捕食性真菌Duddingtonia flagrans捕食马圆线虫幼虫动态观察

为探讨捕食性真菌Duddingtonia flagrans在捕食线虫的过程中产生的细微变化,如捕食器的形成过程,捕获虫体前后的变化等,利用线虫三期幼虫作为诱导物,在玻片培养基中刺激捕食性真菌产生捕食器,然后对其捕食线虫过程进行了适时动态的观察。结果显示,接种有较强活力的线虫三期幼虫2h后捕食器,即菌环开始形成,而后联合形成菌网;4h后虫体被捕食器捕获,24h后线虫即被完全致死;5d后,虫体发生皱缩直至完全消解。D.flagrans还可以自发产生捕食器(菌环),显示了其较强的捕食线虫的效率和能力。本研究为阐明捕食性真菌捕食机制提供了重要的参考依据。     

捕食线虫性真菌Duddingtonia flagrans冻干制剂临床模式

本试验选用耐高温高压的菌种袋作为玉米粒培养基的培养容器,对Duddingtonia flagrans厚壁孢子进行批量培养,然后经孢子洗脱液将孢子洗脱后,将其制备成冻干制剂,并在内蒙古地区的呼和浩特市和林格尔、包头市萨拉齐、呼伦贝尔西旗等地养殖场共165头绵羊中进行了临床应用研究。通过设立不同的试验组和对照组,使用常用驱虫药伊维菌素进行驱虫试验,同时配合口服捕食线虫性真菌Duddingtonia flagrans厚壁孢子,在不同时间进行动物直肠采粪,对粪便进行第三期幼虫培养,然后检测比较不同组别粪便中感染性幼虫的数量,评价捕食线虫性真菌临床应用模式效果。结果显示,将伊维菌素与D.flagrans冻干制剂联合应用于绵羊的寄生性线虫病防治,可使粪便中幼虫数量降低100%,效果优于单独用药组。结果表明,捕食线虫性真菌Duddingtonia flagrans冻干生物制剂与驱虫药物联合使用的临床应用模式,可以取得较好的家畜线虫病临床防控效果,值得进一步在生产实践中进行深入研究和推广。     

食线虫真菌防治寄生线虫的研究概况

文章综述了近年来食线虫真菌防治寄生线虫的研究概况，包括食线虫真菌的捕食机制和食线虫真菌防治线虫实践以及今后研究方向。从此文还可以看出，Arthrobotrys oliosopra和Duddingtonia flagrans两株真菌是最有发展潜力的食线虫真菌，且D．flagrans比A.oligosopra的捕食效果更好，研究者已把D．flagrans厚壁孢子制成片剂和团块，饲喂后，能显著减少羊的寄生性线虫的幼虫。     

嗜线虫节丛孢(Arthrobotrys flagrans)用于生物防治动物寄生线虫的研究进展

<正>捕食线虫性真菌Arthrobotrys flagrans首次由Duddington于1948年从腐烂的蔬菜里发现^[1]，并将该菌命名为Trichothecium flagrans。Cooke于1969年重新定义后为该种建立了一个新属(Duddingtonia属)^[2]，称为Duddingtonia flagrans，是该属唯一的种。Scholler等于1999年通过分析捕食线虫性真菌的18S r DNA和内部转录间隔区(ITS)的基因序列，认为捕食线虫性真菌的捕食器官类型在其属的分类上有重要意义。因此将捕食线虫性真菌划分为4个属，即Arthrobotrys、Monacrosporium、Dactylella及Gamsylella属^[3]。     

捕食性真菌Duddingtonia flagrans基因组DNA的提取及基因组survey分析

采用透析膜培养法结合DNA柱式提取法进行基因组DNA提取,同时利用第2代高通量测序技术,分析了捕食性真菌Duddingtonia flagrans基因组概况。结果显示:透析膜培养结合DNA柱式提取法所提取的基因组DNA质量及浓度均能达到测序标准;捕食性真菌Duddingtonia flagrans基因组约为37.6 Mb,GC含量为44.95%,基因组杂合度较低。结果表明:透析膜培养法结合柱式法提取其基因组DNA的方法简便可行,为丝状真菌DNA提取,提供了新的思路;基因组survey分析为捕食性真菌Duddingtonia flagrans全基因组精细图谱的绘制奠定了基础。     

少孢节丛孢菌捕食器表面聚合物的诱导与提取方法研究

为研究捕食线虫性真菌——少孢节丛孢菌(Arthrobotrys oligospora)捕食器表面聚合物的生化性质与作用,建立一种高效的表面聚合物诱导与提取方法尤为关键。试验首先用大豆蛋白胨液体培养基在氨基酸诱导作用下,产生大量含捕食器菌丝,然后分别用了5种不同化学性质溶液提取表面聚合物,经透析和浓缩后,测定了其中蛋白质的含量及其对线虫幼虫的毒性作用,同时还进行了提取液对真菌菌丝生长及其捕食作用的研究。通过综合分析得出,用5 mol/L LiCl溶液提取表面聚合物最为有效,其提取的表面聚合物溶液对线虫没有明显的杀线虫活性,但与捕食性真菌黏附捕获线虫过程密切相关;并通过比较源自含捕食器菌丝和普通营养菌丝的表面聚合物物质之间的差别,找到了参与黏附捕获线虫过程的物质,这为今后深入开展对捕食线虫性真菌表面聚合物的研究,进而阐明其作用机理提供了有价值的资料。     

食线虫真菌防治家畜胃肠道线虫研究进展

在自然界,食线虫真菌本身作为动物胃肠线虫的天敌而存在,它们在世界各地各种各样的环境中被发现。目前,利用食线虫真菌对动物寄生线虫的生物防治已取得初步成果,论文介绍了食线虫真菌在家畜胃肠道线虫生物防治方面的研究进展,重点阐述了捕食线虫性真菌对家畜寄生线虫的体内外杀虫试验和田间试验的现状。诸多研究表明,鞭式达丁屯氏菌（Duddingtonia flagrans ）由于能通过动物的消化道,因而对控制家畜胃肠道线虫病最具应用潜力。建议今后在规模化生产生防制剂关键技术上取得突破,使其尽快用于畜牧业生产。     

捕食性真菌Duddingtonia flagrans原生质体的快速制备与再生

为快速获得大量捕食性真菌Duddingtonia flagrans原生质体并使其再生,本试验研究了酶质量浓度、酶解温度、菌龄、酶解时间对D.flagrans原生质体产生的影响,以确定D.flagrans原生质体最佳快速制备条件。结果表明,在溶壁酶2mL/L、蜗牛酶8g/L、纤维素酶8g/L时,35℃恒温条件下,酶解菌龄为2d的菌丝7h,捕食性真菌D.flagrans产生原生质体数量最多且能够再生。这为下一步转化并标记和克隆捕食性相关基因奠定了重要基础。     

食线虫真菌Duddingtonia flagrans冻干制剂低温保存后体外杀虫效果研究

测定食线虫真菌Duddingtonia flagrans冻干制剂在4 ℃贮存2年期间对绵羊捻转血矛线虫感染性幼虫(L3)杀虫活性的变化。用食线虫真菌D.flagrans 当地分离株(SDH035)进行厚垣孢子培养,经真空冷冻干燥后制备成冻干制剂。三批冻干制剂分别在4 ℃保存1年和2年时取样,加入含有捻转血矛线虫虫卵的绵羊粪便为试验组,对照组的粪便不加真菌,然后进行粪便培养、L3分离和计数,计算L3的平均数和百分减少率。     

真菌D.flagrans对放牧幼驹预防圆形线虫感染的作用

马胃肠线虫危害严重 ,尤其是圆形线虫。过去控制大型线虫和小型线虫主要是使用驱虫药。由于广泛使用化学药物 ,抗药性在毛线虫普遍存在 ,主要是抗苯并咪唑。各种放牧的实践措施和机械化放牧卫生要求用于控制该病 ,但收效甚微。用食线虫真菌生物防治自由生活阶段的线虫 ,可能会改变和调节现存的防治方法。食线虫真菌Duddingtonia flagrans能减少反刍动物自由生活阶段的线虫已被一系列试验所证实。用Arthrobotrys oligospora捕食马的毛线虫和捕食反刍动物寄生线虫效果一致。D.flagrans用于生物防治动物寄生线虫有不容置疑的潜力。Larsen等 (…     

Interaction between copper oxide wire particles and Duddingtonia flagrans in lambs

An experiment was completed to determine if copper oxide wire particles (COWP) had any effect on the activity of the nematode-trapping fungus Duddingtonia flagrans in growing lambs. COWP has been used recently as a dewormer in small ruminants because of nematode resistance to anthelmintics. D. flagrans has been used to control free-living stages of parasitic nematodes in livestock. Katahdin and Dorper lambs, 4 months of age, were administered no or 4 g COWP (n=24/dose) in early October 2003. Haemonchus contortus was the predominant gastrointestinal parasite during the trial, which was acquired naturally from pasture. Half the lambs from each COWP group were supplemented with corn/soybean meal with or without D. flagrans for 35 days. Fecal egg counts (FEC) and packed cell volume (PCV) were determined weekly between days 0 (day of COWP administration) and 35. Feces from lambs in each treatment group were pooled and three replicates per group were cultured for 14 days at room temperature. Larvae (L3) were identified and counted per gram of feces cultured. Treatment with COWP was effective in decreasing FEC, which remained low compared with FEC from lambs not treated with COWP. This led to an increase in PCV in these lambs (COWP x day, P<0.001). Number of larvae was decreased in feces from lambs treated with COWP and D. flagrans between days 14 and 35 compared to the other groups of lambs (COWP x D. flagrans x day, P<0.003). Percentage of larvae identified as H. contortus decreased in feces collected from lambs treated with COWP and D. flagrans between days 14 and 28 compared with other treatments (COWP x D. flagrans x day, P<0.05). Other trichostrongyles were present and remained less than 7% in feces collected from control lambs. There was no adverse effect of COWP on the ability of D. flagrans to trap residual larvae after COWP treatment. With fewer eggs being excreted due to the effect of copper on H. contortus, and the additional larval reducing effect exerted by the nematode destroying fungus D. flagrans, the expected result would be a much lower larval challenge on pasture when these two tools are used together in a sustainable control strategy.     

Absence of obvious short-term impact of the nematode-trapping fungus Duddingtonia flagrans on survival and growth of the earthworm Aporrectodea longa

The nematode-trapping fungus Duddingtonia flagrans may be used in biological control of parasitic nematode larvae in faeces of domestic host animals after feeding the hosts with fungal chlamydospores. In this experiment a possible undesirable fungal impact on earthworms, of the species Aporrectodea longa, was investigated. As earthworms eat animal faeces, D. flagrans may come into contact with earthworms both in their alimentary tract and on their body surface. However during the experimental period of 20 days, when earthworms were living in soil and eating cattle faeces that were heavily infested with viable chlamydospores of D. flagrans there were no indications of internal or external mycosis among the earthworms.     

The efficacy of two isolates of the nematode-trapping fungus Duddingtonia flagrans against Dictyocaulus viviparus larvae in faeces.

A series of experiments was carried out to examine the effects of two different isolates of the nematode-trapping fungus Duddingtonia flagrans to reduce the number of free-living larvae of the bovine lungworm, Dictyocaulus viviparus. A laboratory dose-titration assay showed that isolates CI3 and Troll A of D. flagrans significantly reduced (P < 0.05 to P < 0.001) the number of infective D. viviparus larvae in cultures at dose-levels of 6250 and 12,500 chlamydospores/g of faeces. The larval reduction capacity was significantly higher for Troll A compared to CI3 when lungworm larvae were mixed in faecal cultures with eggs of Cooperia oncophora or Ostertagia ostertagi and treated with 6250 chlamydospores/g of faeces. Both fungal isolates showed a stronger effect on gastrointestinal larvae than on lungworm larvae. Two plot trials conducted in 1996 and 1997 involved deposition of artificial faecal pats containing free-living stages of D. viviparus and C. oncophora on grass plots. Herbage around the pats was collected at regular intervals and infective larvae recovered, counted and identified. These experiments showed that both D. flagrans isolates reduced the number of gastrointestinal as well as lungworm larvae in faecal pats. During both plot trials, the transmission of C. oncophora larvae, but not D. viviparus, from faecal pats to the surrounding herbage was clearly affected by climatic conditions. After collection of faecal pats from the grass plots one month after deposition, the wet and dry weight of pats as well as organic matter content were determined. No differences were found between the fungus-treated and non-treated control pats. This indicated that the rate of degradation of faeces was not affected by the addition of the fungus.     

Efficiency of feeding Duddingtonia flagrans chlamydospores to grazing ewes on reducing availability of parasitic nematode larvae on pasture

Gastrointestinal nematodes are of concern in sheep production because of production and economic losses. Control of these nematodes is primarily based on the use of anthelmintic treatment and pasture management. The almost exclusive use of anthelmintic treatment has resulted in development of anthelmintic resistance which has led to the need for other parasite control options to be explored. The blood sucking abomasal parasitic nematode Haemonchus contortus causes severe losses in small ruminant production in the warm, humid sub-tropic and tropics. This study evaluated the effectiveness of a nematode trapping fungus, Duddingtonia flagrans, in reducing availability of parasitic nematode larvae, specifically H. contortus, on pasture. Chlamydospores of D. flagrans were mixed with a supplement feed which was fed daily to a group of crossbred ewes for the duration of the summer grazing season. A control group was fed the same supplement feed without chlamydospores. A reduction in infective larval numbers was observed in fecal cultures of the fungus-fed group. Herbage samples from the pasture grazed by the fungus-fed group also showed a reduction in infective larvae. There were no significant (P > 0.05) differences in overall fecal egg count, packed cell volume or animal weight between fungus-fed and control groups. Tracer animals were placed on the study pastures at the end of the study to assess pasture infectivity. Although tracer animals were only two per group, those that grazed with the fungus-fed group had substantially reduced (96.8%) nematode burdens as compared to those from the control group pasture. Results demonstrated that the fungus did have activity against nematode larvae in the feces which reduced pasture infectivity and subsequently nematode burdens in tracer animals. This study showed that D. flagrans, fed daily to grazing ewes, was an effective biological control agent in reducing a predominantly H. contortus larval population on pasture.     

Biological control of helminths in grazing animals using nematophagous fungi

For the last 50 years the control of gastro-intestinal nematodes (GIN) in grazing animals has almost entirely been alleviated by the use of anthelmintics. Due to development of resistance against the drugs, especially in the GIN of sheep and goats it has become necessary to develop new, innovative strategies such as the use of nematode destroying fungi. Despite experiments to employ various species against plant and animal parasitic nematodes were already attempted in the 1930's, it was not until the 1990's when selection by simulating passage through the gastro-intestinal tract of cattle led to isolation of the fungus Duddingtonia flagrans that a major breakthrough was achieved. This fungus, producing sticky three-dimensional network and now isolated world-wide, is special due to its capacity to prolifically produce high numbers of thick-walled resting spores, chlamydospores. These spores survive passage through the gastro-intestinal tract of grazing livestock and are capable of growing and subsequently trap nematodes, including larval stages of parasitic nematodes. The great potential of this fungus as a biological control agent has been demonstrated through numerous trials with cattle, sheep, horses, and pigs. But these trials have also pointed towards some potential limitations in the activity spectrum of the fungus (Dictyocaulus, Nematodirus) beside the whole group of parasites spreading through infective stages protected inside resistant eggs (e.g. Ascaris, Trichuris). So far, in the few reported studies conducted, no negative environmental impact has been found, but it is important that further studies are conducted on this important issue. Although the potential use of D. flagrans chlamydospores has been verified through numerous trials it is necessary to develop practical delivery systems such as slow release devices, feed-blocks or similar to be able to implement this tool in future integrated control strategies. Such control strategies could include the use of biological control, grazing management, smart use of existing drugs, parasite resistant animal breeds, bioactive forages, and possibly vaccines.     

Environmental Impact on Soil Nematodes Following the Use of the Ivermectin Sustained-release Bolus or the Nematophagous Fungus Duddingtonia flagrans to Control Nematode Parasites of Cattle in Sweden

Management of nematode parasites of grazing livestock is essential, but there is concern about the potential environmental risk from agents used in their control. In this experiment, parasites in young cattle were controlled, using ivermectin boluses or the predacious fungus Duddingtonia flagrans , over 3 years. Treatment differences were sought among the soil nematodes recovered from soil samples collected from the paddocks on which treated animals grazed, and compared with similar samples taken from an untreated control paddock. Analysis of the soil nematode fauna on 21 occasions failed to demonstrate any impact of parasite management on soil nematodes. In 0-22 mm soil total nematode abundance averaged 524 000 m -2 , with 18 nominal taxa and H' diversity 2.41. There were both underlying paddock and year-to-year, climate-related, differences. The results of this trial not only confirm the lack of any adverse environmental impact of D. flagrans , a promising biological control agent, on soil nematodes, but also fail to show any impact arising from the use of ivermectin boluses.     

Effect of nematophagous fungus Duddingtonia flagrans and energy supplementation on the epidemiology of naturally infected kids

Gastrointestinal (GI) nematode infection is a major constraint for grazing livestock production. The increasing prevalence and severity of anthelmintic-resistant nematodes in many parts of the world has led to a search for non-chemical control options. Under experimental conditions, the nematophagous fungus Duddingtonia flagrans is emerging as an alternative to chemotherapy for the control of GI nematode infection in biological production systems. Also, recent information points to the role of energy nutrition to increase the immune response against GI nematode infection. In this study the effect of D. flagrans and energy supplementation on the epidemiology of GI nematode infections is explored on grazing kids. Four groups of 10, 4-month old goats were turned out on infected pasture in the early spring and allocated to four separate paddocks where they were rotationally grazed for 16 weeks. One of these groups (F) received 0.5 x 10(6) D. flagrans spores/kg BW/d. Another group (S) was supplemented with 100 g barley grain per day. A third group (F+S) received both nematophagous fungi and barley supplement treatments simultaneously while the fourth group (C) was used as a non-treated control. Both nematophagous fungi and barley supplement had a significant effect (P<0.01) on reducing pasture infectivity, faecal egg excretion and worm burdens at slaughter that was particularly evident for Trichostrongylus colubriformis. The combination of both treatments showed a synergistic effect on the control of gastrointestinal nematode infections. At slaughter, the average total post-mortem worm count of the F+S group was reduced by 65% compared with the non-treated control. The results herein show that D. flagrans can act as an efficient biological control agent against kid GI nematode infections on pasture, which could further improve carcass characteristics. While small amounts of energy supplement can also reduce kid infection, the effect of D. flagrans as a biological control agent appeared clearly enhanced both in magnitude and duration by energy supplementation. This has clear implications for grazing animals and provides an efficient method for the practical control of parasitic nematodes in biological production systems.     

Nematophagous fungi as a biological control agent for nematode parasites of small ruminants in Malaysia: a special emphasis on Duddingtonia flagrans

Approximately 2,800 fresh dung samples from animals, mainly ruminant livestock, were screened for the presence of nematophagous fungi in Malaysia. Arthrobotrys spp. was noted on numerous occasions, but only one isolate of Duddingtonia flagrans was made. For the purposes of producing sufficient quantities of this fungus for feeding trials in sheep, various, commonly available, cheap plant materials were tested as possible growth substrates. This showed that cereal grains (wheat, millet and rice) were the best media for fungal growth. Pen feeding trials were carried out using sheep, both naturally and experimentally infected with nematode parasites (predominantely Haemonchus contortus), to test the efficiency of D. flagrans when administered either in a grain supplement, or incorporated into a feed block. These showed that the fungus survived gut passage in sheep and that dose rates of approximately 1 x 10(6) D. flagrans spores / animal / day, reduced the percentage of infective larvae developing in faecal cultures by more than 90%. These results indicate that using D. flagrans as a biological control agent of nematode parasites, is a promising alternative to nematode parasite control of small ruminants in Malaysia, where anthelmintic resistance is now a major problem.     

低能氮离子束诱变选育捕食线虫性真菌——少孢节丛孢菌

为获取理想的生物防治捕食线虫性真菌菌株,达到利用自然天敌有效控制家畜寄生性线虫病的目的,作者利用低能氮离子注入诱变捕食线虫性真菌——少孢节丛胞菌(Arthrobotrys oligospora)的分生孢子,测定分生孢子萌发后菌丝的生长速度、产孢能力及菌株对线虫幼虫的捕食率,筛选正突变株。结果表明,低能氮离子束具有诱变少孢节丛孢菌的潜能,诱变后的1株少孢节丛孢菌的生长速度、产孢量和对线虫幼虫的捕食率呈现出了正突变效果。