Bioactive sorbicillinoids from a rhizospheric soil-derived Paecilomyces sp. KMU21009.

A new hybrid sorbicillinoid named paeciureallin (1) and a new monomeric sorbicillinoid named paecillyketide (2), along with six known analogues (3-8), were isolated from the rhizospheric soil-derived fungus Paecilomyces sp. KMU21009 associated with Delphinium yunnanense. Their structures were elucidated by extensive spectroscopic analysis and comparison with literature values. Paeciureallin (1) is the first example of hybrid sorbicillinoids possessing a rare sorbicillinoid urea unit and containing a ß-D-ribofuranose functionality. In pharmacological studies, compounds 1 and 2 were evaluated for in vitro anti-inflammatory and cytotoxic activities. Paeciureallin (1) exhibited moderate cytotoxicity against SW480 and A549 cell lines, and the IC50 values were 32.0 ± 0.1 and 34.4 ± 2.0 µM, respectively.

Evaluation of antileishmanial activity of harzialactone a isolated from the marine-derived fungus Paecilomyces sp.

Fractionation of extracts from the culture broth of the marine-derived fungus, Paecilomyces sp. 7A22, resulted in the isolation of the harzialactone A (HA), a known compound previously isolated from fungi of marine environments. The chemical structure of HA was determined by spectroscopic analyses. Upon evaluation of HA on antileishmanial assays against Leishmania amazonensis, HA exhibited significant activity against promastigotes forms with IC50 of 5.25 µg mL-1 and moderate activity against intracellular amastigotes with IC50 of 18.18 µg mL-1. This is the first report on the antileishmanial activity of HA, and the effects of HA presented in this work suggest that this class of compounds are suitable for future biological in vitro and in vivo studies for the search of natural products with activity against Leishmania spp. Furthermore, the present results corroborate marine-derived fungi as a promising source of natural products with antiparasitic activity.

Effect of Paecilomyces tenuipes Extract on Testosterone-Induced Benign Prostatic Hyperplasia in Sprague-Dawley Rats.

: Benign prostatic hyperplasia (BPH) is one of the major public health concerns, which has a high prevalence rate and causes significant decline in men's quality of life. BPH is highly related to sexual hormone metabolism and aging. In particular, dihydrotestosterone (DHT), to which testosterone is modified by 5α-reductase (5AR), has a significant effect on BPH development. DHT binds to an androgen receptor (AR) and steroid receptor coactivator 1 (SRC-1); then, it induces the proliferation of a prostate cell and expression of prostate specific antigen (PSA). Paecilomyces tenuipes (P. tenuipes) is a mushroom that has been popularized by the artificial cultivation of fruiting bodies based on silkworms by researchers from the Republic of Korea. In a previous study, we identified the effect of PE on PSA mRNA expression in LNCaP cells. This suggests that PE may have an inhibitory effect on androgen signaling. Therefore, we confirmed the expression of androgen signaling-related factors, such as AR, SRC-1, and PSA in LNCaP. Furthermore, we confirmed the androgen signaling inhibitory effect of PE using the testosterone propionate (TP)-induced BPH rat model. A BPH rat model was established with a four-week treatment of daily subcutaneous injections of testosterone propionate (TP, 3 mg/kg) dissolved in corn oil after castration. The rats in the treatment group were orally gavaged P. tenuipes extract (PE), finasteride (Fi), or saw palmetto extract (Saw) with TP injection. DHT induced an increase in the expression levels of AR, SRC-1, and PSA proteins in LNCaP cells. On the contrary, the PE treatment reduced the expression levels. In vivo, the BPH group showed an increase in prostate size compared with the control group. The PE gavaged group showed a decrease in prostate size compared with the BPH group. In addition, the protein expressions of AR, 5AR2, and PSA were significantly lower in the PE gavaged group than BPH group in prostate tissue. These results suggest the beneficial effects of PE on BPH via the modulation of AR signaling pathway.

Potential therapeutic effects of Cordyceps cicadae and Paecilomyces cicadae on adenine-induced chronic renal failure in rats and their phytochemical analysis.

BACKGROUND: Natural Cordyceps cicadae (C. cicadae) has been utilized extensively in traditional Chinese medicine to treat chronic renal diseases, heart palpitations, infantile convulsions, and dizziness. However, given its slow growth and immoderate exploitation, C. cicadae resources have been severely depleted. By contrast, Paecilomyces cicadae (P. cicadae), as the anamorph stage of C. cicadae, is easy to cultivate, and this kind of cultivated P. cicadae has good and controllable quality. PURPOSE: This study aimed to compare the therapeutic effects of C. cicadae and P. cicadae on adenine-induced chronic renal failure (CRF) rats. In accordance with the aforementioned studies, our work subsequently analyzed the intrinsic relationships between the efficacy and pharmacodynamic substances of C. cicadae and P. cicadae to conclude whether or not P. cicadae could be used as an alternative to C. cicadae in treating CRF. METHODS: Rats were administered with C. cicadae (1.0 g/kg) or P. cicadae (1.0 g/kg) by gavage for 4 weeks. Furthermore, we applied Fourier transform infrared spectroscopy, gas chromatography-mass spectrometry, liquid chromatography-tandem mass spectrometry, and ultraviolet spectrophotometry to comprehensively detect and analyze the chemical constituent differences from ten batches each of C. cicadae and P. cicadae. RESULTS: This study revealed that both C. cicadae and P. cicadae exerted obvious therapeutic effects on CRF and were more consistent with their chemical compositions. CONCLUSION: P. cicadae can be used as an alternative to C. cicadae for treating CRF to cater to market demands.

Extract of Paecilomyces hepiali mycelia induces lipolysis through PKA-mediated phosphorylation of hormone-sensitive lipase and ERK-mediated downregulation of perilipin in 3T3-L1 adipocytes.

BACKGROUND: Cordyceps sinensis has been used for centuries in China as one of the most valued herbal medicine and tonic food. Paecilomyces hepiali, a fungal strain isolated from natural C. sinensis, has been used widely as a substitute of C. sinensis in medicine and health food. P. hepiali has been reported to have various pharmaceutical benefits, including triglyceride-lowing activity. However, its effects on triglyceride metabolism in adipocytes remain unknown. The purpose of the present study was to evaluate the effect of P. hepiali mycelia on adipocyte lipolysis and to clarify the underlying mechanisms. METHODS: The fully differentiated 3T3-L1 adipocytes were treated with methanol extract of Paecilomyces hepiali mycelia (PHME). Contents of glycerol released into the culture medium and intracellular triglyceride were measured as indices of lipolysis using glycerol assay kit and Oil red O staining, respectively. Then, effects of PHME on the main lipases or kinases involved in lipolysis regulation were investigated. Protein expression of adipose triglyceride lipase (ATGL) and perilipin, as well as phosphorylation of hormone-sensitive lipase (HSL), AMP-activated protein kinase (AMPK), and mitogen-activated protein kinases (MAPKs) were determined by western blotting. Moreover, nucleosides, important constituents of PHME, were analyzed using high performance liquid chromatography (HPLC). RESULTS: Treatment with PHME led to a significant increase in glycerol release thereby reduced intracellular triglyceride accumulation in fully differentiated adipocytes. PHME upregulated protein kinase (PK) A-mediated phosphorylation of HSL at serine residues of 563 and 660. Meanwhile, PHME treatment also upregulated phosphorylation of extracellular signal-regulated kinase (ERK), and downregulated the protein level of perilipin. Pretreatment with the PKA inhibitor, H89, blunted the PHME-induced lipolysis and the phosphorylation of HSL (Ser 563 and 660). Moreover, pretreatment with ERK inhibitor, PD98059, weakened the PHME-caused glycerol release and downregulation of perilipin expression. HPLC analysis indicated there were adenosine, cordycepin, uridine and vernine in PHME. CONCLUSIONS: Our results showed that PHME significantly induced lipolysis in 3T3-L1 adipocytes, which is mainly mediated by activation of HSL through PKA pathway and by downregulation of perilipin through activation of ERK pathway.

Antioxidant Activity and Infrared Spectroscopy Analysis of Alcoholic Extracts Obtained from Paecilomyces hepiali (Ascomycetes).

This research studies the influence of substrate on the antioxidant activity of alcohol extracts of Paecilomyces hepiali. We used corn, rice, millet, and peas as substrates. Antioxidant activity was measured with the DPPH radical scavenging method. Concentrations of extracts (6.25, 3.12, 1.56, 0.78, and 0.39 mg/mL) were applied in all evaluations. Overall antioxidant activity was expressed as the concentration of substrate that decreased DPPH radical levels by 50% (IC50DPPH) for 7 methanol and 7 ethanol extracts. A comparison of IC50DPPH allowed us to conclude that the methanol extracts are more active in scavenging stable DPPH radicals than are the ethanol extracts. The substrate with antioxidant properties most suitable for cultivation of P. hepiali was rice supplemented with non-defatted soy flour. The extract most effective in scavenging stable radicals was the methanol extract of sample 4 (IC50DPPH = 2.33 mg/mL) cultivated on rice with nondefatted soy flour. The methanol extract of sample 7 cultivated on peas was less effective (IC50DPPH = 11.50 mg/mL). By crystallizing these extracts, we managed to obtain sufficient quantities of 6 samples in a solid state, for which infrared spectra were measured and confirmed the presence of amino acids in the extracts.

Paecilomyces tenuipes extract prevents depression-like behaviors in chronic unpredictable mild stress-induced rat model via modulation of neurotransmitters.

The medicinal fungus Paecilomyces tenuipes exhibits a variety of pharmacological effects, including antidepressive effects. The chronic unpredictable mild stress (CUMS)­induced rat model has served an important role in studies involving antidepressants screening. The aim of the present study was to evaluate the antidepressant­like activity of P. tenuipes N45 aqueous extract (PTNE) in a CUMS­induced rat model of behavioral despair depression. Following 4 weeks of PTNE treatment, behavioral tests were conducted to investigate the antidepressant­like activities, and the levels of neurotransmitters and hormones in blood and hypothalamus were measured. The results demonstrated that PTNE treatment significantly increased movement in the forced running test, whereas the immobility time was reduced in the hotplate test and the forced swim test in depression­model rats. PTNE treatment was able to normalize the levels of hormones and neurotransmitters in serum and hypothalamus of CUMS rats. The data demonstrated that PTNE treatment may be a potential pharmaceutical agent in treatment­resistant depression, and the effects of PTNE may be partly mediated through normalizing the levels of neurotransmitters.

Total Phenolic Content and Antioxidant Activity of Mycelial Extracts from the Medicinal Fungus Paecilomyces hepiali (Ascomycetes).

Paecilomyces hepialid, a fungus originating in the Tibetan Plateau, has been used as a substitute for Ophiocordyceps sinensis because the primary chemical compounds and pharmacological effects of P. hepialid are similar to those of O. sinensis. P. hepialid has been developed into a dietary supplement and pharmaceutical products. The antioxidant activity of extracts using 2 solvents (water and ethanol) from mycelia obtained from 2 cultivation modes (solid-state and submerged cultivation) were evaluated in this study. Four strains of P. hepialid obtained from Qinghai, Sichuan, and Yunnan Provinces were included; the total phenolic content and in vitro antioxidant activity of mycelial extracts were compared. The total phenolic content and antioxidant activity of strains were found to be affected by the cultivation mode and extraction solvent. The ethanol extracts of solid-state cultivation of strain 2138, obtained from Sichuan Province, exhibited the highest antioxidant activity. The results showed that different strains might require different cultivation modes and extraction solvents for better antioxidant activity. However, solid-state cultivation and ethanol extraction are generally recommended based on the analyses conducted. Strain 2138 may be a good candidate for the purpose of producing functional foods. The results suggest that strain selection is important when P. hepialid is used to manufacture pharmaceutical products.

Free-Radical Scavenging Activities of Cultured Mycelia of Paecilomyces hepiali (Ascomycetes) Extracts and Structural Characterization of Bioactive Components by Nuclear Magnetic Resonance Spectroscopy.

Current research is focused on testing the cultivation of Paecilomyces hepiali mycelia on various plant substrates and producing fungus or mycelial biomass with qualitatively interesting substances. P. hepiali mycelia was cultivated using solid-state fermentation of different substrates. Mycelial biomass was then analyzed, and antioxidant activity was evaluated using the DPPH radical scavenging method for different ethanolic extracts based on a millet substrate (extract 1) or a chickpea substrate (extract 2). Extract 1 corresponds to a half-maximal DPPH radical inhibitory concentration of 1.73 mg/mL; the inhibitory concentration of ethanol extract 2 was almost 4.5 times higher at 7.92 mg/mL. Extracts 1 and 2 were separated into fractions by column chromatography and the chemical structures were determined for the substances that formed the most effective fraction of sample 1. The chemical structures of all compounds in the most active fraction of sample 1 were analyzed by 1H, 13C, distortionless enhancement by polarization transfer, correlation spectroscopy, heteronuclear single-quantum correlation spectroscopy, and heteronuclear multiple-bond correlation spectra.

Anti-nociceptive effects of Paecilomyces hepiali via multiple pathways in mouse models.

Paecilomyces hepiali (PH), a well-known medicinal fungus, has various pharmacological efficacies. In our study, the antinociceptive effects of PH and underlying mechanisms were evaluated using various mouse models. An acetic acid-induced writhing test, hot plate test, and formalin test were employed to evaluate the antinociceptive activities of PH. The levels of neuronal nitric oxide synthase (nNOS) in the hypothalamus and monoamine neurotransmitters in the serum and hypothalamus of experimental mice were examined. Additionally, hot plate tests using mice pretreated with various antagonists were used to determine the mechanisms of PH-mediated antinociception. The PH-enhanced latency period of mice in the hot plate test was significantly blocked by pretreatment with atropine and glibenclamide. PH shortened the phase I and phase II reaction times of formalin-treated mice. Strongly reduced writhing and stretching induced by acetic acid were observed in PH-treated mice, indicating that PH mainly exerts antinociceptive activity on neurogenic pain. After thermal pain stimulation for 30 s, compared to control mice, 7-day PH-treated mice had lower nNOS and dopamine levels, and increased levels of serotonin in both the serum and hypothalamus. Collectively, our data showed that PH mediated antinociceptive activities via multiple pathways, including monoamines, nNOS/ATP-sensitive K+ channels, and M-type acetylcholine receptors.

Paecilonic acids A and B, bicyclic fatty acids from the jellyfish-derived fungus Paecilomyces variotii J08NF-1.

Two new bicyclic fatty acids, paecilonic acids A and B (1 and 2), were isolated from the culture broth of the marine fungus Paecilomyces variotii derived from the jellyfish Nemopilema nomurai. Compounds 1 and 2 share the same molecular formula and possess a 6,8-dioxabicyclo[3.2.1]octane core skeleton. The planar structures of compounds 1 and 2 were established by spectroscopic analysis, which included NMR and ESI-MS/MS. Relative and absolute configurations were determined by analyzing coupling constants, NOESY correlations, and optical rotations.

Anti-diabetic activities of Paecilomyces tenuipes N45 extract in alloxan-induced diabetic mice.

Due to the limitations of existing anti-diabetic drugs, the treatment of diabetes mellitus remains a significant challenge. The present study aimed to investigate the hypoglycemic, hypolipidemic and antioxidant effects of Paecilomyces tenuipes N45 extracts on alloxan-induced type I diabetes mellitus in mice. Diabetic Kunming mice were orally administered with water extract (WE) at doses of 2.50, 0.25 and 0.05 g/kg) or alcohol extract (AE) at doses of 2.00, 0.20 and 0.04 g/kg, for 3 weeks, following which the levels of factors associated with blood glucose, lipids and free radicals were determined. The anti-diabetic activities of AE and WE were further confirmed via an oral glucose tolerance test. Similar to the effects of metformin, Paecilomyces tenuipes N45 extracts led to a significant reduction in blood glucose levels, increase in serum insulin concentration and normalization in the densities of low-density lipoprotein cholesterol and high density lipoprotein cholesterol. The Paecilomyces tenuipes N45 extracts exerted antioxidative effects, indicated by regulation in the levels of superoxide dismutase, malondialdehyde and glutathione peroxidase. Taken together, the results of the present study demonstrated that Paecilomyces tenuipes N45 extract, a safe pharmaceutical agent, exerted anti-diabetic and anti-nephropathic activities and, thus, offers potential as a novel therapeutic agent in the treatment of diabetes.

[Determination of 5 nucleosides components in culture of Paecilomyces hepialid by HPLC].

The concentration of 5 nucleosides, uracil, uridine, guanidine, adenine and adenosine in culture of Paecilomyces hepialid was determined by the developed method of HPLC. The HPLC method was performed on a Waters SunFire C18 (4.6 mm x 250 mm, 5 µm) column with methanol-water gradient elution as the mobile phase. The detection wavelength was 260 nm and the colunmn temperature was controlled at 30 °C. The linear range was 10.00-200.00 mg · L(-1) (r = 0.9994) for uracil, 10.10-202.00 mg · L(-1) (r = 0.9992) for uridine, 10.00-200.00 mg · L(-1) (r = 0.9991) for guanidine, 10.30-206.00 mg · L(-1) (r = 0.9992) for adenine and 10.45-209.00 mg · L(-1) (r = 0.9991) for adenosine, respectively. The RSD of precision was 0.032%, 0.035%, 0.039%, 0.049%, 0.00080%, respectively. The average recoveries of uracil, guanidine, adenine, and adenosine were 97.34%, 99.10%, 101.6%, 98.61% and 100.2% with RSD of 1.3%, 2.1%, 0.96%, 0.95%, and 1.3% respectively. The method showed high sensitivity, good selectivity, linearity and repeatability, which was suitable for the content analysis of 5 nucleosides components in P. hepialid and its extracts.

Dimeric Octaketide Spiroketals from the Jellyfish-Derived Fungus Paecilomyces variotii J08NF-1.

Paeciloketals (1-3), new benzannulated spiroketal derivatives, were isolated from the marine fungus Paecilomyces variotii derived from the giant jellyfish Nemopilema nomurai. Compound 1 was present as a racemate and was resolved into enantiopure 1a and 1b by chiral-phase separation on a cellulose column. Compounds 2 and 3, possessing a novel benzannulated spiroketal skeleton, were rapidly interconvertible and yielded an equilibrium mixture on standing at room temperature. The relative and absolute configurations of compounds 2 and 3 were determined by NOESY analysis and ECD calculations. Compound 1 showed modest antibacterial activity against the marine pathogen Vibrio ichthyoenteri.

Acute and subchronic toxicity studies on safety assessment of Paecilomyces tenuipes N45 extracts.

Paecilomyces tenuipes, one of the commonly used Chinese medicinal fungus, has received much attention over the world, which possesses various active compounds and biological activities. However, little toxicological information is available. Therefore, the present study evaluated the potential toxicity of aqueous and ethanol extracts of Paecilomyces tenuipes N45 via acute and subchronic administration in mouse and rat, respectively. For improving the extraction rate of aqueous extract, response surface methodology (RSM) was employed to optimize the extraction condition first in this paper. The obtained optimal extract conditions were temperature 80 °C, liquid-solid ratio 50 mL·g-1 and time 3 h. In the acute toxicity test, aqueous and ethanol extracts caused neither mortality nor toxicological signs, and the maximum tolerance dose was estimated over 15 g/kg. No mortality or adverse effects was observed in subchronic toxicity studies. No significant difference in bodyweight, relative organ weight or hematological parameters was noted during the experiment. Comparing with nontreated rats, ALT, K and BUN levels were changed in experimental group detecting via biochemical analysis. No abnormality of internal organs was noted between treatment and control groups in gross and histopathological examinations. Our present study suggested that the tolerance dose of the Paecilomyces tenuipes N45 extracts were more than 15 g/kg and no-observed-adverse-effect level (NOAEL) of the extracts for both male and female rats after 90-day adminstation. Additionally, the extracts may possess renal-protective and hepato-protective effects.

Paecilomycies japonica reduces repeated nicotine-induced neuronal and behavioral activation in rats.

BACKGROUND: Many studies have demonstrated that repeated injections of nicotine can produce progressive increases in locomotor activity and enhanced expression of c-fos and tyrosine hydroxylase (TH) in brain dopaminergic areas. Paecilomyces japonica (PJ) is a herbal medicine that is commonly used to treat opiate and other addictions in Eastern Asia. However, its influence on nicotine addiction has not been examined. This study was carried out to investigate the effects of PJ on repeated nicotine-induced behavioral sensitization of locomotor activity and c-Fos and TH expression in the rat brain using immunohistochemistry. METHODS: Rats were pretreated with PJ (10, 25, 50, 100, and 200 mg/kg, intraperitoneally) 30 min before repeated injections of nicotine (0.4 mg/kg, subcutaneously, twice daily for 7 days). Locomotor activity was measured in rats during 7-day nicotine treatments. On the seventh day, c-Fos and TH expression were assessed. RESULTS: Pretreatment with PJ decreased the development of nicotine-induced sensitization, c-Fos expression in the nucleus accumbens and striatum, and TH expression in the ventral tegmental area. PJ decreased nicotine-induced locomotor activity by modulating brain dopaminergic systems. CONCLUSION: The results of the present study suggest that PJ may be a useful agent for preventing and treating nicotine addiction.

Potent anti-calmodulin activity of cyclotetradepsipeptides isolated from Isaria fumosorosea using a newly designed biosensor.

Seven cyclotetradepsipeptides, namely beauverolides C (1), F (2), I (3), Ja (4), L (5), M (6), and N (7), were isolated from the entomopathogenic fungus Isaria fumosorosea. The beauverolides were evaluated as potential calmodulin (CaM) inhibitors using the newly designed CaM biosensor hCaM M124C-AF350; these peptides displayed high affinity to the protein with dissociation constants (Kd) ranging from 0.078 µM to 3.44 µM. Beauverolide Ja, the only one containing a tryptophan residue in its structure, showed the highest affinity. The docking study predicted that beauverolides could bind to CaM in the same site of interaction as chlorpromazine, a well-known calmodulin ligand.

Huperzine A production by Paecilomyces tenuis YS-13, an endophytic fungus isolated from Huperzia serrata.

Huperzine A (HupA), a naturally occurring alkaloid in the plant family Huperziaceae, has drawn great interest for its potential application in Alzheimer disease therapy. Our primary objective was to identify alkaloid- and HupA-producing fungi from the Chinese folk herb, Huperzia serrata. We established a rapid and efficient model for screening HupA-producing endophytic fungal strains. The presence of HupA in Paecilomyces tenuis YS-13 was analysed by thin-layer chromatography, high-performance liquid chromatography and mass spectrometry. The fermentation yield of HupA was 21.0 µg/L, and the IC50 of the crude extract of YS-13 fermentation broth was 1.27 ± 0.04 mg/mL. This is the first report of P. tenuis as a HupA-producing endophyte isolated from Huperziaceae.

Optimization for production of exopolysaccharides with antitumor activity in vitro from Paecilomyces hepiali.

In the present study, optimal medium for the growth of mycelia and the production of exopolysaccharides from Paecilomyces hepiali HN1 (PHEPS) in submerged culture was investigated. As a result, the maximum production of mycelia (12.98 ± 0.14 g/L) and PHEPS (5.33 ± 0.11 g/L) were achieved under the optimal medium of sucrose 46.08 g/L, yeast extract 4.71 g/L, (NH4)2SO4 5.72 g/L, KH2PO4 1.70 g/L, CaCl2 0.50 g/L, MgSO4 0.50 g/L, potato extract 1% and malt extract 1%. Furthermore, the antitumor activity of PHEPS in vitro was evaluated by using three cell lines of human liver tumor HepG2 cells, breast cancer MCF-7 cells and cervical cancer Hela cells. It was found that PHEPS exhibited relative higher anti-proliferative activity against HepG2 cells than MCF-7 cells and Hela cells. At a concentration of 500 µg/mL and 72 h treatment, the inhibition rate of PHEPS on HepG2 cells reached to 62.58%. All these results suggested that PHEPS could be explored as novel natural antitumor agent with great potential application.

Paeciloside A, a new antimicrobial and cytotoxic polyketide from Paecilomyces sp. strain CAFT156.

Paeciloside A (1) and eight known compounds, acremoauxin A (2), farinosones A (3) and B (4), 1,5-dideoxy-3-C-methyl-arabitol (5), ergosterol, ergosterol peroxide, cerebroside C, and adenosine, were isolated from cultures of Paecilomyces sp. CAFT156, an endophytic fungus occurring in Enantia chlorantha Oliv (Annonaceae) leaves. The structure of the new compound 1 was elucidated using MS, UV, 1D and 2D NMR experiments, while its absolute configuration was established by subsequent single-crystal X-ray diffraction analysis using copper Kα radiation and invariom nonspherical scattering factors. Paeciloside A (1) and compounds 2, 4, and 5 displayed inhibitory effects on two gram-positive bacteria, Bacillus subtilis and Staphylococcus aureus at a concentration of 40 µg per paper disk and moderate cytotoxicity towards brine shrimp larvae (Artemia salina). This study presents the first report on an endophytic fungus isolated from E. chlorantha Oliv and its natural products.