In vivo anti-aging properties on fat diet-induced high fat Drosophila melanogaster of n-butanol extract from Paecilomyces hepiali

The purpose of this study was to explore the potential of the development and application of Paecilomyces hepiali,a fungus with edible and medicinal value,as a foodborne antioxidant and anti-aging agent.Its n-butanol extract(PHE)from rice cultures was selected for anti-aging experiment because of significant free radical scavenging activity in vitro.In vivo,PHE could significantly prolong the mean lifespan,50%survival days,and the maximum lifespan of Drosophila on a high-fat diet.It is amazing that the mean lifespan increased from 19.1 days to 32.9 days,50%survival days increased from 15.7 days to 34.3 days,and the maximum lifespan extended from 44.7 days to 52.7 days,when the high-fat female Drosophila model was fed with 10μg/mL PHE.Further research showed that PHE reduced the accumulation of peroxide products and increased the activity of antioxidant enzymes.Then,through antioxidant activity tracking,dimerumic acid(compound 1,the IC_(50) value of 3.4μg/mL on DPPH free radicals scavenging activity),4,5-dihydroxy-3-methoxypentanoic acid(compound 2,new compound),and thymidine(compound 3)were isolated from PHE.It is worth mentioning that dimerumic acid,the major antioxidant compound of PHE(content up to 3%),was discovered in P.hepiali for the first time.It was concluded that PHE showed excellent anti-aging activity at a very low concentration on fat diet-induced high fat Drosophila melanogaster,and dimerumic acid may be its main material basis.These results indicated that PHE had the potential to be developed as antioxidant and anti-aging agent in the healthcare industry.

Artificial simulated saliva,gastric and intestinal digestion and fermentation in vitro by human gut microbiota of intrapolysaccharide from Paecilomyces cicadae TJJ1213

In this study,the in vitro digestion and fermentation of two intra-polysaccharide fractions(IPS1 and IPS2)from Paecilomyces cicadae TJJ1213 were investigated.The constituent monosaccharides of IPS1 and IPS2 were not changed after simulated saliva,gastric and small intestinal digestion.However,they can be hydrolyzed and utilized by gut microbiota,and short-chain fatty acids(SCFAs)level were increased after IPS1 and IPS2 treatments.Furthermore,16 S r RNA sequencing analysis of fermentation samples were performed.Alpha-diversity,beta-diversity and taxonomic composition differences analysis revealed that IPS1 and IPS2 promoted the proliferation of beneficial bacteria and modulated the overall structure of gut microbiota.Taxonomic comparison analysis found that IPS1 increased the relative abundances of beneficial bacteria including Megamonas,Bifidobacterium and Lactobacillus,while IPS2 could increase the abundance of Bacteroides,Parabacteroides and Phascolarctobacterium.In addition,they can also decrease the levels of pathogenic bacteria containing Escherichia-Shigella,Klebsiella and Fusobacterium.These results indicated that IPS from Paecilomyces cicadae TJJ1213 could be used as potential candidates for new functional foods.

红树林共生真菌Paecilomyces sp· Tree 1-7代谢产物的研究

用硅胶柱色谱及重结晶等方法,并根据理化性质和光谱数据从一株红树林共生真菌Paecilomycessp.Tree 1-7中分离鉴定了四个化合物,分别为secalon ic ac id A(1),tenellic ac id A(2),大黄素(3),大黄酚(4)。并首次对化合物1和2进行了抗人的肝癌细胞hepG2的活性测试。结果显示,化合物1有很强的抗肿瘤活性,IC50=2.0μg/m l,化合物2的抗肿瘤活性IC50=62.1μg/m l。

红树林共生真菌Paecilomyces sp. Tree 1-7代谢产物的研究

Paecilomyces sp.Tree 1-7是一株采自台湾海峡的红树林共生真菌。用硅胶柱色谱及重结晶等方法,并根据理化性质和光谱数据对该菌的代谢产物进行了研究,从菌丝体中分离鉴定出三个化合物,分别为alatern in(1),5,8-二羟基-4-甲基-香豆素(2),3-羟基-3,6-二甲基-2,5-哌嗪(3)。对1进行了抗人的肝癌细胞hepG2及抑制乙酰胆碱酯酶(AChE)的活性测试,结果显示,化合物1有较强的抗肿瘤活性(IC50=7.0μg/mL),及很好地抑制AChE活性(IC50=0.87μg/mL)。

Effect of Neoseiulus cucumeiris Carrying Paecilomyces fumosoroseus on the Prevention of Aphids and Spider Mites in Eggplant

Using predatory mites Neoseiulus cucumeris Oudermans dusted with en- tomopathgen Paecilomyces fumosoroseus （Wize） Brown ＆ Smith to control aphids and spider mites is a new idea of biological control. Some studies were conducted to confirm the feasibility of this idea. The germination condition of P. fumosoroseus conidia was determined by the method of different saturated salt solutions applying different humidity in small containers, and the pathogenicity test and the pot experi- ment were also conducted, under a suitable condition. There were 6 treatments for pot experiment as following： （1） CK; （2） releasing 50 N. cucumeds, （3） spraying 20 ml of 10^7 spores/ml conidial suspension; （4） releasing 50 N. cucumeris after spraying 20 ml of 10^7 spores/ml conidial suspension; （5） releasing 20 N. cucumeris dusted conidia of P. fumosoroseus; and （6） releasing 50 N. cucumeris dusted with conidia of P. fumosoroseus. The result showed that the germination condition of P. fu- mosoroseus conidia was 25-30 ℃ and 95%-100% RH, and under this condition, the 10s spores/ml conidia suspension caused a mortality of 86.52% in aphids and 41.78% in predatory mites after ten days. From the pot experiment, the numbers of aphids of treatment （3）, （4）, （6） were close to 0 at the 8th day. This study proved that the predatory mites N. cucumeris dusted with the entomogenous P. fu- mosoroseus could provide satisfactory control on both aphids and spider mites in potted eggplants under suitable temperature and humidity. The predatory mite vec- toring entomopathogen system could provide an economic and labor saving way for aphid control.

Optimization of V_A and V_E Production by a Paecilomyces marquandii Strain

In order to further improve the yield of VA and VE produced by a Paecilomyces marquandii strain, using PDA liquid medium as the basic medium, the effects of adding nitrogen source, pH value and Fe2＋ on the VA and VE production were compared with an orthogonal experiment. The results showed that the VA yield was significantly increased when sodium nitrate was added as nitrogen source, pH value of medium was 6, but Fe2＋ was not added; the addition of peptone into the basic medium as a nitrogen source contributed to the improvement of VE yield in mycelia; the PDA liquid medium, as a basic medium, was more conducive to the VA production by the Paecilomyces marquandii strain.

White-spot disease of Chinese soft-shelled turtles (Trionyx sinens) caused by Paecilomyces lilacinus

Chinese soft-shelled turtles （Trionyx sinens） in culture farms using an artificial warming system in Zhejiang, China, often show typical signs of white-spot disease such as white spots on their bodies, skin lesions, anorexia and eventually death. The sick turtles were mostly 5-80 g in weight. A suspected fungal pathogen was isolated from the sick turtles and verified as Paecilomyces lilacinus by sequence analysis of the internal transcribed spacer （ITS） of its ribosomal DNA （rDNA）. Detailed morphological examinations were also conducted to confirm the white-spot disease.

Antidepressant effect of bioactive compounds from Paecilomyces tenuipes in mice and rats

A bioactive compound from Paecilomyces tenuipes（BCPT）has an inhibitory effect on monoamine oxidase A（MAO-A）in vitro.Researchers have thought that BCPT may be a potential antidepressant.The MAO-A suppressor moclobemide served as a control,and this study investigated the mechanisms of BCPT as an antidepressant.Results demonstrated that BCPT induced significantly increased sucrose intake in chronic unpredictable stressed rats,shortened immobility time in forced swimming mice,improved the scores of blepharoptosis and akinesia in reserpine-treated mice,increased the number of 5-hydroxy tryptophan-induced head-twitches,remarkably enhanced the expression of hippocampus mineralcorticoid receptor and glucocorticoid receptor mRNA,decreased the ratio of mineralcorticoid receptor to glucocorticoid receptor and raised the levels of dopamine,norepinephrine and 5-hydroxytryptamine,while decreasing hydroxyindole acetic acid levels or dihydroxy-phenyl acetic acid in chronic unpredictable stressed rats.Behavioral test results suggested that BCPT potentially had antidepressant-like activity.Meanwhile,BCPT increased the levels of neurotransmitters,and mineralcorticoid receptor and glucocorticoid receptor mRNA in the hippocampus,which may be an important mechanism of its antidepressant effect.

Mycelial Growth of <i>Paecilomyces hepiali</i>in Various Agar Media and Yield of Fruit Bodies in Rice Based Media

Growth of?Paecilomyces hepiali?in various agar media and yield of fruit bodies in rice based media were?studied. The best growth in agar media was obtained at 25℃?(61.86 mm colony diameter in 14 days). The initial agar media pH range?from?6 to 8 was found to be?the?most favourable for mycelial growth. This study found that agars made with powders of cereal grains alone do not support good mycelial growth of?P. hepiali. Addition of peptone improved mycelial growth significantly. The most favourable carbon sources were Mannose, Fructose and Glucose. Organic nitrogen sources were found to be?the?most preferred. The results demonstrated that brown rice is better than polished rice in yield of fruit bodies. Addition of peptone was found to be quite significant in enhancing yield of fruit bodies. Peptone, as a supplement, gave a better yield than addition of egg yolk, albumen and a mixture of the two. The medium with?40 g brown rice, 0.325 g glucose, 0.65 g sucrose, 2 g peptone and 65 ml corn steep liquor was found to be?the?most favourable and it yielded 19.3 g of fresh fruit bodies.

Separation of Insecticidal Ingredient of Paecilomyces cicadae (Miquel) Samson

In this study, the insecticidal active substance of the fruiting body of Paecilomyces cicadae （Miquel） Samson that can kill the larvae of Plutella xylostella （Lepidoptera： Plutellidae） was isolated and purified by combined separation with biology experiments for the first time. By means of bioactive guided isolation of 40% ethanol-H2O, dichloromethane, ethyl acetate, n-butyl ethanol, macroporous adsorption resin column, high performance liquid chromatography, and sephadex A-20 column, the insecticidal active compound was purified,and a white powder compound was obtained. The compound was active against the third instar larvae ofP. xylostella in 24, 48, and 72 h at the concentration of 10.0 mg mL-1 with a corrective mortality of （95.1 ± 1.8）, （97.7 ± 1.6）, and （99.1 ± 1.7）%, respectively. The compound was strongly sensitive to the larvae. It was stable at acid environment, and was not sensitive to pepsin, and other chracteristics were studied.

Advances in research on chemical constituents and pharmacological effects of Paecilomyces hepiali

Paecilomyces hepiali is a fungus with edible and pharmaceutic value,isolated from Ophiocordyceps sinensis.Its main bioactive constituents contain alkaloids,cyclic dipeptides,steroids,organic acids,and polysaccharides.Due to the huge development potential of P.hepiali in the field of food and medicine,it has been developed into a variety of products that cater to the needs of the public.In this paper,the chemical constituents and their pharmacological effects of P.hepiali are reviewed,which can provide reference for the development and application of P.hepiali.

A Comparative Study on Chemical Composition and Pharmacological Effects of <i>Paecilomyces hepiali</i>and Wild <i>Ophiocordyceps sinensis</i>

This study looked at comparison of chemical components and pharmacological activity between wild Ophiocordyceps sinensis and Paecilomyces hepiali. The chemical components investigated included amino acids, vitamins, dietary elements, protein, lipid, ash, carbohydrates, crude fibre, ergosterol and mannitol. Studies on pharmacological activity included anti-platelet aggregation, inhibitory effect on IL-8 gene expression, anti-mutagenic activity, skin whitening effect and impro- vement activity on human skin texture. The results show that P. hepiali has a larger total content of seven essential amino acids (leucine, lysine, methionine, phenylalanine, tryptophan, tyrosine and valine) than O. sinensis, 8580 mg/100g and 6180 mg/100g respectively. The total content of dietary elements analysed (potassium, magnesium, zinc, copper, manganese and selenium) was also higher in P. hepiali (3135 mg/100g) than that in O. sinensis (2445 mg/100g). The total content of four vitamins (B1, B2, B6 and E) was almost equal for both fungi. Paecilomyces hepiali had more content of protein, lipid, ash, carbohydrate, ergosterol and mannitol than O. sinensis. However, the contents of lipid and ash were not significantly different between the two fungi. The hot water extract of P. hepiali has been shown to have a number of pharmacological effects which include 1) inhibition of aggregation on human platelet, 2) inhibition of IL-8 gene expression, 3) anti-mutage- nic activity, 4) inhibition effect on production of melanin, and 5) improvement on human skin texture. Anti-platelet aggregation effects on human platelet, inhibition of IL-8 gene expression, anti-mutagenic activity and improvement effect on human skin texture were greater in P. hepiali treatments than that in wild O. sinensis.

Effects of hot-water extract of <i>Paecilomyces hepiali</i>on hypertension parameters in Spontaneously Hypertensive Rats

In this study, effects of hot water extract of Paecilomyces hepiali mycelia on hypertension parameters in spontaneously hypertensive rats (SHR) were investigated. The tested parameters included blood pressure, blood and urine biochemical composition, renin and angiotensin II levels in the blood. Prior to these tests, the extract was examined for toxicity. The fungus was cultivated in a solid medium composed of 40 g brown rice, 0.32 g glucose, 0.65 g sucrose, 2 g peptone and 65 ml corn steep liquor. No abnormality or harmful effects were observed in the toxicity test. Administration of a continuous-dose, once daily, to SHR for 27 weeks (from 13 weeks of age) decreased the systolic blood pressure (SBP) significantly. Levels of blood urea nitrogen, β-lipoprotein lipid peroxides and low density lipoprotein were significantly lower in the treated groups when compared to the control group. Urinary protein was significantly reduced in the middle and high dose groups. In comparison with the control group (0 mg/kg/10ml/day), significantly higher values were obtained for total cholesterol in groups that were given middle (170 mg/kg/10ml/day) and high (250 mg/kg/10ml/day) dosages. In all dosages (low, middle and high) the values for triglyceride were significantly higher than value found in the control group. In terms of angiotensin II levels, the value in the control group was markedly higher than values in the other groups. The results suggest that oral administration of hot water extract of P. hepiali mycelia has ability to control hypertension in rats.

Effect of paecilomyces hepiall chen on serum inflammatory factors,oxidative stress and renal function in patients with diabetic nephropathy

Objective: To investigate the effect of paecilomyces hepiall chen on serum inflammatory factors, oxidative stress and renal function in patients with diabetic nephropathy. Methods: A total of 124 cases of diabetic nephropathy patients were selected as the research object, based on the random data table they were divided into the control group and the observation group, 62 cases in each group, two groups of patients were given conventional treatment, on this basis, the patients in the control group were treated with valsartan capsules treatment, while patients in the observation group were given valsartan capsules combined with paecilomyces hepiall chen treatment. To compare the levels of blood pressure [systolic blood pressure (SBP), diastolic blood pressure (DBP)], inflammatory factor [tumor necrosis factor -α (TNF-α), high sensitive C reactive protein (hs-CRP)], oxidative stress [superoxide dismutase (SOD), malondialdehyde (MDA)] and renal function index [serum creatinine (Scr), urea nitrogen (BUN)] of the two groups before and after treatment. Results: Before treatment, there were no significantly different of the levels of SBP, DBP, TNF-α, hs-CRP, SOD, MDA, Scr and BUN between the two groups. After treatment, the observation group SBP, DBP, TNF-α, hs-CRP, MDA, Scr and BUN level were (135.71±15.64) mmHg, (84.96±13.97) mmHg, (46.37±5.92) ng/L, (5.99±0.94) mg/L, (20.91±2.38) μmol/L, (83.86±10.16) μmol/L and (6.53±1.37) mmol/L, were significantly lower than before treatment, and after treatment the TNF-α, hs-CRP, MDA, Scr and BUN in observation group were significantly lower than the control group;Compared with the levels in the same group before treatment, the two groups of SOD levels were significantly increased, and the observation group [(93.21±7.06) U/mL] was significantly higher than the control group ((79.84±6.12) U/mL)Conclusion: Paecilomyces hepiall chen on treatment of diabetic nephropathy, can effectively relieve chronic inflammation and oxidative stress, improve renal function, has an important clinical value.

一株蛹拟青霉(Paecilomyces militaris)深层发酵产物中抗肿瘤活性物质分析

【目的】分析蛹拟青霉(Paecilomyces militaris)菌株RCEF0927发酵液中的抗中国仓鼠卵巢瘤(CHO)细胞活性及活性成分的具体组成。【方法】用刃天青(Resazurin)法测定样品对CHO细胞的抑制率;用高效液相色谱-高分辨质谱和活性测定联用的方法进行活性成分分析和鉴定。【结果】活性测定结果表明菌株RCEF0927的发酵液具有较强的抗CHO细胞活性;提取实验结果表明抗肿瘤活性物质能较好地被乙酸乙酯提取出来;液相色谱-质谱-活性测定分析表明提取物中活性组分的可能分子式分别为C10H13N5O3、C22H22O9、C23H24O10、C22H22O10、C15H10O4、C15H10O5、C36H70O11、C36H68O12、C38H74O11、C40H76O13和C44H82O14。【结论】根据细胞毒性、色谱特性、紫外光谱特征和质谱碎片,可初步推断发酵物中抗肿瘤活性成分分别为虫草素、4′-羟基异黄酮-7-O-(4″-O-甲基)-β-D-葡萄糖甙、6-甲氧基-4′-羟基异黄酮-7-O-(4″-O-甲基)-β-D-葡萄糖甙、5,4′,-二羟基异黄酮-7-O-(4″-O-甲基)β--D-葡萄糖甙、7,4′,-二羟基异黄酮、5,7,4′,-三羟基异黄酮、油酸酯聚氧乙烯醚、聚氧乙烯山梨醇酐单硬脂酸酯脱水化合物、聚氧乙烯山梨醇酐单油酸酯,以及含聚氧丙烯或聚氧丁烯结构单元的聚氧乙烯山梨醇酐脂肪酸酯衍生物等化合物。上述化合物对CHO细胞的毒性为首次发现。

Multigene phylogeny of the family Cordycipitaceae (Hypocreales): new taxa and the new systematic position of the Chinese cordycipitoid fungus Paecilomyces hepiali

The phylogeny and systematics of cordycipitoid fungi have been extensively studied in the last two decades.However,systematic positions of some taxa in the family Cordycipitaceae have not yet been thoroughly resolved.In this study,a new phylogenetic framework of Cordycipitaceae is reconstructed using multigene(nrSSU,nrLSU,tef-1α,rpb1 and rpb2)sequence data with large-scale taxon sampling.In addition,ITS sequence data of species belonging to the Lecanicillium lineage in the family Cordycipitaceae are used to further determine their phylogenetic placements.Based on molecular phylogenetic data together with morphological evidence,two new genera(Flavocillium and Liangia),16 new species and four new combina-tions are introduced.In the new genus Flavocillium,one new species F.bifurcatum and three new combinations previously described as Lecanicillium,namely F.acerosium,F.primulinium and F.subprimulinium,are proposed.The genus Liangia is built by the new species Lia.sinensis with Lecanicillium-like asexual morph,isolated from an entomopathogenic fungus Beauveria yunnanensis.Due to the absence of Paecilomyces hepiali,an economically and medically significant fungus,in the earlier phylogenetic analyses,its systematic position has been puzzling in both business and academic communities for a long time.Here,P.hepiali is recharacterized using the holotype material along with seven additional samples.It is assigned to the genus Samsoniella(Cordycipitaceae,Hypocreales)possessing Cordyceps-like sexual morph and Isaria-like asexual morph,and thus a new combination,namely S.hepiali is proposed.An additional nine new species in Samsoniella are described:S.alpina,S.antleroides,S.cardinalis,S.cristata,S.lanmaoa,S.kunmingensis,S.ramosa,S.tortricidae and S.yunnanensis.Four new species in Cordyceps are described:C.chaetoclavata,C.cocoonihabita,C.shuifuensis and C.sub-tenuipes.Simplicillium yunnanense,isolated from synnemata of Akanthomyces waltergamsii,is described as a new species.

Bioactive compounds from Paecilomyces tenuipes regulating the function of the hypothalamo-hypophyseal system axis in chronic unpredictable stress rats

Background A bioactive compound from Paecilomyces tenuipes （BCPT） has an inhibitory effect on monoamine oxidase A （MAO-A） and monoamine oxidase B （MAO-B） in vitro and in vivo, which indicates BCPT may be a potential antidepressant. In this study we aimed to study the antidepressant effects of BCPT in the chronic unpredictable stress （CUS） model in rats and explore underlying mechanisms in the hypothalamic-pituitary-adrenal （HPA） axis. Methods The antidepressant effects of BCPT were studied in the chronic unpredictable stress model in rats. Animals were housed isolated, except the control group. Rats were exposed daily to different random stressors from day 1 to 21. Awarding response was detected by calculating the 24-hour consumption of sucrose water. Cortisol （CORT） and adrenocorticotropic hormone （ATCH） contents in serum and arginine vasopressin （AVP） contents in the pituitary body were detected by radio immunoassays. Total RNA of hippocampus or hypothalamus was extracted and subjected to reverse transcription-polymerase chain reaction （RT-PCR） for the measurement of corticotrophin releasing hormone （CRH） mRNA or mineralocorticoid receptor （MR） mRNA and glucocorticoid receptor （GR） mRNA levels. Statistical analyses were performed using one way analysis of variance （ANOVA） followed by Student-Newman-Keuls （SNK） test. Results Chronic unpredictable stress resulted in reduction of sensitivity to reward and abnormality in the HPA axis in the animal model. BCPT improved the reward reaction as measured by increasing sucrose consumption, remarkably reduced serum CORT and ACTH levels and the AVP content in the pituitary body in the CUS-treated rats, decreased the expression of CRH mRNA, enhanced the expression of hippocampus MR mRNA, GR mRNA and decreased the ratio of MR/GR. Conclusions BCPT has potentially antidepressant-like activity and normalized the HPA axis hyperactivity in a CUS model of depression in rats. This may be an important mechanism of its antidepressant effect.

Prenylated indole alkaloids from the marine-derived fungus Paecilomyces variotii

Two new prenylated indole alkaloids,namely,dihydrocarneamide A（1） and iso-notoamide B（2）,were isolated from the marine-derived endophytic fungus Paecilomyces variotii EN-291.The structures of these metabolites were determined based on comprehensive spectral analysis,together with chiral HPLC analysis of the acidic hydrolysates.Unlike other prenylated indole alkaloids such as asperparalines,notoamides,and versicolamides,compounds 1 and 2 are the rare examples of C-5 prenylation,forming the fused dimethyldihydropyran ring at C-5 and C-6 of the indole ring.The cytotoxic activity of the isolated compounds was evaluated.

Taxonomy characterization and cadmium biosorption of fungus strain

The taxonomy characteriazation and cadmium （Cd） biosorption of the high Cd-resistant fungus M1 were investigated. The internal transcribed spacers （ITS） region and β-tubulin genes of the strain were amplified, sequenced and analyzed by molecular biology technology. The Cd biosorption assay was performed by shaking flask. Fourier transform infrared spectroscopy was used to analyze the mycelium. The similarity of gene sequences and phylogenetic trees show the very close relation between the strain and Paecilomyces lilacinus, and the fungus M1 was identified as P. Lilacinus. The initial pH 6 and Cd concentration about 100 mg/L are optimum. Zn and Mn have a little effect on the Cd biosorption of the strain, while Cu and Pb present obvious effects. FTIR analysis shows that the fungus adsorbs Cd by esters, anhydride, and amide. With the preferable absorption capacity, fungus M1 is considered to have good prospects in bioremediation.

右旋糖酐酶产生菌的筛选、鉴定及发酵

从土壤中筛选1株能产生右旋糖酐酶的菌株DN01，对该菌进行了分类鉴定和发酵条件考察。结合菌落形态、孢子形态特征和菌株的ITS rDNA序列分析进行菌株的鉴定，确定该菌株属于拟青霉（Paecilomyces）。在30℃120r／min摇床振荡培养8d后，发酵液上清中的右旋糖酐酶活力最高达到19．34U／ml。通过单因素试验对发酵条件进行考察，结果表明产生右旋糖酐酶的最适碳源为右旋糖酐，最适氮源为硫酸铵，培养基的起始pH为4．O～5．0，最适温度为30℃。