Pathogen Analysis of Bovine Respiratory Disease Complex

To investigate the pathogen of bovine respiratory disease complex (BRDC) of a dairy farm in Guangxi, a strain of Mycoplasma and a strain of gram-negative pathogenic bacterium were isolated and identified by the means of field surveys, clinic observation, pathological examination, isolation studies and so on.Treatments were taken according to drug sensitivity test results.The Mycoplasma strain, growing on PPLO medium, formed typical "fried egg" colonies.A 448 bp of oppF fragment was amplified by PCR from the strain and had 98.4% nucleotide identity with Mycoplasma bovis reference isolate PG5 of USA.The biochemical features of the gram-negative bacterial isolate were same with Serratia marcescens.The PCR amplified 16S rDNA of the gram-negative pathogenic bacterium strain was 1 400 bp.It shared 99.0% nucleotide identity with other Serratia marcescens reference strains obtained from GenBank.Animal experiment showed that the gram-negative pathogenic bacterium isolate could cause the mice to die.The drug sensitivity tests showed all isolates were sensitive to spectinomycin, azithromycin, amikacin, gentamicin and neomycin.It was effective to treat with dexamethasone and spectinomycin.Pathogen analysis and drug treatment showed that the BRDC was caused by Mycoplasma bovis and Serratia marcescens.     

Pathogen Analysis of Goat's Respiratory Diseases

To investigate the pathogens of goat's respiratory disease from a goat farm in Guangxi province, the epidemiological investigation, clinic observation, pathological examination, bacterium isolation and identification, biochemical test, PCR and animal experiment test were conducted.The preventive and control measures were taken on the basis of pathogen epidemiological characteristics and drug sensitive test results.The results showed that Mycoplasma, influenza virus and parasite were negative by isolation and culture or PCR, however, two strains of gram-negative bacteria named MS1 and MS2 were isolated from lung.MS1 was identified as Serratia marcescens by biochemical test and 16S rRNA sequencing, which shared 99% nucleotide homology with other Serratia marcescens in GenBank.And MS2 was identified as E.coil by the same methods that shared 99% nucleotide homology with other E.coli in GenBank.Animal experiment showed that two strains could cause death in mice.The drug sensitive tests showed two strains were highly sensitive to spectinomycin, amikacin, kanamicin and neomycin.Kanamycin and dexamethasone were used to treat the sick goats, and achieved good results.     

Isolation,Identification and Genetic Evolution Analysis of Mink Serratia marcescens

In order to determine drug resistance, pathogenicity and genetic of the pathogenic bacteria in mink of Heilongjiang province, three strains were isolated from dead minks and identified. Based on colony morphology, microscopic characteristics, biochemical test and 16S rRNA sequencing. The antibiotic susceptibility to 24 antibiotics were detected by using standard K-B paper method, and virulence assay, phylogenetic analysis were carried out at the same time. The result showed that the isolates were highly tolerance to antibacterials, could cause 100% death of mice in experimental group and were closely related to strains from activated sludge and volcanic deposits. It showed that Serratia marcescens infection of mink existed in Heilongjiang province. Strains resistance phenomenon was serious and had a strong pathogenicity, and might be originated from the environment. It was the first time that the pathogenic Serratia marcescens isolated from the mink was reported. The results provided information for the detection, identification, diagnosis, treatment and prevention of related diseases.     

水貂源致病性黏质沙雷氏菌的分离鉴定与遗传进化分析

为确定水貂黏质沙雷氏菌的耐药状况、致病性及其遗传特征,本研究从黑龙江省患病死亡水貂中分离得到3株细菌,根据其形态特征、培养特性、生化试验及16S rRNA序列测定对分离菌进行鉴定;随后用K-B法测试分离株对常见的24种抗菌药的敏感性,并进行了致病性试验和遗传进化分析。结果显示,分离菌为黏质沙雷氏菌,对多种常用抗菌药物具有高度耐药性;可引起试验组小鼠100%死亡;分离株与来自活性污泥与火山堆积物环境的菌株遗传亲缘性最近。表明黑龙江省水貂中存在黏质沙雷氏菌的感染,分离株耐药现象较严重并具有很强的致病性,同时提示该菌株可能来源于环境中。本试验从水貂中分离得到致病性黏质沙雷氏菌为首次报道,研究结果可作为该菌感染水貂导致相关疾病的检测、鉴别和预防依据。     

普城沙雷氏菌BU09的分离鉴定及其对疮痂链霉菌的防治效果

[目的]筛选马铃薯疮痂病的生防菌。[方法]从马铃薯疮痂病发病土壤中取样,经过稀释培养和二次划线纯化获得单克隆菌株,通过抑菌试验最终得到1株对马铃薯疮痂病具有较好抑制效果的细菌BU09,通过16S r DNA测序和系统进化分析对该菌株进行分子鉴定。[结果]细菌BU09属于普城沙雷氏菌,对马铃薯疮痂病具有较好的防治效果。[结论]试验结果为马铃薯疮痂病的田间防治提供了参考。     

一株降解结晶纤维素细菌的分离、筛选、鉴定及其产纤维素酶特性

从广西5个自然保护区内采集原始森林深层腐殖土壤等样品147份,采用平板活性筛选法分离得到3500多株能降解结晶纤维素的细菌,并从中筛选到一株能在pH5.0、28℃条件下可高效降解结晶纤维素的细菌菌株N9-1-1.N9-1-1产纤维素酶水解微晶纤维素和纸浆的最适pH、最适温度分别为:4.0、40 ℃和4.5、35 ℃,但对羧甲基纤维素无水解活性.该菌株所产纤维素酶比较符合纤维素同步糖化共发酵工艺生产乙醇的要求.以菌株N9-1-1的总DNA为模板,对其16S rRNA基因进行PCR扩增和序列分析,结果表明,菌株N9-1-1的16S rRNA基因序列和粘质沙雷氏菌(Serratia marcescens)的同源性最高,为99%.形态特征观察和生理生化检测结果表明,菌株N9-1-1具有粘质沙雷氏菌的鉴别性特征.因此,将N9-1-1鉴定为粘质沙雷氏菌(Serratia marcescens).     

热稳定性过氧化氢酶高产菌的筛选、鉴定及酶学性质研究

从无锡惠山采集的土样中分离获得一株过氧化氢酶产量较高的菌株SYBC-01,根据其形态、生理生化特性和16S rDNA序列分析,鉴定该菌为粘质沙雷氏菌(Serratia marcescens)。初步试验表明,该菌产过氧化氢酶酶活达293.88 U/mg细胞,所产过氧化氢酶的最适反应pH 7,最适反应温度70℃,在pH6~10范围内较稳定,60℃以下热温度性较好,表观Km值为23.71 mmol.L-1,Vmax为34 672 U.mg-1蛋白。     

松毛虫赤眼蜂对柞蚕灰卵的寄生选择与适应性

为明确柞蚕Antheraea pernyi灰卵(工厂化繁育赤眼蜂过程中广泛存在的一种感病寄主卵)对松毛虫赤眼蜂Trichogramma dendrolimi寄生选择与适应性及其繁育子代蜂寄生能力的影响,以柞蚕灰卵作为供试寄主,健康卵作为对照,在无选择和双向选择条件下研究松毛虫赤眼蜂对其的寄生选择与适应性,并比较灰卵和健康卵繁育的子代蜂对0、1、2、3日龄米蛾Corcyra cephalonica卵的寄生能力。结果表明,在无选择条件下,松毛虫赤眼蜂在灰卵上的寄生率、羽化率、单卵出蜂数和总蜂数分别为30.0%、54.1%、39.3头和48.7头,而在健康卵上分别达到96.0%、93.0%、82.5头和96.8头。在双向选择条件下,松毛虫赤眼蜂在灰卵上的寄生率、羽化率、单卵出蜂数和总蜂数分别为33.3%、27.1%、24.7头和52.2头,而在健康卵上分别为68.0%、86.3%、60.6头和74.2头。在无选择和双向选择条件下,松毛虫赤眼蜂在灰卵和健康卵上的发育历期和后代雌性比均不存在显著差异,但灰卵繁育的子代蜂对1、2日龄米蛾卵的寄生数则显著低于健康卵繁育的子代蜂,且后者对米蛾卵的寄生数随着米蛾卵日龄的增加呈逐渐下降趋势,而灰卵繁育的子代蜂对0、3日龄米蛾卵的寄生数要显著高于1、2日龄的。表明柞蚕灰卵会对松毛虫赤眼蜂的寄主选择与适应性以及子代蜂的寄生能力产生不良影响。     

植物根际促生菌普城沙雷菌A21-4对黄瓜生长及土壤微生态的影响

为探讨植物根际促生菌普城沙雷菌A21-4在黄瓜上的应用效果,将A21-4菌悬液灌注到黄瓜根际土壤,调查其对黄瓜生长发育和根际土壤微生态的影响。结果表明,根部灌注A21-4能显著促进苗期和田间生育期黄瓜生长发育,并显著提高根系活力、产量和品质,黄瓜产量比对照增加32.22%,且黄瓜果实中蛋白质、可溶性糖和VC含量均比对照显著增加;同时,A21-4对黄瓜根际土壤具有明显的调节作用,显著提高了黄瓜根际土壤脲酶、磷酸酶、蔗糖酶和过氧化氢酶活性,还提高了黄瓜根际土壤细菌和放线菌的数量及黄瓜根际土壤速效氮、磷、钾的含量。