The Cardiorenal Effects of Piper amalago Are Mediated by the Nitric Oxide/Cyclic Guanosine Monophosphate Pathway and the Voltage-Dependent Potassium Channels.

Piper amalago L. is used in Brazilian traditional medicine to treat inflammation, chest pain, and anxiety. This study aimed to investigate the safety and the renal and cardiovascular effects of the volatile oil (VO) and the aqueous (AE) and hydroalcoholic (HE) extracts from P. amalago. The gas chromatography-mass spectrometry analyses identified 47 compounds in the VO, with ß-cyclogermacrene, spathulenol, ß-phellandrene, and α-pinene standing out. Among the 47 compounds also found in AE and HE by liquid chromatography-mass spectrometry, glycosylated flavones, organic acids, amino acids, and amides were highlighted. Some examples of these compounds are methoxy-methylenedioxy cis-cinnamoyl pyrrolidine, methoxy-methylenedioxy trans-cinnamoyl pyrrolidine, and cyclobutene-2,4-bis-(1,3-benzodioxol-5-methoxy-6-yl)-1,3-dicarboxapyrrolidide. The acute toxicity experiments were conducted on female rats (n = 5). The cardiorenal assays (n = 8) and evaluations of vasodilatory effects on the mesenteric vascular bed (n = 5) were conducted on male rats. In either extract or VO, there were no mortality or changes in relative weights or histopathological analysis of the organs. Urinary volume and renal electrolyte excretion were elevated significantly during repeated dose 7-day treatment with different preparations from P. amalago. None of the preparations induced hypotension or changes in cardiac electrical activity. Only HE promoted significant vasodilatory effects in rats' isolated mesenteric vascular beds. These effects were completely abolished in the presence of L-NAME plus 4-aminopyridine. Therefore, P. amalago leaves are safe and present diuretic activity after acute and repeated dose administration over 7 days. Moreover, the HE induced significant vasodilator response in rats' mesenteric vascular beds by NO/cGMP pathway and voltage-dependent K+ channels activation.

Short-term carcinogenesis evaluation of Casearia sylvestris

Abstract Casearia sylvestris Sw., Salicaceae, is an important medicinal plant widely used in Brazil for the treatment of various cardiovascular disorders. This species was included as of interest by Brazilian Unified Health System. Although preclinical studies described cardiovascular protective effects and apparent absence of toxicity, no studies have evaluated its carcinogenic potential. In this study, we proposed a short-term carcinogenesis evaluation of C. sylvestris in Wistar rats, aiming to check the safety of this species to use it as proposed by Brazilian Unified Health System. C. sylvestris leaves were obtained and the crude extract was prepared by maceration from methanol/water. Wistar rats were orally treated for 12 weeks with 50, 250 or 500 mg kg−1 of crude extract or vehicle. Body weight, daily morbidity and mortality were monitored. Blood and bone marrow samples were collect for micronucleus test, comet assay and tumor markers evaluation. Vital organs were removed to macro and histopathological analyses. The crude extract did not induce mutagenic and genotoxic effects and no alterations were observed in important tumor markers. Finally, no detectable signs of injury through gross pathology or histopathological examinations were observed. Our results certify the absence of the crude extract toxicity, indicating its safety, even at prolonged exposure as proposed by Brazilian Unified Health System.

Estimativas de perdas econômicas causadas por reação granulomatosa local após uso de vacina oleosa contra febre aftosa em bovinos de Mato Grosso do Sul / Estimates of economic losses caused by local granulomatous reaction after use of an oily vaccine against FMD in cattle of Mato Grosso do Sul

Lesões exacerbadas à vacina contra a febre aftosa foram observadas em 1.815 de um total de 5.000 bovinos abatidos em frigorífico com inspeção federal. Essas lesões resultaram na condenação de parte das carcaças de acordo com o Regulamento da Inspeção Industrial e Sanitária de Produtos de Origem Animal. Os bovinos haviam sido vacinados em maio de 2012 e abatidos em setembro do mesmo ano e divididos em Lotes 1 e 2 constituídos respectivamente por 1.500 e 315 bovinos. As lesões de reação vacinal eram caracterizadas por nódulos protuberantes, circunscritos, bem delimitados, firmes e de tamanhos variáveis, distribuídos multifocalmente na musculatura do aspecto lateral do pescoço; ao corte apresentavam exsudato purulento. Amostras da lesão de 28 bovinos foram avaliadas histologicamente e os principais achados foram piogranulomas com espaços claros circulares de tamanhos variados no centro, circundados por intenso infiltrado de neutrófilos íntegros e necróticos, circundados por macrófagos epitelioides com citoplasma vacuolizado, ocasionais células gigantes multinucleadas e, mais externamente, por abundante tecido conjuntivo em meio observavam-se linfócitos e plasmócitos. Culturas bacterianas realizadas em amostras do exsudato da lesão de seis bovinos resultaram negativas. Quando as carcaças afetadas foram submetidas ao toalete foi retirado em média 1,8 e 2,0 kg de músculo da área afetada, respectivamente dos Lotes 1 e 2. O prejuízo econômico da propriedade de origem dos bovinos afetados foi de R$ 20.424,00, considerando o preço pago pela arroba do boi no mês e ano da ocorrência. Esses valores à época seriam suficientes para adquirir 29,17 bezerros desmamados para engorda. Se considerarmos 5.000 bovinos vacinados forem considerados como população sob risco o coeficiente de morbidade seria de 0,36%. Os resultados deste estudo demonstram que perdas por reação vacinal, mesmo quando não provocam sinais clínicos marcantes, podem ocasionar importantes prejuízos econômicos...

Lesions caused by excessive reaction to foot and mouth vaccine were observed in 1,815 out of 5,000 cattle slaughtered in a slaughterhouse under federal meat inspection. Such lesions resulted in condemnation of the affect parts of the carcasses in accordance with the Brazilian Federal Meat Inspection Regulations. The affected cattle had been vaccinated on May 2012 and then slaughtered in September of the same year. They were allotted to two groups with 1,500 (Groups 1) and 315 cattle (Group 2). Post vaccination lesions were characterized as salient, firm variable sized nodules multifocally distributed in the musculature of the lateral neck. Pus oozed at the cut surface of these nodules. Samples of 28 cattle were submitted for histopathological examination; main findings were pyogranulomas with central variable-sized clear spaces surrounded by marked infiltrate of viable and dead neutrophils, which were surrounded by epithelioid macrophages with vacuolated cytoplasm and occasional multinucleated giant cell. This inflammatory reaction was walled by abundant connective tissue infiltrated by lymphocytes and plasma cells. Samples of the lesion of six cattle were submitted to bacteriological culture with negative results. When affected carcasses went through dressing, there was an average of 1.8 and 2.0 kg of muscle tissue cut off from the carcasses respectively of Group 1 and 2. Economic losses in the farm of origin of the affected cattle was R$ 20,424.00, considering the price paid by kg of cattle for slaughter at the time of the occurrence. This amount at that time was enough to buy 29.17 weaned calves for fattening. If one considers theses 5,000 cattle vaccinated on the farm as the population at risk, the morbidity rate would be 0.36%. These results indicate that the losses due to vaccine reaction, even with no overt clinical signs, may cause important economic losses...

Administração repetida de doses não tóxicas de monofluoroacetato de sódio não protege contra a intoxicação por este composto em ovinos / Repeated administration of non-toxic doses of sodium monofluoroacetate does not protect against poisoning by this compound in sheep

Com o objetivo de avaliar se repetidas doses não tóxicas de monofluoroacetato de sódio (MFA) induzem resistência à intoxicação por essa substância, 18 ovinos foram distribuídos aleatoriamente em dois grupos experimentais de nove animais cada. Os ovinos do Grupo 1 ingeriram doses crescentes não letais de MFA por seis períodos: 0,05mg/kg por 5 dias; 0,08mg/kg por 4 dias; 0,08mg/kg por 4 dias; 0,1mg/kg por 3 dias; 0,1mg/kg por 3 dias e 0,25mg/kg por 3 dias. Entre o primeiro e o segundo período de administração e entre o segundo e o terceiro período os animais não receberam o MFA por 10 dias consecutivos; entre o terceiro e o quarto período e dentre os demais períodos de administração, os ovinos permaneceram 15 dias sem ingerir o MFA. Quinze dias após o último período de administração os ovinos foram desafiados com a dose única de 1mg/kg de MFA. O Grupo 2 não foi adaptado a ingestão de MFA, estes ovinos receberam dose única de 1mg/kg de MFA no mesmo período em que o G1 foi desafiado. No desafio sete ovinos do Grupo 1 apresentaram sinais clínicos da intoxicação e um ovino se recuperou. No Grupo 2 todos os animais manifestaram quadro clínico da intoxicação por MFA, no entanto, dois ovinos se recuperaram. Os coeficientes de mortalidade foram de 66,6 por cento para o Grupo 1 e de 77,7 por cento para o Grupo 2. Os resultados deste trabalho sugerem que a administração repetida de doses não tóxicas de MFA não protege contra a intoxicação aguda por este composto, portanto, outras alternativas para a profilaxia da intoxicação por plantas que contêm MFA deverão ser pesquisadas, principalmente a utilização intraruminal de bactérias que hidrolisam MFA.

With the objective to assess whether repeated non-toxic doses of sodium monofluoroacetate (MFA) induce resistance to poisoning by this compound, 18 sheep were randomly divided into two experimental groups of nine animals each. Sheep from Group 1 ingested non-lethal increasing doses of MFA for six periods: 0.05mg/kg for 5 days; 0.08mg/kg for 4 days; 0.08mg/kg for 4 days; 0.1mg/kg for 3 days; 0.1mg/kg for 3 days and 0.25mg/kg for 3 days. Between the first and second period of administration and between the second and third period the animals did not receive MFA for 10 consecutive days, between the third and fourth period and during the remaining periods of administration the sheep were left 15 days without ingesting MFA. Group 2 was not adapted to the ingestion of MFA and received a single dose of 1mg/kg of MFA at the same time that Group 1 was challenged. After challenge, seven sheep of Group 1 showed clinical signs of poisoning and one sheep recovered. In Group 2, all animals showed clinical signs of poisoning by MFA, however two sheep recovered. The mortality rate was 66.6 percent in Group 1 and 77.7 percent for Group 2. These results suggest that repeated administration of non-toxic doses of MFA does not protect against acute poisoning by this compound; therefore other alternatives of prophylaxis for poisoning by plants containing MFA should be searched, mainly the use of intraruminal bacteria that hydrolyze MFA.

Poisoning by Brachiaria brizantha in flocks of naïve and experienced sheep.

The aim of this work was to study the effects of protodioscin ingestion in two different flocks of sheep: a flock of 23 crossbreed Mato Grosso do Sul Native sheep raised on Brachiaria spp. pastures from birth (experienced flock) in the state of Mato Grosso do Sul; and another flock (naïve flock) of 18 crossbred Dorper × Santa Inês sheep raised in the state of Paraná in Paspalum notatum and Lolium multiflorum pastures. The two flocks grazed together in a Brachiaria brizantha pasture during a 140-day period in the rainy season. At the beginning of the experiment and every 14 days thereafter, blood samples were collected for determination of serum activities of gamma-glutamyl transferase (GGT) and aspartate aminotransferase (AST), and for determination of the icterus index. On the same days, samples of young, mature and old B. brizantha leaves were collected for protodioscin quantification. Naïve sheep were more susceptible to poisoning by B. brizantha than experienced sheep. Six sheep in the naïve flock were poisoned, and two of these died. Two sheep in the experienced flock were poisoned, and one of them died. The mean activities of serum GGT and AST were significantly higher in the naïve flock, also evidencing a higher susceptibility to the poison. These results suggest that flocks of sheep include animals with different degrees of resistance to Brachiaria spp. poisoning and that culling the susceptible animals may considerably increase of the resistance of the flock. The clinical signs and the lesions were similar to those previously reported. However, in sheep with black coats, the main clinical sign was weight loss without photosensitization-mediated dermatitis. One sheep from the experienced flock presented cirrhosis, with clinical signs of exercise intolerance. The protodioscin concentration (% DM) ranged from 0.87% to 2.58% (mean ± SD: 1.64 ± 0.58) in young leaves, 1.16%-2.53% (1.67 ± 0.44) in mature leaves and 0.98%-2.07% (1.52 ± 0.37) in old leaves. A negative relationship was found between saponin concentration and total cumulative precipitation.

Intoxicação espontânea por Senna obtusifolia em bovinos no Pantanal Sul-Mato-Grossense / Spontaneous poisoning by Senna obtusifolia in cattle of the southern Pantanal

Descreve-se um surto de intoxicação por Senna obtusifolia em bovinos no estado de Mato Grosso do Sul, na região do Pantanal. Em um lote de 313 novilhas, 165 adoeceram e morreram (coeficientes de morbidade de 52,7% e de letalidade de 100%). Os bovinos permaneceram no piquete infestado pela planta por 37 dias. Os sinais clínicos consistiram em relutância em movimentar-se, andar trôpego (incoordenação), decúbito esternal permanente, diminuição do tônus de língua, estado comportamental em alerta, mioglobinúria caracterizada por urina castanho-escuro e fezes ressecadas com ou sem muco (ocasionalmente diarreicas com estrias de sangue). As principais alterações macroscópicas observadas nos 12 bovinos necropsiados estavam nos músculos esqueléticos dos membros pélvicos e foram caracterizadas por graus variáveis de palidez nos grupos musculares. Histologicamente, a lesão mais relevante encontrada foi degeneração e necrose segmentar multifocal nos músculos estriados esqueléticos (miopatia degenerativa tóxica multifocal polifásica). O diagnóstico da intoxicação baseou-se na epidemiologia (massa de forragem e de planta tóxica, análise da lotação do piquete e análise da precipitação pluviométrica), no quadro clínico dos animais e nos achados de necropsia e histopatologia.

Senna sp. poisoning in livestock has been reported in several occasions in Brazil usually from southern Brazil and involving S. occidentalis as the culprit. The objective of this study is to report the occurrence of an outbreak of S. obtusifolia poisoning in cattle in the Pantanal Region of Mato Grosso do Sul, Brazil. In a herd of 313 heifers, 165 were affected and died (morbidity rate of 52.7% and lethality rate of 100%). The 313 heifers remained in the paddock infested by S. obtusifolia for 37 days. Clinical signs consisted of reluctance to move, incoordination, permanent sternal recumbency, decrease in the tonus of the tongue, alertness, myoglobinuria characterized by dark brown urine, and dry stools with or without mucus; or occasionally diarrhea with streaks of blood. The main gross findings in 12 necropsied cattle were in the skeletal muscles of the hind limbs, and were characterized by varying degrees of paleness of muscle groups. Histologically, the most relevant lesion was segmental multifocal degeneration and necrosis in striated skeletal muscles (multifocal lypolyphasic toxic degenerative myopathy). The epidemiological, clinical and pathological data allowed to conclude for the diagnosis of poisoning by S. obtusifolia in this outbreak.

Intoxicação experimental por Simarouba versicolor (Simaroubaceae) em ovinos e indução de resistência ao consumo da planta / Experimental poisoning by Simarouba versicolor (Simaroubaceae) in sheep and induced resistance to consumption of the plant

Simarouba versicolor é uma árvore semidecídua pertencente à família Simaroubaceae. Um surto de intoxicação por S. versicolor em bovinos por brotos da planta presente no pasto em Mato Grosso do Sul e sua reprodução experimental foram descritos. Esse estudo teve por objetivos verificar experimentalmente se os ovinos podem ser utilizados como modelo clínico-patológico no estudo da intoxicação por Simarouba versicolor St. Hil. (fam. Simaroubaceae), determinar se há indução de resistência pela ingestão de pequenas e repetidas doses e, se a planta mantém sua toxicidade quando dessecada. Foram realizados dois experimentos, sendo o primeiro com folhas verdes ou folhas dessecadas e trituradas de S. versicolor em doses únicas de 5g/kg, 5g/kg e 3g/kg a três ovinos (Ovino 1, 2 e 3 respectivamente). O experimento 2, foi realizado com diferentes doses diárias de folhas dessecadas e trituradas de S. versicolor em quatro ovinos que receberam 1,5g/kg, 0,75g/kg, 0,6g/kg e 0,3g/kg e, com um ovino que recebeu 3g/kg como controle positivo (Ovino 4). A administração foi suspensa quando os ovinos apresentaram sinais clínicos da intoxicação. Após doze dias de recuperação, os animais sobreviventes foram desafiados com a mesma dose diária da planta ingerida anteriormente para avaliar o desenvolvimento de resistência. Os sinais clínicos observados nos dois experimentos caracterizaram-se por anorexia, mucosas oculares congestas, polidipsia, sialorreia, fezes pastosas que evoluíram para diarreia líquida fétida esverdeada, decúbito lateral e morte para os Ovinos 1 a 7. As principais lesões histológicas observadas foram necrose do tecido linfoide (linfonodos, baço, placas de Peyer) e enterite necrosante. Com os resultados obtidos, pode-se concluir que os ovinos podem ser utilizados como modelo experimental clínico-patológico na intoxicação por S. versicolor. Com o método utilizado, não houve resistência ao consumo diário de folhas da planta pelos ovinos e, as folhas mantiveram sua toxicidade quando dessecadas.(AU)

Simarouba versicolor St. Hil. is a semideciduous tree belonging to the Simaroubaceae family. An outbreak of poisoning in cattle by shoots of S. versicolor present in the pasture in Mato Grosso do Sul and experimental reproduction of the poisoning was described. This study aimed to verify experimentally whether sheep could be used as a clinical-pathological model in the study of the poisoning caused by S. versicolor, to determine if there develops resistance induced by ingestion of small and repeated doses of the leaves, and if the plant keeps its toxicity when dried. Two experiments were conducted: Experiment 1 with green leaves or dried and powdered leave of S. versicolor, given in single doses of 5g/kg, 5g/kg and 3g/kg to three sheep (Sheep 1, 2 and 3 respectively). Experiment 2 was made with different daily doses of dried and powdered leaves of S. versicolor; to four sheep was given 1.5g/kg, 0.75g/kg, 0.6g/kg and 0.3g/kg, and the positive control (Sheep 4) received 3g/kg. The administration was suspended when the animals showed clinical signs of poisoning. After twelve days of recovery, the surviving sheep were challenged with the same daily dose given previously, to assess the development of resistance. Clinical signs observed in both experiments were characterized by anorexia, congested ocular mucosa, polydipsia, drooling, loose stools which evolved into fetid greenish watery diarrhea, lateral decumbency and death of Sheep 1 to 7. The main histological lesions observed were necrosis of lymphoid tissue (lymph nodes, spleen, Peyer's patches) and necrotizing enteritis. With the results it can be concluded that sheep can be used as experimental model for the clinic-pathological aspects of poisoning by S. versicolor. The method used has not shown resistance to the daily consumption of the plant by the sheep, and the leaves kept their toxicity when dried.(AU)

Spontaneous and experimental intoxication of cattle by Simarouba versicolor A. St.-Hill (Simaroubaceae).

This study describes an outbreak of Simarouba versicolor intoxication in cattle from Mato Grosso do Sul, Brazil, and reproduces it experimentally. Clinical signs of the affected animals were weakness, tremors, hind limbs incoordination, reluctance to move, sternal and lateral recumbency and death. The main necropsy findings, observed in the abomasum and in segments of the small and large intestines, were diffuse redness and mucosal and serosal swelling. Histological examination revealed necrosis of lymphoid tissues and necrotizing enterocolitis. One experiment was carried out using 3 male calves to test the toxicity of a single dose of S. versicolor leaves at 15 g/kg, 5 g/kg and 2.5 g/kg. Clinical signs, necropsy findings and histological examination of calves receiving 15 g/kg and 5 g/kg leaves were similar to those of cattle from the intoxication outbreak. The calf fed 2.5 g/kg leaves developed clinical symptoms of poisoning and recovered naturally. In a second experiment, two male calves received daily administration of S. versicolor leaves at 1.5 g/kg and 2.5 g/kg for 10 days. They developed clinical signs of intoxication within 24 h and recovered eight to nine days after the leaves were administered. These findings suggest that S. versicolor was responsible for the outbreak studied, although this plant does not have cumulative intoxication effects on cattle.