Comparison of coagulation and fibrinolysis in Irish Wolfhounds and age-matched control dogs using tissue plasminogen activator-augmented viscoelastic testing.

OBJECTIVE: To determine if Irish Wolfhounds (IWs), like other sighthounds, are hyperfibrinolytic compared with nonsighthound dogs using 2 native and tissue plasminogen activator (tPA)-enhanced viscoelastic assays, one that is whole blood-based (viscoelastic coagulation monitor [VCM]) and the other that is plasma-based thromboelastography (TEG). DESIGN: Cohort study. SETTING: University teaching hospital. ANIMALS: A convenience sample of 27 IWs recruited from the Irish Wolfhound Association of New England Specialty and the local community, and 27 healthy, age-matched, large-breed control dogs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Blood samples including CBC, biochemistry, traditional coagulation, and viscoelastic testing were collected from IWs and control dogs. Twelve IWs had viscoelastic testing. IWs had lower fibrinogen concentrations (215.5 ± 57.8 vs 251.4 ± 64.5 mg/dL, P = 0.034) and formed weaker clots on both whole-blood VCM and plasma TEG assays (maximum clot firmness [VCM-MCF] = 39.4 [25.1-48.8] vs 48.5 [34.6-57.3], P = 0.0042; maximum amplitude [TEG-MA] = 22.7 [14.7-33.6] vs 32.2 [26.9-42.0], P < 0.0001). IWs were hyperfibrinolytic compared with control dogs on VCM whole-blood assays, with 25 U/mL tPA (lysis at 30 min [VCM-LI30] = 68.1 [0-100] vs\ 99.9 [63.3-100], P = 0.0009; lysis at 45 min [VCM-LI45] = 31.0 [0-100] vs 98.1 [38.4-100], P = 0.0002) but hypofibrinolytic compared with controls on TEG plasma assays with 50 U/mL tPA (lysis at 30 min [TEG-LY30] = 45.7 [4.6-94.6] vs 93.7 [12.3-96.5], P = 0.0004; lysis at 60 min [TEG-LY60] = 68.7 [29.7-96.8] vs 95.7 [34.4-97.6], P = 0.0003). Minimal fibrinolysis was measured on whole-blood VCM or plasma TEG assays without the addition of tPA, and there were no differences between the 2 groups. CONCLUSIONS: Weaker clots were found in IWs than control dogs. With the addition of tPA, IWs had evidence of hyperfibrinolysis on whole-blood VCM assays and hypofibrinolysis on plasma TEG assays compared with control dogs. Without the addition of tPA, however, both groups of dogs showed minimal fibrinolysis on viscoelastic testing.

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.

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.

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.

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.

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.

Safety, biodistribution and viral shedding of oncolytic vaccinia virus TG6002 administered intravenously in healthy beagle dogs.

Oncolytic virotherapy is an emerging strategy that uses replication-competent viruses to kill tumor cells. We have reported the oncolytic effects of TG6002, a recombinant oncolytic vaccinia virus, in preclinical human xenograft models and canine tumor explants. To assess the safety, biodistribution and shedding of TG6002 administered by the intravenous route, we conducted a study in immune-competent healthy dogs. Three dogs each received a single intravenous injection of TG6002 at 105 PFU/kg, 106 PFU/kg or 107 PFU/kg, and one dog received three intravenous injections at 107 PFU/kg. The injections were well tolerated without any clinical, hematological or biochemical adverse events. Viral genomes were only detected in blood at the earliest sampling time point of one-hour post-injection at 107 PFU/kg. Post mortem analyses at day 35 allowed detection of viral DNA in the spleen of the dog which received three injections at 107 PFU/kg. Viral genomes were not detected in the urine, saliva or feces of any dogs. Seven days after the injections, a dose-dependent antibody mediated immune response was identified. In conclusion, intravenous administration of TG6002 shows a good safety profile, supporting the initiation of clinical trials in canine cancer patients as well as further development as a human cancer therapy.

Diagnóstico de cinomose canina por meio de teste imunocromatográfico e sua correlação com achados clínicos e hematológicos no semiárido da Paraíba / Diagnosis of canine distemper by immunochromatographic test and its correlation with clinical and hematological findings in the semiarid region of Paraíba

Objetivou-se através deste trabalho, determinar a prevalência de cinomose canina no semiárido da Paraíba, através de testes rápidos imunocromatográficos, correlacionando-a com os principais achados clínicos e hematológicos. Foram analisadas 67 fichas de animais em que foram realizados testes rápidos para pesquisa de antígeno em amostras nasais e oculares no período de janeiro a dezembro de 2019. Observou-se que 47% (32/67) dos cães analisados foram positivos para cinomose canina. As variáveis que apresentaram diferença estatística significativa (p<0,05) para a infecção foram animais sem raça definida 60% (21/35), animais não vacinados 70% (29/42), e período seco do ano, sendo o mês de agosto (40%; 13/32), com maior ocorrência. Os principais sistemas afetados foram o respiratório 61% (17/28), oftalmológico 70% (22/31), nervoso 69% (13/19), dermatológico 45% (9/20), e gastrintestinal 42% (6/14). As principais alterações hematológicas foram anemia 66% (23/32), leucopenia 76% (19/25) e linfopenia 48% (15/31). Concluiu-se que foi elevada a ocorrência de cinomose canina em animais com suspeita clínica no Semiárido Paraibano, e animais sem raça definida, não vacinados, no período seco do ano foram mais diagnosticados com a enfermidade.

The objective of this study was to determine the prevalence of distemper canine distemper vírus (CDV) infection in the semi-arid region of Paraíba, using rapid immunochromatographic tests, correlating it with the main clinical and hematological findings. 67 records of animals were analyzed in which rapid tests were performed for antigen research in nasal and ocular from January to December 2019. It was observed that 47% (32/67) of compulsory dogs were positive for canine distemper. The variables that defined difference difference (p <0.05) for infection were mixed breed animals 60% (21/35), unvaccinated animals 70% (29/42), and dry period of the year, being the August (40%; 13/32), with greater occurrence. The main affected systems were the respiratory 61% (17/28), ophthalmological 70% (22/31), nervous 69% (13/19), dermatological 45% (9/20), and gastrointestinal 42% (6/14 )) The main changes were hematological, anemia 66% (23/32), leukopenia 76% (19/25) and lymphopenia 48% (15/31). It was concluded that the occurrence of canine distemper in animals with clinical suspicion in the Semiarid Paraibano was high, and non-vaccinated mixed-breed animals in the dry period of the year were more diagnosed with the disease.

Ectodomain shedding by ADAM17 (a disintegrin and metalloproteinase 17) in canine neutrophils.

ADAM17 is a transmembrane protease expressed by most cells in humans and mice that cleaves cell surface substrates primarily in a cis manner, a process referred to as ectodomain shedding. ADAM17 has numerous substrates and plays a broad role in various physiological processes, including as a key regulator of inflammation. At this time, little is known about ADAM17 expression and function in dogs. A well-established ADAM17 substrate is the leukocyte adhesion protein CD62L (L-selectin). We show that a selective inhibitor of ADAM17, but not an inhibitor of its most closely related family member ADAM10, blocks CD62L shedding upon canine neutrophil activation. We also tested several anti-human ADAM17 monoclonal antibodies (mAbs) for staining canine neutrophils. Although most did not recognize canine neutrophils, the mAbs MEDI3622 and D1(A12) did. They also blocked the downregulation of CD62L upon neutrophil activation. MEDI3622 is a human IgG antibody and we found that a canine chimeric version of this mAb also blocked CD62L shedding by canine leukocytes. Taken together, our findings provide the first direct evidence of ADAM17 expression and sheddase activity in dogs, establishing a potential therapeutic target for various inflammatory disorders.

Comparison between hematological and biochemical changes caused by conventional and metronomic chemotherapies in the treatment of canine transmissible venereal tumor / Comparação entre as alterações hematológicas e bioquímicas ocasionadas pela quimioterapia convencional e quimioterapia metronômica no tratamento do tumor venéreo transmissível canino

This study aimed to report the hematological and biochemical changes caused by conventional and metronomic chemotherapies, using vincristine sulfate to treat canine Transmissible Venereal Tumor (TVT). Twelve dogs were selected, six of them for the group receiving conventional chemotherapy (G1), and six dogs for the group receiving metronomic chemotherapy (G2). The G1 group received vincristine sulfate once a week at the dose of 0.75mg/m² until the tumor had disappeared with treatment, and the G2 group was treated with vincristine sulfate three times a week at the dose of 0.25mg/m2 until the tumor had disappeared. Before and after chemotherapy treatment, hematological and biochemical blood tests were performed to evaluate the main alterations caused by both chemotherapeutic models. Dogs undergoing conventional chemotherapy had significant leukocyte changes (p<0.05), causing neutropenia and leukopenia. In dogs undergoing metronomic chemotherapy, leukocytes remained within the reference range. Half of the dogs in group G1 had normochromic, normocytic anemia. The only biochemical alteration observed was the increase of urea in group G2. Thus, metronomic chemotherapy for the treatment of TVT with vincristine sulfate proved to be an excellent method for treatment, with fewer adverse effects, especially in maintaining the leukogram of dogs within normal range and reducing the number of anemia in animals during treatment.(AU)

Esta pesquisa teve como objetivo relatar as alterações hematológicas e bioquímicas causadas pelo tratamento quimioterápico convencional e pela quimioterapia metronômica, utilizando-se sulfato de vincristina para o tratamento do tumor venéreo transmissível canino(TVTC). Foram selecionados 12 cães, sendo seis para o grupo que recebeu quimioterapia convencional (G1) e seis cães para o grupo que recebeu quimioterapia metronômica (G2). O grupo G1 recebeu sulfato de vincristina, uma vez por semana, na dose de 0,75mg/m2, até o desaparecimento do tumor e o grupo G2 foi tratado com sulfato de vincristina, três vezes por semana, na dose de 0,25mg/m2, até o desaparecimento do tumor. Antes e após o tratamento quimioterápico foram realizados exames hematológicos e bioquímicos sanguíneos para avaliação das principais alterações causadas pelos dois modelos quimioterápicos. Os cães submetidos à quimioterapia convencional tiveram alterações leucocitárias significativas (p<0,05), causando uma leucopenia por neutropenia enquanto nos cães, submetidos à quimioterapia metronômica, os leucócitos mantiveram-se dentro do intervalo de referência. A metade dos cães do grupo G1 tiveram uma anemia do tipo normocítica normocrômica. A única alteração bioquímica observada foi o aumento da ureia no grupo G2. Desta forma, a quimioterapia metronômica para o tratamento do TVT com sulfato de vincristina, demonstrou ser um excelente método para a cura do animal, com menores reduções de efeitos adversos, sobretudo, na manutenção do leucograma dos cães e na redução de animais com anemia.(AU)

Characterization of total adenosine deaminase activity (ADA) and its isoenzymes in saliva and serum in health and inflammatory conditions in four different species: an analytical and clinical validation pilot study.

BACKGROUND: Measurement of adenosine deaminase (ADA) can provide information about cell-mediated immunity. This report's objective was to study the enzymatic activity of total ADA (tADA) and its isoenzymes ADA1 and ADA2 in canine, equine, porcine, and bovine serum and saliva and their changes in different inflammatory situations in each species. Besides, an automated method for ADA2 measurement was developed and validated. RESULTS: tADA was present in serum and saliva of healthy animals of the four species. Erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) concentration of 0.47 mM was needed for ADA1 inhibition in canine and porcine samples (serum and saliva) and bovine saliva, whereas for equine saliva 0.94 mM was needed. ADA2 activity was not detected in bovine serum and was very low or absent in equine serum and bovine saliva. An automated procedure to measure ADA2 consisting of adding EHNA to a commercial reagent for tADA measurement provided repetitive (coefficients of variation < 8.8% in serum and < 10% in saliva) and accurate (linearity of serial sample dilutions with R2 > 0.90) results, being equivalent to a manual incubation of the sample with EHNA at a similar concentration. Salivary tADA, as well as ADA1 and ADA2, were higher in dogs with leishmaniosis, horses with acute abdominal disease and pigs with lameness than in healthy animals. tADA and isoenzymes in saliva showed a positive significant correlation with serum ferritin in dogs (r = 0.602, P < 0.01; r = 0.555, P < 0.05; and r = 0.632, P < 0.01; respectively for tADA, ADA1 and ADA2) and serum C-reactive protein in pigs (r = 0.700, P < 0.01, for both tADA and ADA1; r = 0.770, P < 0.001, for ADA2), whereas salivary ADA2 significantly correlated with serum amyloid A in horses (r = 0.649, P < 0.01). In cows, salivary tADA and ADA1 significantly increased after calving, correlating with total white blood cell count (r = 0.487, P < 0.05, for both tADA and ADA1). CONCLUSIONS: The activity of total ADA and its different isoenzymes, can be measured in serum and saliva of dogs, horses, pigs and cows by a simple and fast procedure described in this report. When measured in saliva, these analytes correlated with other biomarkers of inflammation and it could potentially be used as a biomarkers of inflammation and immune activation in the species of this study.

Immune function and serum vitamin D in shelter dogs: A case-control study.

This study sought to establish a baseline understanding of immune function and its association with serum vitamin D in shelter dogs. Ten apparently healthy shelter dogs housed in the Arizona Humane Society for ≥7 days and 10 apparently healthy, age, breed, and sex-matched control dogs were included. Serum 25-hydroxyvitamin D (25[OH]D), the major circulating vitamin D metabolite, was measured using high performance liquid chromatography. Whole blood samples were stimulated with lipopolysaccharide (LPS), lipoteichoic acid, or phosphate buffer solution, and tumor necrosis factor (TNF)-ɑ, interleukin (IL)-6, and IL-10 were measured using a canine-specific multiplex bead-based assay. Phagocytosis of opsonized-Escherichia coli and E. coli-induced oxidative burst were evaluated with flow cytometry. Shelter dogs had decreased percentages of granulocytes and monocytes (GM) that had phagocytized opsonized-E coli (P = 0.019) and performed E. coli-induced oxidative burst (P = 0.011). There were no significant differences in TNF-α, IL-6, IL-10, or 25(OH)D concentrations between shelter and control dogs. Serum 25(OH)D concentrations had a weak positive association with the intensity of GM E. coli-induced oxidative burst (r2 = 0.23, P = 0.03). There was a moderate inverse association between serum 25(OH)D concentration and LPS-stimulated TNF-ɑ production in shelter dogs (r2 = 0.40, P = 0.04). These results demonstrate immune dysregulation in vitro in shelter dogs housed for ≥7 days when compared to age, breed, and sex-matched control dogs. While serum 25(OH)D concentrations did not differ between shelter and control dogs, significant associations between 25(OH)D concentration and immune function parameters in vitro were identified.

Stem Cell Conditioned Medium Treatment for Canine Spinal Cord Injury: Pilot Feasibility Study.

Spinal cord injury (SCI) involves nerve damage and often leads to motor, sensory and autonomic dysfunctions. In the present study, we have designed a clinical protocol to assess the feasibility of systemic delivery of allogenic canine bone marrow tissue-derived mesenchymal stem cell conditioned medium (BMMSC CM) to dogs with SCI. Four client-owned dogs with chronic SCI lasting more than six months underwent neurological and clinical evaluation, MRI imaging and blood tests before being enrolled in this study. All dogs received four intravenous infusions with canine allogenic BMMSC CM within one month. Between the infusions the dogs received comprehensive physiotherapy, which continued for three additional months. No adverse effects or complications were observed during the one, three and six months follow-up periods. Neither blood chemistry panel nor hematology profile showed any significant changes. All dogs were clinically improved as assessed using Olby locomotor scales after one, three and six months of BMMSC CM treatment. Furthermore, goniometric measurements revealed partial improvement in the range of joint motion. Bladder function improved in two disabled dogs. We conclude that multiple delivery of allogenic cell-derived conditioned medium to dogs with chronic SCI is feasible, and it might be clinically beneficial in combination with physiotherapy.

Efficacy of specific antivenom and fresh frozen plasma in reversing Daboia palaestinae venom-induced hemostatic derangement.

OBJECTIVES: To evaluate the in vitro efficacy of specific Daboia (Vipera) palaestinae (Dp) antivenom or fresh frozen plasma (FFP) against Dp venom-induced hemostatic changes DESIGN: In vitro study. SETTING: Laboratory of a university referral hospital. ANIMALS: Five healthy dogs. INTERVENTIONS: Rotational thromboelastometry (including recombinant tissue factor or kaolin activators [EXTEM and INTEM, respectively]) and conventional hemostatic tests were evaluated in citrated whole blood samples that underwent 4 treatments: (1) no additives (control); (2) 15 µg of Dp venom; (3) 15 µg of Dp venom and 10 µL of specific Dp antivenom; (4) 15 µg of Dp venom and 0.3 mL of FFP. A linear mixed-effects regression model was used to compare results between each treatment and the control. MEASUREMENTS AND MAIN RESULTS: Dp-venom engendered statistically significant (P < 0.05) EXTEM changes in 8/17 variables, all indicative of hypercoagulability, which were negated by antivenom but not with FFP. Similarly, Dp-venom induced hypercoagulable, hyperfibrinolytic changes in 12 of 17 INTEM variables, of which only 5 of 12 were negated by antivenom but not with FFP. Fibrinogen concentration was decreased, and the activated partial thromboplastin time (aPTT) was shortened (P < 0.05 for both) in all treatments compared to the control. CONCLUSIONS: This study demonstrated the ephemeral procoagulant phase of Dp envenomation for the first time. Many venom-induced thromboelastometric changes were reversed by specific antivenom but not with FFP. Prospective clinical studies are warranted to investigate whether the present findings translate to clinical efficacy.

Evaluation of hematologic, biochemical, and blood gas variables in stored canine packed red blood cells, and the impact of storage time on blood recipients.

BACKGROUND: Canine packed red blood cells (pRBCs) can be stored under refrigeration for several days; however, cellular metabolism remains active inside the units, thus producing substances that affect their quality. OBJECTIVES: We aimed to evaluate hematologic, biochemical, and blood gas variable alterations that occur in canine pRBCs during storage, and their effects on recipient clinicopathologic parameters. METHODS: The study was conducted in two phases. In phase I, 15 pRBC units containing CPDA-1 were stored for 28 days; samples were collected weekly from the units of days 0 to 28 to measure the packed cell volume (PCV), pH, partial pressure carbon dioxide (PCO2 ), partial pressure oxygen (PO2 ), concentrations of lactate and potassium, and the percent hemolysis. In phase II, another 22 canine pRBC units stored for different time periods (maximum of 21 days) were transfused, and the recipients were evaluated before and after transfusion for changes in clinical parameters (heart rate, respiratory rate, systolic arterial pressure, and rectal temperature) and hematologic variables (PCV, lactate and potassium concentrations, pH, PCO2 , the ratio of arterial oxygen partial pressure to fractional inspired oxygen [PO2 /FiO2 ] ratio, oxygen saturation [SaO2 ], base excess, and bicarbonate [HCO3 ]). RESULTS: In the pRBC units, the PCV increased from 70% to 78.33%, the lactate concentration increased 627%, the potassium concentration increased 183%, the percent hemolysis reached 0.69%, and the pH decreased 9% after 28 days. However, the dogs who received transfusions were not negatively affected. There was a significant increase in PCVs, and a significant decrease in heart rates. CONCLUSION: Canine pRBCs undergo hematologic, blood gas, and biochemical alterations during storage; however, the transfusion of pRBCs stored for up to 21 days increased PCVs without causing harm to the dogs.

Effect of 7.5% hypertonic saline solution on whole blood coagulation in healthy dogs using thromboelastography.

OBJECTIVE: To evaluate the effects of 7.5% hypertonic saline solution (HSS) on whole blood coagulation in healthy dogs and to compare electrolyte and osmolality measurements between in vivo and in vitro dilution with HSS. DESIGN: Experimental study. SETTING: University teaching hospital. ANIMALS: Twelve adult purpose-bred Beagles. INTERVENTIONS: All 12 dogs received 5 mL/kg 7.5% HSS at 1 mL/kg/min. After a 14-day washout period, 5 of these dogs were randomly selected and received the same volume of 0.9% NaCl. Blood samples were collected before infusion, immediately after infusion, and at 30, 60, and 90 minutes after infusion for the measurement of coagulation using thromboelastography. For comparison of electrolyte concentrations and osmolality between in vitro dilution and in vivo dilution of HSS, 6-mL blood samples were diluted with 7.5% HSS (1:18 ratio) at baseline. MEASUREMENTS AND MAIN RESULTS: None of the thromboelastography variables differed significantly between the 7.5% HSS group and the 0.9% NaCl group. The sodium and chloride levels, and the osmolality, were significantly increased at all postinfusion time points compared to baseline, while those levels were significantly higher with in vitro dilution than all postinfusion time points. However, almost all the values gradually decreased and became similar to baseline values in case of in vivo dilution. CONCLUSIONS: The clinically relevant dose of 7.5% HSS (5 mL/kg) did not affect whole blood coagulation significantly in healthy Beagles. Further studies are necessary to assess the effect of HSS on blood coagulation in canine patients with shock.

Pharmacokinetics of multivesicular liposomal encapsulated cytarabine when administered subcutaneously in dogs.

BACKGROUND: Prolonged cytotoxic concentrations of cytarabine (CA) are required for maximum cytotoxicity. DepoCyt is a human liposomal cytarabine (LC) product that lasts longer in plasma and CSF compared with free CA (FC). The use of LC has not been evaluated in dogs. OBJECTIVES: To perform a LC pharmacokinetic (PK) study when administered SC in dogs. ANIMALS: Five healthy female beagles. METHODS: Three-period, 3-treatment, nonblinded, randomized, and crossover design, including a pilot study. LC was administered at 50 mg/m2 SC and FC was administered at 25 and 50 mg/m2 SC and IV. Plasma CA concentrations were measured until 240, 72, and 8 hours after SC LC, SC FC, and IV FC administration, respectively. CA plasma concentrations were quantitated by ultra-high-performance liquid chromatography with mass spectrometry (MS/MS) detection and concentration-time profiles were evaluated by noncompartmental analysis. RESULTS: Subcutaneous LC administration resulted in a maximum plasma concentration of 26.3 to 59.78 ng/mL, time to reach maximum plasma concentration of 2 hours, area under the concentration-time curve to last measurable concentration of 669.3 to 1126 h × ng/mL, and plasma bioavailability (%F) of 19.6% to 31.3%. The PK profiles of FC after SC and IV administration differed when compared with LC. CONCLUSIONS AND CLINICAL IMPORTANCE: In healthy dogs, SC LC administration at 50 mg/m2 results in measurable plasma CA concentrations, is apparently safe and well tolerated, but does not result in prolonged cytotoxic plasma concentrations. Poor absorption of LC prevented establishment of a complete LC PK profile.