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AIMS: The aims of the study were to (i) improve the evaluation criteria of detoxifying Jatropha curcas L. cake (JCC), (ii) isolate and characterize a JCC tolerant strain, (iii) explore its JCC detoxifying potential. METHODS AND RESULTS: The zebrafish was employed as a survival model to screen the strains capable of detoxifying JCC. A strain identified as Mucor circinelloides SCYA25, which is highly capable of degrading all toxic components, was isolated from soil. Different solid-state fermentation parameters were optimized by response surface methodology. The optimal values for inoculation amount, moisture content, temperature, and time were found to be 18% (1·8 × 106 spores g-1 cake), 66%, 26, and 36 days, respectively, to achieve maximum detoxification of the JCC (92%). Under optimal fermentation conditions, the protein content of JCC was increased, while the concentrations of ether extract, crude fiber, toxins, and anti-nutritional substances were all degraded considerably (P < 0·05). Scanning electron microscopy and Fourier transform infrared spectrometer analysis revealed that the fermentation process could disrupt the surface structure and improve the ratio of α-helix to ß-folding in the JCC protein, which may improve the digestibility when the detoxified JCC is used as a feedstuff. CONCLUSIONS: Our results indicate that M. circinelloides SCYA25 is able to detoxify JCC and improve its nutritional profile, which is beneficial to the safe utilization of JCC as a protein feedstuff. SIGNIFICANCE AND IMPACT OF THE STUDY: The newly identified M. circinelloides SCYA25 detoxified JCC in a safe manner to provide a potential alternative to soybean meal for the feed industry. These results also provide a new perspective and method for the toxicity evaluation and utilization of JCC and similar toxic agricultural by-products.

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The toxic kernel cake of Jatropha curcas (KCakeJ) is an emerging health and environmental concern. Although phorbol esters are widely recognized as the major toxin of KCakeJ, convincing evidence is absent. Here, we show that rather than phorbol esters an isomeric mixture of 11-hydroxy-9E-octadecenoic acid, 12-hydroxy-10E-octadecenoic acid and 12-hydroxy-10Z-octadecenoic acid (hydroxy-octadecenoic acids, molecular formula C18H34O3) is the major toxic component. The toxicities of hydroxy-octadecenoic acids on experimental animals, e.g. acute lethality, causing inflammation, pulmonary hemorrhage and thrombi, allergies, diarrhea and abortion, are consistent with those on human/animals caused by Jatropha seed and/or KCakeJ. The hydroxyl group and the double bond are essential for hydroxy-octadecenoic acids' toxicity. The main pathway of the toxicity mechanism includes down-regulating UCP3 gene expression, promoting ROS production, thus activating CD62P expression (platelet activation) and mast cell degranulation. The identification of the major toxin of KCakeJ lays a foundation for establishing an environmentally friendly Jatropha biofuel industry.

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Edible/non-toxic varieties of Jatropha curcas L. are gaining increasing attention, providing both oil as biofuel feedstock or even as edible oil and the seed kernel meal as animal feed ingredient. They are a viable alternative to the limitation posed by the presence of phorbol esters in toxic varieties. Accurate genotyping of toxic/non-toxic accessions is critical to breeding management. The aim of this study was to identify SNP markers linked to seed toxicity in J. curcas. For SNP discovery, NGS technology was used to sequence the whole genomes of a toxic and non-toxic parent along with a bulk of 51 toxic and 30 non-toxic F2 plants. To ascertain the association between SNP markers and seed toxicity trait, candidate SNPs were genotyped on 672 individuals segregating for seed toxicity and two collections of J. curcas composed of 96 individuals each. In silico SNP discovery approaches led to the identification of 64 candidate SNPs discriminating non-toxic and toxic samples. These SNPs were mapped on Chromosome 8 within the Linkage Group 8 previously identified as a genomic region important for phorbol ester biosynthesis. The association study identified two new SNPs, SNP_J22 and SNP_J24 significantly linked to low toxicity with R2 values of 0.75 and 0.54, respectively. Our study released two valuable SNP markers for high-throughput, marker-assisted breeding of seed toxicity in J. curcas.

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Jatropha curcas is an important oilseed plant, with considerable potential in the development of biodiesel. Although Jatropha seed cake, the byproduct of oil extraction, is a residue rich in nitrogen, phosphorus, potassium, and carbon, with high protein content suitable for application in animal feed, the presence of toxic phorbol esters limits its application in feed supplements and fertilizers. This review summarizes the current methods available for detoxification of this residue, based upon chemical, physical, biological, or combined processes. The advantages and disadvantages of each process are discussed, and future directions involving genomic and proteomic approaches for advancing our understanding of biodegradation processes involving microorganisms are highlighted.

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Jatropha dioica Sessé ex Cerv. is a medicinal plant credited with low cytotoxicity in vitro. Thus, the objective of this work was to evaluate the possible genotoxic and cytotoxic effect in vivo of the J. dioica aqueous extract by means of micronucleus assay in mouse peripheral blood. Four different J. dioica aqueous extract dose-units were evaluated (30, 60, 100, and 300 mg/kg). The extract was administered orally to male Balb-C-strain mice every 24 h during 5 days. Blood samples were taken at 0, 24, 48, 72, 96, and 120 h from the mouse's tail and were performed in duplicate extensions. The number of Polychromatic Erythrocytes (PCE), Polychromatic Micronucleus Erythrocytes (PCEMN), and Micronucleus Erythrocytes (MNE) was determined at the different sampling times in the different study groups. Our results showed that the group that received 60 mg/kg of cyclophosphamide (positive control) presented a significant decrease in the PCE (p = 0.044) proportion and a significant increase in MNE (p = 0.032, p = 0.0001). The groups that received the different J. dioica aqueous extract doses did not present either a PCE decrease or an increase in PCEMN and MNE. J. dioica exerts neither a genotoxic nor a cytotoxic effect on mouse peripheral blood at high doses.

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Jatropha has been highlighted as an oleaginous potential for the production of biofuel. The cake, produced by oil extraction, could be used in animal feed. However, some varieties of jatropha are toxic by limiting their incorporation into animal diets. The objective of this study was to evaluate the hepatotoxicity of diets added with jatropha cake ­ JC (Jatropha curcas) in rats. Thirty-five (35) male Wistar adults rats (Rattus norvegicus) with initial weight of 352.1 ± 26.8 g were used. The animals were fed for 21 days with the diets: control, 10, 25, 40 and 50% JC. In the feeding with 50% JC the animals presented themselves prostrate and with piloerection. Development and survival decreased, since the inclusion of JC in diets increased. In rats submitted to 10 and 25% JC there was an increase of 17.52% in the hepatosomatic index in relation to the control group. Increase of JC in the rat diet promoted an increase in the activity of ALT and AST enzymes. Anatomic-histopathological evaluation demonstrated that, regardless of the levels tested, JC in rat diet causes hypertrophy of the hepatocytes, with a reduction in energy reserves. This study demonstrated that the use of JC resulted in decreased food intake, associated with weight loss due to the clinical pattern of toxicity, demonstrated by biochemical and histopathological changes in the liver. It was concluded that the inclusion of jatropha cake in rat feeding presents high hepatotoxic potential leading to lesions in the liver parenchyma.(AU)

O pinhão-manso tem se destacado como oleaginosa potencial para a produção de biocombustível. A torta, coproduto da extração do óleo, poderia ser utilizada na alimentação animal. No entanto, algumas variedades de pinhão-manso são tóxicas, limitando sua incorporação em dietas animais. Objetivou-se neste estudo avaliar a hepatotoxicidade de dietas acrescidas de torta de pinhão-manso (Jatropha curcas) em ratos. Foram utilizados trinta e cinco (35) ratos Wistar (Rattus norvergicus) machos adultos com peso inicial de 352,1 ± 26,8 g. Os animais foram alimentados por 21 dias com as dietas: controle, 10, 25, 40 e 50% TPM. Na alimentação com 50% TPM os animais apresentaram-se prostrados e com piloereção. O desenvolvimento e a sobrevivência apresentaram diminuição conforme o aumento da inclusão de TPM nas dietas. Em ratos submetidos a 10 e 25% TPM houve aumento de 17,52% no índice hepatossomático em relação ao grupo controle. O aumento de TPM na dieta de ratos promoveu aumento da atividade das enzimas ALT e AST. A avaliação anatomo-histopatológica revelou que, independentemente dos níveis testados, a TPM na alimentação de ratos provoca hipertrofia dos hepatócitos, com redução das reservas energéticas. Este estudo demonstrou que a utilização de TPM resultou em diminuição do consumo de alimento associado à perda de peso devido ao quadro clínico de toxicidade demonstrado pelas alterações bioquímica e histopatológica no fígado. Conclui-se que a inclusão de torta de pinhão-manso na alimentação de ratos apresenta alto potencial hepatotóxico levando a lesões no parênquima hepático.(AU)

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Jatropha gossypiifolia L. (Euphorbiaceae) is widely used in popular medicine. However, further toxicological studies are necessary for its reliable use. The present study aimed to evaluate the cytotoxic, genotoxic, and mutagenic effects of ethanolic and aqueous leaf extracts of J. gossypiifolia, using the test system Allium cepa. In addition, the phytochemical profile of the extracts was also obtained. Seeds of A. cepa were subjected to different concentrations of the two extracts (0.001, 0.01, 0.1, 1, and 10 mg/mL). Distilled water was used for the negative control and methyl methanesulfonate (4 x 10(-4) M) and trifluralin (0.84 ppm) for the positive controls. The values of mitotic index at all concentrations of ethanolic extract and at 0.1, 1, and 10 mg/mL aqueous extract showed a significant decrease. Alterations, such as chromosome adherence, C-metaphases, chromosome bridges, nuclear buds, and micronuclei were verified in both extracts but chromosome loss indicating genotoxic activity was observed only in the ethanolic extract. Presence of micronuclei on administration of the extracts, also indicated mutagenic action at the chromosome level. In the ethanolic extract, aneugenicity seemed to be the main activity, probably as a result of the action of terpenes and/or flavonoids, whereas in the aqueous extract, clastogenic action appeared to be the principal activity, presumably as a consequence of the effect of flavonoids and/or saponins. Thus, we suggest that the extracts of this species should be used with great caution for medicinal purpose.

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Objetivou-se com o presente trabalho, avaliar o efeito anti-helmíntico de Jatropha mollissima por meio de experimentos in vitro e in vivo. Inicialmente foi investigada a concentração de extrato com efeito bioativo, pelo teste de evolução da toxicidade do extrato etanólico de J. mollissima sobre o microcrustáceo Artemia salina, obtendo uma CL50 de 660,80µg/ml, que foi testada em coproculturas contendo larvas infectantes de Haemonchus contortus e em animais para a verificação da redução do OPG. Para o teste in vivo o extrato foi dissolvido em água para se obter as concentrações 660,80µg/ml e 1321,6µg/ml, foram coletadas fezes semanalmente e sangue quinzenalmente. Como resultados dos testes in vitro, o extrato etanólico do caule de Jatropha mollissima mostrou-se tóxico sobre A. salina, com CL50 abaixo de 1000 µg/ml e inibiu a eclosão de ovos e o desenvolvimento de larvas de H. contortus, apresentando uma eficiência de 70,77%. O teste in vivo revelou que o extrato é também eficaz em ovinos, com redução significativa na contagem de OPG após 28 dias de experimento, 47 e 44% de redução nos grupos tratados com o extrato, 7,5% no grupo de animais não tratados e 40,6% com a ivermectina. Mesmo parasitados, os animais permaneceram clinicamente saudáveis e sem anemia. O extrato etanólico do caule de Jatropha mollissima pode representar uma alternativa ao controle da verminose ovina, pois retarda a resistência parasitária.(AU)

This study aimed to evaluate the anthelmintic effect of Jatropha mollissima through in vitro and in vivo experiments. Initially we investigated the concentration of extract with bioactive effect, through the toxicity evolution test of the ethanol extract of J. mollissima on the microcrustacean Artemia salina, obtaining CL50 concentration of 660.80µg/ml, which was tested in fecal cultures containing infective larvae of Haemonchus contortus and in animals for the verification of OPG reduction. For in vivo test, the extract was dissolved in water to obtain concentrations of 660.80µg/ml and 1321.6µg/ml. Feces were collected weekly and blood was collected every fifteen days. As a result of in vitro test, the ethanol extract of the stem of J. mollissima proved toxic on A. salina, with CL50 less than 1000µg/ ml and inhibited the eggs hatching and the development of larvae of H. contortus, presenting an efficiency of 70.77%. in vivo test revealed that the extract is also effective in sheep, with a significant reduction in the count of OPG after 28 days of experiment, 47 and 44% of reduction in the groups treated with the extract, 7.5% in the untreated group of animals and 40.6% with ivermectin. Even parasitized, the animals remained clinically healthy and without anemia. The ethanol extract of the stem of Jatropha mollissima may represent an alternative to the control of sheep worms, because it slows the parasitic resistance.(AU)

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Introducción. Estudios preclínicos evidencian efectos dosis dependiente sobre la analgesia e inflamación y neurotoxicidad de las hojas, corteza y raíz de J. curcas L. El propósito del estudio fue evaluar la actividad analgésica y neurofarmacológica de las fracciones de la semilla de J. curcas L. Métodos. Estudio experimental, preclínico y prospectivo. Se distribuyeron 48 ratones en seis grupos control: ácido acético, diclofenaco, tramadol, agua destilada, diazepam y cafeína. Cuatro grupos experimentales: fracción soluble a 500 mg/kg y fracción no soluble a 250, 500 y 750 mg/kg; se evaluó los efectos sobre la algesia, por medio de la prueba de contorsiones abdominales por ácido acético a 1,5 %, y las manifestaciones neurológicas, mediante la prueba de Irwin. Se realizaron pruebas para el análisis de las variables cuantitativas y para las variables cualitativas. Resultados. La inhibición de las contorsiones fue 62,27 %, 56,86 %, 44,12 % y 42,06 % para los grupos 5, 2, 4 y 3, respectivamente. Las manifestaciones neurológicas de los grupos experimentales mostraron presencia y significancia de las variables excitación, sacudidas de cabeza, rascarse, incoordinación motora, cola de Straub, piloerección y estereotipias. Las variables estereotipias y rascado se presentaron en las dos fracciones. Conclusión. La fracción soluble y la no soluble del extracto etanólico de J. curcas L. presentaron efecto analgésico y efectos tóxicos a nivel del sistema nervioso central.

Introduction. Preclinical studies show the dose dependent effects on the analgesia, inflammation and neurotoxicity of the leaves, bark and root. The purpose of the study was to evaluate the analgesic and neuropharmacological activity of the fractions of the seed J. curcas L. Methods. Experimental study, preclinical and prospective. 48 rats were distributed in 6 control groups: acetic acid, diclofenac, tramadol, distilled water, diazepam, caffeine. Four experimental groups: 500 mg/kg of soluble fraction and 250, 500 y 750 mg/kg of insoluble fraction; the effects on the algesia were evaluated using an exam of abdominal contortions with acetic acid on 1,5%; the neurological manifestations were measured with the Irwin test. To analyze the quantitative and qualitative variables exams were used. Results. The inhibition of the contortions were 62,27 %, 56,86 %, 44,12 % y 42,06 % for the groups 5, 2, 4 y 3, respectively. The neurological manifestations of the experimental groups showed the presence and significance of the variables: excitation, head twitches, scratching, motor incoordination, Straub tail, piloerection and stereotypies. The variables stereotypes and scratching presented themselves in the two fractions. Conclusion. The soluble and insoluble fraction of the ethanoic extract of J. Curcas L. presented analgesic and toxic effects on the central nervous system.

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Jatropha curcas L. (Euphorbiaceae) is important for biofuel production and as a feed ingredient for animal. However, the presence of phorbol esters in the oil and cake renders the seeds toxic. The toxicity of J. curcas oil is currently assessed by testing in animals, leading to their death. The identification of toxic and nontoxic improved varieties is important for the safe use of J. curcas seeds and byproducts to avoid their environmental toxicity. Hence, the aim of this study was to propose a short-term bioassay using a plant as a model to screen the toxicity of J. curcas oil without the need to sacrifice any animals. The toxicity of J. curcas oil was evident in germination, root elongation and chromosomal aberration tests in Lactuca sativa. It was demonstrated that J. curcas seeds contain natural compounds that exert phyto-, cyto- and genotoxic effects on lettuce, and that phorbol esters act as aneugenic agents, leading to the formation of sticky chromosomes and c-metaphase cells. In conclusion, the tests applied have shown reproducibility, which is important to verify the extent of detoxification and to determine toxic doses, thus reducing the numbers of animals that would be used for toxicity tests.

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Curcin, a type I ribosomal inhibiting protein-RIP, encoded by curcin precursor gene, is a phytotoxin present in Jatropha (Jatropha curcas L.). Here, we report designing of RNAi construct for the curcin precursor gene and further its genetic transformation of Jatropha to reduce its transcript expression. Curcin precursor gene was first cloned from Jatropha strain DARL-2 and part of the gene sequence was cloned in sense and antisense orientation separated by an intron sequence in plant expression binary vector pRI101 AN. The construction of the RNAi vector was confirmed by double digestion and nucleotide sequencing. The vector was then mobilized into Agrobacterium tumefaciens strain GV 3101 and used for tissue culture independent in planta transformation protocol optimized for Jatropha. Germinating seeds were injured with a needle before infection with Agrobacterium and then transferred to sterilized sand medium. The seedlings were grown for 90 days and genomic DNA was isolated from leaves for transgenic confirmation based on real time PCR with NPT II specific dual labeled probe. Result of the transgenic confirmation analysis revealed presence of the gene silencing construct in ten out of 30 tested seedlings. Further, quantitative transcript expression analysis of the curcin precursor gene revealed reduction in the transcript abundance by more than 98% to undetectable level. The transgenic plants are being grown in containment for further studies on reduction in curcin protein content in Jatropha seeds.

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Jatropha oil is an emerging feedstock for the production of biodiesels. The increasing use of this nonedible, toxic oil will result in higher potential for accidental exposures. A repeated-dose 28-day oral toxicity study was conducted to provide data for risk assessment. Jatropha oil diluted in corn oil was administered by gavage to male and female rats at 0.5, 5, 50 and 500 mg kg(-1) body weight per day for 28 consecutive days. Control rats were administered corn oil only. The growth rates and consumption of food and water were monitored. At necropsy, organs were weighed and hematological parameters assessed. Serum clinical chemistry and C-reactive protein were measured and histological examinations of organs and tissues were performed. Markedly depressed growth rate was observed in males and females receiving Jatropha oil at 500 mg kg(-1) per day. Decreased white blood cell and lymphocyte counts were detected in females at 50 and 500 mg kg(-1) per day and in males at 500 mg kg(-1) per day. These changes were correlated to mild and reversible histological changes in male and female spleens. In the liver, a mild increase in portal hepatocytes cytoplasm density was observed in males and females, while periportal vacuolation was observed exclusively in females. Mild acinar proliferation was observed in the female mammary glands at all dose levels. It is concluded that Jatropha oil produces adverse effects on female rats starting at 50 mg kg(-1) per day with decreased white blood cell and lymphocyte counts and at 500 mg kg(-1) per day in both genders in term of depressed growth rates.

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A huge amount of kernel cake, which contains a variety of toxins including phorbol esters (tumor promoters), is projected to be generated yearly in the near future by the Jatropha biodiesel industry. We showed that the kernel cake strongly inhibited plant seed germination and root growth and was highly toxic to carp fingerlings, even though phorbol esters were undetectable by HPLC. Therefore it must be detoxified before disposal to the environment. A mathematic model was established to estimate the general toxicity of the kernel cake by determining the survival time of carp fingerling. A new strain (Streptomyces fimicarius YUCM 310038) capable of degrading the total toxicity by more than 97% in a 9-day solid state fermentation was screened out from 578 strains including 198 known strains and 380 strains isolated from air and soil. The kernel cake fermented by YUCM 310038 was nontoxic to plants and carp fingerlings and significantly promoted tobacco plant growth, indicating its potential to transform the toxic kernel cake to bio-safe animal feed or organic fertilizer to remove the environmental concern and to reduce the cost of the Jatropha biodiesel industry. Microbial strain profile essential for the kernel cake detoxification was discussed.

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Jatropha curcas L., a multipurpose shrub, has acquired significant economic importance for its seed oil which can be converted to biodiesel an emerging alternative to petro-diesel. In addition to the commercial value, it is also having medicinal and even high nutritional value to use as animal fodder which is limited due to the toxicity. Development of molecular marker will enable to differentiate non-toxic from toxic variety of J. curcas in a mixed population and also for quality control since the toxic components of J. curcas has deleterious effect on animals. In the present study, the efforts were made to generate the specific SCAR marker for toxic and/or non-toxic J. curcas from RAPD markers. Among the markers specific for toxic and non-toxic varieties, four were selected, purified, cloned, sequenced, and designed primers out of which one set of primers NT-JC/SCAR I/OPQ15-F and R could able to discriminate the non-toxic with toxic Jatropha by giving expected 430 bp size amplification in non-toxic variety. Furthermore, novel multiplex PCR was designed using the nrDNA ITS primers to overcome the false negatives. Present work also demonstrates utility of the conserved regions of nrDNA coding genes in ruling out the artifacts in PCR-like false negatives frequently occur in SCAR due to various reasons. The specific SCAR markers generated in the present investigation will help to distinguish non-toxic from toxic varieties of J. curcas or vice versa, and isolated marker along with designed multiplex protocol has applications in quality control for selective cultivation of non-toxic variety and will also assist in breeding and molecular mapping studies.

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The nutrients intake and histopathological occurrences were evaluated in sheep fed diets with increasing levels of inclusion (0%, 15%, 30%, and 45%) of Jatropha curcas dried and crushed fruit shell in substitution to Mombaça grass hay. Twenty castrated male sheep were kept in metabolic cages and fed diets ad libitum (5 to 10% scraps were allowed). At the end of the experiment, the animals were subjected to clinical examination and blood collection, followed by euthanasia and necropsy. The clinical and pathological disorders were characterized by digestive, lung, and heart alterations, with regressive changes affecting kidney and liver. Presence of fluid in the abdominal and pericardial cavities, intestine with little content or empty, reddish serosa, and hyperemic mucosa with streaks of blood were observed during necropsy; as well as edematous mesenteric lymph nodes; red, swollen, and firm lung (hepatization) with emphysema areas, suffusions in the serous of the rumen, and ulcers in the abomasum. Histopathological analysis revealed congestion and pulmonary edema, congestion and vacuolar degeneration of hepatocytes, edematous mesenteric lymph nodes, tubular degeneration, and the presence of calcium crystals in the kidney. Thus, the collected data reveals that J. curcas fruit shell is toxic when added at different levels in sheep diet.

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A espécie Jatropha gossypiifolia L. (Euphorbiaceae), popularmente conhecida como pião-roxo, entre tantos outros nomes, é um bom exemplo do tênue limiar que separa o efeito terapêutico do tóxico. Apesar de ser classicamente conhecida como planta tóxica possui usos na medicina popular. Alguns desses efeitos têm sido comprovados em estudos experimentais, como os de antimicrobiano, antineoplásico, cicatrizante e hipotensor, sendo possivelmente explicados pela presença de substâncias como alcalóides, terpenóides, flavonóides, lignanas e taninos. Esta revisão aborda aspectos importantes, com ênfase na toxicidade crônica dessa espécie, de modo a servir de fonte de informação aos interessados em avaliar a relação risco/benefício do uso terapêutico de Jatropha gossypiifolia L.

The species Jatropha gossypiifolia L. (Euphorbiaceae), popularly known as bellyache bush, among several other names, is an important example of the tenuous threshold that separates the therapeutic from the toxic effect. Although traditionally known as a toxic plant, it has been used in folk medicine. Some of its effects have been proved by experimental studies as antimicrobial, antineoplastic, healing and hypotensive, likely explained by the presence of substances such as alkaloids, terpenoids, flavonoids, lignans and tannins. This review deals with important aspects, focusing on the chronic toxicity of this species, in order to serve as an information source for those interested in evaluating the risk-benefit ratio of the therapeutic use of Jatropha gossypiifolia L.

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Jatropha is a nonedible oil seed plant belonging to Euphorbiaceae family. Global awareness of sustainable and alternative energy resources has propelled research on Jatropha oil as a feedstock for biodiesel production. During the past two decades, several cultivation projects were undertaken to produce Jatropha oil. In future, the increased cultivation of toxic Jatropha plants and utilization of its agro-industrial by-products may raise the frequency of contact with humans, animals, and other organisms. An attempt was thus made to present known information on toxicity of Jatropha plants. The toxicity of Jatropha plant extracts from fruit, seed, oil, roots, latex, bark, and leaf to a number of species, from microorganisms to higher animals, is well established. Broadly, these extracts possess moluscicidal, piscicidal, insecticidal, rodenticidal, antimicrobial, and cytotoxic properties, and exert adverse effects on animals including rats, poultry, and ruminants. The toxicity attributed to these seeds due to their accidental consumption by children is also well documented. An attempt was also made to identify areas that need further study. The information provided in this review may aid in enhancing awareness in agroindustries involved in the cultivation, harvesting, and utilization of Jatropha plants and its products with respect to the potential toxicity of Jatropha, and consequently in application and enforcement of occupational safety measures. Data on the wide range of bioactivities of Jatropha and its products were collated and it is hoped will create new avenues for exploiting these chemicals by the phamaceutical industry to develop chemotherapeutic agents.

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BACKGROUND: Jatropha curcas, a non-edible oil plant, is being promoted as a biofuel plant in a number of countries in tropical and subtropical regions. The kernel meal left after extraction of the oil is a potentially protein-rich feed ingredient. However, the presence of highly toxic phorbol esters limits its use. Degradation of J. curcas phorbol esters by rumen microbes, using an in vitro rumen fermentation system, has been investigated in this study. RESULTS: The difference between phorbol ester contents in the residues obtained with and without substrates at 0, 24, 48 or 72 h of the incubations was statistically similar. Phorbol esters did not affect either the gas or short chain production in the in vitro rumen fermentation system. CONCLUSIONS: Rumen microbes can not degrade phorbol esters. In addition, the phorbol esters do not adversely affect rumen fermentation. Ruminants are expected to be as prone as monogastric animals to the toxicity of Jatropha seeds.

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Protein rich Jatropha curcas kernel meal is toxic. It was detoxified using heat treatment and solvent extraction. Two duration of detoxification process were investigated: shorter (30 min) and longer (60 min) and the detoxified meals so obtained were designated as J(a) and J(b) respectively. Common carp fingerlings (252 fish; 3.2+/-0.07 g) were fed with the following diets: Control containing fishmeal (FM); S(50,) J(a50) and J(b50): 50% of FM protein replaced by soybean meal (SBM), detoxified Jatropha kernel meal (DJ(a)KM and DJ(b)KM); S(75), J(a75) and J(b75): 75% of FM protein replaced by SBM, DJ(a)KM and DJ(b)KM. White blood cells count, mean cell volume and mean cell haemoglobin concentration, calcium and sodium ions and total bilirubin in blood did not differ significantly among the groups. Higher (P>0.05) RBC count was observed in plant protein fed groups compared to control group. Highest alkaline phosphatase and alanine transaminase activities in blood were observed in J(a75), which were not different (P>0.05) from those in J(a50) group, but were higher than in the other groups. No adverse histopathological changes in liver and muscle of any group were observed, but intestinal mucosa of J(a75) groups showed severe pathological lesions. The results demonstrate that Jb was completely detoxified. Since the performance of J(b50) group was similar to control group and better than the other groups, optimum inclusion level of J(b) is 50% replacement of FM protein.

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Jatropha curcas L., a multipurpose shrub has acquired significant economic importance for its seed oil which can be converted to biodiesel, is emerging as an alternative to petro-diesel. The deoiled seed cake remains after oil extraction is toxic and cannot be used as a feed despite having best nutritional contents. No quantitative and qualitative differences were observed between toxic and non-toxic varieties of J. curcas except for phorbol esters content. Development of molecular marker will enable to differentiate non-toxic from toxic variety in a mixed population and also help in improvement of the species through marker assisted breeding programs. The present investigation was undertaken to characterize the toxic and non-toxic varieties at molecular level and to develop PCR based molecular markers for distinguishing non-toxic from toxic or vice versa. The polymorphic markers were successfully identified specific to non-toxic and toxic variety using RAPD and AFLP techniques. Totally 371 RAPD, 1,442 AFLP markers were analyzed and 56 (15.09%) RAPD, 238 (16.49%) AFLP markers were found specific to either of the varieties. Genetic similarity between non-toxic and toxic verity was found to be 0.92 by RAPD and 0.90 by AFLP fingerprinting. In the present study out of 12 microsatellite markers analyzed, seven markers were found polymorphic. Among these seven, jcms21 showed homozygous allele in the toxic variety. The study demonstrated that both RAPD and AFLP techniques were equally competitive in identifying polymorphic markers and differentiating both the varieties of J. curcas. Polymorphism of SSR markers prevailed between the varieties of J. curcas. These RAPD and AFLP identified markers will help in selective cultivation of specific variety and along with SSRs these markers can be exploited for further improvement of the species through breeding and Marker Assisted Selection (MAS).

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