Extraction and Identification of Alkaloids of the <i>Ipomoea fistulosa</i> (Aguapei or Mandiyurá) of Argentina

The classical toxic effect produced by this Ipomoea fistulosa is due to the accumulation of oligosacharides in various tissues cell cytoplasm, mainly nervous, liver and lymphatic tissues, that leads to the cell vacuolitation. These effects are attributed to the presence of substances called swansonina and calisteginas in the vegetables, which produce inhibition of enzymes liposomales responsible for the metabolic carbohydrates. In natural conditions, the plant is not consumed by the animals. Its consumption occurs alone in determined times of year, for lack of sufficient pasture and it is more frequent in young animals. Since standard literature does not register any data on the chemical analysis of the kind Ipomoea, we report here the results on the extraction, isolation and spectroscopic identification of the present alkaloids in this plant.Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicada

Encefalopatía hepática inducida en cerdos por semillas y vainas de Senna occidentalis

Senna occidentalis es una planta tóxica que afecta a diferentes especies de animales. La lesión hallada en la mayoría de los animales intoxicados es la degeneración muscular. Sin embargo, la encefalopatía hepática (EH) es observada en equinos y en humanos. El objetivo de este trabajo fue determinar si, además de la miodegeneración ya reportada, semillas y vainas de S. occidentalis inducen una EH tóxica en cerdos. Diez animales se dividieron en dos grupos (de cinco animales cada uno), un grupo se alimentó con una ración que contenía el 20% de vainas y semillas de S. occidentalis, y los animales controles recibieron ración comercial durante 14 días. Los animales intoxicados presentaron síntomas de aparición brusca, caracterizados por incoordinación, ataxia, desorientación, presión de la cabeza contra objetos duros, depresión y recumbencia lateral. La aspartato aminotransferasa y creatinquinasa incrementaron junto a la Bilirrubina Total en los animales intoxicados. La evaluación histopatológica de los cerdos alimentados con S. occidentalis evidenció tumefacción hepatocelular y necrosis centrolobulillar en el hígado; mientras que el encéfalo presentó vacuolización de la sustancia blanca y astrocitos Alzheimer tipo II en la corteza cerebral. La microscopía electrónica reveló lesiones mitocondriales en el hígado. Estos resultados muestran que en el presente estudio, la lesión muscular ya reportada no se observó, seguramente debido a que la EH tóxica reproducida en los animales evaluados se produjo antes de que ello ocurra. Por otro lado, los animales del presente estudio desarrollaron signos clínicos y lesiones histológicas que fueron similares a esas observadas en casos de envenenamiento accidental. Además futuros estudios son necesarios para identificar el tóxico responsable de la falla hepática aguda, observada en los animales de este estudio.Senna occidentalis is a toxic plant that affects different animal species. The predominant lesion found in most of the intoxicated animals is skeletal muscle degeneration. However, in horses and humans, this poisoning is primarily characterized by hepatic encephalopathy (HE). The aim of this paper was to determine whether, in addition to this myodegeneration, the seeds and pods of S. occidentalis induce toxic EH in pigs. Ten pigs were divided into two groups (of five animals each), one of which were fed with a ration containing 20 % of S. occidentalis pods and seeds, and the other with a commercial ration (control) for 14 days. Poisoned animals had a sudden onset of symptoms, characterized by incoordination, ataxia, disorientation and head pressing, depression and lateral recumbency. Aspartate aminotransferase and Creatine phosphokinase serum activities increased along with an increasement of serum bilirubin in intoxicated animals with S. occidentalis. Histopathological studies of the poisoned pigs showed hepatocellular swelling and centrilobular necrosis in the liver, vacuolization of the white matter and Alzheimer type II astrocytes in the cerebral cortex of the brain. Electron microscopy revealed mitochondrial lesions in liver. These results showed that in this study, the muscle injury previously reported was not observed, most probably because the toxic EH reproduced in the evaluated animals were produced before skeletal muscle degeneration occurred. On the other hand, the animals of the present study developed clinical signs and histological lesions that were similar to those observed in cases of accidental poisoning. Besides, further studies are needed to identify the specific toxin responsible for acute liver failure, observed in the animals of this study.Facultad de Ciencias VeterinariasConsejo Nacional de Investigaciones Científicas y Técnica

Encefalopatía hepática inducida en cerdos por semillas y vainas de Senna occidentalis

Senna occidentalis es una planta tóxica que afecta a diferentes especies de animales. La lesión hallada en la mayoría de los animales intoxicados es la degeneración muscular. Sin embargo, la encefalopatía hepática (EH) es observada en equinos y en humanos. El objetivo de este trabajo fue determinar si, además de la miodegeneración ya reportada, semillas y vainas de S. occidentalis inducen una EH tóxica en cerdos. Diez animales se dividieron en dos grupos (de cinco animales cada uno), un grupo se alimentó con una ración que contenía el 20% de vainas y semillas de S. occidentalis, y los animales controles recibieron ración comercial durante 14 días. Los animales intoxicados presentaron síntomas de aparición brusca, caracterizados por incoordinación, ataxia, desorientación, presión de la cabeza contra objetos duros, depresión y recumbencia lateral. La aspartato aminotransferasa y creatinquinasa incrementaron junto a la Bilirrubina Total en los animales intoxicados. La evaluación histopatológica de los cerdos alimentados con S. occidentalis evidenció tumefacción hepatocelular y necrosis centrolobulillar en el hígado; mientras que el encéfalo presentó vacuolización de la sustancia blanca y astrocitos Alzheimer tipo II en la corteza cerebral. La microscopía electrónica reveló lesiones mitocondriales en el hígado. Estos resultados muestran que en el presente estudio, la lesión muscular ya reportada no se observó, seguramente debido a que la EH tóxica reproducida en los animales evaluados se produjo antes de que ello ocurra. Por otro lado, los animales del presente estudio desarrollaron signos clínicos y lesiones histológicas que fueron similares a esas observadas en casos de envenenamiento accidental. Además futuros estudios son necesarios para identificar el tóxico responsable de la falla hepática aguda, observada en los animales de este estudio.Senna occidentalis is a toxic plant that affects different animal species. The predominant lesion found in most of the intoxicated animals is skeletal muscle degeneration. However, in horses and humans, this poisoning is primarily characterized by hepatic encephalopathy (HE). The aim of this paper was to determine whether, in addition to this myodegeneration, the seeds and pods of S. occidentalis induce toxic EH in pigs. Ten pigs were divided into two groups (of five animals each), one of which were fed with a ration containing 20 % of S. occidentalis pods and seeds, and the other with a commercial ration (control) for 14 days. Poisoned animals had a sudden onset of symptoms, characterized by incoordination, ataxia, disorientation and head pressing, depression and lateral recumbency. Aspartate aminotransferase and Creatine phosphokinase serum activities increased along with an increasement of serum bilirubin in intoxicated animals with S. occidentalis. Histopathological studies of the poisoned pigs showed hepatocellular swelling and centrilobular necrosis in the liver, vacuolization of the white matter and Alzheimer type II astrocytes in the cerebral cortex of the brain. Electron microscopy revealed mitochondrial lesions in liver. These results showed that in this study, the muscle injury previously reported was not observed, most probably because the toxic EH reproduced in the evaluated animals were produced before skeletal muscle degeneration occurred. On the other hand, the animals of the present study developed clinical signs and histological lesions that were similar to those observed in cases of accidental poisoning. Besides, further studies are needed to identify the specific toxin responsible for acute liver failure, observed in the animals of this study.Facultad de Ciencias VeterinariasConsejo Nacional de Investigaciones Científicas y Técnica

Intoxicación con monensina en un feedlot de búfalos de Argentina

El objetivo de este trabajo fue describir un envenenamiento accidental por monensina en búfalos domésticos (Bubalus bubalis) pertenecientes a un feedlot ubicado en la región nordeste de Argentina. Los signos clínicos incluyeron ansiedad, posición ortopneica con rechinamiento de dientes, diarrea, debilidad muscular, depresión y recumbencia. En los animales expuestos a monensina se observaron aumentos significativos de creatin fosfoquinasa y aspartato aminotransferasa. A la necropsia, los búfalos presentaron áreas pálidas de distribución irregular, hidrotórax, congestión y edema pulmonar. Al examen histológico se observó degeneración y necrosis de músculos cardíaco y esquelético. El análisis del alimento reveló niveles tóxicos de monensina, por lo cual se diagnosticó una toxicosis por ionóforos. A pesar de los efectos positivos de la monensina como fuente de alimento de los animales, se debe requerir un mayor nivel de precaución al alimentar búfalos con ionóforos.The aim of this paper was to describe a case of accidental monensin poisoning of feedlot domestic buffalo (Bubalus bubalis) in northeastern Argentina. Clinical signs included anxiety facie with orthopneic position, teeth grinding, diarrhea, muscular weakness, depression and recumbency. In monensin intoxicated animals, a significant increase of creatine phosphokinase and aspartate aminotransferase was observed. At postmortem examination, buffaloes consistently had pale areas that were irregularly distributed in cardiac and skeletal muscles, hydrothorax, congestion and oedema of the lung. Dege-neration and necrosis of the myocardium and skeletal muscle was observed by histological examination. Feed analysis revealed toxic levels of monensin; therefore, ionophore toxicosis was diagnosed. Accordingly, it is required great caution when feeding buffaloes with monensin used as a feed additive.Fil: Garcia, Enrique Nicolas. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; ArgentinaFil: Pistán, David Guillermo. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; ArgentinaFil: Lertora, Walter Javier. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; ArgentinaFil: Mastrantonio Garrido, Guido Enrique. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Área de Toxicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Cholich, Luciana Andrea. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentin

Intoxicação experimental por Ipomoea carnea subsp. fistulosa em caprinos da Argentina: correlação clínico-patológica, com especial atenção ao sistema nervoso central

Ipomoea carnea subsp. fistulosa, aguapei or mandiyura, is responsible for lysosomal storage in goats. The shrub contains several alkaloids, mainly swansonine which inhibits lysosomal α-mannosidase and Golgi mannosidase II. Poisoning occurs by inhibition of these hydrolases. There is neuronal vacuolation, endocrine dysfunction, cardiovascular and gastrointestinal injury, and immune disorders. Clinical signs and pathology of the experimental poisoning of goats by Ipomoea carnea in Argentina are here described. Five goats received fresh leaves and stems of Ipomoea. At the beginning, the goats did not consume the plant, but later, it was preferred over any other forage. High dose induced rapid intoxication, whereas with low doses, the course of the toxicosis was more protracted. The goats were euthanized when they were recumbent. Cerebrum, cerebellum, medulla oblongata, pons and colliculi, were routinely processed for histology. In nine days, the following clinical signs developed: abnormal fascies, dilated nostrils and abnormal postures of the head, cephalic tremors and nystagmus, difficulty in standing. Subsequently, the goats had a tendency to fall, always to the left, with spastic convulsions. There was lack in coordination of voluntary movements due to Purkinje and deep nuclei neurons damage. The cochlear reflex originated hyperreflexia, abnormal posture, head movements and tremors. The withdrawal reflex produced flexor muscles hypersensitivity at the four legs, later depression and stupor. Abnormal responses to sounds were related to collicular lesions. Thalamic damage altered the withdrawal reflex, showing incomplete reaction. The observed cervical hair bristling was attributed to a thalamic regulated nociceptive response. Depression may be associated with agonists of lysergic acid contained in Ipomoea. These clinical signs were correlated with lesions in different parts of the CNS.Ipomoea carnea subsp. fistulosa, aguapeí ou mandiyura, causa uma doença de depósito lisossomal em caprinos. A planta contém vários alcalóides, principalmente swansonine, que inibe uma α-mannosidase lisossomal e uma mannosidase II do Golgi. A intoxicação ocorre pela inibição dessas hidrolases. Há vacuolização neuronal, disfunção endócrina, lesões cardiovasculares e gastrointestinais e distúrbios imunológicos. No presente trabalho é descrita a intoxicação experimental por Ipomoea carnea subsp. fistulosa em caprinos da Argentina. Cinco cabras receberam folhas frescas e ramos de Ipomoea. No começo eles não consumiram a planta, mas depois ela foi preferida a qualquer outras forrageiras. Altas doses induziram uma Pesq. Vet. Bras. 32(1):37-42, janeiro 2012 38 Elvio E. Ríos et al. intoxicação rápida, enquanto que com doses baixas demora mais tempo. As cabras foram sacrificadas quando ficavam em decúbito. Cérebro, cerebelo, medula oblonga, ponte e colículos foram processados rotineiramente para histologia. Aos nove dias, encontramos: fascie anormal, narinas dilatadas, posturas anormais da cabeça, tremores cefálicos e nistagmo, dificuldade para permanecer na estação. Posteriormente, presentearam a tendência de queda, sempre à esquerda, com convulsões espásticas. Os neurônios de Purkinje e os os núcleos profundos foram danificados. Como conseqüência, emergiu a falta de coordenação de movimentos voluntários. A hiper-reflexo coclear originou postura anormal, movimentos de cabeça e tremores. A retirada produz reflexos hipersensibilidade nos flexores dos quatro membros, depois depressão e letargia. As respostas anormais estão ligadas a lesões do colículo. Injurias no tálamo alteram o reflexo de retirada, mostrando uma reação incompleta. Os pelos eriçados da região cervical podem representar uma resposta nociceptiva também regulada pelo tálamo. A depressão pode estar associada com agonistas do ácido lisérgico presentes na Ipomoea. Esses sintomas estão relacionados com lesões em diferentes partes do SNC.Fil: Ríos, Elvio E.. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias. Instituto de Ictiologia del Nordeste; ArgentinaFil: Cholich, Luciana Andrea. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias. Instituto de Ictiologia del Nordeste; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gimeno, Eduardo Juan. Universidad Nacional de la Plata; ArgentinaFil: Guidi, María G.. Universidad Nacional de la Plata; ArgentinaFil: Acosta de Pérez, Ofelia C.. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias. Instituto de Ictiologia del Nordeste; Argentin

Suspected poisoning in beef cattle from ingestion of Prosopis nigra pods in north-western Argentina

The aim of this paper was to present the first report of Prosopis nigra poisoning of cattle in Argentina. Outbreaks occurred in five farms located in Salta and Santiago del Estero provinces. All animals were examined, euthanized and necropsied. Clinical signs included tongue protrusion, twitches and tremors of muscles of mastication, weight loss and lethargy. Severe atrophy of the masseter, buccinator and lingual muscles was observed, along with neuronal vacuolation in the nuclei of the trigeminal, facial, and hypoglossus nerves. These findings and the clinical signs are consistent with results obtained in animals, spontaneously and experimentally intoxicated with Prosopis juliflora in previous studies. Several species of this genus are native to Argentina. Farmers should be warned about the suspected toxicity by Prosopis nigra, since this species has wide geographical distribution in the country.Facultad de Ciencias Veterinaria

Characterization and cytotoxic activity on glial cells of alkaloid-enriched extracts from pods of the plants prosopis flexuosa and prosopis Nigra (Fabaceae)

Introduction: Prosopis spp. pods have shown to be a potential source of protein and energy in livestock. However, prolonged ingestion of some of these species produces neurological symptoms in ruminants. Objective: In the present study, the alkaloid content and the in vitro neurotoxic activity of alkaloid enriched-extracts from P. flexuosa and P. nigra pods were determined in order to elucidate the mechanism of animal poisoning caused by these species. Methods: The main alkaloids present in both extracts were analysed by high performance liquid chromatography-high resolution mass spectrometry (HPLC-HRMS). The cytotoxic activity of Prosopis alkaloid enriched-extracts in primary mixed glial cell culture was assessed by phase contrast microscopy and using neutral red, and lactate dehydrogenase (LDH) activity assays. Results: Juliprosine and juliprosopine were identified in P. flexuosa pods, while the absence of these alkaloids in P. nigra was confirmed. Both extracts (5-30 μg/mL) induced in a dose dependent manner, morphological alterations, such as swelling, enlargement and detachment from the culture surface. Consistent with this, decrease in cell viability and release of LDH 48 hours after exposure, revealed that P. flexuosa pods was significantly more cytotoxic than P. nigra. Conclusions: In P. flexuosa pods, juliprosine and juliprosopine alkaloids were identified for the first time. Moreover, the present study suggests that the cytotoxic effect displayed by both extracts is due to its alkaloid content. However, the presence of piperidine alkaloids in P. flexuosa could explain the greater cytotoxicity on glial cells with respect to P. nigra that was not shown to contain these alkaloids.Fil: Cholich, Luciana Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; ArgentinaFil: Pistan, Maria Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; ArgentinaFil: Torres, Ana Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; ArgentinaFil: Gardner, Dale R.. No especifíca;Fil: Bustillo, Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentin

Set de imágenes de microscopía óptica y electrónica de barrido de Astragalus garbancillo colonizaos por Alternaria

Plants belonging to the genera Astragalus, Oxytropis, Ipomoea, Sida, and Swainsona often contain the toxin swainsonine (SW) produced by a symbiont fungus. Consumption of these SW-containing plants cause a serious neurological disorder in livestock, which can be fatal. The identity of the symbiont fungus in the plant species A. garbancillo was determined and its localization and distribution in different organs studied. Using PCR, we confirm that the fungus belongs to the genera Alternaria section Undifilum. Furthermore, we show by microscopic examination that the fungus mycelium grows in the (i) pith and cortex of petioles and stems, (ii) the external surface and inside the mesocarp of the fruit and (iii) in the mesotesta, and endotesta layers of the seed coat. Accordingly, SW could be detected in the corresponding tissues of petioles, stems, fruits and seeds. Our observations verify that the SW-producing fungus is vertically transmitted in the host plant A. garbancillo.Fil: Cholich, Luciana Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional del Nordeste; ArgentinaFil: Pistan, Maria Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional del Nordeste; ArgentinaFil: González, Ana María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentin