparar los valores de glucosa en caninos obtenidos mediante un glucómetro portátil de uso humano (Accu-chek® Active, Roche Diagnostic, Mannheim, Alemania); veterinario (aLcose® Vet Glu, jjPlus Corporation, New Taipei, Taiwán) y la prueba estándar de laboratorio, esto nos indicará la fiabilidad de los resultados obtenidos mediante estos métodos. Se realizó la toma de muestras de sangre de 50 caninos clínicamente sanos, de los cuales se obtuvo el resultado de glucemia mediante estos tres métodos. Los resultados de nuestra investigación evidenciaron que las tres formas de evaluación de la glucosa sanguínea en perros brindaban resultados estadísticamente diferentes (p < 0.05). Se obtuvo valores de glucosa diferentes entre los tres métodos de medición, teniendo como promedios finales 84.14 mg/dL, 101.12 mg/dL y 91.12 mg/dL correspondientes al glucómetro portátil de uso humano, veterinario y a la prueba estándar de laboratorio respectivamente. En conclusión, los glucómetros portátiles de uso humano subestiman los valores reales de glucosa, mientras que los de uso veterinario lo sobreestiman, comparados con la prueba estándar de laboratorio.

A medição de glicose nos cães é um procedimento habitual realizado no atendimento clínico. Atualmente este valor pode ser obtido por meio de dispositivos portáteis e testes laboratoriais. Esta pesquisa foi realizada com a finalidade de destacar a importância da medição de glicose, visto que nos últimos anos esta avaliação não tem sido muito valorada entre os testes hematológicos, sendo considerada relevante apenas em relação a patologias como a diabetes e outras doenças metabólicas. O presente estudo teve como objetivo comparar os valores de glicose em cães obtidos com glicômetro portátil de uso humano; veterinário e o teste padrão de laboratório. Esta comparação poderá indicar a confiabilidade dos resultados obtidos mediante os métodos avaliados. Foi realizada a amostragem do sangue de 50 caninos clinicamente sadios os quais foram submetidos a avaliação de glicose mediante os três métodos. Os resultados de nossa investigação evidenciaram que as três formas de avaliação da glicose sanguínea têm resultados estatisticamente diferentes (p < 0,05). Os valores de glicose tiveram medias finais de 84,14 mg/dL, 101,12 mg/dL e 91,12 mg/dL para o glicômetro portátil de uso humano (Accu-chek® Active, Roche Diagnostic, Mannheim, Alemanha), veterinário (aLcose® Vet Glu, jjPlus Corporation, Nova Taipei, Taiwan) e o teste padrão de laboratório, respectivamente. Ao concluir, os glicômetros portáteis de uso humano subestimam os valores reais de glicose e os de uso veterinário os superestimam quando comparados com o teste padrão de laboratório.

The measurement of glucose in canines is a common procedure in daily clinical practice. Currently this value can be obtained by use of portable devices and laboratory tests. This research was carried out in order to provide more knowledge about the importance of glucose measurement, since in recent years it has lost value among the hematological tests to be considered because it is only related to certain pathologies such as diabetes or other metabolic diseases. The present study aimed to compare the glucose values in dogs obtained with a portable glucometer for human use, veterinarian use, and the standard laboratory test. This comparison may indicate the reliability of the results obtained through the evaluated methods. A blood sampling of 50 clinically healthy canines was taken and submitted to glucose evaluation using the three methods. Our investigation showed that the three ways of assessing blood glucose have statistically different results (p < 0.05). Glucose values had final averages of 84.14 mg/dL, 101.12 mg/dL, and 91.12 mg/dL for the portable glucometer for human use (Accu-chek® Active, Roche Diagnostic, Mannheim, Germany), veterinary (aLcose® Vet Glu, jjPlus Corporation, New Taipei, Taiwan) and the standard laboratory test, respectively. In conclusion, portable glucometers for human use underestimate the glucose values, and those for veterinary use overestimate them compared to the standard laboratory test.

Alterações no VCM e suas correlações com os valores de RDW-SD e RDW-CV em caninos e felinos / Variation in MCV and its correlation with RDW-SD and RDW-CV values in dogs and cats

A anemia é uma síndrome caracterizada pela diminuição do número de hemácias, hematócrito e/ou concentração de hemoglobina. Conforme o Volume Corpuscular Médio (VCM), as anemias podem ser classificadas em microcíticas, normocíticas ou macrocíticas. O RDW (Amplitude de Distribuição dos Eritrócitos) também é utilizado para ajudar na classificação das anemias, refletindo a anisocitose da população eritrocitária. Neste estudo retrospectivo objetivou-se determinar a correlação entre o RDW-SD (Desvio Padrão), RDW-CV (Coeficiente de Variação), macrocitose e microcitose em caninos e felinos atendidos na rotina clínica do Hospital Veterinário da Universidade Luterana do Brasil. Para a realização deste estudo, selecionou-se 662 laudos de hemogramas realizados (434 caninos e 228 felinos), com faixa etária de seis meses até 10 anos, foram divididos em dois grupos: Grupo 1 ­ Anemia microcítica (255 caninos e 61 felinos); Grupo 2 ­ Anemia macrocítica (179 caninos e 167 felinos). Posteriormente, correlacionou-se os grupos com os valores de RDW-SD e RDW-CV. As análises de correlação foram realizadas utilizando o teste Spearman, para a análise de significância foi utilizado o T Student, no programa IBM SPSS®Statistics. Na análise estatística do grupo canino, não houve correlação da microcitose com o RDW-SD, enquanto o RDW-CV apresentou uma correlação inversamente proporcional, razoável. No grupo macrocítico canino, a análise de correlação com o RDW-SD foi moderada e diretamente proporcional, e com o RDW-CV foi moderada e diretamente proporcional. No grupo felino, não houve correlação entre microcitose e RDW-SD, e com o RDW-CV houve uma correlação razoável e inversamente proporcional. Entre macrocitose em felinos e o RDW-SD houve uma correlação moderada e diretamente proporcional, já o RDW-CV apresentou uma correlação razoável e diretamente proporcional. Conclui-se que os caninos e felinos do grupo microcítico apresentam uma correlação com o RDW-CV. Contudo, os caninos com macrocitose apresentaram correlação tanto para o RDW-CV quanto para o RDW-SD, e os felinos apresentaram uma maior correlação com o RDW-SD.

Anemia is a syndrome characterized by a low red blood cell count, hematocrit and/or hemoglobin concentration. According to the Mean Corpuscular Volume (MCV), anemias can be classified as microcytic, normocytic or macrocytic. The RDW (Red Cell Distribution Width) is also used to help classify anemias, reflecting the anisocytosis of the erythrocyte population. This retrospective study aimed to determine the correlation between RDW-SD (Standard Deviation), RDW-CV (Coefficient of Variation), macrocytosis and microcytosis in canines and felines treated in the clinical routine of the Veterinary Hospital of Universidade Luterana do Brasil. To carry out this study, 662 blood count reports were selected (434 canines and 228 felines), aged between six months and 10 years, divided into two groups: Group 1 ­ Microcytic anemia (255 canines and 61 felines); Group 2 ­ Macrocytic anemia (179 canines and 167 felines). Subsequently, the groups were correlated with the values of RDW-SD and RDW-CV. Correlation analyzes were performed using the Spearman test, for the analysis of significance the T Student was used, in the IBM SPSS® Statistics program. In the statistical analysis of the canine group, there was no correlation between microcytosis and the RDW-SD, while the RDW-CV showed a reasonable, inversely proportional correlation. In the canine macrocytic group, correlation analysis with RDW-SD was moderate and directly proportional, and with RDW-CV it was moderate and directly proportional. In the feline group, there was no correlation between microcytosis and RDW-SD, and with RDW-CV there was a reasonable and inversely proportional correlation. There was a moderate and directly proportional correlation between macrocytosis in felines and RDW-SD, whereas RDW-CV presented a reasonable and directly proportional correlation. It is concluded that the canines and felines of the microcytic group present a correlation with the RDW-CV. However, canines with macrocytosis showed a correlation for both RDW-CV and RDW-SD, and felines showed a greater correlation with RDW-SD.

Persistent hypercobalaminemia three months after successful gradual attenuation of extrahepatic shunts in dogs: a prospective cohort study.

BACKGROUND: Deficiencies in vitamin A and D and disorders in the vitamin B complex are often present in people with chronic liver diseases. So far, the serum concentrations of these vitamins have not yet been studied in dogs with congenital extrahepatic portosystemic shunts (EHPSS), who also have some degree of liver dysfunction. The objective was to assess serum vitamin concentrations in dogs with EHPSS from diagnosis to complete closure. A prospective cohort study was performed using ten client-owned dogs with EHPSS, closed after gradual surgical attenuation. Serum concentrations of vitamin A, 25-hydroxyvitamin D, folic acid, cobalamin and methylmalonic acid (MMA) were measured at diagnosis prior to institution of medical therapy, prior to surgery, and three months after gradual attenuation and complete closure of the EHPSS. RESULTS: At diagnosis, median serum concentrations of vitamin A, 25-hydroxyvitamin D and folic acid were 18.2 µg/dL (8.8 - 79.5 µg/dL), 51.8 ng/mL (19.4 - 109.0 ng/mL), and 8.1 µg/L (5.2 - 14.5 µg/L), respectively, which increased significantly postoperatively (88.3 µg/dL (51.6 - 182.2 µg/dL, P=0.005), 89.6 ng/mL (49.3 - >150.0 ng/mL, P =0.005), and 14.8 µg/L (11.5 - 17.7 µg/L, P <0.001), respectively). Median serum cobalamin concentrations were 735.5 ng/L (470 - 1388 ng/L) at diagnosis and did not significantly decrease postoperatively (P =0.122). Both at diagnosis and three months postoperatively 7/10 dogs had hypercobalaminemia. CONCLUSIONS: Serum concentrations of vitamin A, 25-hydroxyvitamin D and folic acid significantly increase after surgical attenuation. Nevertheless, persistent hypercobalaminemia is suggestive of ongoing liver dysfunction, despite successful surgery.

A cross-sectional retrospective study of SARS-CoV-2 seroprevalence in domestic cats, dogs and rabbits in Poland.

BACKGROUND: Coronaviruses (CoVs) have long been known to cause infection in domestic and free-living birds and mammals including humans. The zoonotic origin of SARS-CoV-2 and the biological properties of CoVs, including ability to cross interspecies barriers, enable its emergence in populations of various animals, including companion animals (cats, dogs, rabbits) an area requiring further study. To date, several cases of cats and dogs positive for SARS-CoV-2 and/or specific antibodies have been described. The aim of our cross-sectional retrospective study is to determine seroprevalence of SARS-CoV-2 in domestic dog, cat and rabbit population during recent COVID-19 pandemic in Poland. RESULTS: In total, serum samples from 279 cats and 343 dogs and 29 rabbits were used in the study. The seroprevalence of SARS-CoV-2 in cats and dogs reached 1.79% (95% CI: 0.77 - 4.13) and 1.17% (95% CI 0.45 - 2.96), respectively (p ≥ 0.05). Anti- SARS-CoV-2 antibodies were detected in 5 cats (mean S/P% 106 ± 48.23) and 4 dogs (mean S/P% 78.5 ± 16.58). All 29 samples from rabbits were negative for SARS-CoV-2 antibodies. No significant gender or age differences in seroprevalence in dogs and cats (p ≥ 0.05) were found. None of the animals with anti-SARS-CoV-2 antibodies displayed respiratory or gastrointestinal signs at the time of sampling. CONCLUSIONS: Our results confirmed previous findings that SARS-CoV-2 infections in companion animals occurs but are not frequent. Future serological testing of large pet population may provide a comprehensive picture of disease dynamics in companion animals.

High-definition ultrasonography in the evaluation of the reproductive tract of bitches during the follicular phase of the estrous cycle.

The aim was to sonographically evaluate the reproductive tract of bitches during the follicular phase of the estrous cycle using High Density (HD) ultrasonic techniques. Females (n = 8) were evaluated at five different times throughout the follicular phase, as determined by vaginal cytology and blood progesterone concentrations. Ultrasonic exams were performed using the ACUSON S2000/SIEMENS device utilizing a multifrequency HD transducer (5.5-18 MHz). Videos of the ovaries were obtained and recordings were evaluated using a DICOM viewer software for counting and measuring the ovarian structures, which were assigned to groups based on diameter in mm: G1: ≤ 1; G2: from 1.01 to 3.5; G3 from 3.51 to 5.5; G4: from 5.51 to 10. There was a greater uterine thickness with the progression of the follicular phase (P < 0.05). Six distinct regions were identified in the uterine wall. The ovarian dimensions increased (P < 0.05) as stage of the follicular phase advanced. There was fluid detected around the ovaries after ovulation. There was a characteristic fat tissue hyperechogenicity around the ovaries at all timepoints. There was a difference in the number of ovarian structures of each dimension group at each time there were assessments (P < 0.05). There was a difference in diameter of the largest ovarian structure and in average value of wall thickness at all timepoints when there were evaluations (P < 0.05). The HD ultrasonography technique provides for excellent image resolution, allowing for a more precise characterization of the bitch's reproductive structures and changes occurring during the follicular phase of the estrous cycle.

Analytical validation of a point-of-care test and an automated immunoturbidimetric assay for the measurement of canine C-reactive protein in serum.

C-reactive protein (CRP) is an acute phase protein, which is used to evaluate and monitor the response of the innate immune system to a variety of inflammatory processes in the dog. The purpose of this study was to analytically validate a point-of-care assay (IDEXX Catalyst CRP Test) and an immunoturbidimetric assay (Gentian Canine CRP Immunoassay) for the measurement of serum CRP concentrations in dogs. These 2 assays (Catalyst, Gentian) were compared to a previously validated enzyme-linked immunosorbent assay (Tridelta Development EIA Canine CRP Assay). Linearity, precision, reproducibility, and accuracy were assessed using leftover serum samples. Agreement between assays was assessed using leftover serum samples and serum from clinically healthy dogs. Observed to expected ratios (O/E) for dilutional parallelism were 83.9 to 163.1% and 108.3 to 160.6% for the Catalyst and the Gentian assays, respectively. Coefficients of variation for intra-assay variability ranged from 6.4 to 9.5% for the Catalyst assay and 1.5 to 2.6% for the Gentian assay. Coefficients of variation for inter-assay variability ranged from 3.8 to 18.2% for the Catalyst assay and 4.5 to 5.8% for the Gentian assay. The mean O/E for recovery were 97.9% and 98.5% for the Catalyst and Gentian assays, respectively. Correlations between assays were as follows: Catalyst and Tridelta (R 2 = 0.76), Gentian and Tridelta (R 2 = 0.79), and Catalyst and Gentian (R 2 = 0.98). The Catalyst and Gentian assays are both acceptable for measuring CRP in dog serum, but their results are not directly comparable with the Tridelta assay.

La protéine C réactive (CRP) est une protéine de phase aiguë, qui est utilisée pour évaluer et surveiller la réponse du système immunitaire inné à une variété de processus inflammatoires chez le chien. Le but de cette étude était de valider analytiquement un test au point de service (test IDEXX Catalyst CRP) et un test immunoturbidimétrique (Gentian Canine CRP Immunoassay) pour la mesure des concentrations sériques de CRP chez le chien. Ces deux tests (Catalyst, Gentian) ont été comparés à un test immuno-enzymatique précédemment validé (Tridelta Development EIA Canine CRP Assay). La linéarité, la précision, la reproductibilité et l'exactitude ont été évaluées à l'aide d'échantillons de sérum restants. La concordance entre les tests a été évaluée à l'aide d'échantillons de sérum restants et de sérum provenant de chiens cliniquement sains. Les rapports observés/attendus (O/E) pour le parallélisme de dilution étaient de 83,9 à 163,1 % et de 108,3 à 160,6 % pour les tests Catalyst et Gentian, respectivement. Les coefficients de variation pour la variabilité intra-test variaient de 6,4 à 9,5 % pour le test Catalyst et de 1,5 à 2,6 % pour le test Gentian. Les coefficients de variation pour la variabilité inter-test variaient de 3,8 à 18,2 % pour le test Catalyst et de 4,5 à 5,8 % pour le test Gentian. L'O/E moyen pour la récupération était de 97,9 % et de 98,5 % pour les tests Catalyst et Gentian, respectivement. Les corrélations entre les tests étaient les suivantes : Catalyst et Tridelta (R 2 = 0,76), Gentian et Tridelta (R 2 = 0,79) et Catalyst et Gentian (R 2 = 0,98). Les tests Catalyst et Gentian sont tous deux acceptables pour mesurer la CRP dans le sérum de chien, mais leurs résultats ne sont pas directement comparables avec le test Tridelta.(Traduit par Docteur Serge Messier).

Effect of age, sex, and body size on the blood biochemistry and physiological constants of dogs from 4 wk. to > 52 wk. of age.

BACKGROUND: Blood biochemistry and reference intervals help to differentiate between healthy and ill dogs as well as to provide information for the prognosis, evaluation, and monitoring; however, these intervals are often obtained from adult animals. It is essential to understand that puppies and adults are physiologically different, which justifies the need to obtain age-specific biochemical reference intervals. The aim of this research was to assess the potential effect of age, sex, body size, and their interaction on routine biochemical analytes and physiological constants (body temperature, heart rate, and respiratory rate). To carry out the research, we selected 197 healthy dogs of both sexes and different body sizes (small, medium and large) classified by age: group I (4-8 wk), group II (9-24 wk), group III (25-52 wk), and group IV (> 52 wk). The biochemical analysis included the measurement of the enzymatic activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), and the concentrations of cholesterol, triglycerides, total proteins, albumin, globulins, glucose, urea, and creatinine. Statistical analyses used analysis of variance (ANOVA) and a general linear model (GLM), which allows the comparison of multiple factors at two or more levels (p < 0.05). RESULTS: The results of this study showed that ALT, total protein, albumin, globulin, urea, creatinine, and body temperature levels were lower in puppies than in adult dogs of group IV (p < 0.05), while the enzymatic activity of ALP, LDH, glucose concentration, and heart rate were higher. Whereas sex, body size and the interaction did not show a significant effect (p > 0.05). CONCLUSIONS: Some biochemical components are influenced by age. For this reason, this manuscript contributes with additional data for the clinical interpretation of blood biochemical results in puppies.

Hemorrhagic shock and hemostatic resuscitation in canine trauma.

Hemorrhage is a significant cause of death among military working dogs and in civilian canine trauma. While research specifically aimed at canine trauma is limited, many principles from human trauma resuscitation apply. Trauma with significant hemorrhage results in shock and inadequate oxygen delivery to tissues. This leads to aberrations in cellular metabolism, including anaerobic metabolism, decreased energy production, acidosis, cell swelling, and eventual cell death. Considering blood and endothelium as a single organ system, blood failure is a syndrome of endotheliopathy, coagulopathy, and platelet dysfunction. In severe cases following injury, blood failure develops and is induced by inadequate oxygen delivery in the presence of hemorrhage, tissue injury, and acute stress from trauma. Severe hemorrhagic shock is best treated with hemostatic resuscitation, wherein blood products are used to restore effective circulating volume and increase oxygen delivery to tissues without exacerbating blood failure. The principles of hemostatic resuscitation have been demonstrated in severely injured people and the authors propose an algorithm for applying this to canine patients. The use of plasma and whole blood to resuscitate severely injured canines while minimizing the use of crystalloids and colloids could prove instrumental in improving both mortality and morbidity. More work is needed to understand the canine patient that would benefit from hemostatic resuscitation, as well as to determine the optimal resuscitation strategy for these patients.

Canine pyometra: interferences of age and type in blood count and serum biochemistry / Piometra canina: interferências da idade e tipo sobre o hemograma e a bioquímica sérica

Hematological and serum biochemical characteristics and prevalence of microorganisms in the uterine content of bitches with pyometra were described, according to type and age. This study was carried out at the Veterinary Medicine Hospital Renato Rodenburg de Medeiros Netto of the UFBA with 132 patients diagnosed with pyometra through imaging and laboratory tests, divided into two groups: Group 1 - less than eight years (n=72) and Group 2 - more than eight years (n=60). Both groups were submitted to Ovariohysterectomy (OH), in which blood and uterine lumen secretion samples were collected. The highest incidence of pyometra was found in bitches under 8 years (54.5%), Poodle breed (31.8%) and mixed-breed (27.3%). Open pyometra represented 80.3% of all cases. The death rate (6.0%) did not show significant difference between the types. Anemia (73.0%), thrombocytopenia (41.2%) and leukocytosis (77.0%) were observed. Urea and ALP were above the reference values in 41.8% and 52.2% of each group, respectively, while ALT (94.0%) and creatine (82%) remained normal in most animals. Serum biochemistry did not show significant differences (P<0.05). Escherichia coli and Klebsiella sp. grew in 33.3% and 16.7% of the samples, respectively. Therefore, no significant hematological and serum biochemical differences were observed in bitches with pyometra in both age categories.

Descreveu-se características hematológicas, bioquímicas séricas e prevalência de microrganismos no conteúdo uterino de cadelas com piometra, de acordo com o tipo e idade. Realizou-se este estudo no Hospital de Medicina Veterinária Renato Rodenburg de Medeiros Netto da UFBA com 132 pacientes diagnosticadas com piometra, por exames de imagem e laboratoriais, divididas em dois grupos: Grupo 1 – menos de oito anos (n=72) e Grupo 2 – mais de oito anos (n=60), submetidas a Ovariectomia (OH), quando se coletou amostras de sangue e secreção do lúmen uterino. A incidência maior deu-se em cadelas abaixo de 8 anos (54,5%), da raça Poodle (31,8%) e sem raça definida (SRD) (27,3%), sendo 80,3% de piometra aberta. A taxa de óbito (6,0%) não apresentou diferença significativa entre os tipos. Observou-se a ocorrência de anemia (73,0%), trombocitopenia (41,2%) e leucocitose (77,0%). Os valores de ureia e FA estiveram acima dos valores de referência em 41,8% e 52,2% das cadelas respectivamente, enquanto a ALT (94,0%) e a creatina (82%) mantiveram-se dentro dos valores de referência na maioria dos animais. A bioquímica sérica não apresentou diferenças significativas (P<0,05). Observou-se crescimento de Escherichia coli e Klebsiella sp. em 33,3% e 16,7% das amostras, respectivamente. Sendo assim, não se observou diferenças significativas hematológicas e na bioquímica sérica de cadelas com piometra nas categorias de idade.

Comparison of glucose concentrations in canine whole blood, plasma, and serum measured with a veterinary point-of-care glucometer.

Previous studies have determined that, compared to whole blood, serum or plasma used in a portable blood glucometer (PBG) may provide more accurate results. We investigated the accuracy of a veterinary PBG (AlphaTRAK 2; Zoetis) for the measurement of glucose concentrations in serum, plasma, and whole blood compared to plasma glucose concentration measured by a biochemical analyzer. Blood samples from 53 client-owned dogs were collected. Lin concordance correlation coefficient (ρc) and Bland-Altman plots were used to determine correlation and agreement between the results obtained for the different sample types. Glucose concentration in whole blood measured by the veterinary PBG was more strongly correlated with the glucose concentration measured by the biochemical analyzer (ρc = 0.92) compared to plasma and serum glucose concentrations (ρc = 0.59 and 0.57, respectively). The mean differences between the glucose concentrations in whole blood, plasma, and serum measured by the veterinary PBG and the glucose concentration determined by the biochemical analyzer were 1.0, 6.3, and 6.7 mmol/L (18, 113, and 121 mg/dL), respectively. Our findings suggest that, when using this veterinary PBG, the accuracy of a glucose measurement obtained is higher when using whole blood compared to plasma or serum. Use of whole blood allows for more correct assessment and diagnosis, which are necessary for appropriate therapeutic intervention.

Pharmacokinetics of a high-concentration formulation of buprenorphine (Simbadol) in male dogs.

OBJECTIVE: To describe the pharmacokinetics of buprenorphine in dogs following administration of a high-concentration formulation of buprenorphine. STUDY DESIGN: Prospective, randomized, crossover study. ANIMALS: A total of six healthy male intact Beagle dogs, aged 9-13 months and weighing 10.3 ± 1.4 kg (mean ± standard deviation). METHODS: Dogs were randomized to be administered buprenorphine (0.12 mg kg-1; Simbadol, 1.8 mg mL-1) via the intravenous (lateral saphenous) or subcutaneous (dorsal interscapular) route followed by the alternative route of administration after a 14 day interval. Blood was sampled before administration and at set times up to 72 hours after injection. Plasma buprenorphine concentration was measured using liquid chromatography-tandem mass spectrometry. RESULTS: A three-compartment model with zero or biphasic rapid and slow first-order input in (intravenous or subcutaneous data, respectively) and first-order elimination from the central compartment best fitted the data. The rapid first-order input accounted for 63% of the dosage absorption. Typical values (% interindividual variability) for the three compartment volumes were 900 (33), 2425 (not estimated) and 6360 (28) mL kg-1. The metabolic and two distribution clearances were 25.7 (21), 107.5 (74) and 5.7 (61) mL minute-1 kg-1. The absorption half-life for the fast absorption phase was 8.9 minutes with a 0.7 (103) minute delay. The absorption half-life for the slow absorption phase was 347 minutes with a 226 (42) minute delay. Median (range) bioavailability calculated from noncompartmental analysis was 143 (80-239)%. Calculated terminal half-life was 963 minutes. CONCLUSIONS AND CLINICAL RELEVANCE: The high-concentration formulation of buprenorphine administered subcutaneously had a large volume of distribution and a rapid absorption phase followed by slower, delayed absorption. The high estimate of bioavailability should be interpreted with caution as values above 100% are most commonly related to experimental issues.

Safety and immunological effects of recombinant canine IL-15 in dogs.

Interleukin-15 (IL-15) is a pleiotropic cytokine that plays pivotal roles in innate and adaptive immunity. It is also a promising cytokine for treating cancer. Despite growing interest in its use as an immunotherapeutic, its safety and immunological effects in dogs have not been reported. In this study, healthy dogs were given recombinant canine IL-15 (rcIL-15) intravenously at a daily dose of 20 µg/kg for 8 days and monitored for 32 days to determine the safety and immunological effects of rcIL-15. The repeated administration of rcIL-15 was well tolerated, did not cause any serious side effects, and promoted the selective proliferation and activation of canine anti-cancer effector cells, including CD3+CD8+ cytotoxic T lymphocytes, CD3+CD5dimCD21-, and non-B/non-T NK cell populations, without stimulating Treg lymphocytes. The rcIL-15 injections also stimulated the expression of molecules and transcription factors associated with the activation and effector functions of NK cells, including CD16, NKG2D, NKp30, NKp44, NKp46, perforin, granzyme B, Ly49, T-bet, and Eomes. These results suggest that rcIL-15 might be a valuable therapeutic adjuvant to improve immunity against cancer in dogs.

Evaluation of the i-STAT Alinity v in a veterinary clinical setting.

Many point-of-care (POC) analyzers are available for the measurement of electrolytes and acid-base status in animals. We assessed the precision of the i-STAT Alinity v, a recently introduced POC analyzer, and compared it to 2 commonly used and previously validated POC analyzers (i-STAT 1, Stat Profile pHOx Ultra). Precision was evaluated by performing multiple analyses of whole blood samples from healthy dogs, cats, and horses on multiple i-STAT Alinity v analyzers. For comparison between analyzers, whole blood samples from dogs and cats presented to the emergency room were run concurrently on all 3 POC instruments. Reported values were compared by species (dogs and cats only) using Pearson correlation, and all values from all species were analyzed together for the Bland-Altman analysis. Results suggested that the i-STAT Alinity v precision was very good, with median coefficients of variability <2.5% for all measured parameters (except the anion gap), with variable ranges of coefficients of variation. In addition, good-to-excellent correlation was observed between the i-STAT Alinity v and i-STAT 1, and between the i-STAT Alinity v and Stat Profile pHOx Ultra for all parameters in both cats and dogs, respectively. In this cohort, the i-STAT Alinity v had clinically acceptable bias compared to the currently marketed analyzers and can be used for monitoring measured analytes in cats and dogs, although serial measurements in a single animal should be performed on the same analyzer whenever possible.

Vasoactive intestinal peptide in canine portosystemic shunt in the absence of portal hypertension.

Background: The congenital portosystemic shunt (PSS) is a common vascular anomaly in dogs. Vasoactive intestinal peptide (VIP) is produced in various organs (including the small intestine, large intestine, and pancreas), leading to abdominal vasodilation, increased blood flow, increased pancreatic blood flow, and promotion of pancreatic endocrine and exocrine secretions. However, there have been no reports on the concentration of VIP in the portal and peripheral veins in canine PSS. Aim: The aim of this pilot study was to evaluate whether dogs with PSS have a different VIP concentration in their portal system in general. Methods: Six dogs with an extrahepatic portosplenic shunt were included in the study. Blood samples were taken from the saphenous and portal veins during PSS ligation surgery with an amerid constrictor, to evaluate and compare the VIP concentration in both samples. VIP was measured using a commercial canine enzyme-linked immunosorbent assay kit. Results: The breeds included Mongrels (n = 2), Norfolk Terriers (n = 1), Miniature Dachshunds (n = 1), and Maltese (n = 2), and their ages were 9.3 ± 6.5 months; the bodyweight was 3.3 ± 0.8 kg. The concentration of VIP in the saphenous vein was 17.75 ± 13.88 pg/ml; on the contrary, the concentration of VIP in the portal vein was 29.7 ± 20.29 pg/ml. There was no significant difference in the VIP concentration between veins. Conclusion: There was no difference in the VIP concentration between the portal and saphenous veins, suggesting a non-association between VIP and the PSS, in the absence of portal hypertension.

Serum cross-linked N-telopeptide of type I collagen across the canine life span: An investigation in intact and neutered male and female dogs.

In this study, we conducted study to explore the association between serum cross-linked N-telopeptide of type I collagen (NTx), a marker of bone resorption, and age, body weight, and blood biochemical parameters as well as the neutered and intact status in male and female dogs. We targeted 145 healthy dogs (aged 0.33-18.33 years); 70 were males (38 intact, 32 castrated), and 75 were females (31 intact, 44 ovariohysterectomized). We found that the NTx levels were significantly higher in dogs aged ≤2 years than in older dogs. NTx concentration tended to decrease with age in dogs aged ≤2 years, but not significantly, and remained constant in dogs aged >2 years. Accordingly, we investigated sex/sterilization status in two age cohorts (juvenile-to-young-adult, ≤2 years of age; adult-to-geriatric, >2 years of age). In the adult-to-geriatric cohort, NTx concentration was highest in intact males, followed by neutered males, neutered females, and intact females. The intact vs. neutered difference was significant in males, but not in females. Our results suggested that estradiol deficiency may not affect bone metabolism in female dogs, but androgen deficiency may affect bone metabolism in male dogs. Furthermore, age did not affect bone metabolism after 2 years. NTx concentrations were significantly higher in the juvenile-to-young-adult cohort than in the adult-to-geriatric cohort and tended to decrease with age, similar to young humans. This study unveils novel sex differences in canine serum NTx concentrations and suggests the effect of neutering on bone metabolism, showing that serum NTx concentrations change with age.

Serum tT4, fT4 and TSH concentrations in German Shepherd dogs depending on age and type of work.

Serum concentration of thyroid hormones in healthy dogs varies according to age, sex, breed or professional activity. The aim of this study was to determine the influence of both age and dogs' work involvement on TSH and thyroid hormones values. Thyroid-stimulating hormone (TSH), total thyroxine (tT4) and free thyroxine (fT4) were tested in the serum of 57 healthy, German Shepherd dogs. The dogs were divided into study groups according to age: dogs aged 3 to 6 years (A), dogs over the age of 6 years (B) and involvement: police-working dogs (C) and accompanying animals (D). Mean values of TSH, tT4 and fT4 ranged from 0.19 to 0.31 ng/ml, 15.58 to 17.25 nmol/L and 11.83 to 17.89 pmol/L, respectively. The highest values of TSH were in group B and the lowest were in group A, while there was an inverse dependence in case of fT4 concentration. The highest mean values of tT4 were in dogs in group C and the lowest in group B. There were statistically significant differences in TSH (p=0.007) and fT4 (p=0.003) concentrations between the age groups. The results indicate that a dog's age is an important factor in the case of thyroid profile results interpretation.

Hemostatic capacity of canine chilled whole blood over time.

OBJECTIVE: To determine the hemostatic potential of canine chilled whole blood maintained at clinically relevant storage conditions. DESIGN: In vitro experimental study. SETTING: Government blood and coagulation research laboratory and government referral veterinary hospital. ANIMALS: Ten healthy Department of Defense military working dogs. INTERVENTIONS: One unit of fresh whole blood was collected from each of 10 military working dogs using aseptic technique. Blood was maintained in a medical-grade refrigerator for 28 days at 4°C (39°F) and analyzed before refrigeration (day 0) and after (days 2, 4, 7, 9, 11, 14, 21, and 28). MEASUREMENTS AND MAIN RESULTS: Ten units of canine blood were analyzed with whole blood platelet aggregation, thromboelastography, CBC, biochemical analysis, blood gas, and prothrombin/activated partial thromboplastin/fibrinogen assay. Clotting strength of chilled blood was maintained up to 21 days despite significant decreases in platelet aggregation to ADP, collagen, or γ-thrombin, significant prolongation of prothrombin and activated partial thromboplastin times, and reduced speed of clot formation (K time, alpha angle). Fibrinogen concentration, WBC, RBC, and platelet counts did not change over time. CONCLUSIONS: Chilled canine whole blood loses a small percentage of clot strength through 21 days of refrigerated storage. Further research is needed to determine if this hemostatic potential is clinically relevant in hemorrhaging dogs who require surgical intervention or are exposed to traumatic events.

Effects of storage and leukocyte reduction on the concentration and procoagulant activity of extracellular vesicles in canine packed red cells.

OBJECTIVES: To characterize the size and procoagulant activity of extracellular vesicles (EV) that accumulate in canine packed red blood cells (pRBCs) over time and the effect of leukocyte reduction on these characteristics. DESIGN: Prospective cohort study. SETTING: Private small animal specialty referral hospital and university research laboratories. ANIMALS: Ten healthy blood donor dogs. INTERVENTIONS: Five pRBCs units were obtained according to standard protocols, and 5 were leukocyte-reduced prior to processing. Platelet-free supernatant from the pRBC units was collected on days 0, 10, 20, 32, and 42. MEASUREMENTS AND MAIN RESULTS: Nanoparticle tracking analysis was performed to determine the size and concentration of EVs. Thrombin generation associated with phosphatidylserine-positive EVs was determined using a capture assay. Factor Xa generation associated with phosphatidylserine-positive EVs and tissue factor-positive EVs was measured in a subset of EVs isolated by centrifugation of the supernatant at 20,000 × g. R package nparLD and the Mann-Whitney U-test were used to determine the effect of duration of storage and the effect of leukocyte reduction, respectively. Small (mean < 125 nm) procoagulant EVs accumulated over time, with significant increases occurring on or after day 20 in both non-leukocyte reduced and leukocyte-reduced units. The procoagulant activity of the EVs was due to phosphatidylserine, not tissue factor. Increases in EV concentration and procoagulant activity occurred earlier in non-leukocyte reduced units. Extracellular vesicle accumulation and procoagulant activity were not decreased at any individual time point by leukocyte reduction. CONCLUSIONS: Further studies characterizing and determining the clinical relevance of small procoagulant EVs in pRBCs are warranted.

An in vitro study of canine cryopoor plasma to correct vitamin K-dependent coagulopathy in dogs.

OBJECTIVE: To compare the efficacy of fresh frozen plasma (FFP) with cryopoor plasma (CPP) to treat vitamin K-dependent factor deficiency in a canine in vitro setting. DESIGN: In vitro laboratory study. SETTING: University veterinary medical teaching hospital. ANIMALS: Seven units of FFP and 6 units of CPP from unique canine donors from the university veterinary blood bank. INTERVENTIONS: Canine FFP was adsorbed by oral barium sulfate suspension to mimic vitamin K-dependent coagulopathy. A sequential mixing study was completed by adding FPP or CPP to the adsorbed plasma. Measurements of prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen, and factor activities of factors II, VII, and IX (FII, FVII, and FIX) were compared between the 2 treatment groups. MEASUREMENTS AND MAIN RESULTS: When comparing the sequential addition of CPP or FPP to adsorbed plasma, the following had no statistical significance: PT (P = 0.94), aPTT (P = 0.66), FII (P = 0.05), and FIX (P = 0.90). There was a dose-dependent decrease with PT and aPTT and a dose-dependent increase with FII and FIX. In contrast, after the addition of either CPP or FFP, there was a significant difference between the treatment groups for the concentration of fibrinogen (P = 0.005) and activity of FVII (P = 0.044), with FFP resulting in a greater concentration of fibrinogen and CPP resulting in a greater concentration of FVII. Measurements of factor X (FX) were initially included in the study but were later excluded because FX appeared to be continually adsorbed even after the addition of CPP or FFP. CONCLUSIONS: CPP partially corrected the coagulation times and concentration of vitamin K-dependent coagulation factors to the same degree as FFP. CPP, generally less expensive than FFP, may provide an alternative treatment option for vitamin K-dependent coagulopathies, although in vivo testing is needed.

Analysis of blood clotting with the total thrombus analysis system in healthy dogs.

To date, coagulation tests are unable to reflect in vivo coagulation status in the same system, including platelet function, fibrin clot formation, and whole blood flow. The Total Thrombus Analysis System (T-TAS), which is a microfluidic assay that simulates conditions in vivo, measures whole blood flow at defined shear rates under conditions designed to assess platelet function (PL-chip) or coagulation and fibrin clot formation (AR-chip). The T-TAS records occlusion start time, occlusion time, and area under the curve. We evaluated this test in healthy control dogs. We also investigated the effect in vivo of acetylsalicylic acid (ASA), and the effect in vitro of an anticoagulation drug (dalteparin; low-molecular-weight heparin; LMWH). The CV of the AUC of both chips was good (CVs of 6.45% [PL] and 1.57% [AR]). The inhibition of platelet function by ASA was evident in the right-shift in the PL test pressure curve. The right-shift in the AR test pressure curves showed that the administration of LMWH inhibited both platelets and the coagulation cascade. The T-TAS may be useful in the evaluation of canine blood coagulation.