不同培养条件对捕食性真菌Duddingtonia flagrans生长的影响

为了研究不同生长条件对捕食线虫性真菌Duddingtoniaflagrans生长的影响,本研究将带有D.flagrans菌丝的琼脂块分别接种于不同碳氮比及不同pH值(4.0~8.5)的CMA固体培养基中,在不同温度下(10℃~45℃)进行培养,每天定时观察菌株的发育情况,并测量菌丝生长长度。结果表明,D.flagrans生长最适pH值范围为6.5~7.0,其中以pH 7.0中生长最快;最适温度为35℃;最适碳氮比范围为5∶1~10∶1,其中在碳氮比为5∶1时,生长率最高。菌丝呈辐射状生长,纵横交错,无色,分隔,生长后期有缠绕圈和厚壁孢子的形成。上述结果为进一步研究D.flagrans的生长特性及捕食作用条件奠定了基础。     

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。     

Biological control of trichostrongyles in calves by the fungus Duddingtonia flagrans fed to animals under natural grazing conditions

The present study was conducted in the 1993 grazing season with yearling calves exposed to a pasture with a natural mixed trichostrongyle larval infection. It was shown that daily feeding with the microfungus Duddingtonia flagrans during the first 2 months of the season led to a lowered herbage infectivity and a reduced acquisition of Ostertagia sp. and Cooperia sp. later in the season. In addition, the procedure delayed the onset of clinical disease. This was due to the nematode-destroying effects of the fungi in the dung excreted by the fungus-treated calves, as evidenced by results from a parallel in vitro assay on faecal larval cultures. The paper discusses future research needs before practical biological control can be implemented.     

Predacious activity of the nematode-trapping fungus Duddingtonia flagrans against cyathostome larvae in faeces after passage through the gastrointestinal tract of horses

This study was undertaken to examine the potential of the nematode-trapping microfungus Duddingtonia flagrans to survive passage through the gastrointestinal tract of horses and subsequently to destroy free-living stages of cyathostomes in faecal cultures. Three different oral dose levels were tested, two horses being used for each level. Faeces were collected twice daily and the number of parasite eggs per gram of faeces were determined. The numbers of infective third stage larvae which developed in faecal cultures were determined after the cultures had been incubated for 2 weeks at 24°C. Results showed a positive relationship between dose level and reduction in the number of infective larvae. Fungi were recovered in faeces at times which corresponded to high larval reduction.     

嗜线虫真菌中Duddingtonia flagrans培养性状的初步研究

研究嗜线虫真菌中Duddingtonia flagrans在不同生长环境中的培养特性.选择普通琼脂培养基(WA)、玉米琼脂培养基(CMA)、马铃薯琼脂培养基(PA)和面粉琼脂培养基(FA)4种不同培养基培养,结果表明,在WA中真菌生长速度最快,在PA中产孢量最多.从应用的角度考虑,PA为最佳培养基;以10～35 C每隔5℃设置7种不同温度组培养,结果表明,10℃和35℃时不生长,25℃和30℃时菌丝生长率较高,产孢量较多,其中以30℃培养效果最好.     

Capability of the nematode-trapping fungus Duddingtonia flagrans to reduce infective larvae of gastrointestinal nematodes in goat feces in the southeastern United States: dose titration and dose time interval studies

Infection with gastrointestinal nematodes, particularly Haemonchus contortus, is a major constraint to goat production in the southeastern United States. Non-anthelmintic control alternatives are needed due to increasing resistance of these nematodes to available anthelmintics. Two studies were completed in Central Georgia in August 1999, and April–May 2000, using Spanish does naturally infected with Haemonchus contortus, Trichostongylus colubriformis, and Cooperia spp. to evaluate effectiveness of nematode-trapping fungi as a biological control agent. In the first experiment, five levels of Duddingtonia flagrans spores were mixed with a complete diet and fed once daily to the does (three per treatment) in metabolism crates. The treatment concentrations were (1) 5×10⁵, (2) 2.5×10⁵, (3) 10⁵, and (4) 5×10⁴ spores per kilogram body weight (BW), and (5) no spores. Fungal spores were fed for the first 7 days of the 14-day trial, and fecal samples were collected daily from individual animals for analysis of fecal egg count and establishment of fecal cultures. Efficacy of the fungus at reducing development of infective larvae (L3) in the fecal cultures was evaluated. The mean reduction in L3 from day 2 of the treatment period until the day after treatment stopped (days 2–8) was 93.6, 80.2, 84.1, and 60.8% for animals given the highest to lowest spore doses, respectively. Within 3–6 days after termination of fungal spore feedings, reduction in L3 development was no longer apparent in any of the treated animals. In a second experiment, effectiveness of 2.5×10⁵ spores of D. flagrans per kilogram BW fed to does every day, every second day, and every third day was evaluated. Reduction in L3 development by daily feeding was less in the second experiment than in the first experiment. Daily fungal spore feeding provided more consistent larval reduction than intermittant feeding (every second or third day). When fed daily under controlled conditions, D. flagrans was effective in significantly reducing development of L3 and appears to be an effective tool for biocontrol of parasitic nematodes in goats.     

Soil Nematode Populations Beneath Faecal Pats from Grazing Cattle Treated with the Ivermectin Sustained-release Bolus or Fed the Nematophagous Fungus Duddingtonia flagrans to Control Nematode Parasites

The size and composition of the nematode assemblage in soil under faecal pats derived from young cattle treated or untreated with either ivermectin sustained-release boluses, or the nematophagous fungus Duddingtonia flagrans, were studied in each of three years. Soil samples taken 4, 6, 8 and 10 weeks after four deposition dates in 1998 showed significant temporal effects in many taxa and treatment effects in a few genera. In 2000, soil samples taken 10 weeks after deposition in July, August and September showed treatment effects in the plant-associated Tylenchus and Cephalenchus, and the bacterial-feeding Cephalobus 1 and Cephalobus 2 taxa. However, overall it was found that the nematode assemblages were similar below all three types of pat, and the assemblages varied with the season of deposition. D. flagrans, the novel biological control agent being tested against the free-living stages of nematode parasites of cattle, had no detectable impact on the size or the structure of the soil nematode communities under the faecal pats.     

用嗜线虫真菌防治家畜胃肠道线虫病的研究进展

简要论述了嗜线虫真菌种类及其捕捉器官和国内外生物防治家畜胃肠道线虫病的研究进展。国内外防治该病主要是用各种驱虫药物定期驱虫，由此造成寄生虫抗药性、动物组织残留药物、环境污染、线虫反复感染等诸多负作用。用线虫天敌嗜线虫真菌生物防治家畜胃肠道线虫病成为该领域研究的重点课题之一。实验室和野外的大量研究结果证明，嗜线虫真菌在通过动物胃肠道时能不被消化，从而保持活性，在粪便中能有效捕食营自由生活阶段的线虫幼虫，使放牧草场中感染性线虫幼虫明显减少，呈现出良好的生物防治效果。文章重点介绍了嗜线虫真菌Duddingtonia flagrans，对用嗜线虫真菌生物防治家畜胃肠道线虫病的前景充满信心。     

Effect of biological control through the daily application of spores of Duddingtonia flagrans in lambs kept under an evasive grazing system in the Netherlands

In 2004, an experiment was carried out to evaluate the effect of biological control through feeding spores of Duddingtonia flagrans on parasitic gastroenteritis in lambs, kept under an evasive grazing system. In total 66 lambs were used. Forty naturally infected 3-month old ram lambs were weaned in mid June, and divided into four groups of 10 lambs. On 21 June, G1–G4 were moved to four separate virtually clean plots, they were moved after 4 and 8 weeks to similar plots, and housed after 12 weeks to be necropsied 16 days later. The other 26 lambs had been raised helminth-free, and were used as pairs of tracer lambs. All but one of these pairs, were grazed during the last 2 weeks on each plot. The remaining pair (TA) was grazed during the last 2 weeks on pasture (30 August to 13 September) on the plot that had been grazed by G3 between 19 July and 16 August, to study inhibited development in Haemonchus contortus. All lambs were fed 200 g of concentrates daily throughout the whole period, and those of G1 and G2 were also fed 500,000 spores of D. flagrans/kg bodyweight daily.

The faecal cultures demonstrated a high reduction in yield as a result of fungal application. However, no differences between groups were seen in weight gain, faecal egg counts, pasture larval counts, worm counts and tracer worm counts. H. contortus was the dominant species, and it is obvious that the moves at 4-week intervals prevented the development of severe haemonchosis. This is in particular demonstrated by the much higher worm counts in the two TA tracer lambs grazed. Nevertheless, increases to high faecal egg counts 3 weeks after the first and second moves, indicated acquisition of infection before these moves and at least subclinical haemonchosis. This was supported with the worm counts of lambs and tracer lambs. A higher proportion of inhibited early L4 than in other tracers and than in the permanent lambs were found in the pair of TA tracer lambs. This indicates that moves to new pastures in late summer and autumn delays the onset of inhibition.     

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。