RESUMO
Dairy cattle are at the greatest risk of developing diseases around the time of calving because of compromised immune responses and the occurrence of oxidative stress. Both the development of compromised immunity and oxidative stress are influenced directly or indirectly by the metabolism of polyunsaturated fatty acids (PUFA) and fat-soluble vitamins. The cytochrome P450 (CYP450) family of enzymes is central to the metabolism of both classes of these compounds, but to date, the importance of CYP450 in the health of dairy cattle is underappreciated. As certain CYP450 isoforms metabolize both PUFA and fat-soluble vitamins, potential interactions may occur between PUFA and fat-soluble vitamins that are largely unexplored. For example, one CYP450 that generates anti-inflammatory oxylipids from arachidonic acid additionally contributes to the activation of vitamin D. Other potential substrate interactions between PUFA and vitamins A and E may exist as well. The intersection of PUFA and fat-soluble vitamin metabolism by CYP450 suggest that this enzyme system could provide an understanding of how immune function and oxidant status interconnect, resulting in increased postpartum disease occurrence. This review will detail the known contributions of bovine CYP450 to the regulation of oxylipids with a focus on enzymes that may also be involved in the metabolism of fat-soluble vitamins A, D, and E that contribute to antioxidant defenses. Although the activity of specific CYP450 is generally conserved among mammals, important differences exist in cattle, such as the isoforms primarily responsible for activation of vitamin D that makes their specific study in cattle of great importance. Additionally, a CYP450-driven inflammatory positive feedback loop is proposed, which may contribute to the dysfunctional inflammatory responses commonly found during the transition period. Establishing the individual enzyme isoform contributions to oxylipid biosynthesis and the regulation of vitamins A, D, and E may reveal how the CYP450 family of enzymes can affect inflammatory responses during times of increased susceptibility to disease. Determining the potential effect of each CYP450 on disease susceptibility or pathogenesis may allow for the targeted manipulation of the CYP450 pathways to influence specific immune responses and antioxidant defenses during times of increased risk for health disorders.
Assuntos
Doenças dos Bovinos/enzimologia , Sistema Enzimático do Citocromo P-450/metabolismo , Inflamação/veterinária , Animais , Anti-Inflamatórios , Bovinos , Ácidos Graxos Insaturados/metabolismo , Feminino , Inflamação/enzimologia , Estresse Oxidativo/fisiologia , Vitaminas/metabolismoRESUMO
Fatty acids (FA) provide an energy source to the liver during negative energy balance; however, when FA influx is excessive, FA can be stored as liver lipids or incompletely oxidized to ß-hydroxybutyrate (BHB). The objectives of this study were to quantify plasma and liver FA profiles and hepatic gene expression in cows diagnosed with hyperketonemia (HYK; BHB ≥ 1.2 mM) or not (nonHYK; BHB < 1.2 mM) to determine a relationship between FA profile and expression of hepatic genes related to oxidation and gluconeogenesis. Production parameters, blood samples (-28, -3, 1, 3, 5, 7, 9, 11, and 14 d relative to parturition; n = 28 cows), and liver biopsies (1, 14, and 28 d postpartum; n = 22 cows) were collected from Holstein cows. Cows were retrospectively grouped as HYK or nonHYK based on BHB concentrations in postpartum blood samples. Average first positive test (BHB ≥ 1.2 mM) was 9 ± 5 d (± SD). Cows diagnosed with HYK had greater C18:1 and lower C18:2 plasma proportions. Liver FA proportions of C16:0 and C18:1 were related to proportions in plasma, but C18:0 and C18:2 were not. Some interactions between plasma FA and HYK on liver FA proportion suggests that there may be preferential use depending upon metabolic state. Cows diagnosed with HYK had decreased pyruvate carboxylase (PC) expression, but no difference at 1 d postpartum in either cytosolic or mitochondrial isoforms of phosphoenolpyruvate carboxykinase (PCK). The increased PC to PCK ratios in nonHYK cows suggests the potential for greater hepatic oxidative capacity, coinciding with decreased circulating BHB. Interestingly, FA, known regulators of PC expression, were not correlated with PC expression at 1 d postpartum. Taken together, these data demonstrate that HYK cows experience a decrease in the ratio of hepatic PC to PCK at 1 day postpartum prior to HYK diagnosis which, on average, manifested a week later. The differential regulation of PC involved in HYK diagnosis may not be completely due to shifts in FA profiles and warrants further investigation.
Assuntos
Doenças dos Bovinos/enzimologia , Cetose/veterinária , Piruvato Carboxilase/metabolismo , Idade de Início , Animais , Bovinos , Doenças dos Bovinos/metabolismo , Ácidos Graxos/sangue , Ácidos Graxos/metabolismo , Feminino , Cetose/enzimologia , Cetose/metabolismo , Fígado/metabolismo , Estudos RetrospectivosRESUMO
The objective of this study was to evaluate the association between ketonemia and serum paraoxonase-1 (PON1), malondialdehyde (MDA), and other blood components in tail and mammary veins of dairy cows. Forty-two Holstein dairy cows with decreased feed intake were divided into HIGH (≥ 1.2 mM; n = 31) and LOW (< 1.2 mM; n = 11) groups based on the ß-hydroxybutyrate concentration in plasma collected from the tail vein. The HIGH group had a significantly greater plasma non-esterified fatty acid (NEFA) concentration, but significantly lower serum PON1 activity and phospholipid concentration, and a tendency to have a lower cholesterol ester concentration than the LOW group. Serum PON1 activity was not correlated with the MDA concentration but was positively correlated with serum concentrations of cholesterol esters and phospholipids, and negatively correlated with the plasma NEFA concentration. These results suggest that serum PON1 activity is reduced by hyperketonemia and the relevance of PON1 to MDA seems to not be direct, though it is involved.
L'objectif de la présente étude était d'évaluer l'association entre l'acétonémie et la paraoxonase-1 (PON1), le malondialdéhyde (MDA), et d'autres composés du sang dans les veines caudale et mammaire de vaches laitières. Quarante-deux vaches laitières de race Holstein présentant une diminution de l'ingestion d'aliments furent divisées en groupes ÉLEVÉ (≥ 1,2 mM; n = 31) et BAS (< 1,2 mM; n = 11) basés sur la concentration de ß-hydroxybutyrate de plasma prélevé de la veine caudale. Le groupe ÉLEVÉ avait une concentration plasmatique significativement plus grande d'acides gras non-estérifiés (NEFA), mais le sérum présentait une activité PON1 et une concentration de phospholipides significativement réduite, et une tendance à avoir une concentration d'esters de cholestérol plus faible que le groupe BAS. L'activité de PON1 sérique n'était pas corrélée avec la concentration de MDA mais était corrélée positivement avec les concentrations sériques d'esters de cholestérol et de phospholipides, et corrélée négativement avec la concentration plasmatique de NEFA. Ces résultats suggèrent que l'activité de PON1 sérique est réduite par l'hypercétonémie et la pertinence de PON1 envers MDA ne semble pas être directe, bien qu'elle semble impliquée.(Traduit par Docteur Serge Messier).
Assuntos
Arildialquilfosfatase/sangue , Doenças dos Bovinos/enzimologia , Cetose/veterinária , Malondialdeído/sangue , Ácido 3-Hidroxibutírico/sangue , Animais , Análise Química do Sangue/veterinária , Bovinos , Doenças dos Bovinos/sangue , Ésteres do Colesterol/sangue , Colorimetria/veterinária , Ácidos Graxos não Esterificados/sangue , Feminino , Cetose/sangue , Cetose/enzimologia , Glândulas Mamárias Animais/irrigação sanguínea , Fosfolipídeos/sangue , Cauda/irrigação sanguíneaRESUMO
High blood concentrations of nonesterified fatty acids (NEFA) and altered lipid metabolism are key characteristics of fatty liver in dairy cows. In nonruminants, the mitochondrial membrane protein mitofusin 2 (MFN2) plays important roles in regulating mitochondrial function and intrahepatic lipid metabolism. Whether MFN2 is associated with hepatic lipid metabolism in dairy cows with moderate fatty liver is unknown. Therefore, to investigate changes in MFN2 expression and lipid metabolic status in dairy cows with moderate fatty liver, blood and liver samples were collected from healthy dairy cows (n = 10) and cows with moderate fatty liver (n = 10). To determine the effects of MFN2 on lipid metabolism in vitro, hepatocytes isolated from healthy calves were used for small interfering RNA-mediated silencing of MFN2 or adenovirus-mediated overexpression of MFN2 for 48 h, or treated with 0, 0.6, 1.2, or 2.4 mM NEFA for 12 h. Milk production and plasma glucose concentrations in dairy cows with moderate fatty liver were lower, but concentrations of NEFA and ß-hydroxybutyrate (BHB) were greater in dairy cows with moderate fatty liver. Dairy cows with moderate fatty liver displayed hepatic lipid accumulation and lower abundance of hepatic MFN2, peroxisome proliferator-activated receptor-α (PPARα), and carnitine palmitoyltransferase 1A (CPT1A). However, sterol regulatory element-binding protein 1c (SREBP-1c), acetyl CoA carboxylase 1 (ACACA), fatty acid synthase (FASN), and diacylglycerol acyltransferase 1 (DGAT1) were more abundant in the livers of dairy cows with moderate fatty liver. In vitro, exogenous NEFA treatment upregulated abundance of SREBP-1c, ACACA, FASN, and DGAT1, and downregulated the abundance of PPARα and CPT1A. These changes were associated with greater lipid accumulation in calf hepatocytes, and MFN2 silencing aggravated this effect. In contrast, overexpression of MFN2-ameliorated exogenous NEFA-induced lipid accumulation by downregulating the abundance of SREBP-1c, ACACA, FASN, and DGAT1, and upregulating the abundance of PPARα and CPT1A in calf hepatocytes. Overall, these data suggest that one cause for the negative effect of excessive NEFA on hepatic lipid accumulation is the inhibition of MFN2. As such, these mechanisms partly explain the development of hepatic steatosis in dairy cows.
Assuntos
Doenças dos Bovinos/metabolismo , Bovinos/metabolismo , Fígado Gorduroso/veterinária , GTP Fosfo-Hidrolases/metabolismo , Metabolismo dos Lipídeos , Fígado/metabolismo , Ácido 3-Hidroxibutírico/metabolismo , Animais , Bovinos/genética , Doenças dos Bovinos/enzimologia , Doenças dos Bovinos/genética , Diacilglicerol O-Aciltransferase/genética , Diacilglicerol O-Aciltransferase/metabolismo , Ácidos Graxos não Esterificados/metabolismo , Fígado Gorduroso/enzimologia , Fígado Gorduroso/genética , Fígado Gorduroso/metabolismo , Feminino , GTP Fosfo-Hidrolases/genética , Hepatócitos/enzimologia , Hepatócitos/metabolismo , Mitocôndrias/metabolismo , PPAR alfa/genética , PPAR alfa/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismoRESUMO
Dairy cows with ketosis display severe oxidative stress as well as high blood concentrations of non-esterified fatty acids (NEFA) and ß-hydroxybutyrate (BHB). Cytochrome P4502E1 (CYP2E1) plays an important role in the induction of oxidative stress. The aim of this study was to investigate CYP2E1 expression and activity in the liver of clinically ketotic cows (in vivo) and the effects of NEFA and BHB on CYP2E1 expression and activity in hepatocytes (in vitro). Dairy cows with clinical ketosis exhibited a low blood concentration of glucose but high concentrations of NEFA and BHB. Hepatic mRNA, protein expression, and activity of CYP2E1 were significantly higher in cows with clinical ketosis than in control cows. In vitro, both NEFA and BHB treatment markedly up-regulated the mRNA and protein expressions as well as activity of CYP2E1 in cow hepatocytes. Taken together, these results indicate that high levels of NEFA and BHB significantly up-regulate the expression and activity of hepatic CYP2E1, and may be influential in the induction of oxidative stress in cows with clinical ketosis.
Assuntos
Ácido 3-Hidroxibutírico/farmacologia , Doenças dos Bovinos/enzimologia , Citocromo P-450 CYP2E1/efeitos dos fármacos , Ácidos Graxos não Esterificados/farmacologia , Cetose/enzimologia , Fígado/enzimologia , Ácido 3-Hidroxibutírico/sangue , Animais , Bovinos , Doenças dos Bovinos/sangue , China , Citocromo P-450 CYP2E1/genética , Citocromo P-450 CYP2E1/metabolismo , Indústria de Laticínios , Ácidos Graxos não Esterificados/sangue , Feminino , Expressão Gênica/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Hepatócitos/enzimologia , Cetose/sangue , Estresse Oxidativo/efeitos dos fármacos , RNA Mensageiro/análise , Regulação para Cima/efeitos dos fármacosRESUMO
Several evidences have suggested the involvement of enzymes belonging to the phosphotransfer network, formed by creatine kinase (CK), pyruvate kinase (PK) and adenylate kinase (AK), as well the oxidative stress on the pathogenesis of infectious diseases associated with the central nervous system (CNS). Thus, the aim of this study was to evaluate whether listeriosis alters the brain energy metabolism and/or causes oxidative stress in different brain structures of cattle experimentally infected by Listeria monocytogenes. The cytosolic CK activity was inhibited in the cerebral cortex, cerebellum, brainstem and hippocampus of infected animals compared to uninfected animals, while the mitochondrial CK activity was increased. The PK activity was inhibited in all brain structures of infected animals, while the AK activity was unchanged. Na+, K+-ATPase activity decreased in the cerebral cortex, cerebellum and hippocampus of animals infected by L. monocytogenes. Regarding the oxidative strees variables, the cerebellum and brainstem of infected animals showed increased thiobarbituric acid reactive substances, while the catalase activity was inhibited. Glutathione S-transferarase was inhibited in the cerebral cortex and brainstem of infected animals, and it was increased in the cerebellum. L. monocytogenes was quantified in the liver (nâ¯=â¯5/5) and cerebral cortex (nâ¯=â¯4/5) of the infected cattle. Based on these evidences, the nucleocytoplasmic communication between CK isoenzymes was insufficient to avoid an impairment of cerebral bioenergetics. Moreover, the inhibition on brain PK activity caused an impairment in the communication between sites of ATP generation and ATP utilization. The lipid peroxidation and alteration on antioxidant status observed in some brain structures were also involved during the disease. In summary, these alterations contribute to disease pathogenesis linked to CNS during cattle listeriosis.
Assuntos
Adenilato Quinase/metabolismo , Encéfalo/enzimologia , Doenças dos Bovinos/enzimologia , Creatina Quinase/metabolismo , Listeria monocytogenes/fisiologia , Listeriose/veterinária , Piruvato Quinase/metabolismo , Adenilato Quinase/genética , Animais , Antioxidantes/metabolismo , Encéfalo/metabolismo , Encéfalo/microbiologia , Bovinos , Doenças dos Bovinos/metabolismo , Doenças dos Bovinos/microbiologia , Creatina Quinase/genética , Creatina Quinase Mitocondrial/genética , Creatina Quinase Mitocondrial/metabolismo , Metabolismo Energético , Listeriose/enzimologia , Listeriose/metabolismo , Listeriose/microbiologia , Oxidantes/metabolismo , Estresse Oxidativo , Fosforilação , Piruvato Quinase/genéticaRESUMO
Nuclear factor (erythroid-derived 2)-like factor 2 (NFE2L2, formerly Nrf2) is a transcription factor that binds to the antioxidant response element (ARE) in the upstream promoter region of various antioxidant-responsive genes. Hence, at least in nonruminants, the NFE2L2-ARE signaling pathway plays an important role in the cellular antioxidant defense system. Whether oxidative stress in bovine mammary epithelial cells alters NFE2L2 or the NFE2L2-ARE pathway is unclear. Therefore, the objective of this study was to examine the response in NFE2L2- and NFE2L2-ARE-related components in bovine mammary epithelial cells (BMEC) under oxidative stress. An in silico analysis to identify potential phosphorylation sites on NFE2L2 and the protein kinases was performed with Netphos 3.1 (http://www.cbs.dtu.dk/services/NetPhos/) and Scansite (http://scansite.mit.edu) software. Isolated BMEC were exposed to H2O2 (600 µM) for 6 h to induce oxidative stress. In silico analysis revealed ataxia telangiectasia-mutated (ATM) serine/threonine kinase as a key kinase responsible for the phosphorylation of NFE2L2. Thus, after the 6 h incubation with H2O2, BMEC were transiently transfected with ATM-small interfering RNA (siRNA) 1, 2, or 3. Compared with the control, transfection with ATM-siRNA3 resulted in proliferation rates that were 60.7 and 36.2% lower with or without H2O2. In addition, production of reactive oxygen species and malondialdehyde increased markedly, but activities of superoxide dismutase, glutathione peroxidase, catalase, and glutathione-S-transferase decreased markedly in transfected cells without or with H2O2 compared with the control. Transfected cells had markedly lower protein and mRNA expression of NFE2L2 without or with H2O2 compared with the control. In addition, fluorescent activity of the ARE in transfected BMEC indicated that NFE2L2-driven transcriptional activation decreased under oxidative stress. Overall, results indicate that ATM is a physiologically relevant NFE2L2 kinase. Furthermore, inhibition of ATM activity can cause marked alterations in oxidative stress leading to cell death as a result of diminished capacity of BMEC to cope with H2O2-induced cytotoxicity. The relevance of this kinase in vivo merits further study.
Assuntos
Antioxidantes/metabolismo , Ataxia Telangiectasia/veterinária , Doenças dos Bovinos/enzimologia , Fator 2 Relacionado a NF-E2/metabolismo , Proteínas Serina-Treonina Quinases/genética , Ativação Transcricional , Animais , Elementos de Resposta Antioxidante/genética , Ataxia Telangiectasia/enzimologia , Ataxia Telangiectasia/fisiopatologia , Bovinos , Doenças dos Bovinos/fisiopatologia , Proliferação de Células/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/fisiologia , Feminino , Peróxido de Hidrogênio/farmacologia , Glândulas Mamárias Animais/efeitos dos fármacos , Glândulas Mamárias Animais/fisiologia , Fator 2 Relacionado a NF-E2/genética , Estresse Oxidativo , Fosforilação , Proteínas Serina-Treonina Quinases/metabolismo , RNA Interferente Pequeno , Espécies Reativas de Oxigênio/metabolismo , Transdução de SinaisRESUMO
This study was aimed at demonstrating associations between peripheral biochemical parameters, endometrial leukocyte esterase (LE) and myeloperoxidase (MPO), and bacterial detection with the degree of endometrial inflammation, and determining the best time postpartum for diagnosing endometritis to predict subsequent fertility in dairy cows. Plasma albumin, blood urea nitrogen (BUN), total cholesterol (T-cho), NEFA, and BHBA concentrations were analyzed in 43 Holstein cows at 3, 5 and 7 weeks postpartum (W3, W5 and W7). Endometrial samples were collected at W3, W5 and W7 to examine LE and MPO activities, bacterial detection rates, and PMN% profiles. The 43 cows were divided into healthy (HE), subclinical endometritis (SE), and clinical endometritis (CE) groups, classified differently at W3, W5 and W7 based on the definitions of SE and CE for each of the three weeks pp. LE level had an association with PMN% in all weeks pp (P<0.05). Albumin and BUN levels had weak negative associations with endometrial PMN% at W3. Pathogenic bacterial detection rates were higher in the cows with endometritis at W3 and W5. Conception rate at first artificial insemination tended to be lower (P=0.057) in the cows diagnosed with endometritis at W3 than in the healthy cows. In conclusion, associations were found between endometrial LE and endometritis, but not for MPO and endometritis. Diagnosing endometritis in W3 may be the best moment to predict subsequent fertility.
Assuntos
Bactérias/isolamento & purificação , Infecções Bacterianas/veterinária , Hidrolases de Éster Carboxílico/sangue , Doenças dos Bovinos/sangue , Endometrite/veterinária , Peroxidase/sangue , Animais , Infecções Bacterianas/enzimologia , Infecções Bacterianas/microbiologia , Bovinos , Doenças dos Bovinos/enzimologia , Doenças dos Bovinos/microbiologia , Endometrite/sangue , Endometrite/enzimologia , Endometrite/microbiologia , Endométrio/enzimologia , Endométrio/patologia , Feminino , Fertilidade , Período Pós-PartoRESUMO
Exploration of novel drug targets has been the major thrust area in filarial research. In this regard, identification and characterization of oxidative enzymes that play pivotal role in the survival of filarial parasite inside host are of immense importance. In this study, we are reporting the presence of an important redox regulatory enzyme, thioredoxin reductase (TrxR) in the bovine filarial parasite Setaria cervi. TrxR was found to be exists throughout the developmental stages viz. oocyte, microfilaria and adult of the parasite. Since further studies on this enzyme require adequate quantity, influential extraction parameters were optimized statistically using response surface methodology (RSM) employing a seven factors based Box-Behnken design matrix. ANOVA analysis revealed the relative importance of each parameter and a regression equation was eventually developed that could predict the specific activity (SA) of TrxR. Finally the optimized extraction conditions predicted by RSM was 6.1ml of 61.86mM buffer, pH 6.0, with extraction temperature 39.96°C for 180min in addition to 450rpm agitation and 20µl/ml of protease inhibitor. Therefore this study is going to be the maiden report depicting the identity of TrxR in filarial parasite and the optimized extraction conditions for its isolation with better kinetic efficiency.
Assuntos
Doenças dos Bovinos/parasitologia , Setaria (Nematoide)/enzimologia , Tiorredoxina Dissulfeto Redutase/química , Animais , Bovinos , Doenças dos Bovinos/enzimologia , Cinética , Microfilárias/enzimologia , Microfilárias/patogenicidade , Oócitos/enzimologia , Setaria (Nematoide)/patogenicidade , Especificidade por Substrato , Tiorredoxina Dissulfeto Redutase/isolamento & purificaçãoRESUMO
The aim of this study was to evaluate the influence of dictyocaulosis (mild or severe) on enzymes of NTPDase, 5'-nucleotidase, and adenosine deaminase (ADA) of dairy cows naturally infected by Dictyocaulus viviparus. Blood and faeces were collected from 22 dairy cows of the same farm to evaluate NTPDase (ATP and ADP substrate), 5'-nucleotidase, and ADA activities on days 0 (pre-treatment) and 10 (post-treatment). Seric activities of NTPDase (ATP substrate), 5'-nucleotidase, and ADA were lower (P<0.05) in D. viviparus infected animals compared to uninfected cows. The number of D. viviparus larvae per gram of faeces varied among the animals, and they showed different degrees of severity according to respiratory clinical signs of the disease (cough and nasal discharge). Later, these cows were divided into two groups: those with mild (n=10) and severe (n=12) disease. Cows with severe disease showed higher NTPDase activity (ATP substrate) than those with mild disease (P≤0.05). The opposite occurred with NTPDase (ADP substrate), 5'-nucleotidase, and ADA in cows with severe disease, that is, the enzymatic activity of these seric enzymes significantly decreased (P≤0.05) compared to animals with mild disease. Infected animals showed reduced NTPDase activity (ATP and ADP substrate) after treatment. No enzymatic changes were observed for 5'-nucleotidase, and ADA pre- and post-treatment (P>0.05). Based on these results, we conclude that dictyocaulosis alters NTPDase, 5'-nucleotidase, and ADA activities of cow naturally infected by the parasite, in consequence the enzymes act as inflammatory markers.
Assuntos
5'-Nucleotidase/sangue , Adenosina Desaminase/sangue , Biomarcadores/sangue , Doenças dos Bovinos/enzimologia , Infecções por Dictyocaulus/enzimologia , Animais , Bovinos , Doenças dos Bovinos/parasitologia , Dictyocaulus/isolamento & purificação , Infecções por Dictyocaulus/tratamento farmacológico , Infecções por Dictyocaulus/imunologia , Infecções por Dictyocaulus/parasitologia , Fezes/química , Inflamação , Pirofosfatases/sangueRESUMO
The enzymatic activities of NTPDase, 5'-nucleotidase and adenosine deaminase (ADA) are important in regulating the concentration of adenine nucleotides, molecules known to be involved on platelet aggregation. Fasciolosis causes coagulation disorders that have not been completely elucidated. Taking into consideration the association between the purinergic system and hemostasis, this study aimed to evaluate the enzymatic activities of NTPDase (hydrolyze ATP and ADP), 5'-nucleotidase (hydrolyze AMP) and ADA (deamination of adenosine) in platelets from cattle experimentally infected by Fasciola hepatica on days 20, 40, 60 and 80 post-infection (PI). For this study, 10 healthy Friesian steers were separated into two groups: the group A (n = 5) was used as uninfected control, and the group B was composed of steers experimentally infected by F. hepatica (n = 5). The number of platelets did not differ between groups in the periods evaluated. Reduction of NTPDase (p < 0.05) hydrolysing ATP (days 20, 40 and 60 PI), and ADP (days 40, 60 and 80 PI), and on 5'-nucleotidase hydrolyzing AMP (days 40 and 60 PI) was observed. A reduction (p < 0.05) in ADA activity on day 20 PI, as well as an increase (p < 0.05) in ADA activity on days 40 and 60 PI was observed when compared to the control. Based on these results, we can conclude that ATP, ADP and AMP hydrolysis and adenosine deamination were altered in platelets of cattle infected by F. hepatica. Considering the importance of the purinergic system in hemostasis, it is believed that those changes may contribute to the coagulation impairment observed in acute fasciolosis described in the literature.
Assuntos
Adenosina Desaminase/sangue , Plaquetas/enzimologia , Doenças dos Bovinos/parasitologia , Fasciolíase/veterinária , Nucleotidases/sangue , Animais , Bovinos , Doenças dos Bovinos/sangue , Doenças dos Bovinos/enzimologia , Fasciola hepatica/fisiologia , Fasciolíase/sangue , Fasciolíase/enzimologia , Fezes/parasitologia , Fígado/parasitologia , Fígado/patologia , Masculino , Contagem de Ovos de Parasitas/veterinária , Contagem de Plaquetas/veterináriaRESUMO
The enzyme adenosine deaminase (ADA) is critical for modulating the immune system, and in the presence of zinc, its activity is catalyzed. The aim of this study was to evaluate the ADA activity in pancreas of cattle naturally infected by Eurytrema coelomaticum in relation to the results of zinc levels, pathological findings and parasite load. For this study 51 slaughtered cattle were used. The animals were divided into two groups: Group A consisting of animals naturally infected by E. coelomaticum (n=33) and Group B of uninfected animals (n=18). Blood and pancreas were collected of each animal for analysis of zinc and ADA, respectively. Infected cattle showed a reduction on seric levels of zinc, and decreased ADA activity in the pancreas (P>0.05). A positive correlation between zinc levels and ADA activity was observed. Thus, high parasite load and severity of histopathologic lesions affect the ADA activity in pancreas, as well as the zinc levels in serum of infected animals (negative correlation between these variables). Therefore, we can conclude that cattle infected by E. coelomaticum have low ADA activity in pancreas, which can be directly related to zinc reduction, responsible for ADA activation and catalyzes.
Assuntos
Adenosina Desaminase/metabolismo , Doenças dos Bovinos/metabolismo , Dicrocoeliidae/fisiologia , Pâncreas/parasitologia , Carga Parasitária/veterinária , Infecções por Trematódeos/veterinária , Zinco/sangue , Animais , Bovinos , Doenças dos Bovinos/enzimologia , Doenças dos Bovinos/parasitologia , Pâncreas/metabolismo , Espectrofotometria/veterinária , Infecções por Trematódeos/enzimologia , Infecções por Trematódeos/metabolismo , Infecções por Trematódeos/parasitologiaRESUMO
The aim of this study was to evaluate the oxidative stress in serum and liver and adenosine deaminase (ADA) activity of cattle experimentally infected by Fasciola hepatica. The group A consisted of five healthy animals (uninfected), and the group B was composed of five animals orally infected with 200 metacercariae of F. hepatica. On days 20, 40, 60 and 80 post-infection (PI) serum was collected to measure oxidative stress variables. On day 100 PI, animals were humanely euthanized and liver samples were collected. Infected animals showed lower (P < 0·05) seric ADA activities on days 40 and 60 PI but higher (P < 0·05) in the liver tissue compared with uninfected animals. Seric and hepatic reactive oxygen species (ROS) were higher (P < 0·05) in infected compared with uninfected animals. Hepatic thiobarbituric acid reactive substances were higher (P < 0·05) in infected animals. Catalase and glutathione S-transferase activities were lower in liver tissue of infected animals, while glutathione peroxidase was higher compared with uninfected (P < 0·05). In summary, we conclude that oxidative stress occurs in cattle experimentally infected by F. hepatica, mainly due to excessive ROS production in the course of fasciolosis, contributing to hepatic damage, and that increased in hepatic ADA activity may contribute to the inflammatory process.
Assuntos
Adenosina Desaminase/metabolismo , Doenças dos Bovinos/parasitologia , Fasciola hepatica , Fasciolíase/veterinária , Estresse Oxidativo/fisiologia , Adenosina Desaminase/sangue , Adenosina Desaminase/genética , Animais , Biomarcadores , Bovinos , Doenças dos Bovinos/enzimologia , Doenças dos Bovinos/metabolismo , Fasciolíase/enzimologia , Fasciolíase/parasitologia , Regulação Enzimológica da Expressão Gênica , Fígado/enzimologia , Fígado/patologia , Masculino , Espécies Reativas de OxigênioRESUMO
Uridine 5'-diphospho-glucuronosyltransferase (UGT) liver activity was measured using estradiol-17ß as a substrate in dairy cows with follicular cysts. The activity was significantly lower than that in dairy cows with normal estrous cycles (P<0.01). Liver disorders, such as fatty liver and hepatitis, were observed in half cows with follicular cysts, and liver UGT activity was lower than that in cows with normal estrus cycles. In addition, the liver UGT activity was significantly lower in dairy cows with follicular cysts without liver disorders than in dairy cows with normal estrous cycles. Therefore, the cows were divided into those with low, middle and high liver UGT activities, and liver disorder complication rates were investigated. The complication rate was significantly higher in the low- (78.1%) than in the middle- (22.2%) and high-level (8.3%) groups, suggesting that liver disorders are closely associated with the development of follicular cysts in dairy cows and that steroid hormone metabolism is delayed because of reduced liver UGT activity, resulting in follicular cyst formation. We conclude that reduced estradiol-17ß glucuronidation in the liver and liver disorders are associated with follicular cyst occurrence in dairy cows.
Assuntos
Doenças dos Bovinos/etiologia , Estradiol/metabolismo , Cisto Folicular/veterinária , Glucuronosiltransferase/metabolismo , Falência Hepática/complicações , Fígado/enzimologia , Animais , Bovinos , Doenças dos Bovinos/enzimologia , Ciclo Estral/metabolismo , Feminino , Cisto Folicular/enzimologia , Cisto Folicular/etiologia , Falência Hepática/enzimologia , Folículo Ovariano/metabolismoRESUMO
The most important regulators of tissue remodelling during ovarian follicular growth, development, ovulation and atresia are gonadotropins, steroid hormones, growth factors and different proteolytic enzymes. Matrix metalloproteinases (MMPs) such as collagenase or gelatinase (i.e. MMP-1, -8, -2 and -9) and associated tissue inhibitors of metalloproteinases (TIMP-1, -2, -3 and -4) control connective tissue remodelling during follicular rupture. In this study, we hypothesized that an imbalance in the MMP-TIMP system may be an intra-ovarian component that contributes to the pathogenesis of cystic ovarian disease (COD) in cows. Taking into account that the control of MMP activity by TIMPs could determine their effects in both physiological and pathological conditions, MMP and TIMP mRNA and protein expression was examined by real-time polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC), respectively, in ovaries from control cows and cows with COD. Expression of mRNA encoding MMP-2, TIMP-1 and TIMP-2 was lower in follicular cysts than in control pre-ovulatory follicles, while the results by IHC showed this imbalance only for TIMP-2 protein expression. Additional analysis by zymography to evaluate the gelatinase activity of MMP-2 and MMP-9 demonstrated higher MMP-2 activity in follicular fluid (FF) of cysts than in FF of pre-ovulatory follicles. On the other hand, MMP-9 activity was increased in follicular cysts and absent in the FF of pre-ovulatory follicles. These findings suggest that the altered mRNA expression and activities of the MMP-TIMP system may be related to the failure in ovulation and follicular development observed in COD.
Assuntos
Doenças dos Bovinos/enzimologia , Metaloproteinases da Matriz/metabolismo , Cistos Ovarianos/veterinária , Inibidores Teciduais de Metaloproteinases/metabolismo , Animais , Western Blotting , Bovinos , Feminino , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Evidence suggests that the intracellular bacterial pathogen Coxiella burnetii (which causes Q fever) is widespread, with a near global distribution. While there has been increasing attention to Q fever epidemiology in high-income settings, a recent systematic review highlighted significant gaps in our understanding of the prevalence, spatial distribution and risk factors for Q fever infection across Africa. This research aimed to provide a One Health assessment of Q fever epidemiology in parts of Western and Nyanza Provinces, Western Kenya, in cattle and humans. A cross-sectional survey was conducted: serum samples from 2049 humans and 955 cattle in 416 homesteads were analysed for C. burnetii antibodies. Questionnaires covering demographic, socio-economic and husbandry information were also administered. These data were linked to environmental datasets based on geographical locations (e.g., land cover). Correlation and spatial-cross correlation analyses were applied to assess the potential link between cattle and human seroprevalence. Multilevel regression analysis was used to assess the relationships between a range of socio-economic, demographic and environmental factors and sero-positivity in both humans and animals. The overall sero-prevalence of C. burnetii was 2.5% in humans and 10.5% in cattle, but we found no evidence of correlation between cattle and human seroprevalence either within households, or when incorporating spatial proximity to other households in the survey. Multilevel modelling indicated the importance of several factors for exposure to the organism. Cattle obtained from market (as opposed to those bred in their homestead) and those residing in areas with lower precipitation levels had the highest sero-prevalence. For humans, the youngest age group had the highest odds of seropositivity, variations were observed between ethnic groups, and frequent livestock contact (specifically grazing and dealing with abortion material) was also a risk factor. These results illustrate endemicity of C. burnetii in western Kenya, although prevalence is relatively low. The analysis indicates that while environmental factors may play a role in cattle exposure patterns, human exposure patterns are likely to be driven more strongly by livestock contacts. The implication of livestock markets in cattle exposure risks suggests these may be a suitable target for interventions.
Assuntos
Anticorpos Antibacterianos/sangue , Doenças dos Bovinos/sangue , Coxiella burnetii/isolamento & purificação , Febre Q/sangue , Adolescente , Adulto , Animais , Bovinos , Doenças dos Bovinos/enzimologia , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/microbiologia , Criança , Pré-Escolar , Coxiella burnetii/genética , Coxiella burnetii/imunologia , Estudos Transversais , Feminino , Humanos , Lactente , Quênia/epidemiologia , Gado , Masculino , Pessoa de Meia-Idade , Prevalência , Febre Q/epidemiologia , Febre Q/imunologia , Febre Q/microbiologia , Estudos Soroepidemiológicos , Adulto Jovem , Zoonoses/sangue , Zoonoses/epidemiologia , Zoonoses/imunologia , Zoonoses/microbiologiaRESUMO
The aim of this study was to evaluate seric NTPDase and 5'nucleotidase activities of cattle naturally infected by Eurytrema coelomanticum, as well as to correlate them to histopathological lesions in the pancreas and the degree of parasitism. Blood samples and pancreas of 51 bovines were collected on a slaughterhouse in Southern Brazil: 33 from cattle naturally infected by E. coelomanticum (the Group A), and 18 from uninfected animals (the Group B). Infected animals showed an average of 532 parasites per pancreas. In the pancreatic histology, ducts displayed hyperplasia, stenosis, proliferation of fibrous tissue, and interstitial inflammatory infiltration of lymphocytes. The serum from infected animals showed an increase in NTPDase activity when ATP was used as substrate (P<0.001). For the ADP substrate, there was no difference between groups regarding NTPDase activity (P=0.37), as well as 5'-nucleotidase activity (P=0.27). Correlating NTPDase activity (ATP substrate) with the degree of histopathological lesions (rho=0.66, P<0.001) and the parasitic load on the pancreas (rho=0.65, P<0.001), a positive correlation was observed. Similar results were found between the degree of histopathological lesions and NTPDase activity (ADP substrate; rho=0.29, P=0.03), and 5'nucleotidase activity (rho=0.35, P=0.01). Based on the results of NTPDase and 5'nucleotidase enzymes in cattle naturally infected by E. coleomanticum, it is possible to suggest that these enzymes are involved in the modulation of inflammation, and they can act as markers of inflammatory response.
Assuntos
5'-Nucleotidase/sangue , Apirase/sangue , Doenças dos Bovinos/patologia , Dicrocoeliidae , Inflamação/veterinária , Pâncreas/parasitologia , Infecções por Trematódeos/veterinária , Matadouros , Animais , Antígenos CD , Biomarcadores/sangue , Brasil , Bovinos , Doenças dos Bovinos/enzimologia , Doenças dos Bovinos/parasitologia , Inflamação/enzimologia , Inflamação/patologia , Linfócitos , Pâncreas/patologia , Carga Parasitária , Infecções por Trematódeos/enzimologia , Infecções por Trematódeos/parasitologia , Infecções por Trematódeos/patologiaRESUMO
The bovine papillomavirus E1 helicase is essential for viral replication. In dividing cells, DNA replication maintains, but does not increase, the viral genome copy number. Replication is limited by low E1 expression and an E1 nucleocytoplasmic shuttling mechanism. Shuttling is controlled in part by phosphorylation of E1 by cellular kinases. Here we investigate conserved sites for phosphorylation by kinase CK2 within the E1 nuclear localization signal. When these CK2 sites are mutated to either alanine or aspartic acid, no change in replication phenotype is observed, and there is no effect on the subcellular distribution of E1, which remains primarily nuclear. This demonstrates that phosphorylation of E1 by CK2 at these sites is not a factor in regulating viral DNA replication in dividing cells.
Assuntos
Papillomavirus Bovino 1/metabolismo , Caseína Quinase II/metabolismo , Doenças dos Bovinos/enzimologia , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/metabolismo , Sinais de Localização Nuclear/metabolismo , Infecções por Papillomavirus/veterinária , Proteínas Virais/química , Proteínas Virais/metabolismo , Sequência de Aminoácidos , Animais , Papillomavirus Bovino 1/química , Papillomavirus Bovino 1/genética , Caseína Quinase II/genética , Bovinos , Doenças dos Bovinos/genética , Doenças dos Bovinos/virologia , Núcleo Celular/virologia , Proteínas de Ligação a DNA/genética , Dados de Sequência Molecular , Sinais de Localização Nuclear/química , Sinais de Localização Nuclear/genética , Infecções por Papillomavirus/enzimologia , Infecções por Papillomavirus/genética , Infecções por Papillomavirus/virologia , Fosforilação , Transporte Proteico , Proteínas Virais/genéticaRESUMO
Bovine cutaneous fibropapillomas are benign skin tumours characterized by epithelial and dermal proliferation and induced by Bovine papillomaviruses (BPVs). Cyclooxygenase (COX) 1 and 2 are enzymes involved in pathological conditions, such as inflammation and epithelial carcinogenesis. Here we investigated biochemically and immunohistochemically COX-2 expression in bovine cutaneous fibropapillomas. Eight of twelve fibropapillomas (67%) showed COX-2 positive immunosignal mostly in the cytoplasm of the basal cell layer, while the normal skin did not stain. Biochemical analysis confirmed the expression of COX-2 in tumour samples. This study shows COX-2 expression in cutaneous fibropapillomas, suggesting a contribution in epithelial tumour development.
Assuntos
Doenças dos Bovinos/enzimologia , Ciclo-Oxigenase 2/metabolismo , Deltapapillomavirus , Infecções por Papillomavirus/veterinária , Dermatopatias Virais/veterinária , Animais , Bovinos , Doenças dos Bovinos/metabolismo , Doenças dos Bovinos/patologia , Ciclo-Oxigenase 2/genética , Infecções por Papillomavirus/enzimologia , Infecções por Papillomavirus/metabolismo , Infecções por Papillomavirus/patologia , Dermatopatias Virais/enzimologia , Dermatopatias Virais/metabolismoRESUMO
OBJECTIVE: To investigate whether expression of genes associated with inflammation and activity of indoleamine-2,3-dioxygenase (IDO) correlated with disease status and prevalence of bacteremia in post-partum dairy cattle with and without acute puerperal metritis (APM). PROCEDURES: Blood was collected from cattle with APM and control cattle matched by parity and days in milk. Leukocytes were isolated and expression of 6 genes was quantified. Activity of IDO was measured in serum with higher performance liquid chromatography (HPLC). RESULTS: The relative expression of IL-1ß in cattle with APM was significantly lower than that in controls. IDO activity was not significantly different between bacteremic and non-bacteremic cattle CONCLUSIONS AND CLINICAL RELEVANCE: The expression of IL-1ß was lower in cattle with APM. The lower levels of IL-1ß expression in PBMCs of cattle with APM suggest impaired inflammatory responses and may contribute to the development of the disease in this population of animals.