Cholinesterase activities and oxidative stress in cattle experimentally exposed to nitrate/nitrite in cultivated pasture with different fertilization schemes

Background: Nitrate and nitrite poisoning is associated with pasture intake that has high nitrate levels and leads to acute methemoglobinemia. Pasture may accumulate nitrate under certain conditions, such as excessively fertilized soil or environmental conditions that enhance the N absorption (rain preceded by a period of drought). After ingestion of plants, this substrate reaches the rumen and, in physiological conditions, is reduced to nitrite and afterward to ammonia. The aim of this study was to evaluate changes in cholinesterase activities and oxidative stress caused by subclinical poisoning for nitrate and nitrite in cattle fed with Pennisetum glaucum in three different fertilization schemes.Materials, Methods & Results: In order to perform the experimental poisoning, the pasture was cultivated in three different paddocks: with nitrogen topdressing (urea; group 1), organic fertilizer (group 2) or without fertilizer (group 3; control). Nitrate accumulation in forage was evaluated by the diphenylamine test. After food fasting of 12 h, nine bovine were randomly allocated to one of the experimental groups and fed with fresh forage (ad libitum) from respective paddock. In different time points from beginning of pasture intake (0, 2, 4, 6 and 9 h) heart rate and respiratory frequency were assessed, as well as mucous membrane color and behavioral changes. Blood samples from jugular vein into vials with and without anticoagulant were collected. From blood samples, serum nitrite levels, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzyme activity were evaluated, as well as oxidative stress through the following parameters: levels of nitrate/nitrite (NOx ), thiobarbituric acid reactive substances (TBARS) and reactive oxygen species (ROS), beyond the antioxidant system by enzyme activity measurement of catalase (CAT) and superoxide dismutase (SOD).[...]

Cholinesterase activities and oxidative stress in cattle experimentally exposed to nitrate/nitrite in cultivated pasture with different fertilization schemes

Background: Nitrate and nitrite poisoning is associated with pasture intake that has high nitrate levels and leads to acute methemoglobinemia. Pasture may accumulate nitrate under certain conditions, such as excessively fertilized soil or environmental conditions that enhance the N absorption (rain preceded by a period of drought). After ingestion of plants, this substrate reaches the rumen and, in physiological conditions, is reduced to nitrite and afterward to ammonia. The aim of this study was to evaluate changes in cholinesterase activities and oxidative stress caused by subclinical poisoning for nitrate and nitrite in cattle fed with Pennisetum glaucum in three different fertilization schemes.Materials, Methods & Results: In order to perform the experimental poisoning, the pasture was cultivated in three different paddocks: with nitrogen topdressing (urea; group 1), organic fertilizer (group 2) or without fertilizer (group 3; control). Nitrate accumulation in forage was evaluated by the diphenylamine test. After food fasting of 12 h, nine bovine were randomly allocated to one of the experimental groups and fed with fresh forage (ad libitum) from respective paddock. In different time points from beginning of pasture intake (0, 2, 4, 6 and 9 h) heart rate and respiratory frequency were assessed, as well as mucous membrane color and behavioral changes. Blood samples from jugular vein into vials with and without anticoagulant were collected. From blood samples, serum nitrite levels, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzyme activity were evaluated, as well as oxidative stress through the following parameters: levels of nitrate/nitrite (NOx ), thiobarbituric acid reactive substances (TBARS) and reactive oxygen species (ROS), beyond the antioxidant system by enzyme activity measurement of catalase (CAT) and superoxide dismutase (SOD).[...](AU)

Benefits of a prepartum anionic diet on the health of dairy cows in the transition period: prevented subclinical hypocalcemia and minimizing oxidative stress

Background: There are many metabolic diseases that affect dairy cows during the transition period, among them, the postparturient hypocalcemia, also known as milk fever, puerperal paresis syndrome, or fallen cow. This disease may affect animals 72 h after calving due to a nutritional imbalance in the calcium metabolism, which occurs after a sudden demand of this mineral at the beginning of lactation. This problem is more common in animals with high milk production, and usually after the second lactation. Therefore, this study evaluated whether the anionic diet on the prepartum phase would be beneficial for dairy cows in the transition period. Materials, Methods & Results: Eight Holstein cows on their second and third pregnancy with body condition score between 3.5 and 4 were used. During the experiment, the animals remained in paddock with grazing Tifton 85 (Cynodon spp.) and water ad libitum. Two groups of four animals each were used: the group A (control) received a dietary cation-anion difference (DCAD) diet of 2.57 mEq kg-1, and the second group, called the group B (treatment), received a DCAD diet of -112.2 mEq kg-1. To evaluate animal health, we performed gynecological examinations, in addition to blood samplings on days 20, 10 and 5 prepartum and 3, 7, 12 and 20 postpartum to assess seric levels of calcium (Ca), total protein, albumin, globulin, cholesterol, triglyceride, urea, and hepatic function (AST, ALT and GGT). It was also measured the production of free radicals (ROS) and lipid peroxidation (TBARS) in the transition period of the cows. No statistical differences were observed between groups regarding body condition score, uterine tone, dominant follicle diameter, size and presence of the corpus luteum and the presence of genital discharge up to 30 days postpartum. However, the anionic diet prevented a dramatic reduction in calcium levels, unlike what occurred in the control cows (the group A) where a subclinical hypocalcemia was observed.[...]

Benefits of a prepartum anionic diet on the health of dairy cows in the transition period: prevented subclinical hypocalcemia and minimizing oxidative stress

Background: There are many metabolic diseases that affect dairy cows during the transition period, among them, the postparturient hypocalcemia, also known as milk fever, puerperal paresis syndrome, or fallen cow. This disease may affect animals 72 h after calving due to a nutritional imbalance in the calcium metabolism, which occurs after a sudden demand of this mineral at the beginning of lactation. This problem is more common in animals with high milk production, and usually after the second lactation. Therefore, this study evaluated whether the anionic diet on the prepartum phase would be beneficial for dairy cows in the transition period. Materials, Methods & Results: Eight Holstein cows on their second and third pregnancy with body condition score between 3.5 and 4 were used. During the experiment, the animals remained in paddock with grazing Tifton 85 (Cynodon spp.) and water ad libitum. Two groups of four animals each were used: the group A (control) received a dietary cation-anion difference (DCAD) diet of 2.57 mEq kg-1, and the second group, called the group B (treatment), received a DCAD diet of -112.2 mEq kg-1. To evaluate animal health, we performed gynecological examinations, in addition to blood samplings on days 20, 10 and 5 prepartum and 3, 7, 12 and 20 postpartum to assess seric levels of calcium (Ca), total protein, albumin, globulin, cholesterol, triglyceride, urea, and hepatic function (AST, ALT and GGT). It was also measured the production of free radicals (ROS) and lipid peroxidation (TBARS) in the transition period of the cows. No statistical differences were observed between groups regarding body condition score, uterine tone, dominant follicle diameter, size and presence of the corpus luteum and the presence of genital discharge up to 30 days postpartum. However, the anionic diet prevented a dramatic reduction in calcium levels, unlike what occurred in the control cows (the group A) where a subclinical hypocalcemia was observed.[...](AU)

Oxidative stress and changes on the adenosinergic system of cats infected by feline leukemia virus (FeLV)

Background: The feline leukemia virus (FeLV) is clinically important retroviruses that infect domestic and wild feline worldwide, affecting more than 3 million cats representing a great risk for premature death. Some studies have demonstrated that oxidative stress, as well as the adenosinergic system, exert an important role in the pathogenesis of viral diseases. Oxidative stress is considered a disturbance in the antioxidant/oxidant status in favor of the excessive generation or lower removal of free radicals. Therefore, the aim of this study was to evaluate whether changes on the adenosinergic system and oxidative stress occurred in cats positives for feline leukemia virus (FeLV). Materials, Methods & Results: Forty-nine serum samples of cats (between 4 months and 13 years of age) seen at the Veterinary Hospital of the University of Santa Catarina State (UDESC - Lages, SC, Brazil) were used. Blood samples were collected from the jugular vein and stored in tubes without anticoagulant to obtain serum. An aliquot was used to detect both viral infections using the kit SNAP FIV/FeLV, that detects the p27 protein from FeLV and antibodies against the p24 protein from FIV. The test was performed according the manufacture’s recommendations. Based on this result, this study involved 20 FeLV negative cats, 20 FeLV positive symptomatic cats, and 9 FeLV positives asymptomatic cats. Serum samples were used to determine the activities of adenosine deaminase (ADA) and glutathione-S-transferase (GST), as well as thiobarbituric acid reactive substances (TBARS) and reactive oxygen levels (ROS). Seric ADA and GST activities, as well as TBARS (lipid peroxidation) and ROS (free radical) levels were determined spectrophotometrically according to the specific method. Seric ADA activity was increased in symptomatic animals compared to negatives and also to asymptomatic positive animals, and the same was observed for seric TBARS levels.[...]

Oxidative stress and changes on the adenosinergic system of cats infected by feline leukemia virus (FeLV)

Background: The feline leukemia virus (FeLV) is clinically important retroviruses that infect domestic and wild feline worldwide, affecting more than 3 million cats representing a great risk for premature death. Some studies have demonstrated that oxidative stress, as well as the adenosinergic system, exert an important role in the pathogenesis of viral diseases. Oxidative stress is considered a disturbance in the antioxidant/oxidant status in favor of the excessive generation or lower removal of free radicals. Therefore, the aim of this study was to evaluate whether changes on the adenosinergic system and oxidative stress occurred in cats positives for feline leukemia virus (FeLV). Materials, Methods & Results: Forty-nine serum samples of cats (between 4 months and 13 years of age) seen at the Veterinary Hospital of the University of Santa Catarina State (UDESC - Lages, SC, Brazil) were used. Blood samples were collected from the jugular vein and stored in tubes without anticoagulant to obtain serum. An aliquot was used to detect both viral infections using the kit SNAP FIV/FeLV, that detects the p27 protein from FeLV and antibodies against the p24 protein from FIV. The test was performed according the manufactures recommendations. Based on this result, this study involved 20 FeLV negative cats, 20 FeLV positive symptomatic cats, and 9 FeLV positives asymptomatic cats. Serum samples were used to determine the activities of adenosine deaminase (ADA) and glutathione-S-transferase (GST), as well as thiobarbituric acid reactive substances (TBARS) and reactive oxygen levels (ROS). Seric ADA and GST activities, as well as TBARS (lipid peroxidation) and ROS (free radical) levels were determined spectrophotometrically according to the specific method. Seric ADA activity was increased in symptomatic animals compared to negatives and also to asymptomatic positive animals, and the same was observed for seric TBARS levels.[...](AU)

Activities of the enzymes NTPDase, 5'-nucleotidase and adenosine deaminase in platelets of rats experimentally infected with Leptospira interrogans serovar icterohaemorrhagiae

Background: Leptospirosis, a spirochetal zoonotic disease caused by different serovars of Leptospira interrogans, is increasingly recognized as an important cause of hemorrhagic fever. Although the haemorrhagic potential of leptospirosis was noted by Weil (1886) as early as 1886, its pathophysiology is still not clearly elucidated, particularly regarding the cause and mechanisms of bleeding. Studies with ectonucleoside triphosphate diphosphohydrolase (NTPDase; EC 3.6.1.5; CD39), 5'-nucleotidase (EC 3.1.3.5; CD73) and adenosine deaminase (ADA) have demonstrated the involvement of these enzymes in thromboregulation mechanisms, and altered enzymatic activities have been reported in many diseases. Since leptospirosis is a disease increasingly recognized as an important cause of hemorrhagic fever, the aim of this study was to evaluate these enzymes activities and parameters of platelet aggregation in platelets from rats experimentally infected with Leptospira interrogans serovar icterohaemorrhagiae during different periods of experimental infection. Materials, Methods & Results: For this purpose, thirty-six adult male rats were divided into two groups: A, as uninfected control (subgroups A1, A2 and A3); and B, infected (subgroups B1, B2 and B3). Group B was inoculated intraperitoneally (Day 0) with 2 x 108 organisms per rat. Blood samples were collected on days 05 (A1 and B1), 10 (A2 and B2) and 20 (A3 and B3) post-inoculation (PI). In the infected group, platelet count had a decrease on day 10 PI and prothrombin time (PT) had an increase on day 5 PI. In the same group, platelet aggregation decreased (P < 0.01) day 10 PI. The hydrolysis of ATP in platelets was also decreased (P < 0.05) on day 10 PI, when compared to the control group. By the other side, ADP hydrolysis was increased (P < 0.05) on days 5 and 10 PI. 5'-nucleotidase activity was significantly increased on day 5 (P < 0.01) and 20 (P < 0.05) PI. Results of adenosine deamination into inosine by ADA in platelets showed a signifi cant (P < 0.01) increase on days 5 and 10 PI in the infected group. Discussion: Studies with NTPDase, 5'nucleotidase and ADA have demonstrated the involvement of these enzymes in the thromboregulation mechanisms, and altered enzymatic activities have been reported in many diseases. It has been established that extracellular adenosine nucleotides and adenosine are versatile signaling molecules known to participate in an array of platelet functions. For example, the nucleotide ADP is the main promoter of platelet aggregation, while adenosine can act as a vasodilator and an inhibitor of platelet aggregation. In addition, high concentrations of ATP have been shown to inhibit ADP-induced aggregation in vitro, while low concentrations of ATP can significantly enhance platelet aggregation. In our experimental study the coagulation cascade was activated, since when the activities of NTPDase, 5'-nucleotidase and ADA were analyzed is possible to suggest that levels of ATP were decreased, unlike of ADP and AMP levels, supposedly increased during determinate periods of our experiment. Adenosine levels were also enhanced due to the higher levels of its precursors. This cascade activation may be a mechanism of bleeding prevention front to leptospires infection, especially the ones caused by serovar icterohaemorrhagiae.

NTPDase and acetylcholinesterase activities in silver catfish, Rhamdia quelen (Quoy & Gaimard, 1824) (Heptapteridae) exposed to interaction of oxygen and ammonia levels

The effects of various levels of oxygen saturation and ammonia concentration on NTPDase (ecto-nucleoside triphosphate diphosphohydrolase, E.C. 3.6.1.5) and acetylcholinesterase (AChE, E.C. 3.1.1.7) activities in whole brain of teleost fish (Rhamdia quelen) were investigated. The fish were exposed to one of two different dissolved oxygen levels, including high oxygen (6.5 mg.L-1) or low oxygen (3.5 mg.L-1), and one of two different ammonia levels, including high ammonia (0.1 mg.L-1) or low ammonia (0.03 mg.L-1) levels. The four experimental groups included the following (A) control, or high dissolved oxygen plus low NH3; (B) low dissolved oxygen plus low NH3; (C) high dissolved oxygen plus high NH3; (D) low dissolved oxygen plus high NH3. We found that enzyme activities were altered after 24 h exposure in groups C and D. ATP and ADP hydrolysis in whole brain of fish was enhanced in group D after 24 h exposure by 100 percent and 119 percent, respectively, compared to the control group. After 24 h exposure, AChE activity presented an increase of 34 percent and 39 percent in groups C and D, respectively, when compared to the control group. These results are consistent with the hypothesis that low oxygen levels increase ammonia toxicity. Moreover, the hypoxic events may increase blood flow by hypoxia increasing NTPDase activity, thus producing adenosine, a potent vasodilator.(AU)

No presente estudo, avaliou-se o efeito de diferentes níveis de saturação de oxigênio e amônia sobre a atividade das enzimas NTPDase (ecto-nucleosídeo trifosfato difosfohidrolase, E.C. 3.6.1.5) e acetilcolinesterase (AChE, E.C. 3.1.1.7) em encéfalo total de jundiás (Rhamdia quelen). Os peixes foram expostos a diferentes níveis de oxigênio dissolvido e amônia, níveis altos de oxigênio (6,5 mg/L) ou baixos de oxigênio (3,5 mg/L), e níveis altos de amônia (0,1 mg/L) ou baixos de amônia (0,03 mg/L). Os peixes foram divididos em quatro diferentes grupos: (A) controle ou alto nível de oxigênio dissolvido mais baixo nível de NH3; (B) baixo nível de oxigênio dissolvido mais baixo nível de NH3; (C) alto nível de oxigênio dissolvido mais alto nível de amônia; (D) baixo nível de oxigênio dissolvido mais alto nível de NH-3. As atividades de ambas as enzimas nos grupos C e D somente foram alteradas após 24 horas de exposição. A hidrólise do ATP e ADP em encéfalo total de jundiás foi aumentada após 24h de exposição para 104 por cento e 155 por cento no grupo D quando comparado ao grupo controle, respectivamente. A atividade da AChE apresentou após 24h de exposição um aumento de 37 por cento no grupo C e 27 por cento no grupo D, ambos comparados ao grupo controle. Os resultados obtidos corroboram com a hipótese que baixos níveis de saturação de oxigênio aumentam a toxicidade da amônia. Além disso, os eventos de hipóxia podem aumentar o fluxo sanguíneo, e este evento aumenta a atividade da NTPDase produzindo adenosina, um potente vasodilatador(AU)

NTPDase and acetylcholinesterase activities in silver catfish, Rhamdia quelen (Quoy & Gaimard, 1824) (Heptapteridae) exposed to interaction of oxygen and ammonia levels

The effects of various levels of oxygen saturation and ammonia concentration on NTPDase (ecto-nucleoside triphosphate diphosphohydrolase, E.C. 3.6.1.5) and acetylcholinesterase (AChE, E.C. 3.1.1.7) activities in whole brain of teleost fish (Rhamdia quelen) were investigated. The fish were exposed to one of two different dissolved oxygen levels, including high oxygen (6.5 mg.L-1) or low oxygen (3.5 mg.L-1), and one of two different ammonia levels, including high ammonia (0.1 mg.L-1) or low ammonia (0.03 mg.L-1) levels. The four experimental groups included the following (A) control, or high dissolved oxygen plus low NH3; (B) low dissolved oxygen plus low NH3; (C) high dissolved oxygen plus high NH3; (D) low dissolved oxygen plus high NH3. We found that enzyme activities were altered after 24 h exposure in groups C and D. ATP and ADP hydrolysis in whole brain of fish was enhanced in group D after 24 h exposure by 100 percent and 119 percent, respectively, compared to the control group. After 24 h exposure, AChE activity presented an increase of 34 percent and 39 percent in groups C and D, respectively, when compared to the control group. These results are consistent with the hypothesis that low oxygen levels increase ammonia toxicity. Moreover, the hypoxic events may increase blood flow by hypoxia increasing NTPDase activity, thus producing adenosine, a potent vasodilator.(AU)

No presente estudo, avaliou-se o efeito de diferentes níveis de saturação de oxigênio e amônia sobre a atividade das enzimas NTPDase (ecto-nucleosídeo trifosfato difosfohidrolase, E.C. 3.6.1.5) e acetilcolinesterase (AChE, E.C. 3.1.1.7) em encéfalo total de jundiás (Rhamdia quelen). Os peixes foram expostos a diferentes níveis de oxigênio dissolvido e amônia, níveis altos de oxigênio (6,5 mg/L) ou baixos de oxigênio (3,5 mg/L), e níveis altos de amônia (0,1 mg/L) ou baixos de amônia (0,03 mg/L). Os peixes foram divididos em quatro diferentes grupos: (A) controle ou alto nível de oxigênio dissolvido mais baixo nível de NH3; (B) baixo nível de oxigênio dissolvido mais baixo nível de NH3; (C) alto nível de oxigênio dissolvido mais alto nível de amônia; (D) baixo nível de oxigênio dissolvido mais alto nível de NH-3. As atividades de ambas as enzimas nos grupos C e D somente foram alteradas após 24 horas de exposição. A hidrólise do ATP e ADP em encéfalo total de jundiás foi aumentada após 24h de exposição para 104 por cento e 155 por cento no grupo D quando comparado ao grupo controle, respectivamente. A atividade da AChE apresentou após 24h de exposição um aumento de 37 por cento no grupo C e 27 por cento no grupo D, ambos comparados ao grupo controle. Os resultados obtidos corroboram com a hipótese que baixos níveis de saturação de oxigênio aumentam a toxicidade da amônia. Além disso, os eventos de hipóxia podem aumentar o fluxo sanguíneo, e este evento aumenta a atividade da NTPDase produzindo adenosina, um potente vasodilatador(AU)

Presence of multiple acid phosphatases activity in seedlings of cucumber, radish and rocket salad / Presença de atividade de múltiplas fosfatases ácidas em plântulas de pepino, rabanete e rúcula

Acid phosphatases (3.1.3.2) are a group of enzymes widely distributed in nature, which catalyze the hydrolysis of a variety of phosphate esters in the pH range of 4-6. We confirmed the presence of acid phosphatases in seedlings of cucumber (Cucumis sativus), radish (Raphanus sativus) and rocket salad (Eruca vesicaria) under different assay conditions using a rapid and simple preparation. The results showed that the optimum pH and temperature used for all species were close to 5.5 and 35ºC, respectively. The enzyme was inhibited by molybdate, fluoride, azide, levamisole, orthovanadate, Zn2+ and Cu2+. Suramin had no effect on enzyme activity. The acid phosphatase from cucumber, radish and rocket salad hydrolyzed a wide variety of phosphate esters and the highest activity was observed with PPi, ATP and GTP. These results demonstrate that the enzyme investigated in this study is different from well known ester phosphate cleaving plant enzymes (apyrase and inorganic pyrophosphatases) and this preparation could be a useful tool to future toxicological studies and to study initially all isoforms of acid phosphatase.(AU)

As fosfatases ácidas (3.1.3.2) são um grupo de enzimas amplamente distribuídas na natureza, as quais catalisam a hidrólise de uma variedade de ésteres de fosfato com uma variação de pH entre quatro e seis. Foi confirmada a presença de fosfatases ácidas em plântulas de pepino (Cucumis sativus), rabanete (Raphanus sativus) e rúcula (Eruca vesicaria) sob diferentes condições de ensaio usando uma preparação rápida e simples. Os resultados mostraram que o pH e a temperatura ótimos para todas as espécies foram 5,5 e 35ºC, respectivamente. A enzima foi inibida por molibdato, fluoreto, azida, levamisole, ortovanadato, Zn2+ e Cu2+. O inibidor suramim não afetou a atividade enzimática. As fosfatases ácidas de pepino, rabanete e rúcula hidrolisaram uma ampla variedade de ésteres de fosfato e a maior atividade foi observada com PPi, ATP e GTP para pepino e rabanete e PPi, frutose-6-fosfato e GTP para rúcula. Esses resultados demonstraram que a enzima investigada neste estudo é diferente das conhecidas enzimas de plantas que clivam ésteres de fosfato (apirase e pirofosfatases inorgânicas). Desse modo, esta preparação pode ser uma ferramenta útil para futuros estudos toxicológicos e para se estudar inicialmente todas as isoformas das fosfatases ácidas.(AU)