桑树芽条枯萎病病原研究 I.病原分离及致病性测定

笔者对陕西省安康地区桑园春季发生的一种梢条和嫩芽枯萎病害的病原作了初步研究,从感病材料各典型病部分离得到桑Ⅰ号菌和桑Ⅱ号菌,经LOPAT部分项目测定及常规致病性测定,究明桑Ⅰ号菌(丁香假单胞杆菌)为该病害主要致病细菌,桑Ⅱ号菌(草生欧氏杆菌)可以引起感染,其症状不如Ⅰ号菌典型,但两菌混合接种有加重病情的现象,故与本病害有密切的关系.     

Bacterial infection of the lower respiratory tract in 34 horses

OBJECTIVE: To investigate associations between the bacteriology and aspects of history, clinical presentation, outcome and pathology of lower respiratory tract disease of 34 horses. PROCEDURE: Detailed aerobic and anaerobic bacteriological investigations were performed on clinical specimens from horses with pneumonia, lung abscessation and necrotic pneumonia with or without pleurisy in an attempt to identify those bacteria that might contribute to the initiation and progression of infection. RESULTS: Bacteria were cultured from 33 of the 34 horses. In ten cases, only aerobic/facultatively anaerobic isolates were cultured while aerobic/facultatively anaerobic bacteria and obligately anaerobic bacteria were isolated in the other 23 cases. Moderate to large numbers of anaerobic bacteria were isolated only when the estimated duration of illness was at least five days. Bacteria were not cultured from 12 of the pleural fluid samples but were always cultured from pulmonary samples (either transtracheal aspirates from live horses or pulmonary lesions at necropsy). Streptococcus equi subsp zooepidemicus was isolated in the three cases where only one bacterial species was cultured. In the other 30 cases, multiple species were isolated. These included most often and in greatest numbers, Streptococcus equi subsp zooepidemicus, Pasteurellaceae, Escherichia coli, anaerobic cocci, Eubacterium fossor, Bacteroides tectum, Prevotella heparinolytica, Fusobacterium spp, and pigmented members of the genera Prevotella and Porphyromonas. Aerobic/facultatively anaerobic organisms were isolated from 97% of horses, while obligately anaerobic organisms were cultured from 68% of horses. CONCLUSION: There was no association between the isolation of any specific bacterium and the outcome of disease. However, obligately anaerobic bacteria (such as anaerobic cocci, Bacteroides tectum, P heparinolytica and Fusobacterium spp) and the facultatively anaerobic species Escherichia coli, were recovered more commonly from horses that died or were euthanased than from those that survived. There was an association between failure of horses to recover from pleuropneumonia and delay in diagnosis and initiation of treatment.     

蒙古马肠道细菌总DNA提取方法的比较

本试验采集蒙古马新鲜粪便样品,采用酚－氯仿抽提法和2种细菌基因组DNA提取试剂盒法进行样品中细菌基因组DNA的提取。通过DNA浓度和纯度的测定,并将其作为模板扩增细菌16S rDNA V3区特异性片段对提取效果进行比较,从而找出更适合蒙古马粪便中细菌基因组DNA的提取方法。结果显示,3种提取方法得到的基因组DNA均能用于PCR扩增的模板,其中B试剂盒法提取的DNA数量较多,且纯度高,更适合用于提取蒙古马肠道微生物细菌基因组DNA及后续的蒙古马肠道微生物多样性等分子生物学试验。     

柳树叶饲料化调制加工技术研究

为了探索垂柳(Salix babylonica)树叶的饲料化加工技术,在研究6个生育期(展叶期、开花初期、飞絮期、青绿期、枯黄期、落叶期)柳树叶营养物质含量变化的基础上,分别将其调制成青贮饲料,测定并比较不同生育期柳树叶制备青贮饲料的营养物质含量及发酵品质指标,并对枯黄期和落叶期树叶进行添加乳酸菌菌剂青贮对比试验。结果表明,根据原料营养物质判断,展叶期柳树叶营养品质与优质豆科牧草品质近似,但是随着生育期的延迟,柳树叶营养物质含量和饲用价值显著下降。将不同生育期柳树叶直接青贮之后,无品质改善效果,青贮饲料发酵品质差,饲用价值不大。但是,添加乳酸菌菌剂后调制的柳树叶青贮饲料发酵品质得到了显著改善。综上表明,柳树叶如果直接饲喂利用,最好选择在较为幼嫩的展叶期;如果开发利用其他生育期或枯落后的柳树叶,以添加乳酸菌菌剂调制成青贮饲料为好。     

甜高粱根际溶磷菌溶磷能力及菌株特性研究

利用蒙金娜培养基,从甜高粱根际分离具有溶磷能力的菌株,通过溶磷圈法筛选出5株溶磷能力较强的菌株,利用钼锑抗比色法和Salkowski比色法分别对溶磷特性与分泌IAA能力进行测定,结果表明,筛选的菌株分解磷酸钙的能力存在显著差异,溶磷量在107.26~233.95 μg/mL之间,各菌株均具有分泌IAA能力,IAA分泌量在15.23~27.79 μg/mL之间,接种溶磷菌后甜高粱株高、茎粗、干重都比对照明显提升。因此, 溶磷能力和分泌IAA能力较强的菌株(WD₅₁、WD₂₀) 可作为研制微生物肥料的优良菌株。     

低温条件下混合乳酸菌制剂对芦苇发酵品质的影响

为探讨低温条件下混合乳酸菌制剂对芦苇(Phragmites australis Cav.Trin.ex Sterd.)发酵品质的影响,对含水量为40%的芦苇添加混合乳酸菌制,分别设计4℃和0℃的发酵温度,密封发酵,分别取3,4,5,6,7和8周的发酵样品分析。结果表明:在0℃条件下,发酵后第5周pH值出现显著下降(P<0.05),第8周NDF含量显著降低(P<0.05)。在4℃条件下,发酵第3周起pH值一直保持在4.20以下。在相同发酵时间内,4℃条件比0℃条件时的pH值显著降低(P<0.05),乳酸菌数量、乳酸产量显著提高(P<0.05),乙酸产量显著降低(P<0.05)。在低温条件下,对每克干物质添加6.63(lg CFU·g^-1DM)的混合乳酸菌,可使含水量为40%的芦苇启动发酵。为保证得到优良发酵质量,发酵时间应在6周以上。     

实时荧光定量PCR对瘤胃纤维分解菌定量方法的构建

试验构建了白色瘤胃球菌、黄色瘤胃球菌及产琥珀酸丝状杆菌等3种瘤胃纤维分解菌实时荧光绝对定量PCR的标准品及标准曲线,以用于纤维分解菌的定量测定。提取瘤胃微生物总DNA,以各菌特异性引物进行PCR扩增,回收纯化PCR产物,与pMD18-T Vector连接并转化到大肠杆菌。用氨苄青霉素培养基筛选阳性重组质粒,提取含目的片段质粒DNA,通过PCR及测序鉴定重组质粒。根据OD值确定浓度,将梯度稀释的质粒作为模板,进行荧光定量PCR反应做出标准曲线。结果表明:所构建的3条标准曲线具有很高的相关性(R2>0.999),并获得了高扩增效率产物(白色瘤胃球菌为101%、黄色瘤胃球菌为98.0%、产琥珀酸丝状杆菌为97.7%),说明成功构建了瘤胃纤维分解菌实时绝对定量PCR的标准品和标准曲线,为定量研究瘤胃纤维分解菌奠定基础。     

Fatty acid composition of ruminal bacteria and protozoa,and effect of defaunation on fatty acid profile in the rumen with special reference to conjugated linoleic acid in cattle

Objectives of this study were to compare fatty acid (FA) composition of ruminal bacterial (B) and protozoal (P) cells, and to investigate effect of protozoa on FA profile in the rumen of cattle. Three cows were used to prepare ruminal B and P cells. Four faunated and three defaunated cattle (half‐siblings) were used to study effect of protozoa on ruminal FA profile. Proportions of C16:0 and C18:0 in total fatty acids in B cells were 20.7% and 37.4%, whereas those in P cells were 33.4% and 11.6%, respectively. Proportions of trans‐vaccenic acid (VA) and cis‐9, trans‐11 conjugated linoleic acid (CLA) in B cells were 3.9% and 1.0%, and those in P cells were 5.5% and 1.6%, respectively, being higher in P cells. Proportions of C18:1, C18:2 and C18:3 in P cells were two to three times higher than in B cells. Proportions of unsaturated fatty acids, VA and CLA in B cells of faunated cattle were higher than those of defaunated. VA and CLA in the ruminal fluid of faunated were also 1.6 to 2.5 times higher than those of defaunated. This tendency was similar for cell‐free fraction of ruminal fluid. These results indicate that protozoa contribute greatly in VA and CLA production in the rumen.     

Susceptibility pattern of bacterial isolates in equine ulcerative keratitis: Implications for empirical treatment at a university teaching hospital in Sydney

Corneal ulceration is a common ophthalmic condition in horses. It is frequently caused by trauma to the corneal surface, followed by secondary infection by commensal or pathogenic organisms including Streptococcus equi subspecies zooepidemicus, Pseudomonas aeruginosa and Staphylococcus spp. Emerging antimicrobial resistance amongst these organisms has raised the need for appropriate antimicrobial therapy selection, to optimise treatment efficacy while minimising further antimicrobial resistance. Medical records of 38 horses presented at the University Veterinary Teaching Hospital Camden for ulcerative keratitis between 2010 and 2020 were reviewed to identify those with positive bacterial cultures and antimicrobial susceptibility profiles (13/38). Common susceptibility patterns were identified and used to guide the empirical treatment of equine bacterial corneal ulcers. Pseudomonas spp. (64.3%), Streptococcus equi subspecies zooepidemicus (14.3%) and Actinobacillus spp. (14.3%) were most commonly identified. Susceptibility to amikacin, gentamicin and ciprofloxacin was observed in 100%, 66.7% and 85.7% Pseudomonas spp. isolates respectively. Resistance to polymyxin B and neomycin occurred in 85.7% and 71.4% of Pseudomonas spp., respectively. All Streptococcus equi subspecies zooepidemicus organisms in this study were susceptible to ceftiofur, cephalexin, penicillin and ampicillin, while they were all resistant to gentamicin, neomycin, enrofloxacin and marbofloxacin. Predominating in this study, Pseudomonas spp. maintained overall aminoglycoside susceptibility despite some emerging resistance, and good fluoroquinolone susceptibility. High resistance to Polymyxin B could have arisen from its common use as first-line therapy for bacterial corneal ulcers. Although further research is required, these new findings about predominant bacteria in equine corneal ulceration in the Camden region and their antimicrobial susceptibility patterns can be used to guide the empirical treatment of bacterial corneal ulcers in horses.     

The Effect of pH and Potassium Phosphate Buffer on the Toxicity of Cadmium for Bacteria

An increase in the pH of Plate Count Agar led to increased toxicity of cadmium (Cd) for Micrococcus luteus ATCC 9341, Staphylococcus aureus ATCC 12600, Clostridium perfringens ATCC 8798, Escherichia coli ATCC 11775 and Pseudomonas aeruginosa ATCC 10145. The effect of the pH on toxicity was not clearly observed with Streptococcus bovis ATCC 9809 and was absent with Bacillus subtilis ATCC 8473.When potassium phosphate buffer was added to the growth medium, the Cd toxicity for M. luteus and B. subtilis was enhanced. The toxicity of Cd for S. bovis decreased when potassium phosphate buffer was added to the medium. When the pH increased at differing phosphate concentrations, the sensitivity of M. luteus to Cd decreased.In the authors’ opinion, the effect of pH on Cd toxicity for bacteria is due to a continuously increasing negative charge towards an alkaline value by most bacteria which increases the affinity of cations towards the cell wall.