Effect of dietary supplementation with discardednutrient medium of Cordyceps militaris on layingperformance and diversity of intestinal flora in laying hens

The aim of the experiment was to explore the feasibility of discarded nutrient medium of Cordyceps militaris as feed in the production of laying hens. 100 g/kg, 200 g/kg, 300 g/kg of discarded nutrient medium of Cordyceps militaris were added to the basal diet of laying hens. The results showed that the optimal addition of discarded nutrient medium of Cordyceps militaris in the diet of laying hens was 10%. According to the results of measuring the conventional indicators of eggs, the weight of eggs produced by laying hens fed with discarded nutrient medium of Cordyceps militaris was higher than that of laying hens fed with ordinary laying hens. The content of interleukin-1(IL-1) in the experimental group was significantly higher than that in the control group, and the concentration of IL-1 increased by 141.5 pg/mL, which indicated that the application of discarded nutrient medium of Cordyceps militaris effectively improved the immunity of laying hens. The high-throughput analysis of the intestinal contents of the two groups of laying hens showed that the microbial population abundance of the intestinal tract of the experimental group was greater than that of control group, and the application of discarded nutrient medium of Cordyceps militaris increased the diversity of bacteria in the intestinal tract of laying hens. In addition, the sensitivity of some pathogenic bacteria in the intestinal tract of chickens to drugs was also increased, thereby reducing the use of antibiotics. The secondary utilization of discarded nutrient medium of Cordyceps militaris has great development and utilization prospects, which provided a scientific reference and basic theoretical basis for the development of discarded nutrient medium of Cordyceps militaris as feed in the production of laying hens.

Optimization of Rice Medium for Cultivation of Cordyceps militaris in the Alpine Region

In this study,six Cordyceps militaris strains introduced to Tibet were used as experimental materials to screen the dominant C.militaris strain by comparing mycelial growth rate,fruiting body yield and growth status.The composition of rice medium for C.militaris cultivation was optimized by orthogonal experiment.According to the results,although C.militaris strain ＂TAAAS1＂ did not exhibit the highest mycelial growth rate,it had uniform fruiting body formation and higher fruiting body yield than other strains,which suggested that C.militaris strain ＂TAAAS1＂ was most suitable for cultivation and production of C.militaris in the alpine region.The optimal rice medium for C.militaris cultivation consisted of 4 g/L pupa powder,20 g/L glucose,1.5 g/L KH_2PO_4 and 2 g/L MgSO_4.This study provided the theoretical basis for cultivation and production of C.militaris in the alpine region.

Extraction, Purification and Anti-tumor Activity of Polysaccharide from Mycelium of Mutant Cordyceps Militaris

A water-soluble polysaccharide（named MCMP） was isolated from the mycelium with high yield mutation Cordyceps militaris by hot-water extraction, deproteinization by sevage, alcohol precipitation, anion-exchange and gel filtration chromatography CL-6B. The polysaccharide contained mannose, rhamnose, galactose and glucose in a molar ratio of 59.36：1：8.31：39.50, of which the average molecular weight is 8100. In our research, Hep-G2 cells, Hela cells and mesangial cells were chosen to determine the anti-tumor activity of the polysaccharide. The results of MTT assay show that polysaccharides of the mutant strain presented inhibitory activity on the cells proliferation after 48 h incubation.

The protoplast two-way fusions and fusant characteristics of Antrodia cinnamomea and Cordyceps militaris

This study generated two fused protoplasts of Antrodia cinnamomea and Cordyceps militaris in two ways.The protoplasts of A.cinnamomea were inactivated by heat to inactivate biochemical processes and enzymatic activities in the cytoplasm,and the protoplasts of C.militaris were inactivated by UV radiation to invalidate their genome function,then they were fused under optimal conditions to get a fusion rate as(7.42±0.8)×10^(-6) fusants/mL;the new fusants were abbreviated as Ac-Cm.On the other hand,when A.cinnamomea and C.militaris were treated with heat and UV oppositely using similar experiments,the fusion rate was(9.70±0.68)×10^(-5) fusants/mL,and the new fusants were abbreviated as Cm-Ac.We selected each of two best-growing fused colonies Ac-Cm-1,Ac-Cm-2,Cm-Ac-1,and Cm-Ac-2,together with parental A.cinnamomea and C.militaris,and studied their morphology,growth antagonism tests,and genetic relationships by 18 S rRNA sequencing.In comparison with the initial cultures of 4 fusants,the yields of adenosine,biomass,cordycepic acid,cordycepin,total polysaccharide,and total triterpenoids were increased up 1.305-50.1563 times in the optimal medium conditions.For gene stability tests,those of the four fusants and their outputs were stabilized within 10 generations.

Procoagulant constituents from Cordyceps militaris

Cordyceps militaris,belongs to Clavicipitaceae family,was investigated for its chemical compounds,and six compounds were isolated and purified by silica gel column chromatography,Sephadex LH-20 and recrystallization,and their structures were elucidated by spectral techniques and physicochemical properties as ergosterol(1),adenosine(2),cordycepin(3),ergosterol peroxide(4),tetracosanoic acid 2,3-dihydroxypropylester(5),mannitol(6).Procoagulant activity was screened by assaying the activated partial thromboplastin time(APTT),prothrombin time(PT),thrombin time(TT)and fibrinogen(FIB)in vitro.The results indicated that ergosterol,adenosine,cordycepin,ergosterol peroxide and mannitol showed strong procoagulant activity.

Effects of Different Cultivation Media and Periods on the Content of Main Active Components of Cordyceps militaris Strain QC04

In this study,Cordyceps militaris strain QC04 was cultivated in oat,rice,and wheat media for different time periods.We studied the effects of different cultivation media and periods on the fruiting body biomass and active components of C.militaris,aiming to provide reference for the production and utilization of C.militaris QC04.The results showed that the dry weight of the fruiting body of C.militaris was the highest in the wheat medium,moderate in the oat medium,and the lowest in the rice medium.The content of cordycepin and adenosine in the fruiting body and residual medium was higher in the oat and rice media than in the rice medium.The content of cordycepin and adenosine in fruiting body and residual medium peaked on day 55.Furthermore,the content of cordycepin in the fruiting body was lower than that in the residual medium,while the content of adenosine showed an opposite trend.The content of cordycepic acid in the rice medium was generally higher than that in the oat and wheat media.As the cultivation period extended,the fruiting body biomass declined and the content of cordycepic acid in the fruiting body increased slightly.

Inhibitory Effects of Cordyceps militaris( Bombyx mori )Extract against Respiratory Syncytial Virus

[ Objective] This study aimed to investigate the inhibitory effects of 95%, 70% ethanol extracts and water extract of Cordyceps militaris （Bombyx mori）, on cells infected with respiratory syncytial virus （RSV）. [Method] Human embryonic kidney cells （HEK293, RSV virus-sensitive cells） were infected with respiratory syncytial virus and incubated with tissue cell culture method, to analyze the inhibitory effects of different extracts of Cordyceps militaris （Bombyx mori） on proliferation of RSV-infected cells by cytopathic effect （CPE） observation and tetrazolium salt reduction assay （ MTF method）. [ Result] Different extracts of Cordyceps militaris （Bombyx mori） were non-toxic to HEK293 cells. CPE occurred successively in various sample groups after 48 h, and the degree of CPE was reduced gradually with the increasing concentration. To be specific, 95 % ethanol extract （100 μg/ml）, 70% ethanol extract （100, 25 μg/ml） and water extract （100, 25 μg/ml） of Cordyceps militaris （BombTx mori） exhibited inhibitory effects against RSV subtype A; 70% ethanol extract （100, 25 μg/ml） and water extract （100, 25 μg/ml） of Cordyceps militaris （Bombyx mori） exhibited inhibitory effects against RSV subtype B; the effects of water extract （100 μg/ml） of Cordyceps militaris （Bombyx mori） on RSV subtypes A and B were similar to positive control group. However, different extracts of Cordyceps militaris （Bombyx mot/） with low concentrations （6.25, 1.56 μg/ml） exhibited no inhibitory effects against respiratory syncytial virus. [ Conclusion ] Cordyceps militaris （Bombyx mori） exhibited significant inhibitory effects against syncytial virus, and the inhibitory effects were related to the water soluble components.

Optimization of Extraction Process and Anti-ultraviolet Activity of Polysaccharide from Cordyceps militaris

Extraction process of Cordyceps militaris polysaccharide(PS)was optimized with single factor experiment and orthogonal test.Then,the anti-UVB ability of the polysaccharide was investigated to develop anti-UV sun-screening agent through determining the cell viability after UVB irradiation.The results showed that the optimum were solid-liquid ratio at 1∶40(g/ml)at 80℃ for 4 h,and the extract ratio was 27.4%according to the verification test.The PS showed no cytotoxicity to HaCaT cells and pretreatment with the PS significantly decreased UVB-induced cytotoxicity in a dose-dependent relationship.It suggests that the PS has strong anti-UVB irradiation ability and has a good application prospect in developing sunscreen agent.

Optimization of the Cultivation Device for Cordyceps militaris and Development of Its Characteristic Product

In order to better promote the development of the Cordyceps militaris industry,the C.militaris cultivation device was optimized.Multiple layers of bearing plates are arranged on a support frame,and six cultivation boxes are placed on each layer of bearing plate.The optimized device improves the cultivation method of C.militaris,improves the space utilization rate,and can greatly increase the output of C.militaris.Meanwhile,combining with local special R.roxburghii in Guizhou,a C.militaris and R.roxburghii compound drink were developed,which extends the industrial chain of C.militaris,which meets the current social needs and has good market prospects.The research results can promote rural industrial revolution and further help the revitalization of rural industries.

Antioxidant Activities of Aqueous Extract from Cultivated Fruit-bodies of Cordyceps militaris （L.） Link In Vitro

Biological antloxldants extracted from plants and fungi have potential abilities to scavenge free radicals and Inhibit lipid peroxldatlon, playing Important roles in preventing diseases, for example, cancer, and aging Induced by reactive oxygen species, which may cause oxidative damage to DNA, proteins and other macromolecules. The antloxldant potency of cultivated fruit-bodies of Cordyceps militarls （L.） Link was investigated In this study. Five established In vitro systems were employed, including the 1,1-dlphenyl-2- plcryldrazyl （DPPH） free radical scavenging, hydroxyl radical eliminating, iron chelating, Inhibition of Ilnolelc acid lipid peroxldatlon and reducing power. The aqueous extract from cultivated fruit-bodies was subjected to the test of amino acid, polysaccharlde and mannitol. Ascorblc acid （Vc）, butylated hydroxytoluene （BHT） and ethylenedlamlnetetraacetlc acid （EDTA） were used as positive controls for comparisons. Among the assays, the aqueous extract of C. mllltarls frult-bodles shows a significant scavenging effect on DPPH, eliminating the capability on hydroxyl radicals and the chelating effect on ferrous Iron. The extract also shows positive results of Inhibiting Ilnoleic acid lipid peroxldatlon and reducing power.

Research progress on Cordyceps militaris polysaccharides

Cordyceps militaris is a famous medicinal and edible fungus,whose polysaccharides are important biologically active substances.In recent years,the molecular structure and pharmacological functions of C.militaris polysaccharides were fully studied.However,the potential mechanism of their pharmacological functions and their chemical modification have not been systematically elucidated.Moreover,the obtention and usage of C.militaris polysaccharides present several issues,including low polysaccharide yields and poor pharmacological functions in industrial production.This review presents the latest results on the pharmacological mechanism of C.militaris,including information on how to improve C.militaris industrial production through stabilization of the strain to prevent degeneration and chemical modifications of its polysaccharides.In addition,this article examines C.militaris fermentation conditions,the polysaccharides extraction methods developed for industrial production,and the strategies to increase the yield of polysaccharides and improve their pharmacological functions.This review could provide a theoretical basis for future research on C.militaris polysaccharides,and also serve as an important reference value in the field of functional foods and medicine.

Biotransformation of soy whey into a novel functional beverage by Cordyceps militaris SN-18

Soy whey,a liquid nutritional by-product of soybean manufacture,is rich in proteins,oligosaccharides and isoflavones.Soy whey can be used to produce functional beverages,instead of discarding it as a waste.In this study,unfermented soy whey(USW)and Cordyceps militaris SN-18-fermented soy whey(FSW)were investigated and compared for their physicochemical and functional properties by high performance liquid chromatography(HPLC)and DNA damage assay.Results show that C.militaris SN-18 fermentation could increase the contents of essential amino acids,total phenolic and flavonoid and isoflavone aglycones and eliminate the oligosaccharides in soy whey.Furthermore,C.militaris SN-18 could significantly enhance the ABTS radical scavenging ability,reducing power and ferric reducing power of soy whey,and its fermented products could prominently attenuate Fenton reaction-induced DNA damage.These findings indicate that soy whey can potentially be converted into a novel soy functional beverage by C.militaris SN-18 fermentation.

Polysaccharides Purified from Wild Cordyceps Activate FGF2/FGFR1c Signaling

Land animals as well as all organisms in ocean synthesize sulfated polysaccharides. Fungi split from animals about 1.5 billion years ago. As fungi make the evolutionary journey from ocean to land, the biggest changes in their living environment may be a sharp decrease in salt concentration. It is established that sulfated polysaccharides interact with hundreds of signaling molecules and facilitate many signaling transduction pathways, including fibroblast growth factor （FGF） and FGF receptor signaling pathway. The disappearance of sulfated polysaccharides in fimgi and plants on land might indicate that polysaccharides without sulfation might be sufficient in facilitating protein ligand/receptor interactions in low salinity land. Recently, it was reported that plants on land start to synthesize sulfated polysaccharides in high salt environment, suggesting that fungi might be able to do the same when ex- posed in such environment. Interestingly, Cordyceps, a fungus habituating inside caterpillar body, is the most valued traditional Chi- nese Medicine. One of the important pharmaceutical active ingredients in Cordyceps is polysaccharides. Therefore, we hypothesize that the salty environment inside caterpillar body might allow the fungi to synthesize sulfated polysaccharides. To test the hypothesis, we isolated polysaccharides from both lava and sporophore of wild Cordyceps and also from Cordyceps militaris cultured without or with added salts. We then measured the polysaccharide activity using a FGF2/FGFRlc signaling-dependent BaF3 cell proliferation assay and found that polysaccharides isolated from wild Cordyceps activated FGF2/FGFR signaling, indicating that the polysaccha- rides synthesized by wild Cordyceps are indeed different from those by the cultured mycelium.

Cultivated Cordyceps: A Tale of Two Treasured Mushrooms

Ophiocordyceps sinensis and Cordyceps militaris both contain many bioactive compounds that confer potential therapeutic benefits. This review discusses the possible use of cultivated C. militaris as an effective substitute for native O. sinensis in the face of ever-increasing prices of O. sinensis because of its short supply. On the one hand, cultivated C. militaris contains higher levels of cordycepin when compared with that of wild-type O. sinensis and cultivation of C. militaris has been shown to be capable of reducing the risk of heavy metal contamination. On the other hand, there is a paucity of robust in vivo studies and randomized controlled tests comparing the pharmacology and use of C. militaris and O. sinensis. For extraction of cordycepin as western-style tablets, the use of cultivated C. militaris rather than O. sinensis represents the most appropriate future approach. For many other purposes, comparative pharmacology and clinical trials are in urgent needs.

野生蛹虫草菌株培养特性及子实体多糖含量比较分析

蛹虫草(Cordyceps militaris)是一种药食两用真菌,为获得高产优质的菌种资源,以采集分离的5株蛹虫草野生菌株(XY002、XY008、XY011、XY029、XY032)为研究对象,通过对5株菌株的分子鉴定、交配型基因分子检测、培养特性及子实体多糖含量测定,确定5株菌株皆为蛹虫草菌株,除XY008只含有MAT1-1-1交配型基因外,其他菌株都含有两种交配型基因MAT1-1-1和MAT1-2-1,5株菌株在菌丝生长速度、分生孢子数量、子实体形态、产量及子实体多糖含量均存在较大差异。综合培养特性及多糖含量分析的结果表明,菌株XY011子实体产量较高达29.60 g/瓶,多糖含量最高达100.79 mg/g,子实体长度最长达11.41 cm,而且子实体粗壮,发育周期短,确定为优势菌株,具有较好的开发价值。

北虫草CNZ虫草酸提取方法的筛选和优化

为获得北虫草(Cordyceps militaris)CNZ虫草酸最适的提取方法,比较了6种虫草酸提取方法,结果表明超声辅助醇提法虫草酸的得率最高,达113.31 mg/g,可作为初始提取方法。进一步通过单因素试验和正交试验优化提取次数、固液比、超声时间、乙醇浓度等条件,获得北虫草CNZ虫草酸提取条件为:乙醇浓度20%(体积分数),固液比1∶10(g∶mL),超声时间20 min,提取次数3次;在优化的提取条件下,北虫草CNZ虫草酸得率为161.06 mg/g,与初始提取条件相比提高了42.1%。