Assessment of endophyte-derived tremorgenic compounds in Ipomoea asarifolia using mouse models.

Ipomoea asarifolia has been associated with a tremorgenic syndrome in livestock. Recently indole diterpene compounds were identified in I. asarifolia, some of which have been shown to cause a tremorgenic syndrome. In this study, the tremorgenic nature of I. asarifolia was assessed using a mouse model. Adult mice were fed rodent chow containing 10, 15, 20 and 25% endophyte infected (E+), or 25% endophyte free (E-), I. asarifolia for 14 days. The mice fed E+ chow developed a tremorgenic syndrome as characterized by visually observed muscle tremors and an inability to traverse a balance beam, whereas the mice fed E- chow did not develop tremors and had similar muscle coordination to control mice. A lactating mouse model was also used to determine if the compounds can be transferred to nursing pups via the milk. Nursing pups were exposed via their mother's milk for 21 days, from post-natal day 0-21. The pups from dams exposed to E+ chow developed a similar tremorgenic syndrome. Data presented in this study demonstrate that the tremorgenic compounds in I. asarifolia are endophyte derived. Additionally, both adult mice and nursing pups are good models for studying the tremorgenic nature of I. asarifolia and related plants.

Detection of swainsonine and calystegines in Convolvulaceae species from the semiarid region of Pernambuco / Detecção de swainsonina e calisteginas em espécies de Convolvulaceae da região semiárida de Pernambuco

Numerous plant species worldwide including some Ipomoea (Convolvulaceae) and Sida (Malvaceae) species in Brazil cause lysosomal storage disease in herbivores and are known to contain swainsonine and calystegines as the main toxic compounds. The aim of this work was to determine swainsonine and calystegines concentrations in species of Convolvulaceae from the semiarid region of Pernambuco. Seven municipalities in the Moxotó region were visited and nine species were collected and screened for the presence of swainsonine and calystegines using an HPLC-APCI-MS method. The presence and concentration of these alkaloids within the same and in different species were very variable. Seven species are newly reported here containing swainsonine and/or calystegines. Ipomoea subincana contained just swainsonine. Ipomoea megapotamica, I. rosea and Jacquemontia corymbulosa contained swainsonine and calystegines. Ipomoea sericosepala, I. brasiliana, I. nil, I. bahiensis and I. incarnata contained just calystegines. The discovery of six Ipomoea species and one Jacquemontia species containing toxic polyhydroxy alkaloids reinforces the importance of this group of poisonous plants to ruminants and horses in the semiarid region of Pernambuco. Epidemiological surveys should be conducted to investigate the occurrence of lysosomal storage disease associated to these new species.(AU)

Numerosas espécies de plantas em todo o mundo, incluindo algumas espécies de Ipomoea (Convolvulaceae) e Sida (Malvaceae) no Brasil, causam doença de armazenamento lisossomal em herbívoros e são conhecidas por conterem swainsonina e calisteginas como princípios tóxicos. O objetivo deste trabalho foi determinar a concentração de swainsonina e calisteginas em espécies de Convolvulaceae da região semiárida de Pernambuco. Sete municípios na região do Sertão do Moxotó foram visitados, onde foram coletadas amostras das folhas de nove espécies de Convolvulaceae para avaliação da presença de swainsonina e calisteginas utilizando-se cromatografia líquida com espectrometria de massa. A presença e concentração destes alcaloides nas folhas de plantas da mesma espécie e dentre as espécies foram muito variáveis. Seis novas espécies de Ipomoea e uma espécie de Jacquemontia contendo swainsonina e/ou calisteginas são relatadas neste estudo. Ipomoea subincana continha apenas swainsonina. Ipomoea megapotamica, I. rosea e Jacquemontia corymbulosa continham swainsonina e calisteginas. Ipomoea sericosepala, I. brasiliana, I. nil, I. bahiensis e I. incarnata continham apenas calisteginas. A descoberta de novas espécies de Ipomoea e Jacquemontia contendo alcaloides polihidroxílicos tóxicos reforçam a importância deste grupo de plantas tóxicas para ruminantes e equinos na região semiárida de Pernambuco. Pesquisas epidemiológicas devem ser realizadas para investigar a ocorrência de doença de depósito lisossomal associada a essas novas espécies.(AU)

Identification of Indole Diterpenes in Ipomoea asarifolia and Ipomoea muelleri, Plants Tremorgenic to Livestock.

Ipomoea asarifolia has been associated with a tremorgenic syndrome in livestock in Brazil and was recently reported to contain tremorgenic indole diterpenes. Ipomoea muelleri has been reported to cause a similar tremorgenic syndrome in livestock in Australia. Ipomoea asarifolia and I. muelleri were investigated by high-performance liquid chromatography-high-resolution mass spectometry (HPLC-HRMS) and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) for indole diterpene composition. The high-resolution mass spectrometric data in combination with MS/MS fragmentation mass spectral data provided valuable information for indole diterpene characterization. The previous report of indole diterpenes in I. asarifolia was confirmed and expanded; and the presence of indole diterpenes in I. muelleri is reported for the first time. Two new indole diterpenes were isolated and their structures determined by 1D and 2D NMR spectroscopy and given the names 11-hydroxy-12,13-epoxyterpendole K and 6,7-dehydroterpendole A. The presence of terpendole K and terpendole E in I. asarifolia is unequivocally demonstrated for the first time. This is the first detailed MS analysis of known indole diterpenes and possible isomers in I. asarifolia and I. muelleri.

Analysis of Swainsonine and Swainsonine N-Oxide as Trimethylsilyl Derivatives by Liquid Chromatography-Mass Spectrometry and Their Relative Occurrence in Plants Toxic to Livestock.

There are limited data concerning the occurrence of swainsonine N-oxide in plants known to contain swainsonine and its relative impact on toxicity of the plant material. A liquid chromatography-mass spectrometry method based on a solvent partitioning extraction procedure followed by trimethylsilylation and analysis using reversed phase high-pressure liquid chromatography-mass spectrometry was developed for the analysis of swainsonine and its N-oxide. The concentrations of each were measured in several swainsonine-containing taxa as well as two endophytic isolates that produce swainsonine. In vegetative samples the relative percent of N-oxide to free base ranged from 0.9 to 18%. In seed samples the N-oxide to free base ratio ranged from 0 to 10%. The measured concentrations of swainsonine N-oxide relative to swainsonine only slightly increases the actual toxicity of the various plant samples in a combined assay of both compounds.

Haematologic alterations caused by Ipomoea carnea in experimental poisoning of guinea pig.

Ipomoea carnea subsp. fistulosa (Convolvulaceae) causes poisoning of goats, sheep and cattle in many tropical and subtropical countries. The pathophysiology of this poisoning mainly involves an abnormal glycoprotein metabolism. The aim of this study was to describe the potential toxicity of I. carnea in a guinea pig model through its effect on hematopoiesis in a time course study of 40 days. Experimental poisoning was achieved by feeding animals with "small balls" prepared with milled leaves of I. carnea mixed with commercial crushed pellets for rodents. Hematologic and biochemical parameters, bone marrow and spleencellularities, histopathologic evaluations and lectin-histochemistrywere performed during the scheduled time of the study.The treatment with "small balls" caused significant changes in the weight of spleen, a notable decrease in peripheral red blood cells, and concomitantwith morphological and histopathologicalalterationsin hematopoietic tissues. Overall, the present study suggested that 20 days ofthis treatmentcouldbe enough to develop bone marrow hypoplasia and vacuolation of white cells of spleen, blood and lymph nodes with a transient erythropoietic contribution of the splenic niche.Moreover, this work provides a cheap and simple method for detecting preclinical cases of intoxication by I. carnea in livestock.

90 Days toxicological assessment of hydroethanolic leaf extract of Ipomoea asarifolia (Desr.) Roem. and Schult. (Convolvulaceae) in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Ipomoea asarifolia (Convolvulacae), commonly known as "morning glory" is found across West Africa. Preparations of the plant are used traditionally for the treatment of diverse ailments including diabetes, neuralgia, arthritic pain and stomach ache. This study was designed to assess the safety profile of the hydroethanolic leaf extract of I. asarifolia through a 90-day subchronic toxicity study in rats. MATERIALS AND METHODS: I. asarifolia was administered p.o. at doses of 40, 200 and 1000mg/kg to separate groups of rats for 90 days. Distilled water was given p.o. to rats in the control group. Some set of rats in each group were left for additional 30 days without administration of the extract for reversibility study. Animals were weighed weekly and relevant parameters were assayed at the end of the main and reversibility study periods. RESULTS: There was no significant change (p>0.05) in the body weight of rats, and food and water intake in I. asarifolia treated groups compared with control. I. asarifolia (40-1000 mg/kg) significantly but reversibly reduced (p<0.05, 0.001) sperm motility and count. The extract did not generally cause significant change (p>0.05) in the weight of vital organs and haematological parameters except in the case of reversible reduction in the level of haemoglobin and red blood cell count (p<0.01; 40 mg/kg). The level of biochemical parameters and electrolytes were not significantly changed (p>0.05) except for the reversible reduction in the level of aspartate aminotransferase (AST; p<0.0001; 200 and 1000 mg/kg) and increase in the level of Na(+) (p<0.01; 200 mg/kg). The level of kidney reduced glutathione (GSH) was reversibly increased (p<0.01; 1000 mg/kg) while the level of enzymatic and non-enzymatic in vivo antioxidants was generally comparable and not significantly different (p>0.05) from control in respect of all other vital organs. Histological presentations were generally normal in respect of the liver, kidneys, brain, heart, lungs, pancreas, spleen and testes. CONCLUSIONS: The findings in this study suggest that the hydroethanolic leaf extract of I. asarifolia is relatively safe administered orally for an extended period with potential renal in vivo antioxidant activities. However, the extract may cause reversible male sterility, anaemia and hypernatraemia.

Poisoning by Ipomoea asarifolia in lambs by the ingestion of milk from ewes that ingest the plant.

Two experiments, each with 10 pregnant ewes (8 treated and 2 controls) were performed to determine if nursing lambs of lactating ewes become intoxicated when the ewes ingest Ipomoea asarifolia but do not show clinical signs themselves. In the first experiment the sheep grazed I. asarifolia in the field while in the second, sheep were maintained in individual bays consuming dry I. asarifolia at 10% and 20% into their ration. In both experiments the lambs remained confined, consuming only their mother's milk. Four of 8 lambs in the grazing experiment and the 4 nursing lambs from the ewes given 20% I. asarifolia showed signs of I. asarifolia poisoning. These results confirm that the tremorgenic compound of I. asarifolia or its toxic metabolites are eliminated in milk and can intoxicate nursing lambs.

A review on Ipomoea carnea: pharmacology, toxicology and phytochemistry.

Phytomedicines are increasingly being established in modern medical science. The shrub Ipomoea carnea has been used traditionally for thousands of years. However, there are few scientific studies on this medicinal plant, and most of the information are scattered. In this review, we have summarized the existing knowledge and recent progress on the medicinal importance of I. carnea. Different extracts of I. carnea plant possess anti-bacterial, anti-fungal, anti-oxidant, anti-cancer, anti-convulsant, immunomodulatory, anti-diabetic, hepatoprotective, anti-inflammatory, anxiolytic, sedative and wound healing activities. However, some toxicological effects have been also reported. Some of the major phytochemicals associated with the bioactivity of I. carnea have been characterized, which have been discussed in this study too. This review article might be beneficial for phytotherapy research, as I. carnea can be a good source for drug development.

Experimental intoxication of guinea pigs with Ipomoea carnea: behavioural and neuropathological alterations.

Ipomoea carnea is a toxic plant that affects goats, with symptoms being characterised by nervous disorders and death. Swainsonine and calystegines are the principal toxic components isolated from I. carnea, which also yields lysergic acid derivatives. The aim of this study was to improve the clinical characterisation of experimental intoxication by I. carnea in guinea pigs through the evaluation of behavioural changes and to perform a thorough histopathological analysis of the affected CNS. Leaves of I. carnea were administered to guinea pigs. Open-field gait analysis and monoamine levels were measured. The poisoned animals exhibited increased vocalisation, lethargy, and a reduction in the locomotion frequency after the fourth week of intoxication, as demonstrated in the open-field test. Significant differences were observed in hind-limb gait width by the last week of intoxication. After 65 days, the guinea pigs were euthanised, necropsied, and examined using light and electron microscopy. At the end of the experiment, plasma serotonin decreased. In contrast, dopamine decreased, and noradrenaline increased in urine. Brain sections were evaluated with conventional histological methods and immunohistochemistry (IHC), as well as by transmission electron microscopy (TEM). Vacuoles were observed throughout the brain, but they were particularly prominent in the brainstem. In addition, there were PAS-negative regions, and the Nissl substance was dispersed or absent, which was confirmed with the Kluver-Barreda stain. Moderate microgliosis was observed by immunohistochemistry. In the medulla oblongata, numerous ubiquitin-positive spheroids together with neuronal degeneration were observed in the nucleus gracilis/cuneatus. Furthermore, vacuoles were observed in astrocytes, oligodendrocytes, and endothelial cells by TEM. Our results showed that the behavioural effects may have been caused by alterations in the brain in conjunction with changes in monoamine levels. This research confirms the utility of this model for studying the pathogenesis of plant-induced lysosomal storage diseases.

Doença de depósito lisossomal induzida pelo consumo de Ipomoea verbascoidea (Convolvulaceae) em caprinos no semiárido de Pernambuco / Lysosomal storage disease induced by the consumption of Ipomoea verbascoidea (Convolvulaceae) in goats at northeastern Brazil

The aim of this paper was to reproduce the poisoning of Ipomoea verbascoidea in goats and describe the epidemiological, clinical and pathological aspects of spontaneous poisoning by this plant in Pernambuco. For this, we studied the epidemiology of the disease in seven municipalities in the semiarid region of the State. Three spontaneously poisoned goats were examined and then euthanized and necropsied (Group I). To reproduce the disease, the dried leaves of I. verbascoidea containing 0.02% swainsonine were supplied at doses of 4g/kg (0.8mg swainsonine/kg) to two groups of three animals. The goats in Group II received daily doses of the plant during 40 days and were euthanized on the 41st day of the experiment. Goats from Group III received daily doses of the plant during 55 days and were euthanized on the 120th day of the experiment. Other three goats constituted the control group (Group IV). In experimental groups, the brain lesions were evaluated by histopathology; additionally the cerebellar lesions were evaluated by morphometry, by measuring the molecular layer thickness, the number of Purkinje cells and the area of the cell bodies of these cells. The main clinical signs and microscopic lesions in goats poisoned were similar to those reported by swainsonine containing plants. In goats of GII and GIII, the first nervous signs were observed between 22th and 29th days; clinically, the disease developed by these animals was similar to the spontaneous cases. The goats of GIII did not recover from the neurologic signs. These results show that the consumption of the plant by 26-28 days after observation of the first clinical signs is enough to cause irreversible damage. By morphometric analysis, the molecular layer of the cerebellum of the goats of Group I and III were thinner than those of goats in the control group, and Purkinje neurons were atrophic. It is suggested that these changes are responsible for the neurological picture observed in goats that stop eating the plant and have sequelae of poisoning.

O objetivo deste trabalho foi reproduzir a intoxicação por Ipomoea verbascoidea em caprinos e descrever os aspectos epidemiológicos, clínicos e histopatológicos da intoxicação espontânea por essa planta no Estado de Pernambuco. Para isso, realizou-se o acompanhamento da epidemiologia da doença em sete municípios do semiárido pernambucano. Três caprinos espontaneamente intoxicados foram examinados e, em seguida eutanasiados e necropsiados (Grupo I). Para reproduzir experimentalmente a doença, as folhas secas de I. verbascoidea contendo 0,02% de swainsonina, foram fornecidas na dose de 4g/kg (0,8mg de swainsonina/kg) a dois grupos de três animais. Os caprinos do Grupo II receberam a planta diariamente por 40 dias e foram eutanasiados no 41º dia de experimento. Os caprinos do Grupo III receberam a planta diariamente por 55 dias e foram eutanasiados no 120º dia de experimento. Outros três caprinos constituíram o grupo controle (Grupo IV). Nos grupos experimentais, as lesões encefálicas foram avaliadas por histopatologia e adicionalmente avaliaram-se as lesões cerebelares por morfometria, mediante mensuração da espessura da camada molecular, do número de neurônios de Purkinje e da área dos corpos celulares dessas células. Os principais sinais clínicos e lesões microscópicas foram semelhantes aos previamente reportados em animais intoxicados por plantas que contem swainsonina. Nos caprinos do GII e GIII, os primeiros sinais clínicos foram observados entre o 22º e 29º dia de experimento; clinicamente a doença desenvolvida por esses animais foi semelhante aos casos espontâneos. Nenhum dos caprinos do GIII se recuperou dos sinais neurológicos. Esse resultado evidencia que o consumo da planta por 26-28 dias após a observação dos primeiros sinais clínicos é suficiente para provocar lesões irreversíveis. Pela análise morfométrica, a camada molecular do cerebelo dos caprinos do Grupo I e III eram mais delgadas que às dos caprinos do grupo controle, e os neurônios de Purkinje estavam atróficos. Sugere-se que essas alterações sejam responsáveis pelo quadro clínico neurológico observado nos caprinos que deixam de ingerir a planta e apresentam seqüelas da intoxicação.

Aversão alimentar condicionada no controle de surtos de intoxicações por Ipomoea carnea subsp. fistulosa e Turbina cordata em caprinos / Conditioned food aversion to control outbreaks of poisoning by Ipomoea carnea subsp. fistulosa and Turbina cordata in goats

A aversão alimentar condicionada é uma técnica que pode ser utilizada em animais para evitar a ingestão de plantas tóxicas. A técnica foi utilizada em uma fazenda para controlar a intoxicação por Turbina cordata e em outra para Ipomoea carnea subsp. fistulosa. Os caprinos eram presos à noite, e na manhã do dia seguinte lhes era ofertada a planta verde, recém-colhida, por dez minutos. Os caprinos que ingerissem qualquer quantidade da planta eram identificados, pesados e tratados com LiCl na dose de 175mg/kg peso vivo através de sonda esofágica. No rebanho da fazenda na que havia T. cordata a técnica foi aplicada a cada dois meses durante o período em que a planta é encontrada. Durante todo o experimento, de dezembro de 2009 a abril de 2011 não ocorreu nenhum novo caso de intoxicação no rebanho e diminuiu gradualmente o número de animais avertidos e a quantidade de planta que ingeriam os mesmos durante o processo de aversão. Na fazenda na que ocorria intoxicação por I. carnea a maioria de rebanho foi avertido em dezembro de 2010, 15-20 dias antes do início das chuvas, e os animais não ingeriram a planta espontaneamente no campo até setembro-outubro de 2011, durante o período da seca, quando havia extrema carência de forragem e iniciaram a ingerir a planta no campo. Posteriormente, apesar de três tratamentos aversivos com 21 dias de intervalo, os animais continuaram a ingerir a planta e ocorreram casos clínicos. A técnica de aversão alimentar condicionada demonstrou ser eficiente e viável para o controle da intoxicação por T. cordata. Para a intoxicação por I. carnea a técnica impediu a ingestão da planta somente durante a época de chuvas, mas não durante a seca, quando há pouca disponibilidade de forragem. A diferença nos resultados com as duas plantas é, aparentemente, resultante das condições epidemiológicas diferentes nas que ocorrem as intoxicações. T. cordata desaparece durante a maior parte do período de seca. A planta rebrota e fica verde durante o fim de seca, quando diminui a oferta de forragem, por curto espaço de tempo, permanecendo verde durante a época de chuvas. I. carnea, por crescer próximas as fontes de água, em áreas húmidas, permanece verde durante todo o período da seca, quando é maior a escassez de forragem, favorecendo desta forma a ingestão da planta pelos animais.

Conditioned food aversion is used to train livestock to avoid the ingestion of toxic plants. This technique was used to control Turbina cordata poisoning in goats in one farm, and to control Ipomoea carnea subsp. fistulosa poisoning in another farm. The goats were penned at night and the next morning the green plants were offered for 10 minutes. Goats that ingested any amount of the plant were treated through a gastric tube with 175mg of LiCl/kg body weight. In the flock in which the poisoning by T. cordata was occurring, the goats were averted every two months during the period that the plant was found in the pastures. During the experiment, from December 2009 to April 2011, new cases of poisoning were not observed, and there was a progressive decrease in the number of goats that ingested the plant and were averted. In the farm where I. carnea poisoning was occurring, most of the goats were averted in December 2010, 15-20 days before the first rains. The goats of this flock did not ingest the plant spontaneously in the field until September-October 2011, when, due to the dry season, there was a severe forage shortage, and the goats started to ingest the plant in the field. Later, despite three aversive treatments with 21 days intervals, the goats continued to ingest the plant and some animals became poisoned. In conclusion, conditioned food aversion was effective in to control intoxication by T. cordata. The technique was also effective in conditioning goats to avoid consuming I. carnea during the rainy season, but not during the dry season, with low forage availability in the field. The differences in these results seem to be due to the epidemiology of both poisonings: T. cordata is senescent and unavailable during most of the dry period, and green biomass is typically available either at the very end of the dry season, for a short period of time, and during the rainy season when there is no shortage of forage. In contrast, I. carnea grows in wet areas near water sources, and stays green during the dry period when there is a lack of other forage.

The use of ultrasonography to study teratogenicity in ruminants: evaluation of Ipomoea carnea in goats.

BACKGROUND: Ipomoea carnea (I. carnea) is a poisonous plant found in Brazil and other tropical countries that often poison livestock. The plant contains the alkaloids calystegines and mainly swainsonine, which inhibit cellular enzymes and cause systematic cell death. The objective of this study was to evaluate the perinatal effects of I. carnea in goats. METHODS: Forty-seven pregnant goats were randomly allocated into 5 treatment groups and given the following doses (g/kg BW) of I. carnea: 0 (IC0), 1.0 (IC1), 3.0 (IC3), 5.0 (IC5) and 7.5 (IC7). The treatment animals were given fresh I. carnea from day 27 of gestation to parturition. Weight gains and serum biochemistry were evaluated. Fetuses were evaluated using ultrasonographic measurements. RESULTS: Goats from the IC7 group showed clinical signs of poisoning. Ultrasound examination revealed that I. carnea feeding in all treatment groups reduced fetal movement compared to the controls. There was an increase in the total number of birth defects (retrognathia and arthrogyposis) in the IC7 and IC5 groups compared to the controls. CONCLUSION: The results show that I. carnea has teratogenic potential in goats. In addition, ultrasounds were useful in evaluating fetotoxicity and teratogenicity.

Towards a better understanding of Ipomoea asarifolia toxicity: evidence of the involvement of a leaf lectin.

Natural intoxication of livestock by ingestion of Ipomoea asarifolia leaves has been reported to occur widely in Brazil. Previous studies carried out by our research group provided strong evidence that a lectin could be involved with the toxic properties of I. asarifolia. To reinforce this hypothesis, a lectin-enriched fraction (LEF) was isolated from I. asarifolia leaves and its toxic effects were assessed. Leaves of I. asarifolia were excised from plants growing widely in the field, mechanically wounded and maintained in a chamber at 25 ± 3 °C for 72h in the dark, under near 100% relative humidity. The leaf proteins were extracted, ammonium sulfate precipitated, chromatographed on DEAE-cellulose and Phenyl-Sepharose to produce LEF that under SDS-PAGE showed a molecular mass of 44.0 kDa and after N-terminal amino acid analysis a primary sequence composed of AGYTPVLDIGAEVLAAGEPY. The in vivo toxicity of LEF assessed by intraorbital injection in mice showed induced severe uncoordinated movements without death. LEF reduced the muscular contraction in a dose depend way and at 29.8 µg/mL (CE(50)) it produces 50% inhibition of contraction, suggesting that LEF blunts autonomic neurotransmission. Isolated rat kidneys were perfused with LEF and no effects on the perfusion pressure or renal vascular resistance were observed, but urinary flow and glomerular filtration rate increased. Moreover, the percentage of tubular transport of Na(+), K(+) and Cl(-) decreased. Histological examination of the kidneys perfused with LEF exhibited little alterations. These toxic effects observed above were concomitant with the increase of LEF hemagglutination activity, which strongly suggest that one of the toxic principles of I. asarifolia is a lectin present in its leaves.

Effects of prepartum ingestion of Ipomoea carnea on postpartum maternal and neonate behavior in goats.

Ipomoea carnea is a toxic plant that grows in tropical areas, and is readily consumed by grazing goats. The plant contains the alkaloids swainsonine and calystegines, which inhibit cellular enzymes and cause systematic cell death. This study evaluated the behavioral effects on dams and kids of prenatal ingestion of this plant. Freshly harvested leaves of I. carnea (10 g/kg body weight) were fed daily to nine pregnant goats from the fifth to the 16th week of gestation; five pregnant goats were controls. Dam and kid behavior were evaluated during 2-hr postpartum. Further evaluation of the offspring was performed using various tests after birth: (1) reaching and discriminating their dam from an alien doe (two tests at 12-hr postpartum), and (2) navigating a progressive maze (2, 4, and 6 days postpartum). Postnatal (n = 2) and fetal (n = 2) mortality were observed in the treated group. Intoxicated kids had difficulty in standing at birth, and only one was able to suckle within 2 hr of birth. Treated kids were slower than controls to arrive at their dam in the discrimination test; treated kids often (seven of nine completed tests) incorrectly chose the alien dam (controls: 0/10 tests). During some runs on days 2, 4, and 6 postpartum, treated kids were slower to leave the starting point of the maze, and were slower to arrive at the dam on all test days. This study suggests that the offspring of pregnant goats given I. carnea during gestation have significant behavioral alterations and developmental delays.

Serum and kidney metabolic changes of rat nephrotoxicity induced by Morning Glory Seed.

Previous nephrotoxicology study on Morning Glory Seed (MGS) mainly focused on whole animal level, using creatinine and BUN as biochemical indicators to evaluate renal function. This work was designed to delineate the comprehensive metabolic syndromes of MGS-induced nephrotoxicity. An ultra liquid chromatography coupled with mass spectrometry (UPLC/MS) metabonomic approach was employed to characterize the metabolic profile of plasma and kidney tissue extract from rats treated with MGS at a single nephrotoxic dose of 15 g/kg/day for 14 consecutive days. Acquired data were subjected to principal component analysis (PCA) for differentiating the model and the control groups. The results indicated that certain metabolic pathways, such as lysophosphatidylcholines formation and sphingolipids cycle were accelerated, while the phenylalanine level in serum was decreased. We believe that metabonomic approach is helpful to further understanding and clinical diagnosis of traditional Chinese medicine (TCM) induced nephrotoxicity.

Plantas tóxicas para ruminantes e eqüídeos no Norte Piauiense / Toxic plants for ruminants and equidae in Northern Piauí

Este trabalho teve por objetivo realizar um levantamento sobre as plantas tóxicas para ruminantes e equídeos na Mesorregião Norte do Piauí. Foram feitas 71 entrevistas a médicos veterinários, engenheiros agrônomos, técnicos agrícolas e produtores de 16 municípios, entrevistando pelo menos quatro pessoas por município. As plantas comprovadamente tóxicas que foram apontadas com maior frequência na região estudada foram Ipomoea asarifolia, que causa intoxicações em pequenos ruminantes em todas as áreas visitadas. Stryphnodendron coriaceum pelas mortes que ocasiona é, aparentemente, a planta que causa maiores perdas econômicas na mesorregião estudada. Enterolobium contortisiliquum também foi citada como causa importante de sinais digestivos, abortamentos e fotossensibilização em bovinos da região. Os entrevistados confirmaram a ocorrência de surtos de intoxicação em bovinos por Thiloa glaucocarpa no inicio do período chuvoso. Manihot spp. e Piptadenia macrocarpa são plantas cianogênicas apontadas como causa de mortes superagudas em bovinos. Outras plantas relatadas como tóxicas pelos entrevistados, mas sem que haja comprovação de sua toxicidade, foram Buchenavia tomentosa, Caesalpinia sp., Brunfelsia sp., Luetzelburgia sp., Hybantus ipecaconha, Phisalys angulata e Spondias luta. De acordo com os entrevistados os frutos de Buchenavia tomentosa causam sinais digestivos e abortos em caprinos, ovinos e bovinos. Produtores relatam surtos de intoxicação em caprinos que apresentam sinais digestivos e morte após a ingestão de favas de Luetzelburgia sp. Brunfelsia sp. é relatada como causa de alterações nervosas, no começo das chuvas, quando os animais ingerem as folhas e flores e os asininos são aparentemente mais afetados. Os frutos de Spondias luta foram mencionados como causa de diarréia em caprinos. Experimentos não publicados demonstraram a toxicidade de Brunfelsia sp. em ovinos e de Luetzelburgia sp. como causa de sinais digestivos e mortes ...

The objective of this study was to survey toxic plants for ruminants and equidae in northern Piauí. Seventy one persons were interviewed, including farmers, veterinary practitioners, agronomists, and agrarian technicians from 16 municipalities, performing at least four interviews in each municipality. The most common plant mentioned as a cause of poisoning was Ipomoea asarifolia, which is a well known cause of tremogenic disease in ruminants. Stryphnodendron coriaceum which causes digestive signs was referred as a common cause of death, and is probably the plant that causes most cattle deaths in the region. Enterolobium contortisiliquum was also mentioned as a frequent cause of digestive signs, abortion and photosensitization in cattle. Outbreaks of nephrosis caused by Thiloa glaucocarpa are frequent at the beginning of the raining season. Poisoning by the cyanogenic plants Manihot spp. e Piptadenia macrocarpa are a cause of peracute deaths. Other plants mentioned as toxic were Buchenavia tomentosa, Caesalpinia sp., Brunfelsia sp., Luetzelburgia sp., Hybantus ipecaconha, Phisalys angulata, and Spondias luta. Farmers report that goats are poisoned by the ingestion of the pods of Luetzelburgia sp., which causes digestive signs and death. The ingestion of the fruits of Buchenavia tomentosa is associated with digestive signs and and abortion in ruminants. Brunfelsia sp. is mentioned as a cause of nervous signs at the start of the raining season and donkeys are apparently more affected. The consumption of the fruits of Spondias luta are associated with diarrhea in goats. Recent unpublished experiments demonstrated the toxicity of Brunfelsia sp. as a cause of nervous signs and of Luetzelburgia sp. as a cause of digestive signs in goats. Experiments with other plants are necessary to confirm their toxicity.

Plantas tóxicas para ruminantes e eqüídeos no Norte Piauiense / Toxic plants for ruminants and equidae in Northern Piauí

Este trabalho teve por objetivo realizar um levantamento sobre as plantas tóxicas para ruminantes e equídeos na Mesorregião Norte do Piauí. Foram feitas 71 entrevistas a médicos veterinários, engenheiros agrônomos, técnicos agrícolas e produtores de 16 municípios, entrevistando pelo menos quatro pessoas por município. As plantas comprovadamente tóxicas que foram apontadas com maior frequência na região estudada foram Ipomoea asarifolia, que causa intoxicações em pequenos ruminantes em todas as áreas visitadas. Stryphnodendron coriaceum pelas mortes que ocasiona é, aparentemente, a planta que causa maiores perdas econômicas na mesorregião estudada. Enterolobium contortisiliquum também foi citada como causa importante de sinais digestivos, abortamentos e fotossensibilização em bovinos da região. Os entrevistados confirmaram a ocorrência de surtos de intoxicação em bovinos por Thiloa glaucocarpa no inicio do período chuvoso. Manihot spp. e Piptadenia macrocarpa são plantas cianogênicas apontadas como causa de mortes superagudas em bovinos. Outras plantas relatadas como tóxicas pelos entrevistados, mas sem que haja comprovação de sua toxicidade, foram Buchenavia tomentosa, Caesalpinia sp., Brunfelsia sp., Luetzelburgia sp., Hybantus ipecaconha, Phisalys angulata e Spondias luta. De acordo com os entrevistados os frutos de Buchenavia tomentosa causam sinais digestivos e abortos em caprinos, ovinos e bovinos. Produtores relatam surtos de intoxicação em caprinos que apresentam sinais digestivos e morte após a ingestão de favas de Luetzelburgia sp. Brunfelsia sp. é relatada como causa de alterações nervosas, no começo das chuvas, quando os animais ingerem as folhas e flores e os asininos são aparentemente mais afetados. Os frutos de Spondias luta foram mencionados como causa de diarréia em caprinos. Experimentos não publicados demonstraram a toxicidade de Brunfelsia sp. em ovinos e de Luetzelburgia sp. como causa de sinais digestivos e mortes em caprinos.

The objective of this study was to survey toxic plants for ruminants and equidae in northern Piauí. Seventy one persons were interviewed, including farmers, veterinary practitioners, agronomists, and agrarian technicians from 16 municipalities, performing at least four interviews in each municipality. The most common plant mentioned as a cause of poisoning was Ipomoea asarifolia, which is a well known cause of tremogenic disease in ruminants. Stryphnodendron coriaceum which causes digestive signs was referred as a common cause of death, and is probably the plant that causes most cattle deaths in the region. Enterolobium contortisiliquum was also mentioned as a frequent cause of digestive signs, abortion and photosensitization in cattle. Outbreaks of nephrosis caused by Thiloa glaucocarpa are frequent at the beginning of the raining season. Poisoning by the cyanogenic plants Manihot spp. e Piptadenia macrocarpa are a cause of peracute deaths. Other plants mentioned as toxic were Buchenavia tomentosa, Caesalpinia sp., Brunfelsia sp., Luetzelburgia sp., Hybantus ipecaconha, Phisalys angulata, and Spondias luta. Farmers report that goats are poisoned by the ingestion of the pods of Luetzelburgia sp., which causes digestive signs and death. The ingestion of the fruits of Buchenavia tomentosa is associated with digestive signs and and abortion in ruminants. Brunfelsia sp. is mentioned as a cause of nervous signs at the start of the raining season and donkeys are apparently more affected. The consumption of the fruits of Spondias luta are associated with diarrhea in goats. Recent unpublished experiments demonstrated the toxicity of Brunfelsia sp. as a cause of nervous signs and of Luetzelburgia sp. as a cause of digestive signs in goats. Experiments with other plants are necessary to confirm their toxicity.

Intoxicação por Ipomoea asarifolia em ovinos e bovinos na Ilha de Marajó / Poisoning by Ipomoea asarifolia in cattle and sheep in the Marajó Island, Northern Brazil

Para estudar as intoxicações por plantas que ocorrem na Ilha de Marajó foram visitadas 7 fazendas. Em todas as fazendas visitadas as pastagens eram constituídas de campo nativo, tinham pouco ou nenhuma disponibilidade de forragem e estavam severamente invadidas por Ipomoea asarifolia. Nas 5 propriedades foi relatada a ocorrência da intoxicação por esta planta em ovinos. Animais jovens são mais afetados do que adultos. Em duas dessas propriedades foram observados ovinos com sinais clínicos. Em 4 fazendas, os proprietários relataram a ocorrência da intoxicação por I. asarifolia em bovinos e, em uma dessas, foram observados bovinos com sinais clínicos em duas visitas realizadas, uma no período seco e outra no início do período chuvoso. Em bovinos as mortes ocorrem principalmente quando são afetados bezerros. A doença é observada todos os anos, com morbidade variável e baixa mortalidade. Os sinais clínicos foram característicos de uma doença tremogênica. Um bovino e um ovino, que apresentavam sinais clínicos acentuados e foram eutanasiados e necropsiados, não apresentavam alterações macroscópicas nem histológicas de significação. Em todas as fazendas visitadas eram criados bubalinos juntamente com bovinos e/ou ovinos e em 3 propriedades eram criados caprinos, mas nenhum proprietário relatou a ocorrência da intoxicação nestas duas espécies.(AU)

To determine plant poisonings occurring on Marajo Island, state of Pará, northern Brazil, 7 farms were visited. All farms had native pastures with low forage availability severely infected by Ipomoea asarifolia. In 5 farms poisoning in sheep by the plant was reported. The disease occurs mainly during the dry period and lambs are more frequently affected than adult sheep. Sheep with clinical signs were observed in 2 farms. In 4 establishments farmers reported the occurrence of I. asarifolia poisoning in cattle. In one farm affected cattle were observed during two visits, one during the dry season and another at the start of the raining season. Deaths are observed mainly in calves. In both species the poisoning occurs every year with variable morbidity and low mortality. One bovine and one sheep with severe clinical signs were euthanized and post-mortem examination was performed. No gross or histologic significant lesions were observed. In all farms buffaloes were raised together with cattle and sheep, and in 3 farms goats were also raised. None of the farmers reported the intoxication in these species.(AU)

Intoxicação experimental por Ipomoea asarifolia (Convolvulaceae) em caprinos e ovinos / Experimental poisoning by Ipomoea asarifolia (Convolvulaceae) in goats and sheep

Ipomoea asarifolia causa uma síndrome tremorgênica em ovinos, caprinos, bovinos e búfalos. Este experimento teve como objetivos determinar a toxicidade para caprinos de I. asarifolia verde, colhidas nas épocas de chuva e de estiagem, e da planta seca triturada, determinar a toxicidade da planta para ovinos, e determinar se o princípio ativo da planta é eliminado pelo leite em doses tóxicas para os cordeiros. No primeiro experimento a planta fresca colhida na época de estiagem e na época de chuvas foi administrada a 16 caprinos. A planta colhida na estiagem foi tóxica na dose diária de 5 e 10g por kg de peso animal (g/kg). A planta colhida na época de chuva foi tóxica na dose diária de 20 e 30g/kg, demonstrando que a planta é mais tóxica durante o período seco. A planta seca, colhida na época de estiagem foi administrada a 9 caprinos em doses diárias de 1.7, 2, 3.4 e 5.1g/kg. Doses de 3, 4 e 5.1g/kg causaram sinais clínicos, demonstrando que a planta mantém a toxicidade após a secagem. A planta fresca colhida na época de estiagem e na época de chuvas foi administrada a 10 ovinos. A planta colhida na estiagem foi tóxica na dose diária de 5g/kg e na época de chuva foi tóxica nas doses de 10 e 20g/kg. Estes resultados sugerem a maior susceptibilidade dos ovinos à intoxicação do que os caprinos. Como alguns produtores mencionam que cordeiros lactentes que não estão pastando se intoxicam através do leite, I. asarifolia foi administrada diariamente nas doses de 2.5, 5 e 10g/kg a 5 ovelhas, a partir do dia do parto (2 ovelhas), do último dia de prenhez (1 ovelha) e 60 dias antes da parição (2 ovelhas). As ovelhas, mas não os cordeiros, apresentaram sinais clínicos, sugerindo que o princípio ativo da planta não é eliminado no leite ou colostro em doses tóxicas para os cordeiros. Em um ovino eutanasiado não foram observadas lesões macroscópicas nem histológicas. Os achados ultra-estruturais mais significativos foram encontrados nos dendritos das...(AU)

Ipomoea asarifolia causes a tremogenic syndrome in sheep, goats, cattle and buffaloes. The objectives of the experiments were (1) to determine the toxicity to goats of fresh I. asarifolia collected during the raining and the dry season, and the toxicity of the dried plant, and (2) to determine the toxicity of the plant to sheep, and if the active principle is eliminated through the milk. In the first experiment the plant collected in the dry season and in the raining season was fed to 16 goats. The plant collected during the dry season caused clinical signs at the daily doses of 5g and 10g/kg body weight. The plant collected during the raining season was toxic at daily doses of 20g and 30g/kg, indicating that the plant is more toxic during the dry season. The plant collected in the dry season and dried was fed to 9 goats at doses of 1.7g, 2.0g, 3.4g, and 5.1g per kg. Daily doses of 3.0g, 4.0g and 5.1g/kg caused clinical signs, showing that the plant maintains its toxicity after being dried. In the second experiment the fresh plant collected in the dry and in the raining season was fed to 10 sheep. The plant collected in the dry season was toxic at the dose of 5g/kg, and the plant collected in the raining season was toxic at the doses of 10g and 20g/kg. The experimental results suggest that sheep are more susceptible to the poisoning than goats. As some farmers mentioned that suckling non-grazing lambs are poisoned by milk ingestion, I. asarifolia was fed at daily doses of 2.5g, 5.0g and 10g/kg for variable periods to 5 sheep from the day of parturition (2 sheep), after the last day of pregnancy (1 sheep) and 60 days before parturition (2 sheep). The sheep but not the lambs showed clinical signs of intoxication suggesting that the active principle is not eliminated through the milk at doses toxic for the lambs. In one euthanized sheep no gross or histologic lesions were detected. The main ultra-structural findings were found in Purkinje...(AU)

Embryotoxic effects of prenatal treatment with Ipomoea carnea aqueous fraction in rats / Efeitos embriotóxicos do tratamento pré-natal com extrato aquoso de Ipomoea carnea em ratos

The embryotoxic effects of prenatal daily exposure to 0.0, 0.7, 3.0 or 15.0 mg/kg of the aqueous extract (AQE) from Ipomoea carnea (I. carnea) dried leaves on gestational days 5û21 were studied in rats. Maternal reproductive performance, skeletal and visceral abnormalities, and malformations were evaluated. Moreover, anatomopathological findings in dams following the treatment were recorded. Regarding the dams, our results show that body weight, weight gain, food and water consumption, and reproductive performance were all unaffectedby exposure to the different doses of the AQE. Nonetheless, dams treated with AQE presented a dose-dependent cytoplasmic vacuolation in the liver, kidneys, thyroid and adrenal glands. Fetal examination did not show external abnormalities or malformations. Evidences of several skeletal and visceral abnormalities were found, particularly after the higher dose of AQE. A reduced ossification centers were also detected. The present data show that prenatal ingestion of the I. carnea AQE in rats induces embryotoxicity. These effects are attributed to an active principle from I. carnea acting on maternal homeostasis, or directly in the conception.

Os efeitos embriotóxicos da exposição diária pré-natal a 0,0, 0,7, 3,0 ou 15,0mg/kg do extrato aquoso da I. carnea nos dias 5 a 21 de gestação foram estudados. Foram avaliados a performance reprodutiva materna, anormalidades esqueléticas e viscerais e malformações. Além disso, após o tratamento foram encontrados achados anatomopatológicos. Em relação às ratas mães, nossos resultados mostraram que a exposição às diferentes doses não afetou o peso corporal, ganho de peso, consumos de água e ração e performance reprodutiva. Apesar disso, apresentaram vacuolização citoplasmática de forma dose-dependente em fígado, rins, tireóide e glândula adrenal. Exames fetais não mostraram anormalidades externas ou malformações, sendo somente encontradas evidências de anormalidades esqueléticas e viscerais após altas doses do extrato. Foi observada redução dos centros de ossificação. Os presentes dados mostram que a ingestão prenatal do extrato de I. carnea induz embriotoxicidade. Estes efeitos são atribuídos à ação na homeostase maternal ou diretamente na concepção.