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Low temperatures are a key condition for various phytopathogenic bacteria, by favouring their pathogenic potential and the development of plant diseases. So far, the signal transduction pathways related to low temperatures in phytopathogenic bacteria are little understood. The occurrence of intracellular oxidative stress under this condition has been suggested as a molecular event belonging to this signalling pathway. In this study, using the Pseudomonas savastanoi pv. phaseolicola model bacteria, we evaluated intracellular redox state under low‐temperature (18°C) conditions. Generation of an oxidative‐stress biosensor (pKL1) and fluorometry analyses with the P. savastanoi pv. phaseolicola‐roGFP2 bioreporter strain were performed. At low temperatures (18°C) the response capacity or roGFP2 oxidation diminished in relation to 28°C. Likewise, changes in the intracellular redox potential of P. savastanoi pv. phaseolicola during its growth at 28°C and 18°C were observed, with greater oxidation degree in cells grown at 18°C. These results demonstrated that low temperatures induce intracellular oxidative stress in P. savastanoi pv. phaseolicola. From this study, the oxidative stress is established as part of the cellular response to low‐temperature conditions in this bacteria. This is the first report of the relation of oxidative stress with low‐temperature conditions in phytopathogens bacteria. [ABSTRACT FROM AUTHOR]

Phaseolotoxin is an antimetabolite toxin produced by diverse pathovars of Pseudomonas syringae which affects various plants, causing diseases of economic importance. Phaseolotoxin contributes to the systemic dissemination of the pathogen in the plant, therefore it is recognized as a major virulence factor. Genetic traits such as the Pht cluster, appear defining to the toxigenic strains phaseolotoxin producers. Extensive research has contributed to our knowledge concerning the regulation of phaseolotoxin revealing a complex regulatory network that involves processes at the transcriptional and posttranscriptional levels, in which specific and global regulators participate. Even more, significant advances in understanding how specific signals, including host metabolites, nutrient sources, and physical parameters such as the temperature, can affect phaseolotoxin production have been made. A general overview of the phaseolotoxin regulation, focusing on the chemical and physical cues, and regulatory pathways involved in the expression of this major virulence factor will be given in the present work. [ABSTRACT FROM AUTHOR]

Heavy metal pollution is a severe concern worldwide, owing to its harmful effects on ecosystems. Phytoremediation has been applied to remove heavy metals from water, soils, and sediments by using plants and associated microorganisms to restore contaminated sites. The Typha genus is one of the most important genera used in phytoremediation strategies because of its rapid growth rate, high biomass production, and the accumulation of heavy metals in its roots. Plant growth-promoting rhizobacteria have attracted much attention because they exert biochemical activities that improve plant growth, tolerance, and the accumulation of heavy metals in plant tissues. Because of their beneficial effects on plants, some studies have identified bacterial communities associated with the roots of Typha species growing in the presence of heavy metals. This review describes in detail the phytoremediation process and highlights the application of Typha species. Then, it describes bacterial communities associated with roots of Typha growing in natural ecosystems and wetlands contaminated with heavy metals. Data indicated that bacteria from the phylum Proteobacteria are the primary colonizers of the rhizosphere and root-endosphere of Typha species growing in contaminated and non-contaminated environments. Proteobacteria include bacteria that can grow in different environments due to their ability to use various carbon sources. Some bacterial species exert biochemical activities that contribute to plant growth and tolerance to heavy metals and enhance phytoremediation. [ABSTRACT FROM AUTHOR]

Plant-associated bacteria in heavy-metal-contaminated environments could be a biotechnological tool to improve plant growth. The present work aimed to isolate lead- and cadmium-tolerant endophytic bacteria from the roots of Typha latifolia growing in a site contaminated with these heavy metals. Endophytic bacteria were characterized according to Pb and Cd tolerance, plant-growth-promoting rhizobacteria activities, and their effect on T. latifolia seedlings exposed and non-exposed to Pb and Cd. Pb-tolerant isolates were identified as Pseudomonas azotoformans JEP3, P. fluorescens JEP8, and P. gessardii JEP33, while Cd-tolerant bacteria were identified as P. veronii JEC8, JEC9, and JEC11. They all exert biochemical activities, including indole acetic acid synthesis, siderophore production, and phosphate solubilization. Plant–bacteria interaction assays showed that P. azotoformans JEP3, P. fluorescens JEP8, P. gessardii JEP33, and P. veronii JEC8, JEC9, JEC11 promote the growth of T. latifolia seedlings by increasing the root and shoot length, while in plants exposed to either 5 mg/L of Pb or 10 mg/L of Cd, all bacterial isolates increased the shoot length and the number of roots per plant, suggesting that they are plant-growth-promoting rhizobacteria that could contribute to T. latifolia adaptation to the heavy metal polluted site. [ABSTRACT FROM AUTHOR]

Phaseolotoxin is a major virulence factor of the bean pathogen bacterium P. savastanoi pv. phaseolicola. This toxin plays a key role in the development of the halo blight disease in bean plants. So far, the signal transduction pathways involved in the synthesis of phaseolotoxin have not been elucidated. The influence of regulation mechanisms related to the oxidative stress response, in particular the OxyR protein, it has been suggested to be involved in this process.. In this study we evaluated the role of OxyR in P. savastanoi pv. phaseolicola, mainly compared to the synthesis of phaseolotoxin and the virulence of this phytopathogen. Generation of the oxyR-mutant, pathogenicity and virulence tests, and analyses of gene expression by RT-PCR assays were performed. The results showed that OxyR exerts an effect on the synthesis of phaseolotoxin and positively influences the expression of the Pht and Pbo cluster genes. Likewise, OxyR influences the production of pyoverdine by the control of the expression of the genes encoding the PvdS sigma factor, involved in the synthesis of this pigment. This study is the first report on members of the OxyR regulon of P. savastanoi pv. phaseolicola NPS3121. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) is one of the most toxic heavy metals for plant physiology and development. This review discusses Cd effects on auxin biosynthesis and homeostasis, and the strategies for restoring plant growth based on exogenous auxin application. First, the two well-characterized auxin biosynthesis pathways in plants are described, as well as the effect of exogenous auxin application on plant growth. Then, review describes the impacts of Cd on the content, biosynthesis, conjugation, and oxidation of endogenous auxins, which are related to a decrease in root development, photosynthesis, and biomass production. Finally, compelling evidence of the beneficial effects of auxin-producing rhizobacteria in plants exposed to Cd is showed, focusing on photosynthesis, oxidative stress, and production of antioxidant compounds and osmolytes that counteract Cd toxicity, favoring plant growth and improve phytoremediation efficiency. Expanding our understanding of the positive effects of exogenous auxins application and the interactions between bacteria and plants growing in Cd-polluted environments will allow us to propose phytoremediation strategies for restoring environments contaminated with this metal. [ABSTRACT FROM AUTHOR]

The Typha genus comprises plant species extensively studied for phytoremediation processes. Recently, Pseudomonas rhodesiae GRC140, an IAA-producing bacterium, was isolated from Typha latifolia roots. This bacterium stimulates the emergence of lateral roots of Arabidopsis thaliana in the presence and absence of cadmium. However, the bacterial influence on cadmium accumulation by the plant has not been determined. Moreover, the P. rhodesiae GRC140 effect in Cd phytoextraction by T. latifolia remains poorly understood. In this work, an axenic hydroponic culture of T. latifolia was established. The plants were used to evaluate the effects of cadmium stress in axenic plants and determine the effects of P. rhodesiae GRC140 and exogenous indole acetic acid (IAA) on Cd tolerance and Cd uptake by T. latifolia. Biomass production, total chlorophyll content, root electrolyte leakage, catalase activity, total glutathione, and Cd content were determined. The results showed that Cd reduces shoot biomass and increases total glutathione and Cd content in a dose-dependent manner in root tissues. Furthermore, P. rhodesiae GRC140 increased Cd translocation to the shoots, while IAA increased the Cd accumulation in plant roots, indicating that both treatments increase Cd removal by T. latifolia plants. These results indicate that axenic plants in hydroponic systems are adequate to evaluate the Cd effects in plants and suggest that T. latifolia phytoextraction abilities could be improved by P. rhodesiae GRC140 and exogenous IAA application. [ABSTRACT FROM AUTHOR]

Several aflatoxin inhibitors can modulate the antioxidant system in fungi. In this work, the effect of the ethanolic extract of Capsicum chinense and Piper nigrum fruits, capsaicin, and piperine on the expression of the aflE, aflG, aflH, aflI, aflK, aflL, aflO, aflP, and aflQ genes involved in the aflatoxin biosynthetic pathway in Aspergillus parasiticus were studied by qRT-PCR analysis. As well as, the effect on the expression of fungal antioxidant genes (sod1, catA, and cat2) and enzymatic activity of catalase (CAT) and superoxide dismutase (SOD). Results reveal that the highest (p < 0.05) radial growth inhibition (68 and 86%) and aflatoxins production inhibition (73 and 80%) was observed with capsaicin and piperine respectively, at 300 µg/mL, instead of the ethanolic extract at the same concentration. The qRT-PCR analysis showed that compounds and extracts at 300 µg/mL induced a down-regulation of aflatoxin genes and an up-regulation on the fungal antioxidant genes. CAT activity increased by 23.15, 36.65, 51.40, and 65.50%, in the presence of C. chinense and P. nigrum extract, capsaicin, and piperine exposure, respectively. While SOD activity was not significantly impacted (p > 0.05). In conclusion, the capsaicin and piperine, two antifungal and anti-aflatoxigenic compounds produce an up-regulation of antioxidant defense genes accompanied by an enhancement of catalase enzymatic activity in A. parasiticus. [ABSTRACT FROM AUTHOR]

Affinin present in Heliopsis longipes roots has been identified as an anti-aflatoxin molecule. However, its mechanism of action has yet to be clarified. Aflatoxins biosynthesis involves not less than 27 enzymatic reactions. In this work, the genes aflG, aflH, aflI, aflK, aflL, aflM, aflO, aflP, and aflQ of the aflatoxins cluster and the aflS gene encoding an internal regulatory factor involved in aflatoxins biosynthesis in Aspergillus parasiticus, were studied by qRT-PCR. Results demonstrated that ethanolic extract of H. longipes roots and affinin inhibit aflatoxin biosynthesis and fungal growth in a dose-dependent manner. At 300 µg/mL, ethanolic extract and affinin presented the highest inhibition of radial growth (86% and 94%) and aflatoxin production (68% and 80%). The qRT-PCR analysis demonstrated that nine tested genes were down-regulated by affinin and ethanolic extract. The most down-regulated was the aflK, a gene that encodes an enzyme cyclase with double function during the aflatoxin biosynthesis. While no significant down-regulation was obtaining for aflH gene. Exposure to affinin also resulted in decreased transcript levels of the internal regulator factor aflS. Based on our results, a model showing the regulatory mechanism in aflatoxin biosynthesis and its role in gene expression was proposed. In conclusion, affinin modulates the expression of several aflatoxin biosynthetic genes, leading to mycotoxin biosynthesis inhibition. Therefore, H. longipes roots is a suitable candidate to developed control strategies via lowering gene expressions as a future perspective in reducing aflatoxin contamination. [ABSTRACT FROM AUTHOR]

Abstract Background Pseudomonas syringae pv. phaseolicola, the causal agent of halo blight disease in beans, produces a toxin known as phaseolotoxin, in whose synthesis participate a group of genes organized within the genome in a region known as the "Pht cluster". This region, which is thought to have been acquired by horizontal gene transfer, includes 5 transcriptional units, two monocistronic (argK, phtL) and three polycistronic (phtA, phtD, phtM), whose expression is temperature dependent. So far, the regulatory mechanisms involved in phaseolotoxin synthesis have not been elucidated and the only well-established fact is the requirement of low temperatures for its synthesis. In this work, we searched for regulatory proteins that could be involved in phaseolotoxin synthesis, focusing on the regulation of the phtD operon. Results In this study we identified the global regulator IHF (Integration Host Factor), which binds to the promoter region of the phtD operon, exerting a negative effect on the expression of this operon. This is the first regulatory protein identified as part of the phaseolotoxin synthesis system. Our findings suggest that the Pht cluster was similarly regulated in the ancestral cluster by IHF or similar protein, and integrated into the global regulatory mechanism of P. syringae pv. phaseolicola, after the horizontal gene transfer event by using the host IHF protein. Conclusion This study identifies the IHF protein as one element involved in the regulation of phaseolotoxin synthesis in P. syringae pv. phaseolicola NPS3121 and provides new insights into the regulatory mechanisms involved in phaseolotoxin production.

Abstract Background Pseudomonas syringae pv. phaseolicola is a Gram-negative plant-pathogenic bacterium that causes "halo blight" disease of beans (Phaseolus vulgaris L.). This disease affects both foliage and pods, and is a major problem in temperate areas of the world. Although several bacterial genes have been determined as participants in pathogenesis, the overall process still remains poorly understood, mainly because the identity and function of many of the genes are largely unknown. In this work, a genomic library of P. syringae pv. phaseolicola NPS3121 was constructed and PCR amplification of individual fragments was carried out in order to print a DNA microarray. This microarray was used to identify genes that are differentially expressed when bean leaf extracts, pod extracts or apoplastic fluid were added to the growth medium. Results Transcription profiles show that 224 genes were differentially expressed, the majority under the effect of bean leaf extract and apoplastic fluid. Some of the induced genes were previously known to be involved in the first stages of the bacterial-plant interaction and virulence. These include genes encoding type III secretion system proteins and genes involved in cell-wall degradation, phaseolotoxin synthesis and aerobic metabolism. On the other hand, most repressed genes were found to be involved in the uptake and metabolism of iron. Conclusion This study furthers the understanding of the mechanisms involved, responses and the metabolic adaptation that occurs during the interaction of P. syringae pv. phaseolicola with a susceptible host plant.

In the present study, α-amylase and α-glucosidase inhibitory effect and antioxidant activity of capsaicin and piperine from the ethanolic extract of Capsicum chinense (EECch) and Piper nigrum (EEPn) fruits were investigated. Results revealed that EECch exhibited the highest phenolic (154 mg GAE/100 g of tissue) and flavonoid content (75 mg RtE/100 g of tissue) in comparison with EEPn. The predominant compound detected in EECch and EEPn by GC-EIMS analysis was the capsaicin and piperine, respectively. The capsaicin and piperine showed the highest α-amylase and α-glucosidase inhibitory effect and antioxidant activity rather than extracts. The EEPn (IC50= 216 µg/mL) and piperine (IC50= 105 µg/mL) present a highest α-amylase inhibitory effect, while the EECch (IC50= 225 µg/mL) and capsaicin (IC50= 117 µg/mL) showed highest anti-α-glucosidase activity. Molecular docking established that capsaicin and piperine bind at the α-glucosidase and α-amylase through hydrophobic interactions, hydrogen bond, and charge interactions with amino acid residues. The enzyme inhibitory activity and antioxidant properties exhibited by EECch and EEPn could be attributed to the capsaicin and piperine content and other compounds present such as phenolic compounds and flavonoids. These fruits are potential sources of natural antioxidant agents and α-amylase and α-glucosidase inhibitors. [ABSTRACT FROM AUTHOR]

Aflatoxins produced by Aspergillus parasiticus are toxic and carcinogenic metabolites. The biosynthesis of this mycotoxins is a complex process and involves at least 30 genes clustered within an approximately 82 kB gene cluster. In the present study, the effect of Capsicum chinense and Piper nigrum fruits on Aspergillus parasiticus growth and aflatoxin production were studied in relation to the expression of aflD, aflM, aflR, and aflS four; key genes of aflatoxins biosynthesis pathway. GC-EIMS analysis identified capsaicin (66,107 µg g−1) and piperine (1,138 µg g−1) as the most abundant compounds in C. chinense and P. nigrum fruits, respectively. The antifungal and anti-aflatoxigenic assays showed that C. chinense, P. nigrum, capsaicin, and piperine inhibited A. parasiticus growth and aflatoxins production in a dose-dependent manner. The piperine at 300 µg mL−1 produced higher radial growth inhibition (89%) and aflatoxin production inhibition (69%). The expression of aflatoxin biosynthetic genes was evaluated by quantitative real-time PCR (qRT-PCR) and revealed that aflatoxin inhibition occurring via downregulating the aflS and aflR, and subsequently aflD and aflM genes. These results will improve our understanding of the mechanism of aflatoxin regulation by C. chinense, P. nigrum, capsaicin, and piperine, and provides a reference for further study. [ABSTRACT FROM AUTHOR]

In the present study, ethanolic extract from Heliopsis longipes roots and affinin/spilanthol against Aspergillus parasiticus growth and aflatoxins production were studied in relation to the expression of aflD and aflR, two key genes of aflatoxins biosynthetic pathway. Phytochemical analysis of the ethanolic extract by GC-EIMS identified affinin/spilanthol (7.84 ± 0.27 mg g−1) as the most abundant compounds in H. longipes roots. The antifungal and anti-aflatoxigenic assays showed that affinin/spilanthol at 300 µg mL−1 produced the higher inhibition of radial growth (95%), as well as, the higher aflatoxins production inhibition (61%) in comparison to H. longipes roots (87% and 48%, respectively). qRT-PCR revealed that the expression of aflD and aflR genes showed a higher downregulation in affinin/spilanthol at 300 µg mL−1. The expression ratio of alfD was suppressed by affinin/spilanthol in 79% and aflR in 84%, while, a lower expression ratio suppressed by H. longipes was obtained, alfD (55%) and aflR (59%). Affinin/spilanthol possesses higher antifungal and anti-aflatoxigenic activity against A. parasiticus rather than H. longipes roots, and this anti-aflaxotigenic activity occurring via downregulation of the aflD and aflR genes. Thus, H. longipes roots and affinin/spilanthol can be considered potent antifungal agents against aflatoxigenic fungus, especially, affinin/spilanthol. [ABSTRACT FROM AUTHOR]

Bacillus sp. strain MA04 a plant growth-promoting rhizobacteria (PGPR) showed hemolytic activity on blood agar plates, and the supernatant from liquid culture in nutrient broth at 24 h exhibited emulsification activity, suggesting the production of biosurfactants. In antagonist assays, the supernatant showed antifungal activity against phytopathogenic fungi such as Penicillium expansum, Fusarium stilboides, Sclerotium rolfsii y Rhizoctonia solani, finding a reduction of mycelial growth of all fungi tested, ranging from 35 to 69%, this activity was increased with time of culture, accomplishing percentages of inhibition up to 85% with supernatants obtained at 72 h. Then, the crude biorsurfactant (CB) was isolated from the supernatant in order to assay its antagonistic effect on the phytopathogens previously tested, finding an increase in the inhibition up to 97% at 500 mg/L of CB. The composition of CB was determined by infrared spectroscopy, identifying various functional groups related to lipopeptides, which were purified by high-performance liquid chromatography and analyzed by MALDI-TOF/TOF-MS, revealing a mixture of fengycins A and B whose high antifungal activity is been widely recognized. These results show that PGPR Bacillus sp. MA04 could also contribute to plant health status through the production of metabolites with antimicrobial activity. [ABSTRACT FROM AUTHOR]

This study evaluated the role of oxidative stress on the expression of Pht cluster genes involved in phaseolotoxin synthesis in Pseudomonas syringae pv. phaseolicola. Results demonstrate that the expression of Pht cluster genes is regulated by oxidative stress in a manner dependent of the ROS present in the cell. The presence of H2O2 and Paraquat, influences on the expression of the Pht cluster genes in function of the compound and of the concentration evaluated, demonstrating that expression of Pht genes is part of the oxidative stress response in P. syringae pv. phaseolicola NPS3121. [ABSTRACT FROM AUTHOR]

The recently discovered type VI secretion system (T6SS) is implicated in the pathogenic and/or virulence processes of diverse bacteria. The expression pattern of the T6SS differs among different organisms and also depends on several environmental factors. We initiated a study of the conditions that influence T6SS gene expression in Pseudomonas syringae pv. phaseolicola NPS3121. Our results indicate that low temperatures and plant extracts impact the expression of T6SS genes and that the process is subject to regulation by the GacS/GacA two-component system. [ABSTRACT FROM AUTHOR]

Background: Low temperatures play key roles in the development of most plant diseases, mainly because of their influence on the expression of various virulence factors in phytopathogenic bacteria. Thus far, studies regarding this environmental parameter have focused on specific themes and little is known about phytopathogenic bacteria physiology under these conditions. To obtain a global view regarding phytopathogenic bacteria strategies in response to physiologically relevant temperature changes, we used DNA microarray technology to compare the gene expression profile of the model bacterial pathogen P. syringae pv. phaseolicola NPS3121 grown at 18°C and 28°C. Results: A total of 236 differentially regulated genes were identified, of which 133 were up-regulated and 103 were down-regulated at 18°C compared to 28°C. The majority of these genes are involved in pathogenicity and virulence processes. In general, the results of this study suggest that the expression profile obtained may be related to the fact that low temperatures induce oxidative stress in bacterial cells, which in turn influences the expression of iron metabolism genes. The expression also appears to be correlated with the profile expression obtained in genes related to motility, biofilm production, and the type III secretion system. Conclusions: From the data obtained in this study, we can begin to understand the strategies used by this phytopathogen during low temperature growth, which can occur in host interactions and disease development [ABSTRACT FROM AUTHOR]

Background: Pseudomonas syringae pv. phaseolicola, the causal agent of halo blight disease in beans, produces a toxin known as phaseolotoxin, in whose synthesis participate a group of genes organized within the genome in a region known as the "Pht cluster". This region, which is thought to have been acquired by horizontal gene transfer, includes 5 transcriptional units, two monocistronic (argK, phtL) and three polycistronic (phtA, phtD, phtM), whose expression is temperature dependent. So far, the regulatory mechanisms involved in phaseolotoxin synthesis have not been elucidated and the only well-established fact is the requirement of low temperatures for its synthesis. In this work, we searched for regulatory proteins that could be involved in phaseolotoxin synthesis, focusing on the regulation of the phtD operon. Results: In this study we identified the global regulator IHF (Integration Host Factor), which binds to the promoter region of the phtD operon, exerting a negative effect on the expression of this operon. This is the first regulatory protein identified as part of the phaseolotoxin synthesis system. Our findings suggest that the Pht cluster was similarly regulated in the ancestral cluster by IHF or similar protein, and integrated into the global regulatory mechanism of P. syringae pv. phaseolicola, after the horizontal gene transfer event by using the host IHF protein. Conclusion: This study identifies the IHF protein as one element involved in the regulation of phaseolotoxin synthesis in P. syringae pv. phaseolicola NPS3121 and provides new insights into the regulatory mechanisms involved in phaseolotoxin production. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Bidens odorata Cav (Asteraceae) is used for the empirical treatment of inflammation and pain. Aim of the study This work evaluated the in vitro and in vivo toxicity, antioxidant activity, as well as the anti-inflammatory and antinociceptive effects of an ethanol extract from Bidens odorata leaves (BOE). Materials and methods The in vitro toxicity of BOE (10–1000 µg/ml) was evaluated with the comet assay in PBMC. The in vivo acute toxicity of BOE (500–5000 mg/kg) and the effect of BOE (10–1000 µg/ml) on the level of ROS in PBMC were determined. The in vivo anti-inflammatory activity of BOE was assessed using the TPA-induced ear edema in mice. The antinociceptive activities of BOE (50–200 mg/kg p.o.) were assessed using the acetic acid and formalin tests. The antinociceptive mechanism of BOE was determined using naloxone and glibenclamide. Results BOE lacked DNA damage, and showed low in vivo toxicity (LD 50 > 5000 mg/kg p.o.). BOE inhibited ROS production (IC 50 = 252.13 ± 20.54 µg/ml), and decreased inflammation by 36.1 ± 3.66%. In both antinociceptive test, BOE (200 mg/kg) exerted activity with similar activity than the reference drugs. Conclusion B. odorata exerts low in vitro and in vivo toxicity, antioxidant effects, moderate in vivo anti-inflammatory activity, and antinociceptive effects mediated by ATP-sensitive K + channels. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Costus pulverulentus C. Presl (Costaceae), a species endemic to Mexico, is used for the empirical treatment of cancer, pain, and inflammation. Aim of the study The objective of this study was to evaluate the toxicity, as well as the cytotoxic, antinociceptive, anti-inflammatory and sedative effects of an ethanol extract from Costus pulverulentus stem (CPE). Materials and methods The chemical characterization of CPE was performed by Gas chromatography–mass spectrometry (GC–MS). The toxicity of CPE was evaluated using the comet assay (10–1000 µg/ml during 5 h) and the acute toxicity test (500–5000 mg/kg p.o. and i.p. during 14 days). The cytotoxic effect of CPE (1–250 µg/ml) on human cancer cells was evaluated using the MTT assay. The antinociceptive effects of CPE (50–200 mg/kg p.o.) were evaluated using thermal-induced nociception tests (hot plate and tail flick) and the chemical-induced nociceptive tests (acetic acid and formalin). The sedative activity of CPE (50–200 mg/kg p.o.) was evaluated using the ketamine-induced sleeping time test. Results CPE showed the presence of compounds such as campesterol, stigmasterol β-sitosterol, vanillic acid, among others. In the comet assay, CPE at 200 µg/ml or higher concentrations induced DNA damage. In the acute toxicity test, the LD 50 estimated for CPE was>5000 mg/kg p.o. or i.p. CEP showed moderate cytotoxic effects on prostate carcinoma cells PC-3 cells (IC 50 =179±23.2 µg/ml). In the chemical-induced nociception models, CPE (100 and 200 mg/kg p.o.) showed antinociceptive effects with similar activity to 100 mg/kg naproxen. In the thermal-induced nociception tests, CPE tested at 200 mg/kg showed moderate antinociceptive effects by 28% (hot plate test) and by 25% (tail flick test). In the ketamine-induced sleeping time test, CPE showed no sedative effects. Conclusions C. pulverulents exerts moderate cytotoxic effects in human cancer cells, moderate anti-inflammatory and antinociceptive effects. C. pulverulentus induces antinociceptive effects without inducing sedation. [ABSTRACT FROM AUTHOR]