RESUMO
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.
Assuntos
Adenosina Desaminase/metabolismo , Bovinos/metabolismo , Cães/metabolismo , Cavalos/metabolismo , Inflamação/veterinária , Saliva/metabolismo , Suínos/metabolismo , Adenina/análogos & derivados , Adenosina Desaminase/sangue , Inibidores de Adenosina Desaminase , Animais , Automação , Biomarcadores/sangue , Biomarcadores/metabolismo , Bovinos/sangue , Ensaios Enzimáticos Clínicos/métodos , Ensaios Enzimáticos Clínicos/veterinária , Cães/sangue , Cavalos/sangue , Inflamação/sangue , Inflamação/enzimologia , Isoenzimas/sangue , Isoenzimas/metabolismo , Suínos/sangueRESUMO
Amostras de sangue foram obtidas de 138 equinos registrados da raça Campeiro, com idade média de 9,7±5,4 anos, sendo 14 machos (10,15%) e 124 fêmeas (89,85%). Observaram-se valores médios da atividade sérica de GGT: 13,73±4,08 U/L; AST: 246,34±72,87U/L; ALT: 5,13±1,18U/L; FA: 284,32±53,33UI/L; CK: 132,54±72,25U/L; LDH: 511,38±143,65U/L; e das concentrações séricas de ureia: 38,65±12,62mg/dL e de creatinina: 1,24±0,24mg/dL. Os valores médios de AST, FA, ALT, ureia e creatinina foram semelhantes aos propostos na literatura para outras raças. Os valores de CK, GGT e LDH foram superiores aos comumente utilizados como referências. Não houve diferença nas concentrações de nenhum dos componentes séricos avaliados nas diferentes idades analisadas. Maiores valores médios nas concentrações de ureia e menores de creatinina foram observados nas fêmeas em relação aos machos, assim como a maior atividade sérica de FA em fêmeas gestantes em relação às éguas vazias. O perfil bioquímico sérico de equinos sadios da raça Campeiro apresenta variações peculiares que devem ser levadas em consideração na interpretação de exames laboratoriais.(AU)
Blood samples were obtained from 138 equines of the Campeiro breed with mean ages of 9.7±5.4 years, including 14 males (10.15%) and 124 mares (89.85%). Mean values of serum of GGT: 13.73 ± 4.08U/L; AST: 246.34±72.87U/L; ALT 5.13±1.18U/L; FA: 284.32±53.33IU/L; CK: 132.54±72.25U/L; LDH: 511.38±143.65U/L; and serum urea: 38.65±12.62mg/dL and creatinine: 1.24±0.24mg/dL were observed. The mean values of AST, FA, ALT, urea and creatinine were similar to those proposed in the literature for other breeds. The CK values, GGT, and LDH were higher than those commonly used as references. There was no difference in the concentrations of any of the serum components evaluated at different ages analyzed. Further average values in the lower urea concentrations and creatinine were observed in females compared to males, as well as the higher serum activity of FA in pregnant mares in relation to non-pregnant. Serum biochemical profile of healthy horses of the Campeiro breed presents peculiar variations that must be taken into consideration in the interpretation of laboratory tests.(AU)
Assuntos
Animais , Masculino , Feminino , Gravidez , Ensaios Enzimáticos Clínicos/veterinária , Cavalos/sangue , Técnicas de Laboratório Clínico/veterináriaRESUMO
A indoleamina 2,3-dioxigenase (IDO) é uma enzima responsável por catabolizar o aminoácido triptofano. Sua presença no ambiente uterino placentário está relacionada à tolerância imunológica ao semi-aloenxerto, pois impede a proliferação de células imunológicas maternas, seja pela falta do aminoácido, ou pela ação de alguns catabólitos oriundos da quebra do triptofano, como o ácido quinolínico, que é tóxico principalmente para os linfócitos T. Pouco se conhece sob a influência de substâncias (hormônios e citocinas) presentes na interface materno fetal e a expressão dessa enzima. Por esta razão, formulou-se a hipótese de que hormônios e interleucinas presentes na região uteroplacentária poderiam exercer algum efeito na expressão da IDO. Células oriundas da interface materno fetal de ratas Wistar foram mantidas em cultivo, onde receberam suplementação com estradiol e interferon-γ. A expressão da enzima foi avaliada pela técnica de citometria de fluxo nos períodos de 4, 24 e 48 horas e confirmação da presença proteica por imuno-histoquímica. Os resultados mostraram um aumento na expressão de IDO após a adição de estrógeno (9,03±0,81/11,25±0,25) e interferon-γ (9,03±0,81/20,43±0,60). O efeito do interferon-γ já era esperado como relatado na literatura, contudo, a elevação da expressão da IDO pela adição do estrógeno constitui nova informação sobre possíveis mecanismos envolvidos na ativação da enzima. O melhor esclarecimento desses achados poderia contribuir para uma melhor compreensão da participação dessa enzima na tolerância materno-fetal e para uma futura modulação terapêutica da mesma...
The indoleamine 2,3-dioxygenase (IDO) is an enzyme responsible for catabolizing the tryptophan. Its presence in the placental uterine environment is related to immunological tolerance to the semi-allograft because it prevents proliferation of maternal immune cells, either by the lack of this amino acid or by the action of its catabolites, such as the quinolinic acid, which is particularly toxic for T lymphocytes. Little is known regarding the influence of hormones and cytokines on the expression of IDO in the maternal fetal interface. Therefore, the hypothesis that some hormones and interleukins present in uteroplacental region could have an effect on the expression of IDO on cultured cells was tested. Cells derived from the fetal maternal interface from Wistar rats were kept in culture and supplemented with estradiol and interferon-γ. Expression of the enzyme was assessed by flow cytometry at periods of 4, 24 and 48 hours and confirmation of the presence of protein by immunohistochemistry. The results showed an increasing of IDO expression after the addition of estrogen (9.03±0.81 to 11.25±0.25) and interferon-γ (9.03±0.81 to 20.43±0.60). The effect of interferon-γ was expected as reported in the literature, however, elevated IDO expression by estrogen represents new information on possible mechanisms involved in the enzyme activation. These findings could provide a better understanding of IDO contribution on maternal-fetal tolerance and may collaborate to future therapeutic modulation of this enzyme...
Assuntos
Animais , Feminino , Gravidez , Cobaias , Estrogênios , Interferon gama , Ratos Wistar/embriologia , Citometria de Fluxo/veterinária , Ensaios Enzimáticos Clínicos/veterinária , Imuno-Histoquímica/veterinária , PlacentaRESUMO
The pancreas remains a difficult organ to evaluate using laboratory methods alone. No single laboratory test is diagnostic of pancreatitis (chronic or acute) without other diagnostic modalities concurring with the diagnosis or ruling out other diseases. The diagnosis of pancreatitis is particularly difficult in cats, and pancreatitis often occurs with other diseases. The use of pancreatic cytology may be useful in diagnosing both inflammation and neoplasia. Exocrine pancreatic insufficiency (EPI) can be relatively easily diagnosed when clinically manifested by the measurement of trypsinlike immunoreactivity. Diagnosis is more difficult when EPI is subclinical.
Assuntos
Testes de Química Clínica/veterinária , Insuficiência Pancreática Exócrina/veterinária , Pancreatopatias/veterinária , Testes de Função Pancreática/veterinária , Animais , Gatos , Testes de Química Clínica/tendências , Ensaios Enzimáticos Clínicos/tendências , Ensaios Enzimáticos Clínicos/veterinária , Diagnóstico Diferencial , Cães , Diagnóstico Precoce , Insuficiência Pancreática Exócrina/etiologia , Pancreatopatias/diagnóstico , Pancreatopatias/metabolismo , Pancreatopatias/fisiopatologia , Elastase Pancreática/sangue , Testes de Função Pancreática/tendênciasRESUMO
The leukocyte esterase test pad on a urine dipstick has been used as a preliminary test for bacterial peritonitis in humans but has not previously been evaluated in dogs. Here, free abdominal fluid from 60 dogs was tested on the leukocyte esterase test pad and results were compared with culture and microscopic analysis. Depending on the 'gold standard' comparator, the dipstick had sensitivity of ~60-75%, specificity of ~91-92%, positive predictive value of ~69%, and negative predictive value of ~87-94%. Based on these data, it appears that the leukocyte esterase test pad is most useful for tentative identification of cases in which bacterial infection is unlikely. Therefore a negative test may aid in re-directing clinician attention to alternative diagnoses in dogs with free abdominal effusion, whereas a positive result implies the necessity for further diagnostic tests.
Assuntos
Ascite/veterinária , Hidrolases de Éster Carboxílico , Ensaios Enzimáticos Clínicos/veterinária , Doenças do Cão/diagnóstico , Peritonite/veterinária , Fitas Reagentes , Urinálise/veterinária , Animais , Ascite/microbiologia , Doenças do Cão/microbiologia , Cães , Peritonite/complicações , Peritonite/diagnóstico , Peritonite/microbiologia , Sensibilidade e EspecificidadeRESUMO
A new assay for the determination of lactosylceramide-2,3-sialyltransferase (SAT I, EC 2.4.99.9) and monosialoganglioside sialyltransferase (SAT IV, EC 2.4.99.2) is described. The assay utilised the commercially available fluorophore labelled sphingolipids, boron dipyrromethene difluoride (BODIPY) lactosylceramide (LacCer), and BODIPY-monosialotetrahexosylganglioside (GM1) as the acceptor substrates, for SAT I and SAT IV, respectively. HPLC coupled with fluorescence detection was used to analyse product formation. The analysis was performed in a quick and automated fashion. The assay showed good linearity for both BODIPY sphingolipids with a quantitative detection limit of 0.05 pmol. The high sensitivity enabled the detection of SAT I and SAT IV activities as low as 0.001 µU, at least 200 fold lower than that of most radiometric assays. This new assay was applied to the screening of SAT I and SAT IV activities in ovine and bovine organs (liver, heart, kidney, and spleen). The results provided evidence that young animals, such as calves, start to produce ganglioside sialyltransferases as early as 7 days after parturition and that levels change during maturation. Among the organs tested from a bovine source, spleen had the highest specific ganglioside sialyltransferase activity. Due to the organ size, the greatest total ganglioside sialyltransferase activities (SAT I and SAT IV) were detected in the liver of both bovine and ovine origin.
Assuntos
Bovinos/metabolismo , Ensaios Enzimáticos Clínicos/métodos , Ovinos/metabolismo , Sialiltransferases/análise , Animais , Antígenos CD , Compostos de Boro , Configuração de Carboidratos , Sequência de Carboidratos , Bovinos/crescimento & desenvolvimento , Ensaios Enzimáticos Clínicos/veterinária , Gangliosídeos , Rim/enzimologia , Lactosilceramidas , Fígado/enzimologia , Microssomos/enzimologia , Dados de Sequência Molecular , Miocárdio/enzimologia , Especificidade de Órgãos , Ovinos/crescimento & desenvolvimento , Baço/enzimologia , Especificidade por SubstratoRESUMO
Avaliaram-se a concentração de lactato e a glicemia em sete cadelas em trabalho de parto submetidas à cesariana e em seus respectivos neonatos. O lactato materno foi mensurado utilizando-se medidor portátil e pelo método enzimático colorimétrico. Nos neonatos avaliou-se também o peso ao nascimento. Os valores de lactato materno e neonatal foram estatisticamente diferentes, sendo mais elevado nos neonatos. Ao nascimento, as cadelas e os neonatos encontravam-se normoglicêmicos, e não se observou correlação entre a glicemia de ambos. Os valores de lactato obtidos pelo o analisador portátil (3,5±0,6mmol/L) foram mais altos do que os obtidos pelo método enzimático colorimétrico (1,6±0,6mmol/L). Observou-se correlação significativa entre o peso do neonato e a concentração de lactato, isto é, os filhotes de menor peso apresentaram maior concentração de lactato.
This work evaluated the concentration of lactate and blood glucose in seven bitches in labor subject to caesarean section and in their newborns. The lactate concentration was measured using a portable analyzer and enzymatic colorimetric method. In neonates, the birth weight was evaluated. The maternal and neonatal lactate values were statistically different, being higher in newborns. At birth, the bitches and the newborns had normal glucose status, and no correlation was observed between blood glucose in either. The lactate concentration obtained by the portable analyzer (3.5±0.6mmol/L) was higher than that obtained with the enzymatic colorimetric method (1.6±0.6mmol/L). There was significant correlation between the neonate weight and lactate concentration and in puppies with lighter weight there was a higher concentration of lactate.
Assuntos
Animais , Feminino , Cães , Ácido Láctico/análise , Cesárea/efeitos adversos , Cesárea/veterinária , Glicemia/análise , Animais Recém-Nascidos , Ensaios Enzimáticos Clínicos/veterinária , Índice Glicêmico , Hipóxia Fetal/veterináriaRESUMO
BACKGROUND: In people, lysosomal storage diseases (LSD) can be diagnosed by assaying enzyme activities in dried blood spots (DBS). OBJECTIVE: The aim of this study was to evaluate the feasibility of using DBS samples from dogs and cats to measure lysosomal enzymatic activities and diagnose LSD. METHODS: Drops of fresh whole blood collected in EDTA from dogs and cats with known or suspected LSD and from clinically healthy dogs and cats were placed on neonatal screening cards, dried, and mailed to the Metabolic Laboratory, University Children's Hospital, Frankfurt, Germany. Activities of selected lysosomal enzymes were measured using fluorescent substrates in a 2-mm diameter disk (~2.6 µL blood) punched from the DBS. Results were expressed as nmol substrate hydrolyzed per mL of blood per minute or hour. RESULTS: Reference values were established for several lysosomal enzyme activities in DBS from dogs and cats; for most enzymes, activities were higher than those published for human samples. Activities of ß-glucuronidase, N-acetylglucosamine-4-sulfatase (arylsulfatase B), α-mannosidase, α-galactosidase, α-fucosidase, and hexosaminidase A were measureable in DBS from healthy cats and dogs; α-iduronidase activity was measureable only in cats. In samples from animals with LSD, markedly reduced activity of a specific enzyme was found. In contrast, in samples from cats affected with mucolipidosis II, activities of lysosomal enzymes were markedly increased. CONCLUSIONS: Measurement of lysosomal enzyme activities in DBS provides an inexpensive, simple, and convenient method to screen animals for suspected LSD and requires only a small sample volume. For diseases in which the relevant enzyme activity can be measured in DBS, a specific diagnosis can be made.
Assuntos
Doenças do Gato/diagnóstico , Ensaios Enzimáticos Clínicos/veterinária , Doenças do Cão/diagnóstico , Teste em Amostras de Sangue Seco/veterinária , Doenças por Armazenamento dos Lisossomos/veterinária , Animais , Coleta de Amostras Sanguíneas/veterinária , Doenças do Gato/sangue , Gatos , Doenças do Cão/sangue , Cães , Feminino , Alemanha , Glucuronidase/sangue , Hexosaminidase A/sangue , Iduronidase/sangue , Doenças por Armazenamento dos Lisossomos/sangue , Doenças por Armazenamento dos Lisossomos/diagnóstico , Lisossomos/enzimologia , Masculino , N-Acetilgalactosamina-4-Sulfatase/sangue , Valores de Referência , Especificidade da Espécie , alfa-Galactosidase/sangue , alfa-L-Fucosidase/sangue , alfa-Manosidase/sangueRESUMO
In veterinary medicine, serum creatine-kinase (CK) activity is mostly used to assess skeletal muscle damage. This retrospective study aimed to evaluate the prevalence of increased CK activity in a large, ill-cat population and to characterise associated diseases, clinical and laboratory findings and its prognostic value. Cats with a complete serum biochemistry analysis were consecutively enrolled, divided into two CK activity-based groups (within and above reference interval) and compared. The study included 601 cats. Median serum CK was 402 U/l (range 16-506870). Increased CK (>250 U/l) was observed in 364 (60%) cats, and>30-fold its upper reference limit in 43 (7%). Cats with increased CK had greater (P < or = 0.05) body weight, and were more likely to have a history of collapse, dyspnoea, abnormal lung sounds, cyanosis, shock and paraplegia, higher median serum alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase activities and total bilirubin and triglyceride concentrations, but lower, median total protein, albumin, globulin and cholesterol concentrations and proportion of anorexia than cats with normal CK. Cardiac diseases, trauma, bite wounds, systemic bacterial infections, prior anaesthesia and intramuscular injections were more common (P < or = 0.05) in cats with increased compared to normal CK activity. The hospitalisation period was longer (P=0.007) and treatment cost and mortality were higher (P<0.005) in cats with increased CK activity. However, CK activity was an inaccurate outcome predictor (area under the receiver operator characteristics curve 0.58). Increased CK activity is very common in ill cats.
Assuntos
Doenças do Gato/diagnóstico , Ensaios Enzimáticos Clínicos/veterinária , Creatina Quinase/sangue , Animais , Análise Química do Sangue/veterinária , Doenças do Gato/sangue , Gatos , Doença Crônica , Feminino , Modelos Logísticos , Masculino , Prognóstico , Valores de Referência , Estudos Retrospectivos , Faculdades de Medicina VeterináriaRESUMO
This study reports the clinical, clinicopathological and ultrasonographic findings from dogs with chronic pancreatitis (CP). Fourteen dogs with clinical signs consistent with CP and histological confirmation of the disease were evaluated. Abdominal ultrasound and clinical pathology results were recorded. Sensitivities of pancreatic enzymes for diagnosis of CP were calculated with two different cut-off values. The mean age of affected dogs was 9.1 years. Spaniels were the most common breed with CP, representing seven of the 14 dogs in this study. CP was histologically severe in nine cases. Most dogs showed chronic low-grade gastrointestinal signs and abdominal pain. Five dogs had exocrine pancreatic insufficiency and five dogs had diabetes mellitus. The sensitivity of elevated trypsin-like immunoreactivity for CP was 17 per cent. The sensitivities of canine pancreatic lipase immunoreactivity, lipase and amylase for CP were 44 to 67 per cent or 14 to 28 per cent depending on the cut-off value used. Cholesterol was elevated in 58 per cent of samples. Liver enzymes were often elevated. The pancreas appeared abnormal on 56 per cent of ultrasound examinations. Ten dogs had died by the end of the study period; only one case was due to CP.
Assuntos
Ensaios Enzimáticos Clínicos/veterinária , Doenças do Cão/diagnóstico por imagem , Doenças do Cão/patologia , Pancreatite Crônica/veterinária , Dor Abdominal/etiologia , Dor Abdominal/veterinária , Amilases/sangue , Animais , Biomarcadores/sangue , Colesterol/sangue , Complicações do Diabetes/veterinária , Diabetes Mellitus/veterinária , Cães , Insuficiência Pancreática Exócrina/complicações , Insuficiência Pancreática Exócrina/veterinária , Feminino , Seguimentos , Gastroenteropatias/etiologia , Gastroenteropatias/veterinária , Lipase/sangue , Masculino , Pancreatite Crônica/diagnóstico por imagem , Pancreatite Crônica/patologia , Estudos Prospectivos , Índice de Gravidade de Doença , Tripsina/sangue , UltrassonografiaRESUMO
Determination of seric levels of adenosine deaminase (ADA), an enzyme produced by monocytes/macrophages and lymphocytes, has been used in the diagnosis of human tuberculosis (TB). In the present study, ADA seric activity was evaluated comparatively to the comparative tuberculin test in the diagnosis of bovine tuberculosis. Two hundred fifty-six cattle were classified by origin and by the comparative tuberculin test as TB-positive animals (n = 52, from herds where the Mycobacterium bovis had previously been isolated), and TB-negative animals (n = 204, TB-free herds). The mean ADA seric value from the TB-positive group (4.45 +/- 2.33 U/L) was significantly lower (p = 0.008) than that observed in sera from the TB-negative group (6.12 +/- 4.47 U/L). When animals from a herd with clinical cases of enzootic bovine leukosis of TB-negative group were withdrawn from analysis, the mean ADA seric values of TB-negative group (5.12 +/- 3.75 U/L) was not significantly different anymore from that of the TB-positive group (p = 0.28). There was no agreement in the diagnosis of bovine TB between comparative tuberculin test and determination of ADA seric values, using two different cutoff points, being 6.12 U/L and 15.0 U/L, (kappa = -0.086 and kappa = -0.082, respectively). In conclusion, the determination of ADA seric activity was not a good auxiliary test for bovine TB, because it was not able to distinguish between TB-positive and TB-negative animals.
Assuntos
Adenosina Desaminase/sangue , Ensaios Enzimáticos Clínicos/veterinária , Mycobacterium bovis , Tuberculose Bovina/diagnóstico , Animais , Biomarcadores/sangue , Bovinos , Sensibilidade e Especificidade , Teste Tuberculínico/veterináriaRESUMO
Determination of seric levels of adenosine deaminase (ADA), an enzyme produced by monocytes/macrophages and lymphocytes, has been used in the diagnosis of human tuberculosis (TB). In the present study, ADA seric activity was evaluated comparatively to the comparative tuberculin test in the diagnosis of bovine tuberculosis. Two hundred fifty-six cattle were classified by origin and by the comparative tuberculin test as TB-positive animals (n = 52, from herds where the Mycobacterium bovis had previously been isolated), and TB-negative animals (n = 204, TB-free herds). The mean ADA seric value from the TB-positive group (4.45 ± 2.33 U/L) was significantly lower (p = 0.008) than that observed in sera from the TB-negative group (6.12 ± 4.47 U/L). When animals from a herd with clinical cases of enzootic bovine leukosis of TB-negative group were withdrawn from analysis, the mean ADA seric values of TB-negative group (5.12 ± 3.75 U/L) was not significantly different anymore from that of the TB-positive group (p = 0.28). There was no agreement in the diagnosis of bovine TB between comparative tuberculin test and determination of ADA seric values, using two different cutoff points, being 6.12 U/L and 15.0 U/L, (kappa = -0.086 and kappa = -0.082, respectively). In conclusion, the determination of ADA seric activity was not a good auxiliary test for bovine TB, because it was not able to distinguish between TB-positive and TB-negative animals.
Assuntos
Animais , Bovinos , Adenosina Desaminase/sangue , Ensaios Enzimáticos Clínicos/veterinária , Mycobacterium bovis , Tuberculose Bovina/diagnóstico , Biomarcadores/sangue , Sensibilidade e Especificidade , Teste Tuberculínico/veterináriaRESUMO
Exposure to dioxin-like chemicals that activate the aryl hydrocarbon receptor (AHR) can result in increased cellular and tissue production of reactive oxygen species (ROS). Little is known of these effects during early fish development. We used the fish model, Fundulus heteroclitus, to determine if the AHR ligand and pro-oxidant 3,3',4,4',5-pentachlorobiphenyl (PCB126) can increase ROS production during killifish development, and to test a novel method for measuring ROS non-invasively in a living organism. The superoxide-sensitive fluorescent dye, dihydroethidium (DHE), was used to detect in ovo ROS production microscopically in developing killifish exposed to PCB126 or vehicle. Both in ovo CYP1A activity (ethoxyresorufin-o-deethylase, EROD) and in ovo ROS were induced by PCB126. In ovo CYP1A activity was inducible by PCB126 concentrations as low as 0.003 nM, with maximal induction occurring at 0.3 nM PCB126. These PCB126 concentrations also significantly increased in ovo ROS production in embryonic liver, ROS being detectable as early as 5 days post-fertilization. These data demonstrate that the pro-oxidant and CYP1A inducer, PCB126, increases both CYP1A activity and ROS production in developing killifish embryos. The superoxide detection assay (SoDA) described in this paper provides a semi-quantitative, easily measured, early indicator of altered ROS production that can be used in conjunction with simultaneous in ovo measurements of CYP1A activity and embryo development to explore functional relationships among biochemical, physiological and developmental responses to AHR ligands.
Assuntos
Embrião não Mamífero/efeitos dos fármacos , Fundulidae/embriologia , Bifenilos Policlorados/toxicidade , Espécies Reativas de Oxigênio/metabolismo , Animais , Ensaios Enzimáticos Clínicos/métodos , Ensaios Enzimáticos Clínicos/veterinária , Citocromo P-450 CYP1A1/análise , Citocromo P-450 CYP1A1/biossíntese , Embrião não Mamífero/química , Embrião não Mamífero/metabolismo , Exposição Ambiental , Feminino , Fígado/química , Fígado/metabolismo , Masculino , Espécies Reativas de Oxigênio/análise , Sensibilidade e Especificidade , Fatores de Tempo , Bexiga Urinária/química , Bexiga Urinária/metabolismo , Poluentes Químicos da Água/toxicidadeRESUMO
BACKGROUND: Urine creatinine concentration often is used in ratios such as urine protein:creatinine to compensate for dilution or concentration of spot urine samples. OBJECTIVE: The purpose of this study was to compare the accuracy of different techniques of urine creatinine measurement currently available for veterinary practitioners. METHODS: In 104 samples of canine urine diluted 1:20 with distilled water, creatinine concentration was measured using a kinetic Jaffé reaction assay, and an enzymatic technique on an automatic analyzer (Elimat) and 3 benchtop analyzers (Reflovet, Scil; Vitros DT2, Ortho-Clinical Diagnostics; Vettest 8008, IDEXX) used in veterinary practice. RESULTS: The Jaffé and enzymatic techniques on the Elimat were not significantly different, and their inaccuracy tested with human control urines was <5%. The benchtop analyzers underestimated creatinine concentration, especially at concentrations >2000 mg/L. Inaccuracy was higher with multilayer slide technology systems (Vitros and Vettest) than with the Reflovet system. Results were approximately 25% and 2% lower, respectively, than with the Elimat at urine creatinine concentrations about 2000 mg/L. CONCLUSION: Inaccuracy in urine creatinine measurements using benchtop analyzers should be taken into account when defining decision thresholds, which should be corrected according to the method used to avoid misinterpretations.
Assuntos
Creatinina/urina , Cães/urina , Laboratórios/normas , Ureo-Hidrolases/metabolismo , Urinálise/veterinária , Animais , Autoanálise/métodos , Autoanálise/normas , Autoanálise/veterinária , Ensaios Enzimáticos Clínicos/métodos , Ensaios Enzimáticos Clínicos/normas , Ensaios Enzimáticos Clínicos/veterinária , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Ureo-Hidrolases/análise , Urinálise/instrumentação , Urinálise/normasRESUMO
Empregando-se difusäo radial enzimática em ágargel, atividades hidrolíticas de amostras de conteúdo do intestino delgado de 38 bovinos adultos de matadouro foram mensuradas em animais agrupados como possuidores de pâncreas hígidos (controle) e parasitados por Eurytrema sp, estes, segundo dois graus de infecçäo reconhecidos. Para tal, foram empregados quatro substratos, dos quais caseinato de cálcio e óleo de milho tiveram de ser desconsiderados porque, no caso deles, näo houve formaçäo de halos ou estes apresentaram limites imprecisos. Apesar desses problemas näo terem ocorrido quando o substrato era leite em pó desnatado ou amido, as atividades hidrolíticas encontradas foram equivalentes, independentemente da condiçäo de parasitismo e grau de infecçäo verificados (P>0,05). Concluiu-se, entäo, que a técnica de difusäo radial enzimática, pelo menos com os substratos empregados, foi incapaz de acusar disfunçöes pancreáticas exócrinas em bovinos infectados por Eurytrema sp.
Assuntos
Animais , Masculino , Feminino , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/parasitologia , Doenças Parasitárias em Animais/complicações , Doenças Parasitárias em Animais/diagnóstico , Pancreatite/diagnóstico , Pancreatite/etiologia , Pancreatite/veterinária , Doença Crônica , Ensaios Enzimáticos Clínicos/veterináriaRESUMO
Profiles of plasma enzymes were compared in two strains of single comb white leghorn laying hens, a normal commercial strain and strain UCD-003, which is highly susceptible to fatty liver-hemorrhagic syndrome. Plasma activity of lactate dehydrogenase (LDH), glutamate dehydrogenase (GDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and creatine kinase (CK) averaged 194 +/- 27, 4.0 +/- 2.8, 146 +/- 20, 1.0 +/- 1.0, and 1041 +/- 268 U/liter, respectively in normal birds. Activities of LDH, GDH, AST, and ALT, but not CK, were significantly higher in UCD-003 than in normal hens. A bimodal distribution of activities of all enzymes was found in the UCD-003 hens, with some birds showing activities comparable with those of the normal hens and others with values that were 2-10 times greater than those found in normal hens. These results are consistent with the extensive hepatic lesions observed in the UCD-003 strain of birds. Average gross hemorrhagic scores from visual inspection (scale of 0-3) were 0.28 +/- 0.45 in normal birds and 1.63 +/- 0.94 in the UCD-003 birds. Even though no clear relationship was found between plasma enzyme activities and the extent of liver hemorrhage in individual birds, the UCD-003 hens consistently had average values significantly higher for plasma enzymes that indicate liver damage. The results suggest that measurement of enzyme activities indicative of liver damage in birds, particularly AST, LDH, and GDH, is a valuable tool in the diagnosis of fatty liver-hemorrhagic syndrome in a flock of layers.
Assuntos
Ensaios Enzimáticos Clínicos/veterinária , Fígado Gorduroso/veterinária , Hemorragia/veterinária , Doenças das Aves Domésticas/diagnóstico , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Galinhas , Creatina Quinase/sangue , Fígado Gorduroso/diagnóstico , Fígado Gorduroso/enzimologia , Feminino , Glutamato Desidrogenase/sangue , Hemorragia/diagnóstico , Hemorragia/enzimologia , L-Lactato Desidrogenase/sangue , Oviposição , Doenças das Aves Domésticas/sangue , Doenças das Aves Domésticas/enzimologia , Valores de Referência , SíndromeAssuntos
Doenças do Gato/diagnóstico , Doenças do Cão/diagnóstico , Hepatopatias/veterinária , Animais , Animais Domésticos , Ácidos e Sais Biliares/sangue , Bilirrubina/sangue , Biomarcadores/sangue , Biópsia/veterinária , Gatos , Colestase/diagnóstico , Colestase/veterinária , Ensaios Enzimáticos Clínicos/veterinária , Cães , Hepatopatias/diagnóstico , Testes de Função Hepática/veterináriaRESUMO
Blood of 24 healthy goats of different breeds in the age between two and six years as well as their 36 lambs in the peripartal period was taken at regular intervals from the day 10 ante partum up to the day 30 post partum. The content of plasma--nonspecific, clinical essential enzymes as well as the selenium content--has been examined. 1. Normal parturition shows an influence on the enzyme activity in plasma. Abundant alterations in concentration (p < or = 0.05-0.001) are detectable in AST, LDH, alpha-HBDH, GGT, GLDH and CK. GGT responds indistinctly to birth. GLDH-concentration is significant higher in the lactation period (p < or = 0.01). 2. The registration of OCT in blood plasma and serum of goats is possible but the activity of the enzyme in plasma and serum is slight. 3. In the blood of new born lambs a high plasma activity of CK and GGT can be found. Up to day 5 post natum the activity of CK descends to post partum adult level and up to day 30 post natum lower enzyme values than those of adult goats are found. GGT activity at day of birth after colostrum uptake is more than 5 times higher than at day 5 post natum. Up to day 30 post natum the activity remains above the mean of adult goats.
Assuntos
Enzimas/sangue , Cabras/sangue , Período Pós-Parto/sangue , Prenhez/sangue , Animais , Ensaios Enzimáticos Clínicos/veterinária , Testes Diagnósticos de Rotina/veterinária , Feminino , Plasma , Gravidez , Selênio/sangueRESUMO
In 837 birds which died or were euthanatized because of their disease, blood could be taken within 12 hours before death. Blood chemistry results were compared with specific necropsy findings of different organs. The parameters aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), creatinine kinase (CK), glutamate dehydrogenase (GLDH), glucose, total protein, uric acid, urea and creatinine were measured. Possible reasons for the bad comparability of blood chemistry results and organ lesions are discussed. Glucose and creatinine were found not to be useful in a routine panel for avian patients.
Assuntos
Doenças das Aves/sangue , Aves/sangue , Análise Química do Sangue/veterinária , Ensaios Enzimáticos Clínicos/veterinária , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Análise Química do Sangue/métodos , Glicemia/análise , Proteínas Sanguíneas/análise , Ensaios Enzimáticos Clínicos/métodos , Creatina Quinase/sangue , Creatinina/sangue , Glutamato Desidrogenase/sangue , L-Lactato Desidrogenase/sangue , Ureia/sangue , Ácido Úrico/sangueRESUMO
In this study three different laboratory procedures were compared as predictors of net energy content of raw materials and compound feeds for ruminants. Laboratory measurements included an enzymatic technique (pepsin-cellulase method) and a rumen fluid technique (Hohenheim gas test). An estimate based on crude nutrients only was also tested. A total of 98 feed samples with known in vivo digestibility were available. By multivariate regression analysis equations for the prediction of the net energy content were calculated. The results suggest that in case of compound feeds for dairy the cellulase technique and the gas test are suitable in vitro techniques. With both procedures the relationships between energy contents estimated in vitro and energy contents measured in vivo showed residual errors of less than 4 percent. However for the evaluation of raw materials the gas test was superior. Compared to the cellulase technique the risk of extreme under- or overestimations is apparently reduced and this was found to be the case for own prediction equations as well as equations from literature. Crude nutrients proved to be very poor predictors for all feed samples.