Effects of photobiomodulation therapy on Bothrops moojeni snake-envenomed gastrocnemius of mice using enzymatic biomarkers.

Bothropic venom contains a range of biologically active substances capable of causing severe local and systemic envenoming symptomatology within its victims. The snake anti-venom is effective against systemic effects but has no neutralizing effect against the fast developing local effects. Herein, mice gastrocnemius injected with Bothrops moojeni venom (40 µg/kg) or saline solution were irradiated with HeNe (632.8 nm) and GaAs (904 nm) lasers (daily energy density of 4 J/cm2; 0.03/0.21 power density; 0.07/0.16 spot size; 1.2/0.04 total energy, 1 cm off contact, for HeNe and GaAs lasers, respectively) and euthanized in periods ranging from 3 h to 21 days. Blood biochemistry for creatine kinase (CK), alkaline phosphatase (ALP), acid phosphatase (AP), lactate dehydrogenase (LDH), aspartate transaminase (AST), and myoglobin and histopathological analysis, for assessing the degree of myonecrosis and regeneration of gastrocnemius, were done at every time interval. GaAs laser promoted faster photobiomodulation therapy (PBMT) effects, and the GaAs group exhibited a better clinical outcome than the HeNe group. Within the GaAs group, the serum levels of CK, LDH, AP, AST, and myoglobin, which were increased by the physiological effects of the venom, were reduced to initial baseline before snake envenomation in less time than those irradiated by the HeNe laser. However, the group receiving irradiation from the HeNe laser returned the levels of ALP activity to baseline faster than those of the GaAs group. Histopathological analysis revealed enhanced muscle regeneration in mice groups treated with both lasers. PBM promoted by GaAs and HeNe showed well-developed centrally nucleate regenerating cells and an increased number of newly formed blood vessels when compared to unirradiated muscle. We therefore suggest that GaAs had the best outcomes likely derived from a deeper penetrating longer wavelength. We conclude that PMBT is a promising, non-invasive approach to be further tested in pre-clinical studies with a goal to further its clinical use in skeletal muscle recovery in snakebite victims.

Antifungal potential and phytochemistry of Tetrapterys multiglandulosa CAV aerial parts / Potencial antifúngico e fitoquímica das partes aéreas de Tetrapterys multiglandulosa CAV

Tetrapterys spp. are considered toxic to grazing animals owing to the presence of a poisonous constituent cardiotonic glycoside. However, other chemicals within the plant have allelopathic, bactericidal, and fungicidal activities. The objective of this study was to search for natural products as alternatives in the control of plant pathogenic fungi, which diminish the crop productivity and cause damage to animal and human health. The phytochemical content of the stems and leaves was investigated alongside the fungitoxic potential of aqueous and ethanol extracts from aerial parts of T. multiglandulosa on Fusarium moniliforme and F. graminearum. The leaves and stems were obtained from 20 arrays of T. multiglandulosa from Batayporã City, south of Mato Grosso do Sul, and classified as young and mature. The botanical material was oven dried, crushed, and its aqueous and 20% ethanol extracts were used for qualitative and quantitative phytochemical assays. For in vitro tests of fungal growth inhibition, extract concentrations of 200, 400, 800, 1600, and 2000 µg L-1 were assayed, with two control groups. Phytochemical tests from aqueous and ethanol extracts of young and mature leaves indicated the presence of phenolic compounds, tannins, flavonoids, saponins, alkaloids, and cardiotonic glycosides. The aqueous extract of young leaves showed potential fungicidal activity against F. graminearum, with values ranging from 21.25% to 33.53% colony growth inhibition.

As espécies do gênero Tetrapterys são consideradas plantas tóxicas para animais de pastagem e um dos constituintes químicos responsáveis por estas intoxicações são os heterosideos cardiotônicos os quais também possuem atividade alelopática, bactericida, fungicida, dentre outras. Estes fatos e a busca por produtos naturais que possam ser empregados na agricultura familiar, como uma alternativa, no controle fungos fitopatogênicos que além de diminuir a produtividade podem causar danos à saúde animal e humana nos levou a investigar o potencial fungitóxico do extrato aquoso e etanólico das partes aéreas de Tetrapterys multiglandulosa sobre Fusarium moniliforme e F. graminearum e a determinar os fitoconstituintes. As folhas e caules foram obtidas de 20 matrizes de T. multiglandulosa na região de Batayporã, Mato Grosso do Sul e classificadas em jovens e adultas. O material botânico foi seco em estufa, triturado e os extratos aquoso e etanólico a 20% submetidos aos ensaios fitoquímicos quali e quantitativos. No teste in vitro de atividade de inibição de crescimento fúngico os extratos foram diluídos nas concentrações de 200, 400, 800, 1600 e 2000 µg mL-1 utilizando-se duas testemunhas. Testes fitoquímicos a partir de extratos aquosos e etanólicos de folhas e caules novos e maduros indicou a presença de compostos fenólicos taninos, flavonóides, alcalóides, saponinas e glicósideos cardiotônicos. O extrato aquoso das folhas jovens apresentou maior potencial fungicida contra F. graminearum com 33,53% de inibição de crescimento da colônia.

Performance of sheep grazing in pastures of Brachiaria decumbens, Brachiaria brizantha, Panicum maximum, and Andropogon gayanus with different protodioscin concentrations.

Brachiaria spp. are the most important grasses for ruminants in central-western Brazil. However, the use of these pastures is limited by their toxicity due to steroidal saponins. This experiment was conducted for 60 days to demonstrate the resistance of sheep raised on Brachiaria spp. pastures to steroidal saponin poisoning. The experiment was composed by 48 animals randomly divided into four groups (n = 12). Among them, 32 4- to 5-month-old castrated male crossbred Santa Inês sheep, originating from flocks that had been grazing on Brachiaria spp. for more than three consecutive years, and 16 were non-adapted (naïve) sheep from flocks that never had prior contact with pastures of Brachiaria spp. were randomly divided into four groups. Each of the four experimental groups was composed by eight adapted and four non-adapted animals. The four experimental groups were introduced into paddocks, each of which contained a single grass: either Brachiaria decumbens, Brachiaria brizantha, Panicum maximum, or Andropogon gayanus. The addition of the naïve sheep to the groups was designed to detect pastures' toxicity to naïve sheep and to adjust the stocking rate to optimize the use of forage. The weight gains of sheep grazing on B. decumbens, B. brizantha, and P. maximum were similar; however, the A. gayanus group showed lower weight gains compared with the other groups (P < 0.05). The mean serum activities of γ-glutamyltransferase in the sheep grazing on B. decumbens were higher than those in the sheep from the other groups. No significant differences among the groups were found in aspartate aminotransferase, creatinine, albumin, or total protein serum concentrations. No clinical signs were observed in the adapted sheep in any of the pastures. Of the four non-adapted sheep introduced into the B. decumbens pasture, two showed clinical signs of steroidal saponin poisoning, and one died. No clinical signs were observed in the non-adapted sheep in the other pastures. The saponin (protodioscin) concentrations in the pastures varied from 3.3 to 12.2 g/kg DM in B. decumbens, from 2.8 to 9.1 g/kg DM in B. brizantha, and from 1 to 1.5 g/kg DM in A. gayanus. No saponins were found in P. maximum. It is concluded that sheep from flocks reared in pastures of B. decumbens and B. brizantha were resistant to steroidal saponin poisoning and showed similar weight gains to those of sheep grazing in other tropical pastures.

Intoxication by Senna occidentalis seeds in pregnant goats: prenatal and postnatal evaluation.

Senna occidentalis is a weed toxic to different animal species. Very little is known about the effects of prolonged exposure to low doses of S. occidentalis on developmental toxicology. Thus, the present study proposes an approach to evaluate the perinatal toxicity of S. occidentalis seeds in goats. Twenty-one pregnant goats were fed rations containing 0% (control), 1% (So1 group), 2% (So2 group) and 4% (So4 group) mature S. occidentalis seeds from pregnancy detection on day 27 after mating until parturition; weight gains and serum biochemistry were evaluated. Fetuses were evaluated using ultrasonographic measurements; neonates were evaluated by body morphometry, weight gains, and serum biochemistry. Fetal resorption occurred in 2 So4 dams and one dam died. Only a few minor alterations in serum biochemistry occurred in dams and kids; even so one So4 group dam had tissue lesions as vacuolations in hepatocytes and kidneys; necrosis in skeletal and cardiac muscles and for the first time lesions were observed in sciatic nerve cells. No relevant alterations in body morphometry were observed. This study suggests that 4% S. occidentalis seeds is toxic for pregnant goats, but levels of seeds less than 4% have little impact on fetal and post birth body development.

Intoxicação por Brachiaria spp. em bovinos no Mato Grosso do Sul / Brachiaria spp. poisoning in cattle in Mato Grosso do Sul, Brazil

Mediante a revisão dos arquivos das fichas de necropsia do Laboratório de Patologia Animal, Universidade Federal do Mato Grosso do Sul, foram estudados a epidemiologia, o quadro clínico e a patologia de 29 surtos de intoxicação por Brachiaria spp., ocorridos em bovinos de corte, no Mato Grosso do Sul, de março de 1996 a novembro de 2009. Os surtos ocorreram em todas as épocas do ano, tanto na seca quanto na chuva. Em 24 dos 29 surtos o principal sinal clínico foi a fotossensibilização e em cinco o principal sinal foi o emagrecimento progressivo. Dos 24 surtos de fotossensibilização, 11 ocorreram em pastagens de B. decumbens, dois em pastagens mistas de B. decumbens e B. brizantha, um em B. brizantha e em 10 surtos não foi informada a espécie de Brachiaria envolvida. A morbidade variou de 0,2 por cento a 50 por cento e a letalidade de 44,4 por cento a 100 por cento. Nos casos de fotossensibilização o edema de barbela foi o sinal clínico mais encontrado em bovinos, seguido de dermatite com pele espessada no flanco e períneo, retração cicatricial auricular, icterícia, corrimento ocular. crostas auriculares e oculares, e ulcerações na parte ventral da língua. Em dois surtos foram observados sinais nervosos e em um, diarréia. Nas necropsias o fígado estava aumentado de tamanho, amarelado, com padrão lobular aumentado e, ocasionalmente, com áreas esbranquiçadas e deprimidas. Os rins estavam acastanhados e a urina escura. No exame histológico do fígado encontrou-se tumefação e vacuolização de hepatócitos, proliferação de células epiteliais dos ductos biliares, retenção biliar, fibroplasia periportal discreta ou moderada e infiltrado mononuclear periportal. Todos os casos de fotossensibilização apresentaram macrófagos espumosos no parênquima hepático e em 21 foram observados cristais birrefringentes nos ductos biliares. Cinco surtos com emagrecimento progressivo dos bovinos afetados foram diagnosticados em pastagens de B. decumbens. A principal lesão macroscópica foi o fígado aumentado de volume e amarelado. No estudo histológico, as lesões foram semelhantes às observadas nos casos de fotossensibilização, sendo que cristais refringentes nos ductos biliares foram observados em três animais. Conclui-se que B. decumbens é mais tóxica que B. brizantha e que a intoxicação ocorre, principalmente, em bovinos jovens nas diferentes épocas do ano. No entanto, pesquisas são necessárias para determinar as variações no conteúdo de saponinas litogênicas em Brachiaria spp. e as diferenças de resistência/susceptibilidade à intoxicação de bovinos de diferentes idades e raças em diferentes regiões.

The epidemiology, clinical signs and pathology of 29 outbreaks of spontaneous poisoning by Brachiaria spp. in beef cattle, which occurred from March 1996 to November 2009, in the state of Mato Grosso do Sul, were retrospectively studied. For this, the files of the Veterinary Pathology Laboratory at the Federal University of Mato Grosso do Sul were reviewed. Outbreaks occurred at different times of the year, including dry and rainy seasons. While photosensitization was the main clinical sign in cattle from 24 out of 29 outbreaks, in five of them the main clinical sign was progressive wasting, without photosensitization. Eleven outbreaks of photosensitization occurred in pastures of Brachiaria decumbens, two in mixed pastures of B. decumbens and B. brizantha, and one in B. brizantha. In 10 outbreaks the species of Brachiaria was not informed. Morbidity ranged from 0.2 percent to 50 percent, and fatality rates were between 44.4 percent and 100 percent. In cases of photosensitization brisket edema was the most frequent clinical sign observed. Dermatitis with thickened skin of the flank and perineal region, scar retraction of the ears, jaundice, ocular discharge, crusts in the eyes and ears, and ulceration of the ventral surface of the tongue were also observed. Nervous signs were observed in two cases and diarrhea in one. At necropsy the liver was enlarged, yellowish, with increased lobular pattern, and occasionally with depressed whitish areas. The kidneys were brownish and the urine dark. Upon histological examination of the liver the hepatocytes were swollen and vacuolized. Individual necrosis of hepatocytes, bile duct cell proliferation, biliary retention, and mild to moderate periportal fibrosis were also observed. In all cases foamy macrophages, most in groups, were observed mainly in the centrilobular region. In 21 cattle birefringent crystals were observed within the bile ducts. Five outbreaks of progressive wasting were observed in cattle grazing B. decumbens pastures. The main lesion was an enlarged yellowish liver. Histologic lesions were similar to those observed in cases of photosensitization. Birefringent crystals were observed in the liver of three cases, but foamy macrophages were found in all animals. It is concluded that B. decumbens is more toxic than B. brizantha and that the poisoning occurs at any time of the year. Nevertheless, research is still necessary to determine variations in the saponin content of Brachiaria species and the probable differences on susceptibility/resistance to the poisoning of cattle of different ages, breeds and from different regions.

Intoxicação experimental por Brachiaria decumbens em ovinos confinados / Experimental poisoning by Brachiaria decumbens in feedlot sheep

Foi realizado estudo experimental de intoxicação por Brachiaria decumbens em ovinos confinados que receberam apenas esta forrageira como alimento. Os animais foram avaliados em três períodos do ano, com duração de 60 dias cada. As análises incluíram exame clínico e amostras de sangue para mensuração da atividade sérica das enzimas gama glutamiltransferase e aspartato aminotransferase com o objetivo de avaliar sua importância e utilidade no diagnóstico de fotossensibilização hepatógena causada pela planta. Foram descritos ainda, achados de necropsia e histopatológicos. Dos vinte e quatro animais confinados, cinco morreram no primeiro período (fevereiro a abril), quatro no segundo (junho a agosto) e dois no terceiro (outubro a dezembro), tendo como principais sinais clínicos anorexia, icterícia, fotofobia, dermatite leve, além de desidratação e apatia. Nos achados de necropsia foram constatados icterícia generalizada, vesícula biliar repleta e distendida, padrão lobular hepático evidente e, em dois casos, opacidade de córnea. As alterações microscópicas foram mais significativas no fígado com bilestase, tumefação e vacuolização de hepatócitos, sinusóides com macrófagos, proliferação de ductos e dúctulos biliares com infiltração linfocítica, que variaram na sua severidade conforme o tempo decorrido da ingestão da planta, além de imagens negativas de cristais no citoplasma de macrófagos presentes nos sinusóides hepáticos e no espaço periportal e cristais refringentes oticamente ativos, ocluindo a luz de ductos biliares. Os achados foram característicos de fotossensibilização hepatógena por B. decumbens, principalmente pela presença de fotofobia, mesmo quando os animais não apresentaram fotodermatite. O aumento da atividade sérica da gama glutamiltransferase teve alta correlação com a morte dos animais. O surgimento da elevação da atividade sérica de gama glutamiltransferase foi, em média, 11 dias antes da constatação dos sinais clínicos, ...

Brachiaria decumbens was fed as exclusive source of food to feedlot sheep. Sheep were evaluated in three periods of the year, of 60 days each, regarding their clinical status and was performed collection of blood samples to measure enzymatic activity of gamma glutamyltransferase and aspartate aminotransferase to evaluate their diagnostic importance and usefulness besides the clinical signs, necropsy and histopathology finds in hepatogenous photosensitization Brachiaria related. Out of 24 feedlot sheep, five died in the first period (February-April), four in the second (June-August) and two in the third (October-December). Main clinical signs were anorexia, jaundice, photophobia and two cases of slight dermatitis, besides dehydration, apathy and corneal opacity. Necropsy revealed generalized jaundice, distended gall bladder and liver with evident lobular pattern. The histological changes varied in severity according the time after the beginning of the plant ingestion; they were more significant in the liver and included bilestasis, swelling and vacuolation of hepatocytes, foammy macrophages, bile duct proliferation and mononuclear inflammatory cell infiltration, further crystal negative images were found within cytoplasm of macrophages present inside of hepatic sinusoid and in the periportal space and also optically active refringent crystal in the lumen of bile tubules. These finds are characteristic of hepatogenous photosensitization, mainly by presence of photophobia, even when the sheep did not present photodermatitis. The results of serum biochemistry activity, during the course of poisoning with strong relation between increased gamma glutamyltransferase activity, and lethality. Onset of arising gama glutamyltransferase activity was 11 days, in average, before beginning of clinical signs. Aspartate aminotransferase had increased activity only close to the beginning of clinical signs. Serum biochemistry results of GGT suggest that ...

Sub-acute intoxication by Senna occidentalis seeds in rats.

Senna occidentalis (So) is a weed that grows in pastures along fences and in fields cultivated with cereals such as corn and soybean, and many reports have been showing intoxication with this plant in different animal species. It is also used in many medicinal purposes. The objective of the present study was to better evaluate the toxic effects of prolonged administration of So seeds to rats. Forty male Wistar rats were divided into four groups of 10 animals each, three of them respectively fed rations containing 1%, 2% and 4% So seeds, and the last one (control) fed commercial ration for a period of 2 weeks. Fourteen rats were also used in a pair-feeding (PF) experiment. The rats of the experimental groups showed lethargy, weakness, recumbency, depression and emaciation. Two rats of the 4% group and two of the PF group died during the experiment. Histopathological study showed fiber degenerations in the skeletal (Tibial, pectoral and diaphragm) and cardiac muscles. In the liver parenchyma, was observed vacuolar degeneration and, in the kidney, mild nefrosis in the proximal convoluted tubules. All of these alterations occurred in a dose-dependent fashion. Moderate to severe degeneration and spongiosis in the central nervous system, especially in cerebellum. Electron microscopy revealed mitochondrial lesions in all analyzed tissues.