Operator impact assessment on qualitative and quantitative parameters of canine platelet-rich plasma.

Platelet-rich plasma (PRP) has emerged as a cornerstone in veterinary regenerative medicine. The present study evaluated the impact of the operator on the qualitative and quantitative features of non-activated PRP derived from canine whole blood. Blood was collected in anticoagulant acid citrate dextrose from twelve healthy adult dogs and PRP was prepared according to the double-spin method. Both operators followed an identical protocol and utilized the same equipment for PRP preparation from the pooled blood samples. The resulting PRP underwent characterization, classification and coding based on minimum reporting standards. The consistency and internal reliability of different parameters were also assessed using the intraclass correlation coefficient and Cronbach's alpha values. Variables such as white blood cell (WBC) concentration, relative WBC composition and mean platelet volume (MPV) showed poor reliability, and WBC concentration and MPV also had unacceptable internal consistency. Significant differences were observed in several qualitative and quantitative parameters of the prepared PRP, highlighting the influence of the operator even when the same protocol and equipment were used. Our study has direct implications to regenerative medicine, reinforcing the urgency to set minimum requirements for reporting PRP in research studies.

Development and validation of an ELISA to measure regenerating island-derived protein 3E in canine blood.

BACKGROUND: Regenerating island-derived proteins (REG) are upregulated in people with sepsis, pancreatitis, and gastrointestinal diseases. One member of the REG family, namely REG3E, was recently identified in pancreatic tissue and plasma of dogs, with high expression in pancreatitis and sepsis. OBJECTIVES: We aimed to develop and validate an ELISA to measure REG3E concentrations in canine blood. METHODS: An indirect sandwich ELISA was developed using recombinant canine REG3E protein and polyclonal anti-canine REG3E antibodies raised in guinea pigs and rabbits. Antibody specificity was assessed using western blot and mass spectrometric analysis of protein purified from canine plasma. Assay validation included evaluation of dilutional linearity, parallelism, spiking recovery, repeatability and reproducibility, stability, interferences, and comparison of serum and heparinized plasma. RESULTS: Antibodies bound specifically to REG3E with no evidence of cross-reactivity with other proteins. The limit of detection of the ELISA was 15 ng/mL, and the lower limit of quantification was 30 ng/mL. The assay demonstrated good to excellent linearity, dilutional and mixing parallelism, and recovery, with mean observed-to-expected ratios of 104%, 107%, 102%, and 92%, respectively, and no evidence of a hook effect. Coefficients of variation were ≤8.5% for repeatability and ≤14.3% for reproducibility at three different levels. Measurements of REG3E in plasma were not significantly influenced by different storage conditions, freeze-thawing cycles, hemolysis, lipemia, or icterus. There was no significant difference between REG3E concentrations in heparinized plasma and serum samples. CONCLUSIONS: The canine REG3E ELISA has acceptable precision, accuracy, linearity, and reproducibility for the measurement of REG3E in canine plasma and serum.

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.

A study on the effects of vitamin D supplementation on hematological parameters and serum 25-hydroxy vitamin D in healthy dogs.

BACKGROUND: Limited studies are available on vitamin D supplementation in dogs. This study evaluates the effect of a commercial vitamin D3 supplement on serum 25-hydroxy vitamin D as well as selected biochemical and hematological parameters in healthy dogs. Eight intact male adult dogs with a mean body weight of 20 kg from mixed breeds were included in the study. After adaptation period, dogs received vitamin D3 supplement at the dose of 50 IU/kg body weight per day. Blood samples were collected on days 0, 14, 28 and 42 of supplementation. Food was used for analysis of vitamin D3 content. RESULTS: Significant increase in serum level of 25-hydroxy vitamin D3 was detected since day 14 of supplementation. Changes in serum 25-hydroxy vitamin D3 concentration during time showed an upward significance (p < 0.05). Vitamin D3 content of the food was 2900 IU/kg dry matter. Changes in serum phosphorus levels were upward significant. No dog showed calcium or phosphorus levels above the highest reference level. Liver and kidney parameters remained in the reference range during the experiment. A gradual significant increase was observed in hemoglobin and hematocrit which was started from day 14. Vitamin D3 supplementation had no significant effect on neutrophils, monocytes and lymphocytes percent during the study. CONCLUSIONS: Vitamin D3 supplementation at 50 IU/kg BW daily, increases serum levels of 25-hydroxy vitamin D in healthy dogs fed with a diet containing proper amount of this vitamin. It also increases hemoglobin and hematocrit levels in a time dependent manner without inducing adverse effects.

Oral cinacalcet administration decreases serum ionized calcium and parathyroid hormone concentrations in healthy dogs.

Cinacalcet is an oral calcimimetic that has potential to non-invasively treat primary hyperparathyroidism in dogs (Canis lupis familiaris). There is minimal data assessing its efficacy in dogs. This study aimed to determine whether a single dose of cinacalcet decreases serum ionized calcium (iCa), total calcium (tCa), and parathyroid hormone (PTH) concentrations. Twelve dogs received a median dose of 0.49 mg/kg (range 0.30-0.69 mg/kg) cinacalcet per os. Venous blood samples were collected at time 0 (before cinacalcet administration), 3, 8, and 24 h following cinacalcet administration. PTH, iCa, and tCa concentrations were measured at each time point and compared to 0 hour concentrations. A significant (50%) decrease in serum PTH occurred at 3 h with a median PTH of 4.6 pmol/L (range 2.7-10.8) at baseline and 1.65 pmol/L (range 0.5-14.7) at 3 h; p = .005. A significant, but not clinically relevant, decrease in serum iCa from a median baseline of 1.340 mmol/L (range 1.32-1.41) to a 3 h median of 1.325 mmol/L (range 1.26-1.39), p = .043, was also observed. tCa concentrations were not different. This study showed that a single dose of cinacalcet leads to transient decreases in iCa and PTH concentrations in healthy dogs.

Evaluation of taurine and carnitine concentrations in whole blood, plasma, skeletal muscle and cardiac muscle in dogs.

Little is known about how plasma and whole blood taurine and plasma carnitine correlate to concentrations in skeletal and cardiac muscle and the effects of diet in dogs. The purpose of this study was to evaluate the correlation among plasma, skeletal and cardiac muscle carnitine and taurine and whole blood taurine and determine the effect of diet. The study protocol was approved by the Pet Food Solutions Institutional Animal Care and Use Committee. Thirty-three mixed-breed hounds and 32 beagles were evaluated at Day 0 then removed from their baseline diet and randomized to a test diet: high animal protein, grain-inclusive (HA-GI), low animal protein, grain-free (LA-GF), low animal protein, grain-inclusive (LA-GI), or high animal protein, grain-free (HA-GF). Blood was drawn every 30 days and endomyocardial (mixed breeds only) and skeletal muscle biopsies were collected at Days 0 and 180. The correlations between plasma and whole blood taurine, or plasma carnitine and skeletal and cardiac muscle concentrations were weak (p < 0.01-0.05). Mixed-breed hounds had increased (p = 0.029) whole blood taurine compared to beagles. Plasma taurine was lower with diet HA-GF, (p = 0.009) however, all diets had increased taurine from Day 0 and were, on average within the laboratory reference range. Dogs fed the HA-GI diet had increased cardiac muscle carnitine esters (p = 0.014). Increased carnitine esters were also appreciated in cardiac muscle in all diets from Day 0 to 180 (p = 0.0001). On Day 180 mixed-breed hounds had increased skeletal total carnitine (p < 0.001) compared to all time points and breeds. This study observed no correlation between plasma, whole blood, skeletal and cardiac muscle taurine concentrations but noted some effects between time, breed and diet.

Association of four differently processed diets with plasma and urine advanced glycation end products and serum soluble receptor for advanced glycation end products concentration in healthy dogs.

Advanced glycation end products (AGEs), formed via the Maillard reaction (MR) during processing of foods, have been implicated in inflammatory and degenerative diseases in human beings. Cellular damage is primarily caused by AGE binding with the receptor for AGEs (RAGE) on cell membranes. An isoform of RAGE, soluble RAGE (sRAGE), acts as a decoy receptor binding circulating AGEs preventing cellular activation. Pet food manufacturing involves processing methods similar to human food processing that may increase dietary AGEs (dAGEs). We hypothesized that diet, plasma and urine AGEs, and serum sRAGE concentrations would differ between thermally processed diets. This study examined the association of four differently processed diets: ultra-processed canned wet food (WF); ultra-processed dry food (DF); moderately processed air-dried food (ADF) and minimally processed mildly cooked food (MF) on total plasma levels of the AGEs, carboxymethyllysine (CML), carboxyethyllysine (CEL), methylglyoxal hydroimidazolone-1, glyoxal hydroimidazolone-1, argpyrimidine, urine CML, CEL and lysinoalanine, and serum sRAGE concentration. Ultra-high-performance liquid chromatography-tandem mass spectrometry was used to measure AGEs. sRAGE concentration was measured using a commercial canine-specific enzyme-linked immunosorbent assay kit. Total dAGEs (mg/100 kcal as fed) were higher in WF than in other diets. Plasma total AGEs (nM/50 µL) were significantly higher with WF, with no difference found between DF, ADF, and MF; however, ADF was significantly higher than MF. Urine CML (nmol AGEs/mmol creatinine) was significantly higher with DF than with WF and MF. There were no significant differences in total urine AGEs or serum sRAGE concentration between diets. In conclusion, different methods of processing pet foods are associated with varied quantities of AGEs influencing total plasma AGE concentration in healthy dogs. Serum sRAGE concentration did not vary across diets but differences in total AGE/sRAGE ratio were observed between MF and WF and, ADF and DF.

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.

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.

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.

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.