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1.
J Vet Diagn Invest ; 33(2): 357-361, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33559534

RESUMO

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.


Assuntos
Anticoagulantes/farmacologia , Aspirina/farmacologia , Testes de Coagulação Sanguínea/veterinária , Coagulação Sanguínea , Dalteparina/farmacologia , Cães/sangue , Técnicas Analíticas Microfluídicas/veterinária , Trombose/veterinária , Animais , Testes de Coagulação Sanguínea/instrumentação , Feminino , Masculino , Técnicas Analíticas Microfluídicas/métodos , Trombose/diagnóstico
2.
Vet J ; 267: 105578, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33375957

RESUMO

Hypervolemia can damage the endothelial glycocalyx, a key regulator of vascular permeability, coagulation and inflammation. A starting peri-operative crystalloid fluid rate of 5mL/kg/h is recommended for healthy dogs undergoing elective procedures but higher rates continue to be commonly used. This study aimed to determine if a higher starting perioperative fluid rate was associated with a greater increase in plasma concentrations of hyaluronan, a marker correlated with glycocalyx damage, in systemically healthy dogs undergoing elective surgical procedures. Based on a sample size calculation, 38 dogs undergoing ovariohysterectomy or castration were randomly assigned to receive lactated Ringer's at a starting perioperative fluid rate of 10mL/kg/h (n=19) or 5mL/kg/h (n=19). Plasma hyaluronan concentrations were measured by ELISA in pre- and post-fluid therapy samples. There were no significant differences between groups in hyaluronan values before (baseline, P=0.52) or after perioperative fluid administration (P=0.62). Compared to respective baseline values, hyaluronan values significantly increased following 5 and 10ml/kg/h fluid administration (P=0.02 for both comparisons). This preliminary study identified an increase in hyaluronan over the course of fluid therapy with both the low and high fluid rate. One possible explanation is that both fluid rates contribute to glycocalyx disruption, but it should be emphasized that hyaluronan is not specific to the glycocalyx. Further studies are needed to determine the origin of the increased circulating hyaluronan and its clinical significance in dogs undergoing elective surgical procedures.


Assuntos
Soluções Cristaloides/administração & dosagem , Cães/sangue , Procedimentos Cirúrgicos Eletivos/veterinária , Hidratação/veterinária , Ácido Hialurônico/sangue , Projetos Piloto , Anestesia/métodos , Anestesia/veterinária , Animais , Castração/métodos , Castração/veterinária , Procedimentos Cirúrgicos Eletivos/métodos , Feminino , Hidratação/métodos , Histerectomia/métodos , Histerectomia/veterinária , Masculino , Ovariectomia/métodos , Ovariectomia/veterinária
3.
Acta Vet Scand ; 62(1): 30, 2020 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-32546243

RESUMO

BACKGROUND: Exposure to anticoagulant rodenticides (ARs) in dogs is among the most common causes of poisoning in small animal practice, but information about toxicokinetic of these rodenticides in dogs is lacking. We analysed blood and faeces from five accidentally exposed dogs and 110 healthy dogs by reversed phase ultra-high performance liquid chromatography-tandem mass spectrometry. The aim of the study was to estimate elimination of brodifacoum, bromadiolone and difenacoum after acute exposure, calculate the half-lives of these rodenticides in dogs, estimate faecal elimination in a litter of puppies born, and further to identify the extent of AR exposure in a healthy dog population. RESULTS: Three dogs were included after single ingestions of brodifacoum; two dogs ingested bromadiolone and one dog ingested difenacoum. Maximum concentrations in faeces were found after day 2-3 for all ARs. The distribution half-lives were 1-10 days for brodifacoum, 1-2 days for bromadiolone and 10 days for difenacoum. Brodifacoum and difenacoum had estimated terminal half-lives of 200-330 days and 190 days, respectively. In contrast, bromadiolone had an estimated terminal half-life of 30 days. No clinical signs of poisoning or coagulopathy were observed in terminal elimination period. In blood, the terminal half-life of brodifacoum was estimated to 8 days. Faeces from a litter of puppies born from one of the poisoned dogs were examined, and measurable concentrations of brodifacoum were detected in all samples for at least 28 days after parturition. A cross-sectional study of 110 healthy domestic dogs was performed to estimate ARs exposure in a dog population. Difenacoum was detected in faeces of one dog. Blood and faecal samples from the remaining dogs were negative for all ARs. CONCLUSIONS: Based on the limited pharmacokinetic data from these dogs, our results suggest that ARs have a biphasic elimination in faeces using a two-compartment elimination kinetics model. We have shown that faecal analysis is suitable and reliable for the assessment of ARs exposure in dogs and a tool for estimating the AR half-lives. Half-lives of ARs could be a valuable indicator in the exposed dogs and provides important information for veterinarians monitoring AR exposure and assessment of treatment length in dogs.


Assuntos
Anticoagulantes/farmacocinética , Cães/metabolismo , Rodenticidas/farmacocinética , 4-Hidroxicumarinas/sangue , 4-Hidroxicumarinas/metabolismo , 4-Hidroxicumarinas/farmacocinética , Animais , Anticoagulantes/sangue , Anticoagulantes/metabolismo , Cromatografia Líquida de Alta Pressão/veterinária , Cães/sangue , Fezes/química , Espectrometria de Massas/veterinária , Rodenticidas/sangue , Rodenticidas/metabolismo
4.
Top Companion Anim Med ; 39: 100408, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32482288

RESUMO

Rapid activation of the hypothalamic pituitary adrenal axis and the sympathetic nervous system are hallmarks of the acute stress response and these systems interact with the immune system by signaling though glucocorticoid and adrenergic receptors on immune cells. There is limited information about the effect of these physiologic responses on immunologic parameters of pet dogs enrolled in clinical studies. The objective of this study was to evaluate how travel, instrumentation, and hospitalization alter immunologic parameters in pet dogs. Blood was collected from healthy dogs in a home environment and from healthy dogs at the time of presentation to the hospital and after instrumentation and 24 hours of hospitalization. We found that lipopolysaccharide (LPS)-induced downregulation of toll like receptor 4 (TLR4) was blunted in dogs exposed to stress. Neutrophil and monocyte major histocompatibility complex class II (MHCII) expression increased after transportation to the veterinary hospital but then became similar to that of the control dogs at the end of hospitalization. Peripheral blood mononuclear cell cytotoxicity function was blunted in dogs exposed to the stress of transportation as well as hospitalization. Neutrophil apoptosis was greater in dogs exposed to stress compared to controls although this effect significantly decreased after hospitalization stress. Conversely, stress did not alter induced or spontaneous cytokine production from leukocytes, neutrophil or monocyte expression of TLR4, LPS-induced downregulation of monocyte TLR4, LPS-induced neutrophil and monocyte expression of MHCII or peripheral blood lymphocyte phenotype. Transportation and instrumentation/hospitalization stress should be considered when interpreting immunologic studies in pet dogs.


Assuntos
Cães/imunologia , Exame Físico/veterinária , Estresse Fisiológico , Transportes , Animais , Cães/sangue , Feminino , Linfócitos/metabolismo , Masculino , Neutrófilos/metabolismo
5.
Braz. J. Vet. Res. Anim. Sci. (Online) ; 57(2): [e161756], mai. 2020. tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1097353

RESUMO

An Andean fox was transferred to the Wildlife Hospital of the Universidad San Francisco de Quito for evaluation of injuries caused by a run over. Clinical signs of hypovolemic shock were detected. Radiographies showed multiple pelvic fractures and free fluid in retroperitoneal cavity. The presumptive diagnosis was hemorrhagic shock caused by blood loss secondary to a pelvis fracture. An emergency xenotransfusion using blood from a domestic dog was performed without acute transfusion reactions observed. This is the first report of successful xenotransfusion between a domestic dog and an Andean fox presenting a procedure that could be applied in emergency situations.(AU)


Uma raposa andina foi levada ao Hospital da Vida Selvagem da Universidad San Francisco de Quito para avaliar os ferimentos causados por um atropelamento. Sinais clínicos de choque hipovolêmico foram detectados. Radiografias mostraram múltiplas fraturas pélvicas e fluido livre na cavidade retroperitoneal. O diagnóstico presuntivo foi um choque hemorrágico causado por perda sanguínea secundária a uma fratura pélvica. Uma xenotransfusão de emergência foi realizada com o sangue de um cão doméstico sem reações agudas transfusionais. Este é o primeiro relato bem sucedido de xenotransfusão entre um cão doméstico e uma raposa andina, demonstrando que é um procedimento que poderá ser considerado em situações de emergência.(AU)


Assuntos
Animais , Cães , Cães/sangue , Raposas/sangue , Choque , Transplante Heterólogo , Transfusão de Sangue/veterinária
6.
Vet Surg ; 49(5): 989-996, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32166777

RESUMO

OBJECTIVE: To determine the ability of cell salvage washing and leukoreduction filtration to remove bacterial contamination from canine whole blood. STUDY DESIGN: Ex vivo nested cohort study. SAMPLE POPULATION: Commercially purchased fresh canine whole blood (n = 33 units). METHODS: Commercially obtained canine whole blood was inoculated with known concentrations of one of three species of bacteria, Escherichia coli (ATCC 25922), Staphylococcus pseudintermedius (quality control strain; Texas A&M University), or Pseudomonas aeruginosa (ATCC 27853). Negative controls were inoculated with sterile saline. The inoculated blood was processed through a cell salvage system and filtered through a series of two leukocyte reduction filters. Samples were aseptically collected at five points during processing (inoculum, prewash, postwash, post-first filtration, and post-second filtration) for bacterial enumeration. RESULTS: Bacterial concentrations were reduced by 85.2%, 91.5%, and 93.9% for E coli, S pseudintermedius, and P aeruginosa, respectively, after washing (P < .0001), and bacterial concentrations were reduced by 99.9%, 100%, and 100%, respectively, after the first filtration (P < .0001). After the second filtration, none of the three species of bacteria could be isolated (100% reduction). No bacterial growth was obtained from negative controls throughout the study. The type of bacteria (P = .29) did not allow prediction of bacterial reduction. CONCLUSION: Cell salvage washing combined with leukoreduction filtration eliminated bacterial contamination of whole dog blood (P < .0001). CLINICAL SIGNIFICANCE: Cell salvage washing and leukoreduction filtration could be applied to intraoperative autotransfusion in clinical animals, especially those treated for trauma or hemorrhage with concurrent bacterial contamination.


Assuntos
Sangue/microbiologia , Cães/sangue , Procedimentos de Redução de Leucócitos/veterinária , Animais , Transfusão de Sangue Autóloga , Estudos de Coortes , Escherichia coli , Filtração/veterinária , Leucócitos
7.
J Sports Med Phys Fitness ; 60(6): 942-946, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32141275

RESUMO

BACKGROUND: This study aimed to describe a test protocol for the determination of the maximal lactate steady state (MLSS) in dogs. METHODS: Ten animals were submitted to a variable number of sessions (3 to 5), with a duration of 30 minutes each. Every session started with a warm-up period of 10 minutes at 7 km/h, at a 0% inclination. The treadmill was then set at a 5% inclination and at a constant speed, maintained for 20 m. In subsequent sessions, speed was increased or decreased 0.5 km/h. Blood samples were collected, and blood lactate (BL), heart rate (HR), rectal temperature (RT) and glycaemia were determined at rest (T0), after the warm-up (T1) and every 5 minutes until the end of the test (T2-T5). RESULTS: Significant variations were observed in BL, HR and RT, when comparing values at rest and T1 with those at the MLSS level (P<0.01). Mean speed at MLSS was 14.6±1.47 km/h and BL was 2.7±0.45 mmol/L, a value below 4 mmol/L. A correlation between BL at MLSS and the visual lactate threshold was observed (0.87). A strong agreement between blood lactate levels at the MLSS level and LTv was also observed. CONCLUSIONS: The present results consist, to the authors' knowledge, in the first description of a protocol to determine the MLSS in dogs. It provides information for exercise monitoring and training planning.


Assuntos
Cães/sangue , Teste de Esforço/métodos , Teste de Esforço/veterinária , Ácido Láctico/sangue , Animais , Cães/fisiologia , Feminino , Frequência Cardíaca , Masculino , Condicionamento Físico Animal
8.
J Vet Emerg Crit Care (San Antonio) ; 30(3): 272-278, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32100470

RESUMO

OBJECTIVE: To evaluate the performances of a manual Nageotte hemocytometer method and commercial fluorescent bead-based flow cytometric assay for quantifying [rWBC] in leukoreduced canine packed red blood cell (pRBC) units. DESIGN: Prospective study. Five, commercially purchased, double leukoreduced canine pRBC units were spiked with canine leukocytes to create 6 pRBC standards with the following [rWBC]: < 0.1, 0.375, 1.5, 3.0, 6.0, and 24.0 WBC/µL. [rWBC] of each pRBC standard was measured with the Nageotte hemocytometer and flow cytometric techniques. Limit of detection (LoD), linearity, and bias were determined for each method. For each standard, accuracy and precision were calculated; the cumulative accuracy and mean precision for measurements between the LoD and 24.0 WBC/µL were also determined. SETTING: University veterinary blood bank and clinical pathology laboratory. MEASUREMENTS AND MAIN RESULTS: The Nageotte hemocytometer method had an LoD = 1.48 WBC/µL, inadequate linearity (R2  = 0.92), and a significant negative proportional bias (slope best-fit line = 0.52 ± 0.03). Between [rWBC] 1.5-24 WBC/µL, the technique demonstrated poor cumulative accuracy (6.7%) but acceptable mean precision (17.3%). Relative to a 2 rWBC/µL threshold, at 1.5 WBC/µL the method was inaccurate (6.7%) with acceptable precision (16.6%). The flow cytometric assay had an LoD = 1.3 WBC/µL, acceptable linearity (R2  = 0.99), and a mild positive proportional bias (slope best-fit line = 1.11 ± 0.01). The technique had acceptable cumulative accuracy (80%) and mean precision (10.7%) for measuring [rWBC] between 1.5 and 24 WBC/µL. At 1.5 WBC/µL, this method was acceptably accurate (86.7%) and precise (16.0%). CONCLUSIONS: The flow cytometric assay demonstrated acceptable performance for quantification of [rWBC] in leukoreduced canine pRBC units. The Nageotte hemocytometer method should be used cautiously due to poor accuracy and significant negative bias.


Assuntos
Cães/sangue , Eritrócitos , Citometria de Fluxo/veterinária , Contagem de Leucócitos/instrumentação , Procedimentos de Redução de Leucócitos/veterinária , Leucócitos , Animais , Citometria de Fluxo/métodos , Humanos , Contagem de Leucócitos/métodos , Procedimentos de Redução de Leucócitos/métodos , Estudos Prospectivos
9.
J Vet Emerg Crit Care (San Antonio) ; 30(3): 255-263, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32077234

RESUMO

OBJECTIVE: To assess the in vitro effects of crystalloid and colloid IV fluids on the thromboelastographic (TEG) variables of canine whole blood. DESIGN: In vitro experimental study. SETTING: Veterinary teaching hospital. ANIMALS: Twenty-two healthy dogs. INTERVENTION: Citrated whole blood samples collected from healthy dogs were diluted with 3.4% hypertonic saline (HTS 3.4), 7% hypertonic saline (HTS 7), and 20% mannitol at 8% and 16% dilutions; hydroxyethyl starch 130/0.4 (HES 130/0.4) at 16% dilution; lactated Ringer's solution (LRS) at 16%, 33%, and 66% dilutions; and HTS 7-HES 130/0.4 at 25% and 50% dilutions. Kaolin-activated TEG analysis was concurrently performed on diluted and control (undiluted) samples. MEASUREMENTS AND MAIN RESULTS: Dilution of canine whole blood with LRS compared to control reduced α angle and MA at both 33% (P = 0.009 and P = 0.011, respectively) and 66% dilution (P < 0.001 and P < 0.001, respectively), and prolonged K time at 66% dilution (P = 0.003). At 16% dilution, HTS 3.4, prolonged R time (P = 0.007), while mannitol, a fluid iso osmolar to HTS 3.4, prolonged K time (P = 0.006), reduced α angle (P < 0.001), MA (P = 0.046), and LY60 (P = 0.015). At 8% dilution, HTS 7, a fluid of high osmolarity and tonicity, prolonged R time (P = 0.009) and reduced MA (P = 0.015), while all measured TEG variables were altered at the 16% dilution (P < 0.01 for all variables). HES 130/0.4 reduced α angle (P = 0.031) and MA (P = 0.001) and increased LY60 (P < 0.001) at 16% dilution. Comparing different fluid types, HES 130/0.4 and HTS 3.4 had no to minor, mannitol intermediate, and HTS 7 profound effects on TEG variables (P < 0.05) when compared to LRS at the same dilution. CONCLUSIONS: In vitro dilution of canine whole blood with commonly used IV fluids leads to thromboelastographic changes consistent with hypocoagulability in a dose dependent manner for all fluid types tested. Viscoelastic changes are also influenced by fluid characteristics, specifically tonicity and osmolarity.


Assuntos
Cães/sangue , Derivados de Hidroxietil Amido/farmacologia , Manitol/farmacologia , Substitutos do Plasma/farmacologia , Lactato de Ringer/farmacologia , Solução Salina Hipertônica/farmacologia , Animais , Coagulação Sanguínea/efeitos dos fármacos , Masculino , Tromboelastografia/veterinária
10.
J Anim Sci ; 98(3)2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-32047902

RESUMO

Pet foods may be formulated with decreased starch to meet consumer demands for less processed diets. Fats and oils may be added to low-starch diets to meet energy requirements, but little is known about its effects on canine health. The study objective was to evaluate the effects of feeding healthy adult dogs low carbohydrate, high-fat diets on apparent total tract digestibility, fecal characteristics, and overall health status. Eight adult Beagles were enrolled in a replicated 4 × 4 Latin Square design feeding trial. Dogs were randomly assigned to one of four dietary fat level treatments (T) within each period: 32% (T1), 37% (T2), 42% (T3), and 47% (T4) fat on a dry matter basis. Fat levels were adjusted with the inclusion of canola oil added to a commercial diet. Each dog was fed to exceed its energy requirement based on NRC (2006). Blood samples were analyzed for complete blood counts, chemistry profiles, and canine pancreatic lipase immunoreactivity levels. Apparent total tract digestibility improved (P < 0.05) as the fat level increased for dry matter, organic matter, fat, and gross energy. Fecal output decreased as levels of fat increased in the diet (P = 0.002). There was no effect of fat level on stool quality or short-chain fatty acid and ammonia concentrations in fecal samples (P ≥ 0.20). Blood urea nitrogen levels decreased with increased fat level (P = 0.035). No significant differences were seen in canine pancreatic lipase immunoreactivity (P = 0.110). All blood parameters remained within normal reference intervals. In summary, increased dietary fat improved apparent total tract digestibility, did not alter fecal characteristics, and maintained the health status of all dogs.


Assuntos
Ração Animal/análise , Dieta Hiperlipídica/veterinária , Gorduras na Dieta/farmacologia , Cães/fisiologia , Amônia/análise , Animais , Nitrogênio da Ureia Sanguínea , Dieta/veterinária , Digestão/efeitos dos fármacos , Cães/sangue , Ácidos Graxos Voláteis/análise , Fezes/química , Feminino , Trato Gastrointestinal/efeitos dos fármacos , Nível de Saúde
11.
BMC Vet Res ; 16(1): 25, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32000762

RESUMO

BACKGROUND: Platelets undergo structural, biochemical and functional alterations when stored, and platelet storage lesions reduce platelet function and half-life after transfusion. The objective of this study was to evaluate stored canine platelet concentrates with platelet aggregation, flow cytometry and biochemistry assays. Twenty-two bags of canine platelet concentrates were obtained by the platelet-rich plasma method and were assessed on days 1, 3 and 5 after collection. Parameters such as platelet counts, residual leukocytes, platelet swirling, glucose, lactate, pH, CD62P expression (platelet activation), JC-1 (mitochondrial function) and annexin V (apoptosis and cell death) were assessed. RESULTS: Over the five days of storage there was a significant decrease in glucose, HCO3, pCO2, ATP, pH, swirling and mitochondrial function, associated with a significant increase in lactate levels and pO2. At the end of storage pH was 5.9 ± 0.6 and lactate levels were 2.8 ± 1.2 mmol/L. Results of the quality parameters evaluated were similar to those reported in human platelets studies. The deleterious effects of storage were more pronounced in bags with higher platelet counts (> 7.49 × 1010/unit), suggesting that canine platelet concentrates should not contain an excessive number of platelets. CONCLUSIONS: Quality parameters of canine platelets under standard storage conditions were similar to those observed in human platelets. Our results have potential to be used for the routine evaluation and quality control in veterinary blood banks.


Assuntos
Bancos de Sangue/normas , Plaquetas/fisiologia , Preservação de Sangue/veterinária , Cães/sangue , Animais , Plaquetas/metabolismo , Ativação Plaquetária , Agregação Plaquetária , Testes de Função Plaquetária/veterinária , Controle de Qualidade
12.
J Vet Emerg Crit Care (San Antonio) ; 30(3): 264-271, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32108424

RESUMO

OBJECTIVE: To design and evaluate a method to purify canine albumin from fresh frozen plasma (FFP) or stored plasma (SP) in a manner that could be applied clinically. DESIGN: In vitro experimental study. SETTING: FDA licensed Blood Bank Laboratory and University biochemistry laboratory. ANIMALS: None. INTERVENTIONS: Using equipment that is typically found in veterinary blood banks, plasma bags were thawed, injected with the heat stabilizing agent, sodium caprylate, and then heated and acidified to denature all but albumin proteins. Albumin-rich supernatant was removed, the pH was neutralized, and then pasteurized and refrigerated. Albumin and total plasma protein concentrations were measured and the product was cultured for bacteria at 0, 7, 14, 30, and 60 days post-processing. MEASUREMENTS AND MAIN RESULTS: Seventeen bags of plasma were analyzed for purity, yield, and sterility of the finished albumin product. Bags were divided into categories based on the age of the frozen plasma. Mean yield of albumin for all bags was 77.3% and mean purity was 91.2%. There was no difference between old stored plasma, new stored plasma, and FFP with regard to yield (P = 0.31) or purity (P = 0.24) based on one-way analysis of variances. Overall 1 of 17 bags of plasma (5.9%) tested positive for bacterial contamination on day 60 after processing. CONCLUSIONS: Sodium caprylate is able to stabilize canine albumin enabling it to withstand heating that denatures other plasma proteins. The resulting albumin product is of sufficient quality to potentially be used therapeutically as a colloidal resuscitative fluid. Further study is needed into its safety and effect in dogs.


Assuntos
Albuminas/química , Cães/sangue , Temperatura Alta , Plasma/química , Desnaturação Proteica , Animais , Proteínas Sanguíneas , Humanos
13.
PLoS One ; 15(2): e0229121, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32101539

RESUMO

Since dogs play a central role in the contamination of humans and livestock with Echinococcus granulosus, the development of an effective vaccine for dogs is essential to control the disease caused by this parasite. For this purpose, a formulation based on biodegradable polymeric nanoparticles (NPs) as delivery system of recombinant Echinococcus granulosus antigen (tropomyosin EgTrp) adjuved with monophosphoryl lipid A (MPLA) has been developed. The obtained nanoparticles had a size of approximately 200 nm in diameter into which the antigen was correctly preserved and encapsulated. The efficiency of this system to deliver the antigen was evaluated in vitro on canine monocyte-derived dendritic cells (cMoDCs) generated from peripheral blood monocytes. After 48 h of contact between the formulations and cMoDCs, we observed no toxic effect on the cells but a strong internalization of the NPs, probably through different pathways depending on the presence or not of MPLA. An evaluation of cMoDCs activation by flow cytometry showed a stronger expression of CD80, CD86, CD40 and MHCII by cells treated with any of the tested formulations or with LPS (positive control) in comparison to cells treated with PBS (negative control). A higher activation was observed for cells challenged with EgTrp-NPs-MPLA compared to EgTrp alone. Formulations with MPLA, even at low ratio of MPLA, give better results than formulations without MPLA, proving the importance of the adjuvant in the nanoparticles structure. Moreover, autologous T CD4+ cell proliferation observed in presence of cMoDCs challenged with EgTrp-NPs-MPLA was higher than those observed after challenged with EgTrp alone (p<0.05). These first results suggest that our formulation could be used as an antigen delivery system to targeting canine dendritic cells in the course of Echinococcus granulosus vaccine development.


Assuntos
Antígenos de Protozoários/administração & dosagem , Células Dendríticas/imunologia , Cães/parasitologia , Equinococose/prevenção & controle , Echinococcus granulosus/imunologia , Vacinas Protozoárias/administração & dosagem , Adjuvantes Imunológicos/administração & dosagem , Animais , Antígenos de Protozoários/genética , Antígenos de Protozoários/imunologia , Linfócitos T CD4-Positivos/imunologia , Diferenciação Celular , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Células Dendríticas/efeitos dos fármacos , Cães/sangue , Cães/imunologia , Portadores de Fármacos/química , Portadores de Fármacos/toxicidade , Equinococose/imunologia , Equinococose/parasitologia , Equinococose/veterinária , Echinococcus granulosus/genética , Imunogenicidade da Vacina , Lipídeo A/análogos & derivados , Lipídeo A/química , Lipídeo A/toxicidade , Ativação Linfocitária/imunologia , Monócitos/fisiologia , Nanopartículas/química , Nanopartículas/toxicidade , Poliésteres/química , Poliésteres/toxicidade , Cultura Primária de Células , Vacinas Protozoárias/genética , Vacinas Protozoárias/imunologia , Testes de Toxicidade Aguda , Tropomiosina/administração & dosagem , Tropomiosina/genética , Tropomiosina/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia
14.
Reprod Domest Anim ; 55(4): 454-459, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31957075

RESUMO

Leptin regulates body weight and several physiological processes including reproduction. We evaluated the circulating levels of leptin in pregnant and non-pregnant bitches as well as their correlation with body weight, food intake and number of foetuses. Nineteen healthy German shepherd bitches were used and divided in two groups (pregnant n = 12 and non-pregnant n = 7). Blood samples were collected every 15 days starting from ovulation (Day 0) throughout pregnancy (pregnant group, P) or throughout luteal phase (non-pregnant group, NP) In pregnant bitches, leptin concentrations increased from the day of ovulation (1.32 ± 0.06 ng/ml) up to day 45 (1.51 ± 0.06 ng/ml; p < .01) and returned to baseline values from day 60 post-ovulation. In non-pregnant bitches, leptin concentrations remained constant throughout the whole observation period (estimated marginal mean ± SE=1.33 ± 0.38 ng/ml). Pairwise comparisons showed significant differences between P and NP at day 45 post-ovulation (p < .05). Multivariable models indicated that, controlling for time and litter size, there was a positive relationship between leptin concentration and BW (p < .05) although Pearson coefficients showed that the correlation between BW and leptin was only significant in NP animals at day 45 (r = 0.76, p < .05). The multivariable approach also suggested that, holding BW and time constant, leptin concentrations tend to increase as the number of puppies increased (p = .06). Our study supports indirectly the contribution of the feto-placental unit to the circulating maternal leptin concentration.


Assuntos
Cães/fisiologia , Leptina/sangue , Tamanho da Ninhada de Vivíparos , Prenhez/fisiologia , Animais , Peso Corporal/fisiologia , Cães/sangue , Feminino , Fase Luteal/fisiologia , Ovulação/fisiologia , Placenta , Gravidez , Prenhez/sangue
15.
Res Vet Sci ; 129: 129-136, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31982776

RESUMO

The objectives of this prospective study were determination of reference intervals (RI) for rotational thromboelastometry (ROTEM) parameters in single use reagents and to evaluate correlations between plasmatic coagulation times and ROTEM parameters. Blood was sampled from a jugular vein in 49 client-owned healthy dogs and ex-tem S, in-tem S, fib-tem S and ap-tem S parameters, prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen, haematology, blood chemistry and venous blood gas analysis was performed. Determination of RI was performed using Excel add-in Reference Value Advisor and correlations between PT, aPTT and fibrinogen with selected ROTEM parameters were determined by Spearman correlation. Ex-tem S maximum clot firmness (MCF) RI are smaller compared to RI in people and liquid ex-tem in dogs while maximum lysis was comparable to those in people but smaller than previously reported in dogs. A strong correlation was found between fibrinogen measured by Clauss and fib-tem S and in-tem S MCF (r = 0.541, P < .001 and r = 0.610, P < .001, respectively). PT showed a significant but moderate correlation with ex-tem S CT (r = 0.340, P = .030), in-tem S CFT (r = 0.433, P = .003), fib-tem S CT (r = 0.426, P = .009) and ap-tem S CT (r = 0.354, P = .015) while aPTT was not significantly correlated with any of the evaluated parameters. In conclusion, this study provides single use reagent ROTEM parameter RIs that are different from RI determined with liquid reagents. Significant correlations between fibrinogen concentrations measured by Clauss and clot firmness of fib-tem S and in-tem S profiles and between PT and clotting times of all reagents were identified.


Assuntos
Testes de Coagulação Sanguínea/veterinária , Coagulação Sanguínea/fisiologia , Cães/sangue , Tromboelastografia/veterinária , Animais , Feminino , Fibrinogênio , Humanos , Masculino , Tempo de Tromboplastina Parcial/veterinária , Estudos Prospectivos , Tempo de Protrombina/veterinária , Valores de Referência
16.
J Vet Intern Med ; 34(1): 186-194, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31725186

RESUMO

BACKGROUND: Serum creatinine and symmetric dimethylarginine (SDMA) are used as surrogate markers of glomerular filtration rate (GFR) in clinical practice. Data pertaining to the correlations between GFR, SDMA, and serum creatinine in client-owned dogs are limited. OBJECTIVES: To describe the relationship between GFR, SDMA, and serum creatinine in a population of client-owned dogs, and to compare clinical utility of SDMA to GFR estimation for detecting pre-azotemic chronic kidney disease. ANIMALS: Medical records of 119 dogs that had GFR estimation performed via serum iohexol clearance between 2012 and 2017. METHODS: Prospective study using archived samples. GFR, SDMA, and serum creatinine results were reviewed and submitting practices contacted for outcome data. All dogs included in the study population were non-azotemic. Correlations between GFR, SDMA, and serum creatinine were determined by regression analysis. Sensitivity, specificity, and positive and negative likelihood ratios of different cutoffs for SDMA and serum creatinine for detecting decreased GFR were calculated, using a 95% confidence interval. RESULTS: Serum creatinine and SDMA were moderately correlated with GFR (R2 = 0.52 and 0.27, respectively, P < .0001) and with each other (R2 = 0.33, P < .0001). SDMA >14 µg/dL was sensitive (90%) but nonspecific (50%) for detecting a ≥40% decrease in GFR. Optimal SDMA concentration cutoff for detecting a ≥40% GFR decrease was >18 µg/dL (sensitivity 90%, specificity 83%). CONCLUSIONS AND CLINICAL IMPORTANCE: In non-azotemic dogs being screened for decreased renal function, using a cutoff of >18 µg/dL rather than >14 µg/dL increases the specificity of SDMA, without compromising sensitivity.


Assuntos
Arginina/análogos & derivados , Creatinina/sangue , Doenças do Cão/diagnóstico , Cães/metabolismo , Iohexol/farmacocinética , Insuficiência Renal Crônica/veterinária , Animais , Arginina/sangue , Meios de Contraste/farmacocinética , Doenças do Cão/sangue , Doenças do Cão/metabolismo , Cães/sangue , Taxa de Filtração Glomerular , Insuficiência Renal Crônica/sangue , Insuficiência Renal Crônica/metabolismo , Julgamento Moral Retrospectivo
17.
J S Afr Vet Assoc ; 90(0): e1-e6, 2019 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-31714111

RESUMO

The Coat-A-Count® radioimmunoassay has been long and widely used to determine the concentration of progesterone in serum or plasma of bitches (progRIA), but was discontinued in 2014. The Immulite® 1000 LKPG1 chemiluminescence immunoassay has gained prominence since 2003 to determine the concentration of progesterone in serum of bitches, but the assay changed in 2012 (Immulite® 1000 LKPW1). This study assessed the feasibility of using Immulite® 1000 LKPW1 (progImm) to estimate the time of clinically relevant events during oestrus and compared progRIA and progImm 2 and 3 days after the first or only day of the luteinizing hormone surge (LH1). ProgImm first exceeded 5.1 nmol/L on the same day that progRIA first exceeded 6 nmol/L, a proxy for the occurrence of the LH surge, or the day before in 28 of 31 (90%) of oestrous periods. ProgImm first exceeded 13.6 nmol/L on the same day that progRIA first exceeded 16 nmol/L (a proxy for the day of ovulation) or the day before in 34 of 35 (97%) oestrous periods. ProgImm first exceeded 5.4 nmol/L on LH1 or the day before in 24 of 25 (95%) of oestrous periods. The median of progImm 2 days after LH1 was 1.2 nmol/L lower than the 10.7 nmol/L of progRIA (p = 0.001). The mean of progImm 3 days after LH1 was 2.2 nmol/L lower than the 19.0 nmol/L of progRIA (p 0.001). In conclusion, the days on which progImm first exceeded 5.1 nmol/L, 13.6 nmol/L and 5.4 nmol/L effectively estimate the days on which progRIA reached 6 nmol/L or 16 nmol/L or LH1.


Assuntos
Cães/sangue , Medições Luminescentes/veterinária , Detecção da Ovulação/veterinária , Progesterona/sangue , Radioimunoensaio/veterinária , Animais , Tomada de Decisão Clínica , Estro/sangue , Feminino , Medições Luminescentes/métodos , Detecção da Ovulação/métodos , Radioimunoensaio/métodos , Reprodução/fisiologia
18.
Vet Clin Pathol ; 48(4): 748-753, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31696557

RESUMO

OBJECTIVES: Ghrelin is a major appetite-stimulating hormone. It circulates as acylated ghrelin (AG) and unacylated ghrelin (UAG), which could have different metabolic actions in obesity. Our objective was to study the analytical performance of two new canine AG and UAG ELISAs using blood samples from healthy, normal-weight dogs. Additionally, the effect of a protease inhibitor (PI) on short-term sample storage was analyzed. METHODS: The intra- and inter-assay precision for low, intermediate, and high AG and UAG concentrations, accuracy, limits of quantification (LQ), and detection limit (DL) of a blank sample were determined in ten healthy dogs. To study the effects of a PI on ghrelin concentrations, and AG and UAG concentrations were compared in five canine plasma samples stored for 1 month with (PI+), without (PI-), and with PI added at sample thawing (PI+th). RESULTS: The intra- and inter-assay coefficients of variation were 1.8%-5.7% and 2.9%-6.4% for the AG assay, and 0.8%-7.5% and 2.8%-13.4% for the UAG assay, respectively. Accuracy analyses showed nonsignificant deviation from linearity for the AG (R2  = .99; Runs test: P = .37) and UAG (R2  = .99; Runs test: P = .42) assays. For the AG assay, the upper LQ was >1261 pg/mL, the lower LQ was 6.2 pg/mL, and the DL was 0.3 pg/mL. For the UAG assay, the upper LQ was >1785 pg/mL, the lower LQ was 16.3 pg/mL, and the DL was 1.8 pg/mL. No differences in the AG (P = .54) and UAG (P = .95) concentrations were detected in the plasma samples subjected to PI+, PI-, and PI+th. CONCLUSION: The AG and UAG ELISA assays had acceptable precision, accuracy, lower LQ, and DLs, but upper LQ could not be established. An influence of the PI on short-term storage was not detectable. Long-term storage ± PI was not evaluated and should be investigated further.


Assuntos
Cães/sangue , Ensaio de Imunoadsorção Enzimática/veterinária , Grelina/sangue , Acilação , Animais , Armazenamento de Medicamentos , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Grelina/química , Masculino , Estudos Prospectivos , Reprodutibilidade dos Testes , Inibidores de Serino Proteinase/farmacologia , Sulfonas/farmacologia
19.
Am J Vet Res ; 80(12): 1074-1081, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31763942

RESUMO

OBJECTIVE: To determine the correlation between glucose concentrations in serum, plasma, and blood measured by a point-of-care glucometer (POCG) and serum glucose concentration measured by an automated biochemical analyzer (ABA; gold standard). SAMPLE: 152 canine and 111 feline blood samples. PROCEDURES: For each sample, the glucose concentration in serum, plasma, and blood was measured by a POCG and compared with the ABA-measured glucose concentration by means of the Lin concordance correlation coefficient. Results were summarized by species for all samples and subsets of samples with hyperglycemia (ABA-measured glucose concentration > 112 mg/dL for dogs and > 168 mg/dL for cats) and pronounced hyperglycemia (ABA-measured glucose concentration > 250 mg/dL for both species). The effect of PCV on correlations between POCG and ABA measurements was also assessed. RESULTS: Hyperglycemia and pronounced hyperglycemia were identified in 69 and 36 canine samples and 44 and 29 feline samples, respectively. The POCG-measured glucose concentrations in serum, plasma, and blood were strongly and positively correlated with the gold standard concentration. The PCV was positively associated with the correlation between the POCG-measured blood glucose concentration and the gold standard concentration but was not associated with the correlations between the POCG-measured glucose concentrations in serum and plasma and the gold standard concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that POCG-measured glucose concentrations in serum, plasma, and blood were strongly correlated with the ABA-measured serum glucose concentration, even in hyperglycemic samples. Given the time and labor required to harvest serum or plasma from blood samples, we concluded that blood was the preferred sample type for use with this POCG.


Assuntos
Análise Química do Sangue/veterinária , Glicemia/análise , Gatos/sangue , Cães/sangue , Animais , Análise Química do Sangue/instrumentação , Análise Química do Sangue/normas , Glucose/análise , Sistemas Automatizados de Assistência Junto ao Leito/normas , Soro/química
20.
J Vet Emerg Crit Care (San Antonio) ; 29(6): 630-634, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31625689

RESUMO

OBJECTIVE: Evaluate and quantify the effects of mucosal corn syrup and 50% dextrose application on blood glucose concentrations in healthy dogs, to assess the effectiveness of a widely used practice for treatment of hypoglycemia. DESIGN: Randomized controlled trial. SETTING: University teaching hospital. ANIMALS: Twelve client-owned dogs that were healthy, >1 year of age, weighing >5 kg, and had normal physical exam and biochemical profiles. INTERVENTIONS: Dogs were fasted overnight for a minimum of 12 hours. Once normal physical exam and biochemical profile were confirmed, an IV catheter was placed in a peripheral vein for serial blood sampling. Each dog served as their own control and received each of 3 treatments, the orders of which were randomized for each dog. Treatments included mucosal application of commercially available corn syrup (Karo light syrup), water (control), and 50% dextrose solution, each at a dose of 1 mL/kg of body weight. Blood glucose was measured using a point-of-care glucometer. Samples were taken immediately prior to each treatment and at 5-, 10-, 15-, 20-, 30-, and 60-minute intervals. RESULTS: All treatments were well tolerated and no adverse events were observed. A statistically significant increase in blood glucose was observed at the 15-, 20-, 30-, and 60-minute time points in the corn syrup and 50% dextrose groups as compared with the control. CONCLUSIONS: A significant effect on the blood glucose concentrations of the treated animals was not observed until 15 minutes after application of concentrated glucose solutions. These findings suggest that, in more severely hypoglycemic patients, parenteral glucose administration may be necessary.


Assuntos
Glicemia/efeitos dos fármacos , Cães/sangue , Glucose/administração & dosagem , Xarope de Milho Rico em Frutose/administração & dosagem , Administração através da Mucosa , Animais , Coleta de Amostras Sanguíneas , Feminino , Infusões Intravenosas , Masculino , Zea mays
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