Sheep naturally infected with Anaplasma ovis - evaluation of cardiac and inflammatory biomarkers

Background: Anaplasmosis, also called gall sickness or tropical bovine ehrlichiosis, is an infectious disease caused by species belonging to the genus Anaplasma in domestic and wild animals in tropical and subtropical regions. Anaplasma ovis and A. phagocytophilum are important pathogens of sheep. A. ovis is considered the most common species affecting sheep. The infection is usually subclinical and progresses with high fever, anaemia, icterus, weight loss and abortions. This study aimed to investigate changes in cardiac damage markers, oxidative stress and antioxidant status, cytokines, and acute phase proteins in sheep naturally infected with A. ovis. Materials, Methods & Results: For this purpose, a total of 40 animals, including 20 healthy sheep and 20 sheep infected with anaplasmosis, were used. A. ovis was diagnosed based on clinical findings and peripheral blood smear. Blood smears were prepared from the ear vein. The smears were stained with Giemsa and examined for the presence of Anaplasma spp. Infection was also confirmed by polymerase chain reaction (PCR) analysis. The genomic DNA was isolated from blood, and the MSP-4 gene region was amplified as A. ovis specific target gene. Twenty clinically healthy sheep of the same age group, reared under the same conditions and testing negative in the molecular assessment were used as controls. Blood samples were collected from the cephalic vein and and centrifuged to obtain serum. The serum stored at -20°C until the analysis stage. Serum samples were used for the analysis of cardiac damage markers [troponin I (cTnI), creatine kinase MB (CK-MB), lactate dehydrogenase (LDH) and aspartate transaminase (AST)], oxidative stress parameters [malondialdehyde (MDA), total antioxidant status (TAS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], cytokines [interleukins IL-6, IL-1ß and IL-10, tumour necrosis factor α (TNF-α), and interferon-γ (IFN-γ)] and acute phase proteins [C-reactive protein (CRP), serum amyloid A (SAA) and haptoglobin (Hp)]. cTnI and CK-MB levels were measured using a chemiluminescent immunoassay. MDA, TAS, SOD, CAT, GPx, TNF-α, IL-1ß, IL-6, IL-10, IFN-γ, SAA and Hp levels were measured by an ELISA reader. LDH, AST and CRP levels were measured in an autoanalyzer. cTnI and LDH levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The concentration of AST was decreased in infected animals. MDA, TAS, SOD, CAT and GPx levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The levels of the inflammatory parameters such as TNF-α, IL-1ß, IL-10 and IFN-γ were significantly increased in the infected animals compared to the healthy ones (P < 0.05). Hp level were significantly increased in the infected group compared with the control group (P < 0.05). However, there was no significant change in CK-MB, SAA and CRP concentrations in the infected animals (P > 0.05). Discussion: Ovine anaplasmosis is an obligate intracellular arthropod disease that causes widespread changes in haematobiochemical, immune response and oxidative stress parameters. Cardiac damage is often overlooked in field conditions due to the lack of adequate knowledge about the pathophysiology of the disease. Our results showed that A. ovis infection leads to significant changes in cardiac biomarkers and that the parasite can cause cardiac dysfunction. This is the first report on cardiac damage markers in Anaplasma-infected sheep. Additionally, the levels of proinflammatory and oxidative stress markers that may cause functional disorders were also found to be increased. Thus, measuring markers of cardiac function, oxidative stress and inflammation can be a useful tool in the early diagnosis of ovine anaplasmosis.

Electrocardiographic, echocardiographic and heart biomarker parameters in sheep experimentally poisoned by Palicourea marcgravii (Rubiaceae) / Parâmetros eletrocardiográficos, ecocardiográficos e de biomarcadores cardíacos em ovinos intoxicados experimentalmente por Palicourea marcgravii (Rubiaceae)

The present study aimed to identify and describe cardiac alterations in sheep experimentally poisoned with Palicourea marcgravii through analysis of serum cardiac biomarkers (serum troponin I and creatine kinase - CK-MB) and electro and echocardiographic assessments to contribute to a better understanding of the poisoning pathophysiology. P. marcgravii is the main plant within a group of 22 species that cause sudden death in Brazil; its toxic principle is sodium monofluoroacetate. Eight healthy crossbreed male sheep, aged between five and twelve months, weighing 14 to 27kg, were evaluated. The animals received 1g kg-1 of P. marcgravii plants orally. The sheep were evaluated before administering the plant (T0) through electro and echocardiography and blood collection to assess cardiac biomarkers (CK-MB and cTnI). Collections and analyses were repeated every four hours until the animal's death. During the study, there was the presence of extravasation of serum troponin I carried out in a qualitative test, with positive values at time T4, and the serum CK-MB biomarker had a peak at T4 and slightly decreased at T8. The electro and echocardiographic examinations showed that the cause of death in these animals was due to acute heart failure characterized by arrhythmias, tachycardia/ventricular fibrillation, drop in cardiac output, left ventricular (LV) systolic dysfunction by the progressive decrease in the LV ejection fraction (EF), decrease in LV fractional shortening (FS), and decrease in aortic flow velocity and aortic flow gradient. This study seems to be the first to evaluate cardiac alterations in sheep poisoned by P. marcgravii through cardiac biomarkers and electro and echocardiographic exams.

O presente estudo objetivou identificar e descrever as alterações cardíacas de ovinos intoxicados experimentalmente por Palicourea marcgravii através das análises de biomarcadores cardíacos séricos (troponina I sérica e a creatinoquinase - MB) e das avaliações eletro e ecocardiográficas contribuindo para o melhor entendimento da fisiopatologia da intoxicação. Palicourea marcgravii é a principal planta dentro de um grupo de 22 espécies que causam "morte súbita" no Brasil e seu princípio tóxico é o monofluoracetato de sódio. Foram utilizados oito ovinos saudáveis machos, sem raça definida, com idades entre cinco e doze meses e peso de 14 a 27kg. Os animais receberam 1g/kg de Palicourea marcgravii por via oral. Os ovinos foram avaliados momentos antes da administração da planta (T0) através de eletro e ecocardiograma e coleta de sangue para avaliação dos biomarcadores cardíacos (CK-MB e cTnI). As análises e coletas foram repetidas a cada quatro horas até o óbito do animal. Durante o estudo observou-se extravasamento de troponina I sérica realizada em teste qualitativo, com valores positivos já em T4, assim como o biomarcador CK-MB sérico teve seu pico de aumento em T4 e em T8 houve uma leve redução. Aos exames eletro e ecocardiográfico foi possível determinar que a causa do óbito nestes animais ocorreu devido à insuficiência cardíaca aguda caracterizada por arritmias, taquicardia/fibrilação ventricular, queda no débito cardíaco, disfunção sistólica do ventrículo esquerdo (VE) pela diminuição progressiva da fração de ejeção (EF) do VE, diminuição na fração de encurtamento (FS) do VE, diminuição da velocidade do fluxo aórtico e do gradiente do fluxo aórtico. Este é o primeiro estudo que avalia as alterações cardiológicas de ovinos intoxicados por P. marcgravii através de biomarcadores cardíacos e exames eletro e ecocardiográficos.

Beneficial role of oleuropein in sepsis-induced myocardial injury: possible involvement of GSK-3beta/NF-kB pathway

Purpose The present study explored the potential therapeutic role of oleuropein in sepsis-induced heart injury along with the role of GSK-3beta/NF-kB signaling pathway. Methods Sepsis-induced myocardial injury was induced by cecal ligation and puncture (CLP) in rats. The cardiac injury was assessed by measuring the levels of cTnI and creatine kinase-MB (CK-MB). Sepsis-induced inflammation was assessed by measuring interleukin-6 (IL-6), IL-10 and HMGB1 levels. The different doses of oleuropein (5, 10, and 20 mg/kg) were given prior to CLP. Oleuropein (20 mg/kg) was administered after 6 hof CLP. The expressions of GSK-3beta, p-GSK-3beta (Ser9) and nuclear factor-B (NF-B) were measured in heart homogenates. Results Cecal ligation and puncture was associated with myocardial injury, an increase in IL-6, a decrease in IL-10 and an increase in HMGB1. Moreover, it decreased the ratio of p-GSK-3beta/GSK-3beta and increased the expression of p-NF-kB. Pretreatment with oleuropein attenuated CLP-induced myocardial injury and systemic inflammation in a dose-dependent manner. Administration of oleuropein after the onset of CLP also attenuated cardiac injury and inflammation. It also restored CLP-induced changes in the HMGB1 levels, the ratio of p-GSK-3beta/GSK-3beta and expression of p- NF-kB. Conclusions Oleuropein attenuates sepsis-induced systemic inflammation and myocardial injury by inhibiting NF-kB and GSK-3beta signaling.(AU)

Liraglutide restores late cardioprotective effects of remote preconditioning in diabetic rats via activation of hydrogen sulfide and nuclear factor erythroid 2-related factor 2 signaling pathway

Purpose The present study explored the influence of liraglutide on remote preconditioning-mediated cardioprotection in diabetes mellitus along with the role of nuclear factor erythroid 2-related factor 2 (Nrf2), hypoxia inducible factor (HIF-1) and hydrogen sulfide (H2S). Methods Streptozotocin was given to rats to induce diabetes mellitus and rats were kept for eight weeks. Four cycles of ischemia and reperfusion were given to hind limb to induce remote preconditioning. After 24 h, hearts were isolated and subjected to 30 min of ischemia and 120 min of reperfusion on Langendorff system. Liraglutide was administered along with remote preconditioning. Cardiac injury was assessed by measuring the release of creatine kinase (CK-MB), cardiac troponin (cTnT) and development of left ventricular developed pressure. After ischemia-reperfusion, hearts were homogenized to measure the nuclear cytoplasmic ratio of Nrf2, H2S and HIF-1 levels. Results In diabetic rats, there was more pronounced injury and the cardioprotective effects of remote preconditioning were not observed. Administration of liraglutide restored the cardioprotective effects of remote preconditioning in a dose-dependent manner. Moreover, liraglutide increased the Nrf2, H2S and HIF-1 levels in remote preconditioning-subjected diabetic rats. Conclusions Liraglutide restores the lost cardioprotective effects of remote preconditioning in diabetes by increasing the expression of Nrf2, H2S and HIF-1.(AU)

Cardiac, energy and hormonal blood markers, and lactatemia in cows with displaced abomasum

Background: Displaced abomasum (DA) is a common and economically important disorder that affects dairy cattle. Nutritional factors and adaptive responses that occur in the peripartum play a central role in the pathogenesis. The measurement of blood metabolites represents a useful tool for monitoring and prognostic determination in affected animals. Therefore, the objective was to evaluate cardiac, energy and hormonal blood markers, lactatemia, and insulin sensitivity in cows diagnosed with right displaced abomasum (RDA) and left displaced abomasum (LDA), comparing them with each other. Materials, Methods & Results: Nineteen cases of abomasum displacement in cows were studied, including 9 cases of RDA and 10 cases of LDA. The diagnosis was established by means of physical examination and measurement of the concentration of chlorides in the ruminal fluid (> 30 mEq/L). After diagnosis, clinical-surgical therapeutic management was instituted. At the time of diagnosis (M1) and at the resolution of the case (M2), blood samples were collected to assess the variables: non-esterified fatty acids (NEFA), beta hydroxybutyrate (βHB), L-lactate, creatine kinase (CK), creatine kinase MB (CK-MB), cardiac troponin I (cTnI), lactate dehydrogenase (LDH), glucose, insulin, and cortisol. In addition, insulin sensitivity was estimated using the Revised Quantitative Insulin Sensitivity Check Index (RQUICKI) and RQUICKI-βHB. The means of the variables were compared, separating the effects of groups (RDA and LDA) and moments (M1 and M2), at the level of 5% probability. The concentrations of NEFA, CK-MB, L-lactate, glucose, insulin, and cortisol were higher at M1 and the RQUICKI and RQUICKI-βHB indices were lower at this moment. L-lactate, CK, and CK-MB were higher in the RDA group, while cTnI, βHB, and LDH did not present a group or moment effect. Cardiac markers correlated with the energy profile metabolites, L-lactate, and cortisol. Discussion: The high...

Involvement of HSP90 in ischemic postconditioning-induced cardioprotection by inhibition of the complement system, JNK and inflammation

Purpose To investigate whether heat shock protein 90 (HSP90) is involved in complement regulation in ischemic postconditioning (IPC). Methods The left coronary artery of rats underwent 30 min of occlusion, followed by 120 min of reperfusion and treatment with IPC via 3 cycles of 30s reperfusion and 30s occlusion. The rats were injected intraperitoneally with 1 mg/kg HSP90 inhibitor geldanamycin (GA) after anesthesia. Eighty rats were randomly divided into four groups: sham, ischemia-reperfusion (I/R), IPC and IPC + GA. Myocardial infarct size, apoptosis index and the expression of HSP90, C3, C5a, tumor necrosis factor (TNF)-alpha, interleukin (IL)-1β and c-Jun N-terminal kinase (JNK) were assessed. Results Compared with the I/R injury, the IPC treatment significantly reduced infarct size, release of troponin T, creatine kinase-MB, and lactate dehydrogenase, and cardiomyocyte apoptosis. These beneficial effects were accompanied by a decrease in TNF-α, IL-1β, C3, C5a and JNK expression levels. However, all these effects were abrogated by administration of the HSP90 inhibitor GA. Conclusion HSP90 exerts a profound effect on IPC cardioprotection, and may be linked to the inhibition of the complement system and JNK, ultimately attenuating I/R-induced myocardial injury and apoptosis.(AU)

GDF11 ameliorated myocardial ischemia reperfusion injury by antioxidant stress and up-regulating autophagy in STZ-induced type 1 diabetic rats

Purpose: To investigate whether GDF11 ameliorates myocardial ischemia reperfusion (MIR) injury in diabetic rats and explore the underlying mechanisms. Methods: Diabetic and non-diabetic rats subjected to MIR (30 min of coronary artery occlusion followed by 120 min of reperfusion) with/without GDF11 pretreatment. Cardiac function, myocardial infarct size, creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH), superoxide dismutase (SOD) 15-F2tisoprostane, autophagosome, LC3II/I ratio and Belcin-1 level were determined to reflect myocardial injury, oxidative stress and autophagy, respectively. In in vitro study, H9c2 cells cultured in high glucose (HG, 30mM) suffered hypoxia reoxygenation (HR) with/without GDF11, hydrogen peroxide (H2O2) and autophagy inhibitor 3-methyladenine (3-MA) treatment, cell injury; oxidative stress and autophagy were assessed. Results: Pretreatment with GDF11 significantly improved cardiac morphology and function in diabetes, concomitant with decreased arrhythmia severity, infarct size, CK-MB, LDH and 15-F2tisoprostane release, increased SOD activity and autophagy level. In addition, GDF11 notably reduced HR injury in H9c2 cells with HG exposure, accompanied by oxidative stress reduction and autophagy up-regulation. However, those effects were completely reversed by H2O2 and 3-MA. Conclusion: GDF11 can provide protection against MIR injury in diabetic rats, and is implicated in antioxidant stress and autophagy up-regulation.(AU)

Biochanin A attenuates myocardial ischemia/reperfusion injury through the TLR4/NF-B/NLRP3 signaling pathway

Purpose: Myocardial ischemia/reperfusion (Ml/R) injury is a leading cause of damage in cardiac tissues, with high rates of mortality and disability. Biochanin A (BCA) is a main constituent of Trifolium pratense L. This study was intended to explore the effect of BCA on Ml/R injury and explore the potential mechanism. Methods: In vivo MI/R injury was established by transient coronary ligation in Sprague-Dawley rats. Triphenyltetrazolium chloride staining (TTC) was used to measure myocardial infarct size. ELISA assay was employed to evaluate the levels of myocardial enzyme and inflammatory cytokines. Western blot assay was conducted to detect related protein levels in myocardial tissues. Results: BCA significantly ameliorated myocardial infarction area, reduced the release of myocardial enzyme levels including aspartate transaminase (AST), creatine kinase (CK-MB) and lactic dehydrogenase (LDH). It also decreased the production of inflammatory cytokines (IL-1, IL-18, IL-6 and TNF-) in serum of Ml/R rats. Further mechanism studies demonstrated that BCA inhibited inflammatory reaction through blocking TLR4/NF-kB/NLRP3 signaling pathway. Conclusion: The present study is the first evidence demonstrating that BCA attenuated Ml/R injury through suppressing TLR4/NF-kB/NLRP3 signaling pathway-mediated anti-inflammation pathway.(AU)

Alprostadil attenuates myocardial ischemia/reperfusion injury by promoting antioxidant activity and eNOS activation in rats

Purpose:To investigate the effect of alprostadil on myocardial ischemia/reperfusion (I/R) in rats.Methods:Rats were subjected to myocardial ischemia for 30 min followed by 24h reperfusion. Alprostadil (4 or 8 μg/kg) was intravenously administered at the time of reperfusion and myocardial infarct size, levels of troponin T, and the activity of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) in the serum were measured. Antioxidative parameters, nitric oxide (NO) content and phosphorylated endothelial nitric oxide synthase 3 (p-eNOS) expression in the left ventricles were also measured. Histopathological examinations of the left ventricles were also performed.Results:Alprostadil treatment significantly reduced myocardial infarct size, serum troponin T levels, and CK-MB and LDH activity (P<0.05). Furthermore, treatment with alprostadil significantly decreased malondialdehyde (MDA) content (P<0.05) and markedly reduced myonecrosis, edema and infiltration of inflammatory cells. Superoxide dismutase and catalase activities (P<0.05), NO level (P<0.01) and p-eNOS (P<0.05) were significantly increased in rats treated with alprostadil compared with control rats.Conclusion:These results indicate that alprostadil protects against myocardial I/R injury and that these protective effects are achieved, at least in part, via the promotion of antioxidant activity and activation of eNOS.(AU)

Cardioprotective effect of lipstatin derivative orlistat on normotensive rats submitted to cardiac ischemia and reperfusion

Purpose: To evaluate in vivo animal model of cardiac ischemia/reperfusion the cardioprotective activity of pancreatic lipase inhibitor of the orlistat. Methods: Adult male Wistar rats were anesthetized, placed on mechanical ventilation and underwent surgery to induce cardiac I/R by obstructing left descending coronary artery followed by reperfusion to evaluation of ventricular arrhythmias (VA), atrioventricular block (AVB) and lethality (LET) with pancreatic lipase inhibitor orlistat (ORL). At the end of reperfusion, blood samples were collected for determination of triglycerides (TG), very low-density lipoprotein (VLDL), low-density lipoprotein (LDL), high-density lipoprotein (HDL), lactate dehydrogenase (LDH), creatine kinase (CK), and creatine kinase-MB (CK-MB). Results: Treatment with ORL has been able to decrease the incidence of VA, AVB and LET. Besides that, treatment with ORL reduced serum concentrations of CK and LDL, but did not alter the levels of serum concentration of TG, VLDL and HDL. Conclusion: The reduction of ventricular arrhythmias, atrioventricular block, and lethality and serum levels of creatine kinase produced by treatment with orlistat in animal model of cardiac isquemia/reperfusion injury suggest that ORL could be used as an efficient cardioprotective therapeutic strategy to attenuate myocardial damage related to acute myocardial infarction.(AU)

Indicadores biológicos séricos em veado-campeiro ( Ozotoceros bezoarticus bezoarticus, Linnaeus 1758) de vida livre do Pantanal Sul-Mato-Grossense / Serum biological indicators in free-living pampas deer (Ozotoceros bezoarticus bezoarticus, Linnaeus 1758) of the Pantanal Sul-Mato-Grossense

O objetivo com este estudo foi estabelecer os intervalos de referência de diferentes analitos séricos em Ozotoceros bezoarticus bezoarticus de vida livre. Foram coletadas amostras de sangue de 54 fêmeas e 14 machos adultos, capturados no Pantanal de Mato Grosso do Sul. A determinação das concentrações de cálcio total, albumina, colesterol total, creatinina, glicose, fósforo, proteína total, triglicerídeos e ureia séricas e a atividade das enzimas alanina aminotransferase, aspartato aminotransferase, creatina quinase fração MB e fosfatase alcalina foi realizada em equipamento automático. Valores de globulinas, cálcio total e proporções (razões) entre analitos foram realizados por meio de cálculo matemático. Esta é a primeira descrição do intervalo de referência de parâmetros bioquímicos séricos em veado-campeiro de vida livre do Pantanal de Mato Grosso do Sul. O tamanho da amostra e a distribuição dos dados indicam que os valores médios obtidos são representativos e podem ser usados para a monitorização da saúde, do estado nutricional e do diagnóstico de enfermidades nesse estrato populacional.(AU)

The objective of the present study is to establish the reference intervals of different serum analytes in free - living Ozotoceros bezoarticus bezoarticus. Blood samples were drawn from the 54 females and 14 males, adults, captured in the Pantanal of Mato Grosso do Sul. A determination of total calcium, albumin, total cholesterol, creatinine, glucose, phosphorus, total protein, triglycerides, and serum urea concentrations and an activity of the enzymes alanine aminotransferase, aspartate aminotransferase, creatine kinase MB fraction and alkaline phosphatase were performed in automated equipment. Values of globulins, total calcium and proportions among analytes were performed by means of mathematical calculations. This is a first description of the reference parameters for serum biochemistry of free - living pampas deer in the Pantanal of Mato Grosso do Sul. Sample size and distribution of data indicate that the mean values obtained are representative and can be used for a health monitoring, nutritional status, and diagnosis of diseases in this population stratum.(AU)

Indicadores biológicos séricos em veado-campeiro ( Ozotoceros bezoarticus bezoarticus, Linnaeus 1758) de vida livre do Pantanal Sul-Mato-Grossense / Serum biological indicators in free-living pampas deer (Ozotoceros bezoarticus bezoarticus, Linnaeus 1758) of the Pantanal Sul-Mato-Grossense

O objetivo com este estudo foi estabelecer os intervalos de referência de diferentes analitos séricos em Ozotoceros bezoarticus bezoarticus de vida livre. Foram coletadas amostras de sangue de 54 fêmeas e 14 machos adultos, capturados no Pantanal de Mato Grosso do Sul. A determinação das concentrações de cálcio total, albumina, colesterol total, creatinina, glicose, fósforo, proteína total, triglicerídeos e ureia séricas e a atividade das enzimas alanina aminotransferase, aspartato aminotransferase, creatina quinase fração MB e fosfatase alcalina foi realizada em equipamento automático. Valores de globulinas, cálcio total e proporções (razões) entre analitos foram realizados por meio de cálculo matemático. Esta é a primeira descrição do intervalo de referência de parâmetros bioquímicos séricos em veado-campeiro de vida livre do Pantanal de Mato Grosso do Sul. O tamanho da amostra e a distribuição dos dados indicam que os valores médios obtidos são representativos e podem ser usados para a monitorização da saúde, do estado nutricional e do diagnóstico de enfermidades nesse estrato populacional.(AU)

The objective of the present study is to establish the reference intervals of different serum analytes in free - living Ozotoceros bezoarticus bezoarticus. Blood samples were drawn from the 54 females and 14 males, adults, captured in the Pantanal of Mato Grosso do Sul. A determination of total calcium, albumin, total cholesterol, creatinine, glucose, phosphorus, total protein, triglycerides, and serum urea concentrations and an activity of the enzymes alanine aminotransferase, aspartate aminotransferase, creatine kinase MB fraction and alkaline phosphatase were performed in automated equipment. Values of globulins, total calcium and proportions among analytes were performed by means of mathematical calculations. This is a first description of the reference parameters for serum biochemistry of free - living pampas deer in the Pantanal of Mato Grosso do Sul. Sample size and distribution of data indicate that the mean values obtained are representative and can be used for a health monitoring, nutritional status, and diagnosis of diseases in this population stratum.(AU)

Effects of Origanum Syriacum Essential Oil on Blood Parameters of Broilers Reared at High Ambient Heat

The biochemical and hematological blood values of broilers fed under heat stress and that were administered Origanum syriacum (OS) essential oil were investigated in this study. In total, 400 male broilers (Ross-308; age, 1 day) were used for the period of 42-days of the study. The experiment included ten groups (n = 50/group; each group divided into five sub-groups): normal heat, 22°C: basal feed (BF)-positive control group (PC), BF + 100 ppm OS; BF + 300 ppm OS; BF + 600 ppm OS). Heat stress, 36°C: BF-positive control group (PC), BF + 100 ppm SOS; BF + 300 ppm OS; and BF + 600 ppm OS). Biochemical and hematological parameters were measured in blood collected into EDTA tubes. Total bilirubin, cholesterol, low-density cholesterol, Na+, Ca2+, and Mg2+ increased, whereas alanine aminotransferase, aspartate aminotransferase, creatine kinase (CK), CK-MB, urea, uric acid, Cl-, and K+ decreased (p 0.05); however, no changes were detected in the other hematological values.(AU)

Effects of Origanum Syriacum Essential Oil on Blood Parameters of Broilers Reared at High Ambient Heat

The biochemical and hematological blood values of broilers fed under heat stress and that were administered Origanum syriacum (OS) essential oil were investigated in this study. In total, 400 male broilers (Ross-308; age, 1 day) were used for the period of 42-days of the study. The experiment included ten groups (n = 50/group; each group divided into five sub-groups): normal heat, 22°C: basal feed (BF)-positive control group (PC), BF + 100 ppm OS; BF + 300 ppm OS; BF + 600 ppm OS). Heat stress, 36°C: BF-positive control group (PC), BF + 100 ppm SOS; BF + 300 ppm OS; and BF + 600 ppm OS). Biochemical and hematological parameters were measured in blood collected into EDTA tubes. Total bilirubin, cholesterol, low-density cholesterol, Na+, Ca2+, and Mg2+ increased, whereas alanine aminotransferase, aspartate aminotransferase, creatine kinase (CK), CK-MB, urea, uric acid, Cl-, and K+ decreased (p 0.05); however, no changes were detected in the other hematological values.

RESPOSTAS AGUDAS HEMATOLÓGICAS, MUSCULARES E CARDÍACAS DE RATOS SOB CONDIÇÃO DE OVERREACHING

A síndrome do overtraining (SOT) é condição resultante do desequilíbrio entre intensidade de treinamento e tempo de recuperação, ocasionando redução do desempenho e fadiga. Objetivou-se investigar biomarcadores séricos musculares e perfil hematológico de ratos Wistar submetidos a um protocolo treino para provocar as condições FOR (functional overreaching) ou NFOR/SOT (nonfunctional overreaching) (PT). A amostragem inicial foi de 49 animais. Estes foram aleatoriamente distribuídos em 3 grupos iniciais: grupo sedentário (S, n=16), que não realizou nenhuma atividade física; grupo semissedentário (SSED, n=16), que realizou programa de condicionamento leve; grupo treinado (TR, n=33), que foi submetido a PT de 12 semanas. Para avaliação da aptidão física, os ratos foram submetidos a sete testes máximos de performance (TMP). No TMP-7, os animais do grupo TR foram subdivididos em dois grupos (FOR: n=19 e NFOR: n=14), identificados pela diferença da inclinação () obtida a partir da regressão linear dos desempenhos individuais nos TMP-2, 3, 4, 5, 6 e 7. Após o TMP-7, 8 ratos de cada grupo foram submetidos à eutanásia para obtenção de amostras. Realizou-se hemograma e citometria de fluxo de hemácias. Quantificaram-se as atividades séricas da creatina fosfoquinase (CK), aspartato aminotransferase (AST), lactato desidrogenase (LHD) e mioglobina. Avaliou-se as atividades da troponina cardíaca I (cTnI), isoenzima MB da creatinaquinase (CK-MB), lactato desidrogenase (LHD) e mioglobina. Foi determinada a relação miocardiossomática. Foram quantificados nos miocárdios a respiração mitocondrial máxima, atividade da citrato sintase (CS), e proteína indicadora de estresse (HSP70). Aplicou-se análise de variância de uma via (ANOVA One Way). Constatou-se valores superiores nos grupos SED e SSED em relação ao FOR para contagem de hemácias (RBC), hemoglobina (Hb) e hematócrito (Hct). O volume corpuscular médio (VCM) foi superior no grupo NFOR em comparação a SED e SSED. A contagem total de linfócitos foi maior no grupo NFOR em relação ao grupo FOR. A citometria de eritrócitos, a população 2 (P2) de RBC do grupo NFOR foi superior ao FOR. Não houve diferença entre os grupos para de CK, LDH e mioglobina. A AST do grupo SED foi menor que os demais grupos. Não houve diferença nas atividades séricas dos biomarcadores cardíacos entre os grupos. A relação miocardiossomática de NFOR foi superior aos demais grupos. O ganho de peso corporal foi semelhante entre FOR e NFOR, sendo este menor em relação a SSED. Não houve diferença da respiração máxima mitocondrial entre os grupos. A CS foi maior grupo NFOR em relação ao SED. A HSP70 foi maior no grupo SED, semelhante entre SSED e NFOR e menor no grupo FOR. A redução das variáveis eritrocitárias nos grupos FOR e NFOR foram associadas à expansão do volume plasmático e hemólise provocada por traumas inerentes ao desafio esportivo. A resposta leucocitária do grupo FOR pode estar relacionada à diminuição da resposta do eixo hipotálamo-hipófise-adrenal a agentes estressantes. A citometria de fluxo das hemácias mostrou-se promissora na diferenciação de FOR e NFOR. O condicionamento prévio pode ter atenuado a resposta dos biomarcadores musculares nos grupos treinados. À bioquímica sérica cardíaca evidenciou uma possível atenuação dos biomarcadores, como mecanismo adaptativo. O aumento da massa cardíaca em NFOR pode estar associado à hipertrofia deste órgão ou ao menor ganho de peso corporal. A avaliação da função mitocondrial evidenciou alterações adaptativas na célula muscular cardíaca em resposta ao protocolo de endurance, encorajando estudos complementares.

Overtraining syndrome (SOT) is a condition resulting from the imbalance between training intensity and recovery time, causing reduced performance and fatigue. The objective was to investigate serum muscle biomarkers and hematological profile of Wistar rats submitted to a training protocol to provoke the FOR (functional overreaching) or NFOR / SOT (nonfunctional overreaching) (PT) conditions. The initial sample was 49 animals. These were randomly assigned into 3 initial groups: sedentary group (S, n = 16), who did not perform any physical activity; semi-sedentary group (SSED, n = 16), which carried out a light conditioning program; trained group (TR, n = 33), who underwent PT for 12 weeks. To assess physical fitness, the rats were subjected to seven maximum performance tests (TMP). In the TMP-7, the animals in the TR group were subdivided into two groups (FOR: n = 19 and NFOR: n = 14), identified by the difference in the slope () obtained from the linear regression of individual performances in the TMP-2 , 3, 4, 5, 6 and 7. After TMP-7, 8 rats from each group were euthanized to obtain samples. CBC and erytrocyte flow cytometry were performed. The serum activities of creatine phosphokinase (CK), aspartate aminotransferase (AST), lactate dehydrogenase (LHD) and myoglobin were quantified. The activities of cardiac troponin I (cTnI), MB isoenzyme creatine kinase (CK-MB), lactate dehydrogenase (LHD) and myoglobin were evaluated. The myocardiosomatic relationship was determined. Maximal mitochondrial respiration, citrate synthase activity (CS), and stress indicator protein (HSP70) were quantified in the myocardium. One-way analysis of variance (ANOVA One Way) was applied. Higher values were found in the SED and SSED groups in relation to the FOR for red blood cell count (RBC), hemoglobin (Hb) and hematocrit (Hct). The mean corpuscular volume (VCM) was higher in the NFOR group compared to SED and SSED. The total lymphocyte count was higher in the NFOR group compared to the FOR group. The erythrocyte cytometry, population 2 (P2) of RBC of the NFOR group was higher than the FOR. There was no difference between groups for CK, LDH and myoglobin. The AST of the SED group was lower than the other groups. There was no difference in the serum activities of cardiac biomarkers between the groups. The myocardiosomatic ratio of NFOR was higher than the other groups. The body weight gain was similar between FOR and NFOR, which was lower in relation to SSED. There was no difference in maximum mitochondrial breathing between groups. CS was the largest NFOR group in relation to SED. HSP70 was higher in the SED group, similar between SSED and NFOR and lower in the FOR group. The reduction of erythrocyte variables in the FOR and NFOR groups was associated with the expansion of plasma volume and hemolysis caused by traumas inherent to the sports challenge. The leukocyte response of the FOR group may be related to the decreased response of the hypothalamic-pituitary-adrenal axis to stressors. RBC flow cytometry has shown promise in differentiating FOR and NFOR. The previous conditioning may have attenuated the response of the muscle biomarkers in the trained groups. Serum cardiac biochemistry showed a possible attenuation of biomarkers as an adaptive mechanism. The increase in cardiac mass in NFOR may be associated with hypertrophy of this organ or with lower body weight gain. The assessment of mitochondrial function showed adaptive changes in the cardiac muscle cell in response to the endurance protocol, encouraging further studies.

Avaliação dos indicadores de lesão miocárdica em cães com leishmaniose visceral / Changes in myocardial injury indicators in naturally infected dogs with visceral leishmaniasis

O objetivo deste estudo foi avaliar os indicadores laboratoriais, eletrocardiográficos e histológicos de lesão cardíaca em diferentes grupos clínicos de cães com leishmaniose visceral. Foram analisados marcadores séricos, traçado eletrocardiográfico e fragmentos de tecido cardíaco de 41 cães naturalmente infectados, distribuídos em três grupos: assintomático, oligossintomático e sintomático. Todos os animais apresentaram aumento na atividade sérica da enzima creatina quinase fração MB. No traçado eletrocardiográfico, o complexo de baixa voltagem foi o distúrbio de condução mais frequente (8/10). Na análise histológica, 75,6% dos cães apresentaram reação inflamatória com predomínio de infiltrados linfo-histiocítico (13/31) de intensidade discreta a moderada e distribuição multifocal. As alterações microscópicas identificadas no miocárdio foram independentes dos achados laboratoriais, eletrocardiográficos e do quadro clínico apresentado pelos animais estudados. A ausência de associação entre alterações histopatológicas e os parâmetros investigados alerta para a dificuldade de identificação de cardiopatia em cães com leishmaniose visceral e ressalta a importância de incluir a leishmaniose visceral no diagnóstico de patologias cardíacas principalmente em regiões endêmicas para o agente.(AU)

The aim of this study was to evaluate the laboratory indicators, electrocardiographic and cardiac histological lesions in different clinical groups of dogs with visceral leishmaniasis. Serum markers were analyzed in conjunction with the electrocardiographic tracing and heart tissue fragments of 41 naturally infected dogs which were divided into three groups: asymptomatic, oligosymptomatic and symptomatic. All animals showed increased activity in serum creatine kinase MB fraction. In the electrocardiographic tracing, low voltage complex was the most frequent conduction disorder (8/12). In the histological analysis, 75.6% of the dogs showed inflammatory reaction with predominance of linfohistiocítico infiltrates (13/31) of mild to moderate intensity and multifocal distribution. Microscopic changes identified in the myocardium were independent laboratory findings, an electrocardiographic and clinical picture presented by the studied animals. The lack of association between histopathological changes and the parameters investigated indicate the difficulty in disease identification in dogs with visceral leishmaniasis and highlights the importance of including visceral leishmaniasis in the diagnosis of heart disease especially in endemic regions to the agent.(AU)

Creatine phosphokinase isoenzyme-MB mass (CK-MB mass) and troponin I (cTnI) in dogs (Canis familiaris) / Creatinafosfoquinase-isoenzima MB massa (CK-MB massa) e troponina I (cTnI) em cães (Canis familiaris)

Cardiac markers, especially CK-MB mass and cTnI, have an essential role in both human and veterinary clinical and surgical cardiology, allowing a more precise and accurate diagnosis in myocardial lesions. The goal of this work was to measure these cardiac markers in veterinary medicine, improve their use and provide information about these laboratory methods that allow non-invasive health monitoring of the myocardial cell. Parameters quantification was obtained from sera of healthy dogs examined during routine procedures at the Small Animal Veterinary Hospital of Universidade Estadual do Norte Fluminense Darcy Ribeiro.  The human chemiluminescence essay turbo kit (IMMULITE Turbo, Siemens ®) test proved to be effective in canine species for both CK-MB mass and cTnI. In addition, the values found for CK-MB will significantly contribute to clinical routine or experimental work with dogs.

Os marcadores cardíacos, sobretudo a CK-MB massa e a cTnI, tem demonstrado um papel essencial seja em cardiologia clínica ou cirúrgica veterinária e humana, permitindo um diagnóstico mais preciso e acurado nas lesões miocárdicas. O objetivo deste trabalho foi mensurar os marcadores cardíacos em tela na Medicina Veterinária, aprimorar sua utilização e obter informações sobre estes métodos laboratoriais não invasivos que permitem o monitoramento da integridade da célula miocárdica. A quantificação dos parâmetros foi feita do soro de cães saudáveis atendidos na rotina do setor de Clínica Médica de Pequenos Animais do Hospital Veterinário da Universidade Estadual do Norte Fluminense Darcy Ribeiro. O kit de ensaio imunométrico por quimioluminescência turbo (IMMULITE Turbo, Siemens®) humano mostrou-se eficaz na avaliação na espécie canina, tanto para CK-MB massa quanto cTnI. Além disso, os valores encontrados para CK-MB massa e cTnI contribuirão significativamente para a rotina clínica ou para os trabalhos experimentais em cães.

Creatine phosphokinase isoenzyme-MB mass (CK-MB mass) and troponin I (cTnI) in dogs (Canis familiaris) / Creatinafosfoquinase-isoenzima MB massa (CK-MB massa) e troponina I (cTnI) em cães (Canis familiaris)

Cardiac markers, especially CK-MB mass and cTnI, have an essential role in both human and veterinary clinical and surgical cardiology, allowing a more precise and accurate diagnosis in myocardial lesions. The goal of this work was to measure these cardiac markers in veterinary medicine, improve their use and provide information about these laboratory methods that allow non-invasive health monitoring of the myocardial cell. Parameters quantification was obtained from sera of healthy dogs examined during routine procedures at the Small Animal Veterinary Hospital of Universidade Estadual do Norte Fluminense Darcy Ribeiro.  The human chemiluminescence essay turbo kit (IMMULITE Turbo, Siemens ®) test proved to be effective in canine species for both CK-MB mass and cTnI. In addition, the values found for CK-MB will significantly contribute to clinical routine or experimental work with dogs.(AU)

Os marcadores cardíacos, sobretudo a CK-MB massa e a cTnI, tem demonstrado um papel essencial seja em cardiologia clínica ou cirúrgica veterinária e humana, permitindo um diagnóstico mais preciso e acurado nas lesões miocárdicas. O objetivo deste trabalho foi mensurar os marcadores cardíacos em tela na Medicina Veterinária, aprimorar sua utilização e obter informações sobre estes métodos laboratoriais não invasivos que permitem o monitoramento da integridade da célula miocárdica. A quantificação dos parâmetros foi feita do soro de cães saudáveis atendidos na rotina do setor de Clínica Médica de Pequenos Animais do Hospital Veterinário da Universidade Estadual do Norte Fluminense Darcy Ribeiro. O kit de ensaio imunométrico por quimioluminescência turbo (IMMULITE Turbo, Siemens®) humano mostrou-se eficaz na avaliação na espécie canina, tanto para CK-MB massa quanto cTnI. Além disso, os valores encontrados para CK-MB massa e cTnI contribuirão significativamente para a rotina clínica ou para os trabalhos experimentais em cães.(AU)

AVALIAÇÃO DA TROPONINA CARDÍACA I E CK-MB EM CAPRINOS COM ACIDOSE LÁCTICA RUMINAL EXPERIMENTAL

Objetivou-se com o presente estudo avaliar a presença de lesão cardíaca em caprinos submetidos à indução da acidose láctica ruminal (ALR) com diferentes doses de sacarose, através da mensuração dos biomarcadores sanguíneos de lesão cardíaca, isoenzima MB da creatina quinase (CK-MB) e troponina cardíaca I (cTnI), além de avaliar a eficácia preventiva da suplementação dietética com monensina sódica e as alterações nos metabólitos sanguíneos e na hemogasometria nestes animais. No primeiro estudo, 20 caprinos foram divididos em dois grupos: grupo controle (GC) e grupo tratamento (GT) suplementado com 33mg/animal/dia de monensina sódica. A acidose ruminal foi induzida através da administração intraruminal de 10g/Kg de peso vivo de sacarose. Antes da indução e 4h, 8h, 12h, 24h, 32h, 48h e 72h pós-indução (PI) realizou-se exame clínico, análise do fluído ruminal e mensuração das variáveis sanguíneas: CK, L-lactato, cortisol, CK-MB e cTnI. Os caprinos apresentaram quadro clínico brando de ALR, com pH ruminal mínimo às 8h PI, sem diferença (P>0,05) entre os grupos. O L-lactato plasmático apresentou valores máximos 8h PI e o cortisol elevou-se entre 8h e 12h PI (P<0,05). As concentrações de CK-MB e cTnI foram mais elevadas (P<0.05) no GC e GT, respectivamente, sem diferença (P>0,05) entre os momentos. No segundo estudo, a enfermidade foi induzida em cinco caprinos através do fornecimento intrarruminal de 15g/Kg de peso vivo de sacarose. Antes da indução e 4h, 8h, 12h, 24h, 48h, 72h e 168h PI realizou-se exame clínico, análise do fluido ruminal, hemogasometria e mensuração de AST, CK, CK-MB, cTnI, GGT, LDH e L-lactato no sangue desses animais. Os caprinos apresentaram quadro clínico agudo de ALR, com redução (P<0,05) do pH das 4h às 48h PI, com menor média às 12h PI. Ocorreu alcalose metabólica transitória, antecedendo um quadro de acidose metabólica, o sódio e o potássio elevaram-se inicialmente e em seguida houve hipocalemia (P<0,05). L-lactato e LDH elevaram-se (P<0,05) e as concentrações dos biomarcadores cTnI e CK-MB não se alteraram após a indução (P>0,05). Os modelos experimentais utilizados causaram manifestações clínicas e metabólicas características da ALR, sendo o quadro clínico mais grave nos animais que receberam 15g/Kg de peso vivo de sacarose. A suplementação com monensina sódica, na dose utilizada, não preveniu a ocorrência da enfermidade. A avaliação da bioquímica sanguínea ratificou o diagnóstico e auxiliou no monitoramento a ALR em caprinos e a intensidade do quadro clínico induzido por ambas as doses do substrato não foi suficiente para causar lesão cardíaca e elevação de CK-MB e cTnI.

The aim of this study was to evaluate the presence of cardiac injury in goats submitted to induction of rumen lactic acidosis (RLA) with different sucrose doses, by measuring blood biomarkers of cardiac injury, creatine kinase MB isozyme (CK-MB) and cardiac troponin I (cTnI), in addition to evaluating the preventive efficacy of dietary supplementation with sodium monensin and changes in blood metabolites and blood gas analysis in these animals. In the first study, 20 goats were divided into two groups: control group (CG) and treatment group (TG) supplemented with 33 mg/animal/day of sodium monensin. Ruminal acidosis was induced by intraruminal administration of 10 g/kg of body weight of sucrose. Before induction and 4h, 8h, 12h, 24h, 32h, 48h and 72h post-induction (PI), clinical examination and ruminal fluid analysis were performed and blood variables were measured: CK, L-lactate, cortisol, CK-MB and cTnI. The goats had a mild clinical picture of RLA, with minimum rumen pH at 8h PI, with no difference (P>0.05) between groups. Plasma L-lactate showed maximum values 8h PI and cortisol increased between 8h and 12h PI (P<0.05). CK-MB and cTnI concentrations were higher (P <0.05) in CG and TG, respectively, with no difference (P>0.05) between the moments. In the second study, the disease was induced in five goats by intraruminal administration of 15g/kg of body weight fo sucrose. Before induction and 4h, 8h, 12h, 24h, 48h, 72h and 168h PI, clinical examination, ruminal fluid analysis, blood gas analysis and measurement of AST, CK, CK-MB, cTnI, GGT, LDH and L-lactate were performed in the blood of these animals. The goats presented acute clinical presentation of RLA, with reduction (P<0.05) of pH from 4h to 48h PI, with lower average at 12h PI. Transient metabolic alkalosis occurred, preceding a condition of metabolic acidosis, sodium and potassium increased initially and then hypokalemia occurred (P<0.05). L-lactate and LDH increased (P<0.05) and concentrations of cTnI and CK-MB biomarkers did not change after induction (P> 0.05). The experimental models used caused clinical and metabolic manifestations characteristics of RLA, being the most serious clinical condition in animals that received 15g/kg of body weight of sucrose. Supplementation with sodium monensin at the dose utilized did not prevent the disease from occurring. The evaluation of blood biochemistry confirmed the diagnosis and helped in the monitoring of goat RLA and the intensity of the clinical condition induced by both substrate doses was not sufficient to cause cardiac injury and elevation of CK-MB and cTnI.

EFEITO TERAPÊUTICO DO CLORETO DE MAGNÉSIO NA INTOXICAÇÃO POR Palicourea marcgravii (RUBIACEAE) EM OVINOS

Palicourea marcgravii é a principal planta dentro de um grupo de 22 espécies que causam morte súbita no Brasil. Seu princípio tóxico é o monofluoracetato de sódio. O presente estudo objetivou verificar a eficácia terapêutica do cloreto de magnésio em ovinos intoxicados experimentalmente por Palicourea marcgravii, e ainda, descrever os achados clínico-patológicos e hematológicos que possam auxiliar no diagnóstico ante mortem precoce dessa intoxicação. Apesar de observado maior tempo de evolução, em relação a outros trabalhos utilizando-se a mesma dose da P. marcgravii em ovinos, todos os ovinos intoxicados vieram à óbito. Os ovinos apresentaram sinais clínicos prévios como apatia, taquicardia, jugulares ingurgitadas, pulso venoso positivo, tosse, movimentos de cavar o solo, perda de equilíbrio, queda, decúbito esternal, posição de auto auscultação, movimentos mastigatórios, ranger de dentes, polaquiuria, inquietação e tremores musculares antes do óbito, além de respiração abdominal, tosse, head pressing e nistagmo, sinais nunca antes descritos, e morreram em poucos minutos. Dentre os resultados hematológicos observou-se alteração de hemoglobina, concentração de hemoglobina corpuscular média, hiperfibrinogenemia, hiperglicemia, leucocitose com neutrofilia, elevação sérica de AST, GLDH, ureia, creatinina, CK, LDH, CK-MB, hipofosfatemia e a detecção do extravasamento da cTnI. Alterações de necropsia como jugular ingurgitada, congestão de grandes vasos, edema pulmonar, líquido pericárdico, edema adjacente à vesícula biliar, fígado com bordos arredondados. Microscopicamente observou-se degeneração hidrópico vacuolar dos túbulos uriníferos contorcidos distais, alterações essas semelhantes as já relatadas em estudos anteriores. Aos exames de eletro e ecocardiograma foram detectadas alterações das quais concluiu-se que a causa do óbito foi uma insuficiência cardíaca aguda. Apesar do cloreto de magnésio não evitar a morte dos animais intoxicados o curso da doença foi mais longo quando comparado a estudos anteriores.

Palicourea marcgravii is the main plant within a group of 22 species that cause sudden death in Brazil. Its toxic principle is sodium monofluoracetate. The present study aimed to verify the therapeutic efficacy of magnesium chloride in sheep experimentally intoxicated by Palicourea marcgravii, and also to describe the clinical-pathological and hematological findings that may assist in the early ante-mortem diagnosis of this intoxication. Despite observing a longer evolution time, in relation to other studies using the same dose of P. marcgravii in sheep, all intoxicated sheep died. The sheep showed previous clinical signs such as apathy, tachycardia, engorged jugulars, positive venous pulse, cough, movements of digging the soil, loss of balance, falling, sternal decubitus, "self-auscultation" position, masticatory movements, teeth grinding, polaquiuria , restlessness and muscle tremors before death, in addition to abdominal breathing, coughing, "head pressing" and nystagmus, signs never before described, and died within minutes. Among the hematological results, changes in hemoglobin, mean corpuscular hemoglobin concentration, hyperfibrinogenemia, hyperglycemia, leukocytosis with neutrophilia, serum elevation of AST, GLDH, urea, creatinine, CK, LDH, CK-MB, hypophosphatemia and the detection of extravasation were observed cTnI. Necropsy changes such as engorged jugular, congestion of large vessels, pulmonary edema, pericardial fluid, edema adjacent to the gallbladder, liver with rounded edges. Microscopically, there was a vacuolar hydropic degeneration of distal contorted urine tubules, changes similar to those already reported in previous studies. Electrocardiogram and echocardiogram exams revealed changes that concluded that the cause of death was acute heart failure. Although magnesium chloride does not prevent the death of intoxicated animals, the disease course was longer when compared to previous studies.