枯草芽孢杆菌替代抗生素对平养和笼养肉鸡的影响研究

 将240只1日龄罗斯308（ROSS 308）肉仔鸡，按照饲养方式（地面平养和笼养）以及添加枯草芽孢杆菌替代抗生素与否（替代和不替代）分为4个处理组（1组：抗生素地面平养组；2组：抗生素笼养组；3组：枯草芽孢杆菌地面平养组；4组：枯草芽孢杆菌笼养组），进行42 d的饲养试验。结果显示：试验全期，肉鸡末体重和饲料/增重比指标，4组显著优于（P<0.05）1组，2组和3组之间无差异（P>0.05）。21日龄和42日龄肉鸡血清生化指标分析显示：4组肉鸡血清白蛋白浓度显著大于1组；且血清尿酸浓度4组小于1组；而2组和3组之间所有血清测定指标均无差异。以上结果证明，无论是地面平养还是笼养方式，枯草芽孢杆菌替代抗生素对改善肉鸡生长性能均有良好效果，其作用机理主要是提高饲料蛋白的营养利用率.     

Screening of field performance of entomopathogenic fungi and nematodes against cabbage root flies (Delia radicum l. and d. floralis (fall.); Diptera,Anthomyiidae)

Finnish isolates of Beauveria bassiana (8 isolates), Metarhizium anisopliae (7), Tolypocladium sp. (2), Paecilomyces farinosus (2), P. fumosoroseus (1), Steinernema feltiae (3) and Bacillus thuringiensis ('Muscabac') were tested for efficacy against mixed populations of Delia radicum and D. floralis under field conditions in 1986–90. All pathogens were applied preventatively, the fungi as aqueous conidial or mycelial suspensions, dry conidia or dry mycelial powder. In only two of the nine experiments did B. bassiana or M. anisopliae give some control. In 1986, B. bassiana SF85–2 and Tolypocladium sp. SF85–4 (both at rate 1.2 x 10⁹ conidia plant‐ 1), and ‘Muscabac’ (25 g 1^?1, 1 dl plant^?1) reduced the number of pupae by 80%, 60% and 50%, respectively, as compared with untreated and chemical (isophenphos) controls. In 1990, M. anisopliae SF86–39 at rate 1.6 x 10⁹conidia plant^?1 and 1.5 x 10⁸ CFU plant^?1 and S. feltiae SFS‐22 (35000 plant^?1), increased the yield of cauliflower 2.2, 1.8, and 2.3‐fold, respectively, as compared with the untreated control, but these yields were only 19%, 15% and 19% of those of the chemical (diazinon) control. Paecilomyces isolates were ineffective in the 1986 experiment in which they were included. Our results suggest that it is difficult to find efficient control agents among the fungal and nematode species tested for use as biopesticides against cabbage root flies, but that the potential of M. anisopliae against these pests deserves further study.     

BP神经网络结合正交试验法优化Bacillus velezensis Z-27菌株培养基组分

将BP人工神经网络(Artificiai neural network)技术与传统的正交试验方法相结合,提出一种新的试验分析和处理方法,利用神经网络特有的自学能力,对主要影响因素进行仿真优化,获得Bacillus velezensis Z-27菌株培养基组分,即玉米粉2.0％、豆饼粉1.5％、MnSO4· H2O 0.07％,起始pH为7.0、接种量为2.0％,应用优化得到的培养基组分进行验证试验,取得了较好的效果.     

甘蔗黑穗病菌拮抗菌HAS的筛选和鉴定

采用传统形态学鉴定及16S rRNA基因序列分析,筛选鉴定对甘蔗黑穗病菌具有拮抗作用的细菌的归属,并采用细菌的通用引物P0、P6扩增16S rRNA基因片段.结果表明:得到1 533 bp的DNA片段,其序列与枯草芽孢杆菌序列的同源性高达99％.其形态学特征:菌体为短杆状,能运动；革兰氏阳性反应；3％KOH溶解性实验呈阴性反应；有鞭毛,鞭毛周生；芽孢中生,椭圆形,孢囊稍膨大,初步将该菌株归属为枯草芽孢杆菌.与病原真菌对峙培养的结果表明,该菌对引起甘蔗和其它作物病害的多个植物病原真菌均具有很好的抑制作用.据此,可将分离的菌株HAS确定为对甘蔗黑穗病菌有较好防治效果的枯草芽孢杆菌.     

解淀粉芽胞杆菌HRH317对串珠镰孢菌菌丝形态和超微结构的影响

为明确新型生防菌解淀粉芽胞杆菌Bacillus amyloliquefaciens HRH317菌株对病原菌串珠镰孢菌Fusarium moniliforme的抑制作用,采用牛津杯法对HRH317菌株抑菌活性进行测定,并采用扫描电子显微镜、透射电子显微镜和荧光显微镜对经HRH317菌株发酵上清液处理后的串珠镰孢菌菌丝形态及超微结构进行观察。结果显示,HRH317菌株发酵上清液对串珠镰孢菌有很好的抑菌活性,抑菌圈平均直径可达33.31 mm。扫描电镜结果显示,HRH317菌株发酵上清液处理24 h时,串珠镰孢菌菌丝体出现断裂现象;处理72 h时,串珠镰孢菌菌丝体断裂较严重,多处裂解;处理96 h时,串珠镰孢菌菌丝体彻底瓦解,且无完整菌丝体。透射电镜结果显示,HRH317菌株发酵上清液处理72 h时,串珠镰孢菌菌丝体细胞形态扭曲变形,细胞内结构紊乱,遭破坏。荧光显微镜结果显示,经PI染料染色处理12 h时,串珠镰孢菌细胞有少数细胞被染成红色,细胞膜通透性受一定程度破坏;处理16 h时,串珠镰孢菌细胞大面积被染红;处理20 h时,串珠镰孢菌细胞被染色面积增大;处理24 h时,串珠镰孢菌细胞膜受破坏程度增加,细胞内大面积被染色。表明解淀粉芽胞杆菌HRH317菌株对串珠镰孢菌菌丝形态和超微结构有破坏作用,能抑制病原菌串珠镰孢菌菌丝体生长。     

枯草芽孢杆菌CN620菌株制剂对板栗烂果病的防治及病原的抑制作用初报

从板栗树体上分离得到枯草芽孢杆菌 [Bacillussubtilis(Ehrenberg)Cohn]CN62 0菌株。室内测定其菌株制剂对Glomerellasp .、Alternariaalternata、Trichotheciumroseum、Pestalotiasp .和Fusariumsp .5种板栗烂果病主要病原的抑制作用均为 1 0 0 % ,而CN62 0菌株 50倍培养液 ,对上述 5种病菌的抑制效果为 86.5%～ 96.6%。其抑菌机理主要是使病菌孢子和菌丝畸变 ,细胞壁溶解 ,原生质泄漏。 1 997年和 1 998年连续两年两点对板栗烂果病进行小区防治试验的防治效果分别为 87.7%和 87.6% ,明显高于化学农药处理的 30 .4 %和 31 .7%的效果     

Effect of Fermented Chinese Medicine Preparation by Bacillus subtilis on Growth Performance,Biochemical Indices,Antioxidant Parameters and Ability of Anti-infection of Common Carp

The objective of this study was to evaluate of the growth performance, serum biochemical indices, liver antioxidant parameters and ability of anti-infection with Aeromonas hydrophila by adding the fermented Chinese medicine by Bacillus Subtilis to the carp fish feed, and to find out the appropriate supplemental level of fermented Chinese medicine in diets. Carp fingerling with an initial body weight of (35±0.5) g were set up to 6 experimental diets supplemented with 0 (control group), 0.1‰, 0.5‰, 1‰, 2‰ and 3‰ fermented Chinese medicine for 56 days, respectively. Each group had 30 fish and set up 3 repetitions. After the feeding trail,the fish were challenged with Aeromonas hydrophila to calculate the relative percentage survival (RPS).The results showed as follows:Supplementing fermented traditional Chinese medicine by Bacillus subtilis, the weight gain rate (WGR), specific growth rate (SGR), condition factor (CF), the activities of lysozyme (LSZ), complement 3 (C3),total protein (TP),total anti-oxidation capacity (T-AOC),total superoxide dismutase (T-SOD) and catalase (CAT) in serum of common carp were enhanced at different degrees, while the feed coefficient (FCR)and malondialdehyde (MDA) in serum were declined at different degrees.When the fermented Chinese medicine supplemental levels were 2‰ and 3‰,the effects were significant (P<0.05)(except for C3, ALP and Cr).The experimental results of the virulence of common carp infected with pathogen Aeromonas hydrophila showed that,the fermented herbal could obviously improve the ability of anti-infection, the immune protective rate of the group with 2‰, 3‰ doses of the fermented Chinese medicine could reach 77.23% and 74.63%, respectively. From what had been discussed above, the group with 2‰ and 3‰ doses of the fermented Chinese medicine had significant effect on the growth performance, serum biochemical indices, liver antioxidant parameters and Aeromonas hydrophila anti-infection ability. Considering the cost of adding fermented Chinese medicine, the recommendations for the best additive quantity was 2‰.     

提高苏云金杆菌防治效果技术的研究

系统介绍了Bt的生理特征、杀虫机理以及在应用中存在的问题,提出了提高Bt防治效果的技术措施。     

腐殖酸水溶肥配施枯草芽孢杆菌对基质青菜生长的影响

为探究腐殖酸水溶肥、枯草芽孢杆菌对基质青菜生长的影响 本研究以抗热605青菜为试验材料,共设计6组施肥试验,分别为CK1(有机肥2.5g/盆)、CK2(有机肥2.5g/盆+枯草芽孢杆菌0.5g/盆)、T1(有机肥2.5g/盆+枯草芽孢杆菌0.5g/盆+腐殖酸水溶肥100倍液)、T2(有机肥2.5g/盆+枯草芽孢杆菌0.5g/盆+腐殖酸水溶肥200倍液)、T3(有机肥2.5g/盆+枯草芽孢杆菌0.5g/盆+腐殖酸水溶肥400倍液)、T4(有机肥2.5g/盆+枯草芽孢杆菌0.5g/盆+腐殖酸水溶肥800倍液),研究结过表明:腐殖酸水溶肥配施枯草芽孢杆菌可在一定程度上增加基质青菜的株高、叶宽、叶长、冠幅及地上鲜重,其中腐殖酸水溶肥起主导作用,枯草芽孢杆菌作用较为微弱,综合各生长指标考量以T2处理效果最佳。     

鸡蛋壳蜡样芽孢杆菌的分离鉴定及耐药性分析

【目的】 分析不同环境下鸡蛋外壳携带蜡样芽孢杆菌的情况和分离菌株的基本特征。【方法】 从养殖场和超市各采集9种新鲜鸡蛋,对鸡蛋表面携带的蜡样芽孢杆菌进行分离鉴定、基因分型、毒力基因筛查和耐药性分析。采用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）技术对分离菌株进行初步鉴定,全基因组测序后进行菌种鉴定并确定基因分型、筛选毒力基因和耐药基因,同时采用微量肉汤稀释法测定分离株对头孢曲松、红霉素、万古霉素等12种抗菌药物的耐药性。【结果】 经MALDI-TOF MS初步鉴定和基因组菌种鉴定,确定从养殖场3种未经清洁加工的鲜蛋中分离出的6个菌株为蜡样芽孢杆菌,其序列分型（ST）较为多样,其中有2株新的ST型。在毒力基因筛查方面,6株蜡样芽孢杆菌均检出了非溶血型肠毒素nheB基因、腹泻型毒力因子hlyⅡ基因和侵袭免疫系统的inhA基因;nheA和nheC基因携带率为83.3%,编码溶血型肠毒素HBL的hblA、hblC和hblD基因携带率均为66.7%;cytK和hlyⅢ基因携带率分别为33.3%和50.0%;肠毒素基因BceT、entFM和致吐毒素基因ces均未检出。耐药性方面,6株菌均对庆大霉素、四环素、氯霉素、利福平、利奈唑胺和环丙沙星敏感,对氨苄西林、头孢曲松和泰妙菌素耐药;对红霉素、克林霉素的中介率分别为83.3%和100%;33.3%的菌株对万古霉素耐药。6株菌均携带有磷霉素耐药基因FosB和万古霉素耐药基因簇vanA的部分基因vanRSYZ,其中5株菌携带大环内酯类药物耐药基因mphL,部分菌株携带β-内酰胺类药物耐药基因或核苷类药物耐药基因。【结论】 本试验从养殖场未经消毒处理的鸡蛋外壳上成功分离到了蜡样芽孢杆菌,并发现其携带有多种毒力基因,存在多重耐药性,有潜在致病性和公共安全风险。