Antimicrobial activities and phytochemicals of Murraya paniculata L. flowers, leaves and bark crude extracts

Aims@#Murraya paniculata (L.) has been widely employed in medicine, has also been modified to serve as an ingredient in health foods and found application in cosmetics. This study was aimed to assess the biological activities of M. paniculata by analyzing the chemical compositions of its flowers, leaves and bark.@*Methodology and results@#Crude extracts drawn from the flowers, leaves and bark of M. paniculata underwent testing to determine the antibacterial properties in terms of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), as well as the overall chemical composition, total phenolic content, flavonoids and antioxidant activity. Crude extract of leaves exhibited the most potent antibacterial activity when tested against Staphylococcus aureus TISTR 1466, Bacillus subtilis ATCC 6633 and Pseudomonas aeruginosa ATCC 27853. The crude extract from bark delivered the most significant antibacterial activity when tested against Micrococcus luteus TISTR 9341, Escherichia coli ATCC 1261, Pseudomonas sp., Streptococcus sp. and Methicilin resistant S. aureus (MRSA). For all crude extracts, the MIC value against M. luteus TISTR 9341 was 12.5 mg/mL. Meanwhile, the MBC value for the crude extract of leaves against B. subtilis ATCC 6633 was 12.5 mg/mL, whereas, for flower and bark crude extracts, the MBC value against S. aureus TISTR 1466 was 25 mg/mL. Antioxidant activity was at its highest for the crude extract from bark (IC50 = 1.36 mg/mL). The highest phenolic content was recorded for the crude extract from bark, while the highest flavonoid content came from the crude extract of leaves (70.81 ± 0.31 mgGAE/g extract and 115.73 ± 1.18 mgQE/g extract, respectively).@*Conclusion, significance and impact of study@#The research findings suggest that the crude extracts of M. paniculata leaves and bark show greater significant levels of bioactivity than was the case for crude extracts from flowers. The research findings could help in exploring the possibilities of using M. paniculata for pharmaceutical purposes and in aquaculture.

Antimicrobial and toxicological evaluation of Origanum vulgare: an in vivo study / Avaliação antimicrobiana e toxicológica de Origanum vulgare: um estudo in vivo

Origanum vulgare has been of great interest in academia and pharma industry due to its antioxidant, antifungal and antitumor properties. The present study aimed to find the anti-MRSA potential and in vivo toxicity assessments of O. vulgare. O. vulgare extract was used to monitor anti-MRSA activity in mice. Following MRSA established infection in mice (Mus musculus), treatment with O. vulgare was continued for 7 days. Autopsies were performed and re-isolation, gross lesion scoring and bacterial load in various organs were measured. Additionally, blood sample was analysed for hematological assays. Toxicity assessment of O. vulgare potential as medicine was done at 200 mg/kg and 400 mg/kg by evaluating liver and kidney functions. Bacterial load and gross lesion in lungs and heart were significantly low compared to positive control following O. vulgare treatment. Likewise, O. vulgare treated groups had hematological, neutrophil and TLC values similar to control groups. Increased AST, ALP and total bilirubin along with marked hepatocellular degeneration and distortion around the central vein, inflammatory cell infiltration, and cytoplasmic vacuolization of hepatic cells was observed at higher dose. It is concluded that crude extract of O. vulgare may contain beneficial secondary metabolites and in future may be explored for curing infectious diseases.

Origanum vulgare tem despertado grande interesse na academia e na indústria farmacêutica devido às suas propriedades antioxidantes, antifúngicas e antitumorais. O presente estudo teve como objetivo encontrar o potencial anti-MRSA e avaliações de toxicidade in vivo de O. vulgare. O extrato de O. vulgare foi usado para monitorar a atividade anti-MRSA em camundongos. Após infecção estabelecida por MRSA em camundongos (Mus musculus), o tratamento com O. vulgare foi continuado por 7 dias. As autópsias foram realizadas e o reisolamento, pontuação das lesões grosseiras e carga bacteriana em vários órgãos foram medidos. Além disso, a amostra de sangue foi analisada para ensaios hematológicos. A avaliação da toxicidade do potencial de O. vulgare como medicamento foi feita com 200 mg / kg e 400 mg / kg, avaliando as funções hepática e renal. A carga bacteriana e as lesões graves nos pulmões e no coração foram significativamente baixas em comparação com o controle positivo após o tratamento com O. vulgare. Da mesma forma, os grupos tratados com O. vulgare apresentaram valores hematológicos, de neutrófilos e de TLC semelhantes aos grupos de controle. Aumento de AST, ALP e bilirrubina total juntamente com degeneração hepatocelular marcada e distorção ao redor da veia central, infiltração de células inflamatórias e vacuolização citoplasmática de células hepáticas foram observados em doses mais altas. Conclui-se que o extrato bruto de O. vulgare pode conter metabólitos secundários benéficos e, no futuro, pode ser explorado para a cura de doenças infecciosas.

Antimicrobial and toxicological evaluation of Origanum vulgare: an in vivo study

Abstract Origanum vulgare has been of great interest in academia and pharma industry due to its antioxidant, antifungal and antitumor properties. The present study aimed to find the anti-MRSA potential and in vivo toxicity assessments of O. vulgare. O. vulgare extract was used to monitor anti-MRSA activity in mice. Following MRSA established infection in mice (Mus musculus), treatment with O. vulgare was continued for 7 days. Autopsies were performed and re-isolation, gross lesion scoring and bacterial load in various organs were measured. Additionally, blood sample was analysed for hematological assays. Toxicity assessment of O. vulgare potential as medicine was done at 200 mg/kg and 400 mg/kg by evaluating liver and kidney functions. Bacterial load and gross lesion in lungs and heart were significantly low compared to positive control following O. vulgare treatment. Likewise, O. vulgare treated groups had hematological, neutrophil and TLC values similar to control groups. Increased AST, ALP and total bilirubin alongwith marked hepatocellular degeneration and distortion around the central vein, inflammatory cell infiltration, and cytoplasmic vacuolization of hepatic cells was observed at higher dose. It is concluded that crude extract of O. vulgare may contain beneficial secondary metabolites and in future may be explored for curing infectious diseases.

Resumo Origanum vulgare tem despertado grande interesse na academia e na indústria farmacêutica devido às suas propriedades antioxidantes, antifúngicas e antitumorais. O presente estudo teve como objetivo encontrar o potencial anti-MRSA e avaliações de toxicidade in vivo de O. vulgare. O extrato de O. vulgare foi usado para monitorar a atividade anti-MRSA em camundongos. Após infecção estabelecida por MRSA em camundongos (Mus musculus), o tratamento com O. vulgare foi continuado por 7 dias. As autópsias foram realizadas e o reisolamento, pontuação das lesões grosseiras e carga bacteriana em vários órgãos foram medidos. Além disso, a amostra de sangue foi analisada para ensaios hematológicos. A avaliação da toxicidade do potencial de O. vulgare como medicamento foi feita com 200 mg / kg e 400 mg / kg, avaliando as funções hepática e renal. A carga bacteriana e as lesões graves nos pulmões e no coração foram significativamente baixas em comparação com o controle positivo após o tratamento com O. vulgare. Da mesma forma, os grupos tratados com O. vulgare apresentaram valores hematológicos, de neutrófilos e de TLC semelhantes aos grupos de controle. Aumento de AST, ALP e bilirrubina total juntamente com degeneração hepatocelular marcada e distorção ao redor da veia central, infiltração de células inflamatórias e vacuolização citoplasmática de células hepáticas foram observados em doses mais altas. Conclui-se que o extrato bruto de O. vulgare pode conter metabólitos secundários benéficos e, no futuro, pode ser explorado para a cura de doenças infecciosas.

Antimicrobial and toxicological evaluation of Origanum vulgare: an in vivo study / Avaliação antimicrobiana e toxicológica de Origanum vulgare: um estudo in vivo

Abstract Origanum vulgare has been of great interest in academia and pharma industry due to its antioxidant, antifungal and antitumor properties. The present study aimed to find the anti-MRSA potential and in vivo toxicity assessments of O. vulgare. O. vulgare extract was used to monitor anti-MRSA activity in mice. Following MRSA established infection in mice (Mus musculus), treatment with O. vulgare was continued for 7 days. Autopsies were performed and re-isolation, gross lesion scoring and bacterial load in various organs were measured. Additionally, blood sample was analysed for hematological assays. Toxicity assessment of O. vulgare potential as medicine was done at 200 mg/kg and 400 mg/kg by evaluating liver and kidney functions. Bacterial load and gross lesion in lungs and heart were significantly low compared to positive control following O. vulgare treatment. Likewise, O. vulgare treated groups had hematological, neutrophil and TLC values similar to control groups. Increased AST, ALP and total bilirubin alongwith marked hepatocellular degeneration and distortion around the central vein, inflammatory cell infiltration, and cytoplasmic vacuolization of hepatic cells was observed at higher dose. It is concluded that crude extract of O. vulgare may contain beneficial secondary metabolites and in future may be explored for curing infectious diseases.

Resumo Origanum vulgare tem despertado grande interesse na academia e na indústria farmacêutica devido às suas propriedades antioxidantes, antifúngicas e antitumorais. O presente estudo teve como objetivo encontrar o potencial anti-MRSA e avaliações de toxicidade in vivo de O. vulgare. O extrato de O. vulgare foi usado para monitorar a atividade anti-MRSA em camundongos. Após infecção estabelecida por MRSA em camundongos (Mus musculus), o tratamento com O. vulgare foi continuado por 7 dias. As autópsias foram realizadas e o reisolamento, pontuação das lesões grosseiras e carga bacteriana em vários órgãos foram medidos. Além disso, a amostra de sangue foi analisada para ensaios hematológicos. A avaliação da toxicidade do potencial de O. vulgare como medicamento foi feita com 200 mg / kg e 400 mg / kg, avaliando as funções hepática e renal. A carga bacteriana e as lesões graves nos pulmões e no coração foram significativamente baixas em comparação com o controle positivo após o tratamento com O. vulgare. Da mesma forma, os grupos tratados com O. vulgare apresentaram valores hematológicos, de neutrófilos e de TLC semelhantes aos grupos de controle. Aumento de AST, ALP e bilirrubina total juntamente com degeneração hepatocelular marcada e distorção ao redor da veia central, infiltração de células inflamatórias e vacuolização citoplasmática de células hepáticas foram observados em doses mais altas. Conclui-se que o extrato bruto de O. vulgare pode conter metabólitos secundários benéficos e, no futuro, pode ser explorado para a cura de doenças infecciosas.

Biosíntesis inducida de fengicina y surfactina en una cepa de Bacillus amyloliquefaciens con actividad oomiceticida sobre zoosporas de Phytophthora capsica / Induced biosynthesis of fengycin and surfactin in a strain of Bacillus amyloliquefaciens with oomyceticidal activity on zoospores of Phytophthora capsici

Resumen La aplicación de metabolitos antimicrobianos biosintetizados por especies de Bacillus es una alternativa potencial para controlar Phytophthora capsici (P. capsici) en hortalizas y podría evitar el uso de productos químicos con acción oomiceticida. El objetivo de este estudio fue evaluar el impacto de la adición al medio de cultivo de distintos agentes (ácido glutámico, hierro, celulosa, quitina y células inactivas de Colletotrichum spp.) sobre la biosíntesis de lipopéptidos en Bacillus amyloliquefaciens KX953161.1 y examinar la capacidad oomiceticida de dichos compuestos in vitro sobre las zoosporas de P. capsici. Los lipopéptidos identificados y cuantificados por cromatografía en capa fina de alta resolución (HPTLC) en los extractos crudos fueron fengicina y surfactina. El cultivo bacteriano adicionado con células inactivas de Colletotrichum spp. demostró la mayor biosíntesis de lipopéptidos: 1.847,02 ±11,8 pg/mL de fengicina y 2.563,45 ± 18,4 pg/mL de surfactina. Los tratamientos con menor producción de estos lipopéptidos fueron aquellos a los que se añadió hierro (608,05 ± 22,6 pg/mL de fengicina y 903,74 ±22,1 pg/mL de surfactina) o celulosa (563,31 ±11,9 y 936,96 ±41,1 pg/mL, igual orden). El extracto con los lipopéptidos presentó una inhibición del 100% en la germinación de zoosporas de P. capsici, se observó enquistamiento, malformaciones en el tubo germinal y degradación celular. Se concluye que los lipopéptidos producidos por B. amyloliquefaciens KX953161.1 podrían contribuir al control de P. capsici, sin embargo, se requieren más estudios a fin de elucidar el modo de acción biológica de estos compuestos y optimizar el perfil de producción y el rendimiento.

Abstract A potential alternative to the use of chemical products with oomyceticidal action for the control of Phytophthora capsici in vegetables is the use of antimicrobial metabolites, biosynthesized in Bacillus species. The objective of this study was to induce the biosynthesis of lipopeptides in Bacillus amyloliquefaciens KX953161.1 by using glutamic acid, iron, cellulose, chitin, or inactive Colletotrichum spp. cells. The in vitro oomyceticidal effect of the bacterial lipopeptides on zoospores of Phytophthora capsici was evaluated. The lipopeptides identified and quantified in the crude extracts by high performance thin layer chromatography (HPTLC) were fengycin and surfactin. The bacterial culture with inactive fungal cells yielded the greatest biosynthesis of lipopeptides, at 1847.02± 11.8 and 2563.45± 18.4 pg/ml of fengycin and surfactin, respectively and the treatments that obtained lower production of these lipopepti-des, were those to which iron and cellulose were added with 608.05 ± 22.6 and 903.74± 22.1; 563.31± 11.9 and 936.96± 41.1 pg/ml for fengicin and surfactin, respectively. The lipopeptide extracted showed 100% germination inhibition on zoospores of P. capsici, revealing encystment, malformations in the germ tube and cellular degradation. Lipopeptides have the potential to control P. capsici; however, the biosynthesis of these lipopeptides requires further study to determine their biological mode of action and optimize lipopeptide performance and profile.

Acaricidal activity of Furcraea foetida leaf extract against engorged female Rhipicephalus (Boophilus) microplus ticks

The Rhipicephalus (Boophilus) microplus tick is a major concern for the livestock market worldwide, as it causes serious economic damage. Plant-derived acaricides are an attractive alternative to control this ectoparasite and limit the development of resistance. Therefore, the aim of this study was to evaluate the acaricidal activity of Furcraea foetida leaf extract against engorged female R. (B.) microplus ticks. Our in vitro bioassays showed that the crude extract of leaves from F. foetida caused hemorrhagic swelling and skin lesions in the ticks, and three days of treatment caused 100% mortality. Dose-response assay indicated that this toxicity effect was dose-dependent. Similar effects were observed when the crude extract from F. foetida leaves was denatured by boiling at 100°C. These results suggest that the toxicity of the leaf extract might be associated with thermostable biomolecules. Together, our results show for the first time that the crude extract of F. foetida leaves has acaricidal activity against engorged female R. (B.) microplus ticks and it acts in a dose-dependent manner.

The acetone crude extract of Quercus infectoria (Olivier) galls alters pH of the digestive vacuole of the malaria parasite, Plasmodium falciparum

@#The reduced efficacy of the mainstay antimalarial drugs due to the widespread of drugresistant Plasmodium falciparum has necessitated efforts to discover new antimalarial drugs with new targets. Quercus infectoria (Olivier) has long been used to treat various ailments including fever. The acetone extract of the plant galls has recently been reported to have a promising antimalarial activity in vitro. This study was aimed to determine the effect of the Q. infectoria gall acetone crude extract on pH of the digestive vacuole of Plasmodium falciparum. A ratiometric fluorescent probe, fluorescein isothiocyanate-dextran (FITC-dextran) was used to facilitate a quantitative measurement of the digestive vacuole pH by flow cytometry. Mid trophozoite stage malaria parasites grown in resealed erythrocytes containing FITC-dextran were treated with different concentrations of the acetone extract based on the 50% inhibitory concentration (IC50). Saponin-permeabilized parasites were analyzed to obtain the ratio of green/yellow fluorescence intensity (Rgy) plotted as a function of pH in a pH calibration curve of FITC-dextran. Based on the pH calibration curve, the pH of the digestive vacuole of the acetone extract-treated parasites was significantly altered (pH values ranged from 6.35- 6.71) in a concentration-dependent manner compared to the untreated parasites (pH = 5.32) (p < 0.001). This study provides a valuable insight into the potential of the Q. infectoria galls as a promising antimalarial candidate with a novel mechanism of action.

Study on Improvement Effects of Ganoderma lucidum Polysaccharide Crude Extract on Estradiol-induced Thymus Atrophy in Mice / 中国药房

OBJECTIVE： To study the imp rovement effects of Ganoderma lucidum polysaccharides crude extract on estradiol-induced thymus atrophy in mice. METHODS ：Totally 60 female ICR mice were randomly divided into normal control group（normal saline ），model group （normal saline ），G. lucidum polysaccharides crude extract high-dose and low-dose groups （400，100 mg/kg，by crude drug ），with 15 mice in each group. Except for normal control group ，other groups were given estradiol intraperitoneally （0.1 mg/mice，6 times）every other day to establish thymic atrophy model. The next day after modeling finished，they were given relevant medicine intragastrically ，once a day ，for consecutive 14 d. Twenty-four hours after last medication，organ（thymus，spleen）index，MDA content and GST activity in plasma were determined. HE staining was adopted to observe the pathological changes of thymus and spleen tissue in mice. The thymus cell apoptosis was examined by TUNEL assay ， and the T cell subsets in peripheral blood were detected by flow cytometry. RESULTS ：Compared with normal control group ，the thymus index ，proportion of CD 3+CD4+T cell in peripheral blood and CD 4+/CD8+ ratio were decreased significantly in model group （P＜0.01）；spleen index ，MDA content in plasma and thymocyte apoptosis level as well as the proportion of CD 3+CD8+T cell in peripheral blood were all increased significantly （P＜0.05 or P＜0.01）. Thymic cortex and medullary boundary of mice was blurred；the intercellular space was enlarged ；some cells were damaged and apoptotic in cortex ；no pathological changes were found in the spleen. Compared with model group ，thymus index and GST activity in plasma as well as proportion of CD 3+CD4+T cell in peripheral blood and CD 4+/CD8+ ratio were all increased significantly in G. lucidum polysaccharides crude extract high-dose group（P＜0.05 or P＜0.01）；while MDA content in plasma ，the apoptosis level of thymocytes were all decreased significantly （P＜0.01 or P＜0.05）；and the pathological changes of thymus were improved significantly. MDA content in plasma was decreased significantly in G. lucidum polysaccharides crude extract low-dose group （P＜0.01），and other indexes/pathological changes were not obvious. CONCLUSIONS ：High dose （400 mg/kg）of G. lucidum polysaccharides crude extract can improve the thymus atrophy induced by estradiol in mice.

Efecto in vitro del extracto crudo de un estreptomiceto causante de sarna común de papa en Sinaloa, México / In vitro effect of the crude extract of a potato common scab streptomycete in Sinaloa, Mexico

Abstract A strain isolated from potato common scab superficial lesions in El Fuerte Valley in northern Sinaloa, Mexico, was identified by 16S rRNA and morphological methods. Moreover, the effects of the crude extract of strain V2 was evaluated on radish and potato. The isolate was similar to Streptomyces acidiscabies in its morphological properties; however, the 16S rRNA gene sequence of strain V2 was neither 100% identical to this species nor to the streptomycetes previously reported in Sinaloa, Mexico. Strain V2 did not amplify any specific PCR products for genes necl and tomA, which have been found and reported in S. acidiscabies. Strain V2 produced a PCR product for the txtAB operon, which is related to the production of thaxtomin. In vitro assays using crude thaxtomin extract and a spore suspension of the organism caused necrotic symptoms on radish and potato, which were highly virulent in potato. This study reports that Streptomyces sp. V2 has a toxigenic region (TR) that is associated with the thaxtomin gene cluster.

Resumen Se aisló una cepa de una lesión superficial de sarna común de la papa en un ejemplar procedente del Valle del Fuerte, en el norte de Sinaloa, México. La cepa fue identificada por secuenciación del gen 16S ARNr, y por sus características morfológicas. Los efectos del extracto crudo de dicha cepa, llamada V2, fue evaluado en papa y rábano. El aislado fue similar a Streptomyces acidiscabies en sus características morfológicas, pero la secuencia del gen 16S ARNr de la cepa V2 no fue 100% idéntica a la de dicha especie, ni tampoco a las de cepas identificadas dentro de este taxón previamente en Sinaloa, México. La cepa V2 no amplificó los productos específicos de PCR de los genes nec1 y tomA, los cuales sí se han reportado en S. acidiscabies. La cepa V2 amplificó el producto de PCR para del operón txtAB, relacionado con la producción de taxtomina. A través de ensayos in vitro usando un extracto crudo de taxtomina y una suspensión de esporas del organismo aislado se verificó la producción de síntomas necróticos en rábano y papa, con mayor virulencia en esta última especie. Este estudio indica que Streptomyces sp. V2 tiene una región toxigénica (TR) asociada con el cluster de genes de taxtomina.

Growth retardation and apoptotic death of tumor cells by Artemisia herba-alba oral administration in Ehrlich solid carcinoma bearing mice

ABSTRACT At present, there is a rapidly growing interest in studying the cytotoxic effects of Artemisia herba alba Asso, Asteraceae, in various cancer cell lines. However, its antitumor effectiveness has not been investigated. Therefore, the current study was conducted to study the effect of A. herba alba extract on the proliferation and growth of solid tumor cells in Ehrlich Solid Carcinoma bearing mice. Oral administration of A. herba alba extract resulted in significant reductions in tumor size, tumor weight and mice body weight, as well as caused concurrent significant increases in the DNA breakages and apoptotic DNA damage induction in a time-dependent manner. A. herba alba extract also raised the expression level of p53 gene and reduced of K-ras expression in a time-dependent manner. Minor histological lesions were observed in the liver and kidney tissues sections of mice administered A. herba alba extract compared with the high histological lesions observed in the liver and kidney tissues of artesunate and cisplatin treated groups. Thus, we concluded that A. herba alba extract exhibited promising potential antitumor efficacy with greater safety than artesunate and the commercially used anticancer drug cisplatin in mice.

Antibacterial activity of medicinal plant extracts against Ralstonia solanacearum (Smith) that causes bacterial wilt in hot pepper (Capsicum annuum L. )

This study was conducted to evaluate the in vitro antibacterial activity of some medicinal plants against Ralstonia solanacearum. Bioactive chemicals were extracted from Burcea antidysenterica, Eucalyptus citriodora, Justicia schimperiana, Lantana camara, Melia azedarach and Ricinus communis leaves using maceration method. The bioassay was evaluated by disc diffusion method. The pathogen was isolated from infected Capsicum annuum plants using Casamino acid-Peptone-Glucose agar (CPG) medium. The isolate was identified using cultural, biochemical characteristics, pathogenicity test and found to be R. solanacearum. The methanol extracts had different composition, percentage extract yield, antibacterial activity and relative percentage inhibition. Unlike others, extracts of E. citriodora and R. communis consisted of all the tested secondary metabolites. All species showed antibacterial activity except M. azedarach. Significant differences were recorded in antibacterial activity among species and test concentrations. The highest antibacterial activity and the lowest bacteriostatic and bactericidal concentrations were found from E. citriodora and R. communis extracts. The higher potency of E. citriodora and R. communis extracts suggested the potential of the two species as a biocide to control bacterial wilt. However, further in vivo studies on these species extracts are compulsory.

Study on Improvement Effect of Selenium-enriched Ganoderma lucidum Crude Extract on Lipid Metabolism ，Liver Function and Inflammatory Response in Type 2 Diabetic Model Rats and Its Mechanism / 中国药房

OBJECTIVE： To study the effects of selenium-enriched Ganoderma lucidum crude extract on lipid metabolism， liver function and inflammatory response in type 2 diabetic model rats. METHODS： Totally 120 rats were randomly divided into normal control group （n＝20， normal saline） and model group （n＝100）. Normal control group was fed with normal diet， and model group was fed with high-fat diet. 4 weeks later， model group was given intraperitoneal injection of Streptozotocin solution （30 mg/kg） to induce T2DM model. After modeling， 90 rats were randomly subdivided into model control group （normal saline）， positive control group （metformin， 200 mg/kg） and selenium-enriched G. lucidum crude extract low-dose， medium-dose and high-dose groups （300， 600， 1 200 mg/kg， calculated by extract）， with 18 rats in each group. They were given medicine intragastrically， once a day， from Monday to Saturday. Half of rats in each group were selected 4， 8 weeks after medication； the serum levels of glucose and insulin were detected， and islet resistance index were calculated. The serum levels of liver function indexes （AST， ALT， AKP）， blood lipid indexes （FFA， TC， TG， LDL-C） and inflammatory factors （TNF-α， IL-6， IL-1β） were detected by ELISA. After HE staining， the histopathological changes of liver tissue were observed by microscopy. mRNA and protein expressions of peroxisome proliferator activated receptor α （PPARα） and peroxidase acyl coenzyme A oxidase 1 （ACOX1） in liver tissue were detected by RT-qPCR and Western blot assay. RESULTS： Compared with normal control group， glucose， insulin serum levels and islet resistance index were significantly increased （P＜0.01）； serum liver function indexes， blood lipid indexes and inflammatory factor levels of model control group were increased significantly in model control group after 4 and 8 weeks medication （P＜0.05 or P＜0.01）. The hepatocyte swelling of model control group was round and the volume was significantly larger than that of blank control group.  The liver had different degrees of steatosis and vacuolization， accompanied by a small amount of inflammatory cell infiltration. mRNA and protein expressions of PPARα and ACOX1 in liver tissue were decreased significantly （P＜0.05 or P＜0.01）. Compared with model control group， except that there was no significant decreased in islet resistunce index and AST， ALT， IL-6， IL-1β serum levels after 4 weeks of medication， and glucose， insulin， ALT serum levels after 8 weeks of medication and the levels of 4 blood lipid indexes after 4 and 8 weeks of medication in selenium-enriched G. lucidum crude extract low-dose group （P＞0.05）， above serum indexes of other groups were decreased significantly after 4 and 8 weeks of medication （P＜0.05 or P＜0.01）. After 4 and 8 weeks of medication， the pathological changes of liver tissue in rats were alleviated in varying degrees. protein and mRNA expressions of PPARα and ACOX1 in liver tissue were increased significantly after 4 and 8 weeks of medication （P＜0.05 or P＜0.01）. CONCLUSIONS： Selenium-enriched G. lucidum crude extract can up-regulate protein and mRNA expressions of PPARα and ACOX1 in liver tissue， promote the excretion of accumulated fatty acid and significantly improve fatty acid metabolism， inflammatory response and liver function in T2DM model rats.

FOLIAR ENDOPHYTE FUNGI AS CANDIDATE FOR BIOCONTROL AGAINST Moniliophthora spp. OF Theobroma cacao (Malvaceae) IN ECUADOR / Hongos endófitos foliares como candidatos a biocontroladores contra Moniliophthora spp. de Theobroma cacao (Malvaceae) en Ecuador

ABSTRACT The adaptability of endophytic fungi to their hosts, the ecological benefits that it provides and the various antagonistic mechanisms against pests make them an alternative for the biological control of diseases. The potential of 17 strains of foliar endophytic fungi (FEF) obtained from healthy Theobroma cacao tissue as candidates for the biological control of Moniliophthora roreri (MR) and M. perniciosa (MP) was determined. We evaluated: i) mycoparasitism of FEF against colonies of Moniliophthora spp., ii) the effects of crude metabolites of FEF on the pathogens' growth, and iii) the ability to recolonize healthy leaves of the host by leaf assays. Three strains of Lasiodiplodia theobromae were the most promising: Ec098, Ec151 and Ec157. These strains inhibited the growth of MR and MP, both in the confrontation of the colonies and by their metabolites and, additionally, recolonized the host between 80-100 % of the time. Other strains showed outstanding values i n one indicator, and not desirable in others. For example, Ec035 (L. theobromae) showed the highest levels of mycoparasitism against both pathogens in the interaction of the colonies, and the second best for its metabolites, but could not reinfect the host. Strain Ec059 (Xylaria feejeensis) reinfected 100 %, but did not show desirable attributes of antagonism. On the other hand, the metabolites of Ec107 (Colletotrichum gloeosporioides s.l.) inhibited MR by 60 %, but also stimulated the growth of MP. No strain achieved all desirable characteristics for a biological control agent.

RESUMEN La adaptabilidad de los hongos endófitos a sus hospedantes, los beneficios ecológicos que le brinda y los diversos mecanismos antagónicos contra plagas que poseen los convierten en una alternativa para el control biológico de enfermedades. Se determinó el potencial de 17 cepas de hongos endofíticos foliares (FEF) obtenidas de tejido sano de Theobroma cacao como candidatas para el control biológico de Moniliophthora roreri (MR) y M. perniciosa (MP). Se evaluaron: i) el micoparasitismo de los FEF frente a colonias de Moniliophthora spp., ii) la acción de los metabolitos crudos de los FEF en el crecimiento, y iii) la habilidad para recolonizar hojas sanas del hospedante mediante ensayos de hojas sueltas. Tres cepas de Lasiodiplodia theobromae fueron las más promisorias: Ec098, Ec151 and Ec157. Estas cepas inhibieron el crecimiento de MR y MP, tanto en el enfrentamiento de las colonias como mediante sus metabolitos y, adicionalmente, recolonizaron el hospedante entre el 80-100 % de las veces. Otras cepas mostraron valores destacados en un indicador, y no deseables en otros. Por ejemplo, la Ec035 (L. theobromae) mostró los niveles más altos de micoparasitismo contra ambos patógenos en la interacción de las colonias, y el segundo mejor por sus metabolitos, pero no pudo reinfectar el hospedante. La cepa Ec059 (Xylaria feejeensis) reinfectó 100 %, pero no mostró los atributos deseados de antagonismo. Por su parte, los metabolitos de Ec107 (Colletotrichum gloeosporioides s.l.) inhibieron a MR en un 60 %, pero también estimularon el crecimiento de MP. Ninguna cepa logró todas las características deseables para un agente de control biológico.

Anti-schistosomal activities of Echinops kebericho Mesfin root and Hagenia abyssinica (Bruce) J.F Gmel flower part crude extracts in Swiss albino mice

Objective: To evaluate the in vivo antischistosomal activities of the crude extracts of Echinops kebericho Mesfin (E. kebericho) root and Hagenia abyssinica (Bruce) J.F. Gmel (H. abyssinica) flower. Methods: Mice were infected with (150 ± 10) Schistosoma mansoni cercariae by paddling technique. Crude extracts were administered orally for five consecutive days at doses of 300, 600 and 1 200 mg/kg/day along with 200 mg/kg/day praziquantel and 3% tween 80 given as a control. Results: E. kebericho root extract showed a statistically significant (P < 0.05) reduction in fecal egg count of 64.44%, 42.96% & 26.82% and worm burden of 65.71%, 47.86% & 31.43% at treatment doses of 1 200 mg/kg/day, 600 mg/kg/day and 300 mg/kg/day, respectively. Similarly, H. abyssinica flower extracts showed a significant (P < 0.05) reduction in fecal egg count up to 84.57%, 77.06% & 63.89% and worm burden of 91.43%, 81.43% & 70.71% at a respective dose levels. In addition, a significant (P < 0.05) reduction in liver granuloma score was observed in all H. abyssinica administered dose groups and E. kebericho at 1 200 mg/kg/day dose group as compared to infected untreated control group. Conclusions: H. abyssinica and E. kebericho crude extracts show a promising antischistosomal activity.

Anti-schistosomal activities of Echinops kebericho Mesfin root and Hagenia abyssinica (Bruce) J.F Gmel flower part crude extracts in Swiss albino mice

Objective:To evaluate the in vivo antischistosomal activities of the crude extracts of Echinops kebericho Mesfin (E. kebericho) root and Hagenia abyssinica (Bruce) J.F. Gmel (H. abyssinica) flower.Methods:Mice were infected with (150 ± 10) Schistosoma mansoni cercariae by paddling technique. Crude extracts were administered orally for five consecutive days at doses of 300, 600 and 1 200 mg/kg/day along with 200 mg/kg/day praziquantel and 3% tween 80 given as a control.Results:E. kebericho root extract showed a statistically significant (P < 0.05) reduction in fecal egg count of 64.44%, 42.96% & 26.82% and worm burden of 65.71%, 47.86% & 31.43% at treatment doses of 1 200 mg/kg/day, 600 mg/kg/day and 300 mg/kg/day, respectively. Similarly, H. abyssinica flower extracts showed a significant (P < 0.05) reduction in fecal egg count up to 84.57%, 77.06% & 63.89% and worm burden of 91.43%, 81.43% & 70.71% at a respective dose levels. In addition, a significant (P < 0.05) reduction in liver granuloma score was observed in all H. abyssinica administered dose groups and E. kebericho at 1 200 mg/kg/day dose group as compared to infected untreated control group.Conclusions:H. abyssinica and E. kebericho crude extracts show a promising antischistosomal activity.

Larvicidal efficacy of crude and fractionated extracts of Dracaena loureiri Gagnep against Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus mosquito vectors

Objective: To evaluate the larvicidal efficacy of crude and fractionated extracts of Dracaena loureiri endocarp against Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus mosquitos. Methods: Larvicidal activity was tested according to World Health Organization standard protocol.The third-stage larvae of each mosquito species were exposed to various concentrations of Dracaena loureiri crude extract and six groups of Dracaena loureiri fractionated extracts (RC-DT 009–014). Larval mortality rates were observed after 24 h and 48 h of exposure.Then, a computerized probit analysis of the mortality data was performed to determine lethal concentration 50 (LC50) and lethal concentration 90 values. Results: Anopheles minimus larvae (24-h LC5077.88 mg/L) had the highest susceptibility to crude extract, whereas others (Aedes aegypti, 24-h LC50224.73 mg/L; Aedes albopictus, 24-h LC50261.75 mg/L; and Culex quinquefasciatus, 24-h LC50282.86 mg/L) were significantly less susceptible. The most effective groups of fractionated extracts were RC-DT 012 and RC-DT 013. The mosquito species most susceptible to fractionated extracts was Culex quinquefasciatus, with 24-h LC50 values of 0.66 and 0.94 mg/L for RC-DT 012 and RC-DT 013, respectively. Conclusions: The larvicidal activity of fractionated extracts is more effective than that of crude extract against all tested mosquito species. For the most effective alternative larvicide, purification and a phytochemical constituent analysis must be performed.

Larvicidal efficacy of crude and fractionated extracts of Dracaena loureiri Gagnep against Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus mosquito vectors

Objective: To evaluate the larvicidal efficacy of crude and fractionated extracts of Dracaena loureiri endocarp against Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus mosquitos. Methods: Larvicidal activity was tested according to World Health Organization standard protocol. The third-stage larvae of each mosquito species were exposed to various concentrations of Dracaena loureiri crude extract and six groups of Dracaena loureiri fractionated extracts (RC-DT 009-014). Larval mortality rates were observed after 24 h and 48 h of exposure. Then, a computerized probit analysis of the mortality data was performed to determine lethal concentration 50 (LC

Antiviral activity of Myracrodruon urundeuva against rotavirus

ABSTRACT Myracrodruon urundeuva Allemão, Anacardiaceae, is a medicinal plant widely found in Brazil, especially in the northern region. In our previous study, the ethanolic extract from leaves of M. urundeuva showed antiviral activity against simian rotavirus SA-11. Here, the crude extract was subjected to fractionations in order to subsequently work with more concentrated and pure bioactive compounds, which were analyzed by TLC and HPLC methods to support a better understanding of their virucidal effect. The antiviral activity was evaluated using a rotavirus infection model in MA-104 cells treated with the maximum non-cytotoxic concentration of the crude extract and its fractions. Data were expressed as the percentage inhibition of viral replication calculated by the inhibition of cytopathic effect in the treated cells compared to untreated controls after 48 h of incubation. First, we conducted a fractionation, generating five fractions (F1–F5) which were submitted to antiviral assay. Then, the fraction that showed the highest virucidal effect (F3, PI = 75%) was subjected to a larger partition, yielding eighteen subfractions, which were submitted to new antiviral assays. Terpenes, flavonoids and tannins were the major secondary metabolites detected by TLC analysis in F3. SF1, a flavonoid-enriched fraction, showed the strongest in vitro activity against rotavirus (PI = 92%), preventing cytopathic effect. Chromatographic profiles were obtained by HPLC for the crude extract and SF1, the most potent subfraction. Overall, our data point to the potential anti-rotavirus activity of flavonoid-enriched fraction (SF1) of M. urundeuva leaves, corroborating the traditional use of this species to treat diarrhea and broadening our perspectives on in vivo assays in mice with SF1 isolated or associated with other fractions.

Anti-biofilm activity of the marine bacterium Pseudoalteromonas ruthenica KLPp3 against Serratia marcescens and Vibrio alginolyticus

Aims: Pseudoalteromonas ruthenica KLPp3 is the marine Gram-negative strain isolated from the surface of mud crab at Pulau Perhentian Malaysia. In this work, the anti-biofilm activity of P. ruthenica supernatant was examined on Serratia marcescen and Vibrio alginolyticus. Methodology and results: The crude extract of P. ruthenica KLPp3 was obtained using ethyl acetate. The subminimum inhibitory concentration (MIC) of the crude extract was determined using the minimum inhibitory test. The subMIC crude extract was tested against two of the S. marcescen virulence factors, which are the swarming ability and production of prodigiosin. The crystal violet assay was used to test the anti-biofilm activity of the sub-MIC crude extract against S. marcescen and V. alginolyticus. The productions of prodigiosin were reduced by 72%. The swarming area was reduced by 56.06%. It inhibits 26.9% and 48.5% of biofilm production in S. marcescens and V. alginolyticus respectively. The crude extract was heat stable. Conclusion, significance and impact of study: Besides combating the S. marcescens virulence factor, P. ruthenica KLPp3 crude extract in sub-MIC reduces the formation of biofilm of S. marcescens and V. alginolyticus, which may find applications in biofilm inhibition and prevention.

Anticancer effects of crude extract from Melia toosendan Sieb. et Zucc on hepatocellular carcinoma in vitro and in vivo / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc and its possible molecular mechanisms in vitro and in vivo.</p><p><b>METHODS</b>Transonic alcohol-chloroform extraction method was used to extract toosendanin from the bark of Melia toosendan Sieb. et Zucc, and the content of toosendanin in the crude extract was measured by high performance liquid chromatography (HPLC). Anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc were investigated in in vivo and in vitro studies. In the in vitro experiment, human hepatocellular carcinoma cell lines SMMC-7721 and Hep3B were co-incubated with toosendanin crude extract of different concentrations, respectively. In the in vivo experiment, BALB/c mice were subcutaneously inoculated with mouse hepatocellular carcinoma H22 cells and treated with crude extract.</p><p><b>RESULTS</b>HPLC revealed the content of toosendanin was about 15%. Crude extract from Melia toosendan Sieb. et Zucc inhibited cancer cells growth in a dose- and time-dependent manner. The 50% inhibitory concentration (IC50, 72 h) was 0.6 mg/L for SMMC-7721 cells and 0.8 mg/L for Hep3B cells. Both high-dose [0.69 mg/(kg d)] and low-dose [0.138 mg/(kg d)] crude extract could markedly suppress cancer growth, and the inhibition rate was greater than 50%. Hematoxylin and eosin staining showed necrotic area in cancers and transmission electron microscopy displayed necrotic and apoptotic cancer cells with apoptotic bodies. Immunohistochemistry showed that the expression of Bax and Fas increased and the expression of Bcl-2 reduced.</p><p><b>CONCLUSIONS</b>Toosendanin extract has potent anti-cancer effects via suppressing proliferation and inducing apoptosis of cancer cells in vivo and in vitro. The mechanism of apoptosis involves in mitochondrial pathway and death receptor pathway.</p>