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.

北冬虫夏草(Cordyceps militaris)的研究应用概况(综述)

主要综述了近二十年来国内外对北冬虫夏草的研究应用概况，详细地介绍了北冬虫夏草及其代谢物质“虫草素”的来源，及在功能、药化、药理、临床应用方面的研究进展，北冬虫夏草的生物学特性和工厂化生产研究情况，以及影响北冬虫夏草人工高产栽培与工厂化生产的关键问题，同时也展示了北冬虫夏草的开发利用前景。

蛹虫草Cordyceps militaris JN168产虫草素液态发酵条件的优化

利用单因素筛选和响应面法对蛹虫草Cordyceps militaris JN168产虫草素的液态发酵培养基进行优化,以确定蛹虫草产虫草素的最佳发酵培养基配方。结果表明,蛹虫草产虫草素的最佳碳源为葡萄糖,最适质量浓度为40 g/L;最佳氮源为牛肉膏,最适质量浓度15 g/L;加入的无机盐及其添加量分别为MgSO40.76 g/L,K2HPO40.63 g/L,CaCl20.66 g/L,Na2HPO40.67 g/L。优化后发酵液中虫草素质量浓度达到633.47 mg/L,是优化前的6倍。

蛹虫草(Cordyceps militaris)对药物引起免疫抑制的拮抗作用

实验研究蛹虫草对环磷酰胺造成的小鼠免疫抑制的作用。实验选用天然冬虫夏草为阳性对照。给药方式为每天灌胃 0 .2 g生药水煎剂 ,7d后测定其生物学活性。实验结果表明 :小鼠腹腔巨噬细胞的吞噬率由对照组的 13.97%提高到 2 2 .16 %;骨髓细胞的微核数目由对照组的 16 .6×10 -3 降低到 12 .3× 10 -3 ;外周血白细胞由对照组的 70 %提高到 83%。结果证实 :蛹虫草具有明显的拮抗由环磷酰胺造成的免疫抑制的作用 ,与冬虫夏草比较有相似的生物学活性 ,某些作用明显高于冬虫夏草 ,为蛹虫草的医学应用提供了一定的理论依据。

蛹虫草Cordyceps militaris JN168菌株的诱变育种及液态发酵产虫草素

利用离子束、亚硝基胍及离子束-亚硝基胍诱变法处理蛹虫草Cordyceps militaris JN168,初步获得了几株较高产虫草素的菌株。通过进一步筛选得到复合诱变菌2,并对该菌的液态发酵培养基组分进行了优化。实验结果表明:蛹虫草液态发酵产虫草素的最佳培养基组分为(g/L),葡萄糖40,酵母浸粉25,Mg SO4·7H2O 0.6,K2HPO4·3H2O 0.6,KH2PO40.6。优化后虫草素产量提高了5倍,最高达1 045.65 mg/L。

蛹虫草(Cordyceps militaris)菌丝体液体培养基的优化和发酵条件的研究

采用二次回归通用旋转组合设计方法，对蛹虫草菌丝体液体发酵培养基的最佳组合进行了研究，建立了培养基的反应模型，并用实验数据验证了数学模型，对培养基最佳优化组合的各种因子、影响产量的反应规律及交互作用进行了探讨，并在此基础上进行了蛹虫草菌丝体液体发酵条件的研究。结果表明，蛹虫草菌丝体液体发酵的最适条件为：接种量６％（ｖ／ｖ），发酵初始ｐＨ７．０，培养基装量１００ｍｌ／５００ｍｌ三角瓶，发酵温度２４℃～２８℃，发酵周期９６ｈ。

Cu^(2+)、Zn^(2+)胁迫对蛹虫草(Cordyceps militaris)抗氧化酶活性及脂氧化水平的影响

研究了Cu2+、Zn2+对蛹虫草菌丝体抗氧化酶活性及脂氧化水平的影响。结果表明:高浓度的Cu2+抑制菌丝体的生长和细胞内抗氧化酶的活性;高浓度的Zn2+对菌丝体的生长也有抑制作用但抗氧化酶的活性变化不大。脂氧化水平随细胞内过氧化状态的变化而变化。蛹虫草菌丝体对Cu2+、Zn2+有较高的耐受度。

北冬虫夏草(Cordyceps militaris)单孢菌株配对后对子实体形成的影响与无性型产孢结构关系

由野生子实体分离的低代菌株“ Ｃ．ｍ．Ｈ９５－１”和“ Ｃ．ｍ．Ｈ９７－Ｇ６”的子囊孢子单孢，配对后产生了较大变异，３１％和５０％的配对菌株具有与亲本菌株相同的遗传特性，而且子实体形成能力及生物产量明显增加。配对菌株“ Ｃ．ｍ．Ｈ９７－２６９３”和“ Ｃ．ｍ．Ｈ９７－２６２７”的子囊孢子再配对后，几乎所有的再配对菌株不能回复到亲本菌株的生长型，其子实体的形成能力及生物产量也低于亲本菌株。实验的结果还表明，在ＰＤＡ组合培养基１上，单孢及配对菌株的无性型产孢结构，对子实体形成有着一定的相关性。

北冬虫夏草(Cordyceps militaris)室内栽培技术研究

2003年从北京北冬虫夏草(Cordyceps militaris)子实体的子座部分经单孢分离出Ⅱ号菌株.2004年从北京、上海北冬虫夏草新鲜子实体中采用组织分离法分离出Ⅰ号、Ⅲ号和Ⅳ号菌株,同时收集到开封试管菌株Ⅴ号.Ⅱ号菌株在不同培养基(5,6,7号)进行瓶栽试验,并与4个菌株(Ⅰ号、Ⅲ号、Ⅳ号、Ⅴ号)在同一培养基进行瓶栽比较试验.经过50d的观察表明,Ⅱ号菌株在5号和6号培养基上所产生子实体及每瓶平均数量,平均长度,平均鲜重和干重都比在7号培养基上良好;Ⅰ号、Ⅲ号、Ⅳ号菌株在5号培养基上生的子实体无论每瓶平均数量、平均长度、平均鲜重和干重都明显比Ⅴ号菌株子实体情况优良.

高产虫草素Cordyceps militaris菌的诱变育种

以一株生长性状优良的Cordyceps militaris FFCC5111为出发菌株,通过复合诱变,筛选高产Cordycepin双重营养缺陷型(Xan-+Gua-)菌株。采用200 W、22kHz超声强度联合1%DES复合诱变,获得双重营养缺陷型突变株7株,经抗菌试验、摇瓶培养/发酵复筛至第16天,突变株C.militaris FFCC5111-c产生的Cordycepin总量为4.142g/L,比亲株提高44.5%。试验证明,筛选Xanthine和Guanine双重营养缺陷型突变株是获得Cordycepin高产菌种的有效方法,Cordycepin的生物合成与Guanine代谢相关。

蛹虫草(Cordycepsmilitaris)无性型的多型现象

在萨氏琼脂和PDA上，蛹虫草Cordycepsmilitaris（Vuill）Fr.无性型，蛹草拟青霉Paecilomycesmilitaris(Kob）Br.＆Sm．的一些单孢子株可自发产生突变，基于多型现象及其它形态特征可分三种类型：（1）具野生型菌株特征；产孢结构拟青霉型（Paecilomyces－type），稳定，菌落通常不自发产生角变，大多数单孢子株属此类群。（2）属此类群的单孢子株，在PDA上可形成亮褐色至橙黄色的角变，它们典型的产孢结构为瓶梗轮生，分生孢子常聚集成头状的轮枝孢型（Verticillium－type）。在查氏培养基上则相反，拟青霉型的产孢结构占优势。单孢子株Cm-71是这一类群的代表。（3）单孢子株Cm－42是在所观察单孢子株中形态特征独特的菌株。在PDA上菌丝生长纤细，蛛网状，产孢结构轮枝孢型。蛹草拟青霉对氯酸钾（KClO3）不敏感，在浓度为6％（W／V）的KMM培养基上，23～24℃14天菌落的直径仅1～2cm，无气生菌丝生长。各单孢子株形成的不利用硝酸盐突变株（nit突变株）的途径和形态特征，也可分成类似于粉被虫草CordycepspruinosaPeteh不利用硝酸盐突变株（nit突变株）的三种类型。

蛹虫草(Cordyceps militaris)的交配型研究

利用可以在人工条件完成整个生活史的蛹虫草菌种,分离、鉴定了2个蛹虫草菌株子囊孢子的单孢分离物,对子囊单孢的配对培养和子实体诱导结果显示,蛹虫草具有典型的二极性异宗配合习性。

北冬虫夏草(Cordyceps militaris)主要菌株的超微结构

对北冬虫夏草主要菌株的超微结构研究表明，子实体形成能力不同的菌株，其体细胞结构和无性繁殖结构没有很大的差别；分生孢子的形成方式为芽生式分生孢子，菌丝体的每个细胞里通常仅含１个单倍体的细胞核。而在有子实体形成能力的野生菌株的有性繁殖结构内．主要是子囊和子囊孢子。它们性过程仅发生在子实体外层的双核细胞里，其子囊孢子的形成方式为细胞游离形成方式。文中选用了４６张显微镜照片并组合为１０个图版，首次系统地介绍了我国北冬虫夏草的超微结构特征。

蚕虫草(Cordyceps militaris)的干燥条件及其对活性成分的影响

研究了蚕虫草的烘干和微波干燥条件及其对主要活性成分的影响。结果表明,采用烘干干燥,随着干燥温度的升高,干燥速率增加,且在各试验温度下均达到极显著差异,75%以上的水分是在前2 h失去的;采用微波干燥,随着微波干燥功率的增加,干燥速率也增加,且在各试验微波干燥功率下均达到极显著差异,75%以上的水分是在前6 min失去的。对于活性成分含量来说,微波干燥方法中,900 W是最好的,极显著高于其他功率;采用烘干方法时,虫草素是在60℃时含量最高,而腺苷则是在70℃时含量最高。在生产中可以根据需要选择不同的干燥条件。

蛹虫草(Cordyceps militaris)LaeA蛋白的生物信息学分析

采用Prot Param、Prot Scale、Signal P 4.1、SOPMA、SWISS-MODEL等在线生物学软件对LaeA蛋白质的理化性质、信号肽、跨膜结构域、蛋白质二级和三级结构进行预测分析。结果表明,蛹虫草LaeA蛋白质是由20种295个氨基酸组成,不含信号肽、无跨膜结构域、稳定的亲水蛋白;LaeA蛋白二级结构主要由α-螺旋、无规则卷曲、伸展链和β-转角组成,其中α-螺旋和无规则卷曲是LaeA蛋白的主要二级结构。通过生物信息学预测分析首次获得了蛹虫草LaeA蛋白的理化性质及各级结构信息,为将来深入研究蛹虫草LaeA蛋白的生物学功能奠定了理论基础。

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 Effect of the Hot Water Extracts of the <i>Paecilomyces hepiali </i>and <i>Cordyceps militaris</i>Mycelia on the Growth of Gastrointestinal Bacteria

The gastrointestinal health is very important aspect concerning human health. It refers to nutrient and vitamin absorption, food digestion and various infectious diseases. The Paecilomyces hepiali and Cordyceps militaris are highly enriched with cordycepin and ergosterol which are considered as anti bacterial substances. Present study finds out comparative effect of hot water extract of particular fungal material on growth of six species of gastrointestinal bacteria that belong to both aerobic and anaerobic and, consist with harmful and commensal categories. The appropriate concentration level of hot water extract of both strains was identified. The individual specific bacterial growing media were prepared and calculated;amounts of bacteria cultures were inoculated by using micro pipettes. The optical density and number of bacterial colonies were measured after 24 hours. The pure mycelial extract of P. hepiali with 2 × 10-3 g/ml of concentration has significant effect on depleting the growth of E. coli, E. faecalis, S. aureus, L. gasseri and B. ovatus bacteria. B. longum has no significant effect by particular extract. Same type of extract of C. militaris has significantly reduced the growth of every bacteria used in this study. Hot water extract of C. militaris cultivated on soy bean has significant growth retardation toward E. coli, E. faecalis, S. aureus and L. gasseri. It has stimulated the growth of B. ovatus and B. longum which are considered as beneficial bacteria for human gut. This study shows that extracts of both mycelia include antimicrobial substances like cordycepin and ergosterol which can be used as food supplements to enhance human gut health.

Development of Functional Foods with Cordyceps militaris as a Raw Material

Cordyceps militaris,as a rare edible fungus,has high medicinal and nutritional value,and has attracted much attention.This paper summarized a series of functional foods developed with C.militaris as a raw material from the perspective of Cordyceps drinks,dry C.militaris products,Cordyceps seasonings,and other Cordyceps foods in four broad categories and ten sub-categories,providing reference for continuous development and utilization.

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.