4株酸奶噬菌体的分离

从上海光明乳业股份有限公司车间发酵罐中分离出4株酸奶噬菌体。1号噬菌体头部呈六角形，平均直径为95.1nm，头部剖面平均面积为8184．8nm^2，尾部的平均直径为18．3nm，平均长度为403．4nm；2号噬菌体头部呈六角形，平均直径为85．4nm，头部剖面平均面积为8046．2nm^2，尾部的平均直径为23．3nm，平均长度为549．4nm；3号噬菌体头部呈六角形，平均直径为95．5nm，头部剖面平均面积为9792．3nm^2，尾部的平均直径为19．8nm，平均长度为450．5nm；4号噬菌体头部呈六角形，平均直径为119．3nm．头部剖面平均面积为12586．1nm^2，尾部的平均直径为23．6nm，平均长度为520．5nm。     

BACTERIAL ENDOCARDITIS WITH THROMBUS FORMATION AND ABSCESSATION IN A HORSE

A 2.5 year old Appaloosa stallion with progressive weight loss and a heart murmur was diagnosed as having vegetative endocarditis of the right atrioventricular valves with a mass in the right atrium. The ultrasonographic appearance of the lesion was characterized by extreme reverberation. On postmortem examination, a 9 cm diameter thrombus was found within the right atrium and auricle, occupying approximately 80% of the chamber volume. Additionally, a lenticular abscess extended throughout the parietal cusp of the right atrioventricular valve. Histologic examination revealed that the lesions were septic with numerous gram positive cocci in short chains, suggestive of Streptococcus equi , and gram negative rods.     

The redox potential of growing cultures of Streptococcus bovis Orla-Jensen compared with other facultative anaerobes.

The changes of redox potential were measured in growing cultures of three strains of Streptococcus bovis, together with three strains of Staphylococcus aureus and one strain of each of Lactobacillus plantaram, Lactobacillus casei, and Eschericia coli. It was found that both S. aureus and E. coli could reduce the redox potential of the growth medium to very low values (between —400 mv and —600 mv), whereas the streptococci and lactobacilli were able to cause only slight or insignificant changes of the redox potential. Respirometric measurements confirmed that the capacity of oxygen consumption of S. bovis was very small compared to that of E. coli and S. aureus. On this basis the authors conclude that S. bovis in all probability is unable to contribute significantly to maintenance of the low redox potential of its natural habitat, the rumen. This function must be carried out by other bacteria, such as enterobacteria or staphylococci, which are capable of performing a true, aerobic respiration.     

Effects of Inulin Chain Length on Fermentation by Equine Fecal Bacteria and Streptococcus bovis

Grass fructans can be fermented by Gram-positive bacteria (e.g., Streptococcus bovis) in the equine hindgut, increasing production of lactic acid and decreasing pH. The degree of polymerization (DP) of fructans has been suggested to influence fermentation rates. The objective of the present study was to determine how DP impacts fermentation by equine fecal bacteria and a model S. bovis. Fecal microbes from three mares were harvested by differential centrifugation, washed, and resuspended in anaerobic media containing short-chain (SC; DP ≤ 10) or long-chain (LC) inulin (DP ≥ 23) from 0% to 2% wt/vol. After 24 hours of incubation (37°C), samples were collected for pH determination. Data were analyzed using the general linear models (GLM) procedure testing for the effect of treatment, concentration, and treatment × concentration (SAS v. 9.3). At all concentrations, the pH was lower in SC fermentations than in LC (P < .0001, in all cases). To determine the effect of DP on S. bovis, cultures were grown (39°C, 9 hours) with 0.1%, 0.5%, or 1.3% SC or LC inulin. Optical density (600 nm) was determined by spectrophotometry. Maximum specific growth rates (μ) were determined by linear regression (2–5 hours). Data were analyzed using the one-way analysis of variance procedure (SAS v. 9.3). The final optical density (600 nm), μ, and yield were higher with SC than with LC fermentation (P < .05). These results indicate that SC inulin may be more available for fermentation than LC inulin by equine fecal bacteria and S. bovis, specifically.     

猪链球菌蛋白表面展示系统的构建及展示蛋白的初步观察

为构建猪链球菌(Streptococcus suis)蛋白表面展示系统,本研究通过序列分析,确定猪链球菌的LPxTG蛋白及其信号肽(SP)和胞壁锚定基序(CWA),通过PCR扩增Peno-SP、GFP、CWA的DNA片段并融合,构建强启动子Peno控制表达编码SP-GFP-CWA融合蛋白的DNA片段,将该重组DNA片段连接pSET2载体,获得蛋白表面展示质粒,转化猪链球菌,构建得到以GFP为报告蛋白的猪链球菌蛋白表面展示系统。结果显示,利用猪链球菌的10个LPxTG蛋白及其SP和CWA序列,构建了10个含有Peno-SP-GFP-CWA融合片段的重组pSET2表面蛋白展示质粒pSsPSD1至pSsPSD10,分别转化猪链球菌05ZYH33,PCR鉴定显示其中7个转化猪链球菌。采用western blot初步检测其展示蛋白,结果显示,7个转化阳性菌株均能有效表达GFP蛋白,以成熟GFP条带为指标,均表现出了一定的外源GFP表面展示水平,分别命名为SsPSD1、SsPSD2、SsPSD4、SsPSD7-SsPSD10,其中SsPSD1、SsPSD4、SsPSD8和SsPSD9表面展示水平相对较好,在猪链球菌表面展示外源蛋白方面具有很好的潜力。本研究首次尝试建立猪链球菌蛋白表面展示系统,为猪链球菌表面递呈外源蛋白或抗原提供了新的策略。     

羊链球菌CVCC55001、CVCC55002株的制备与鉴定

为研究羊链球菌CVCC55001、CVCC55002株的菌种特性,制备了新的菌种批并对其形态及生化特性、培养特性、血清学特性、真空度、纯粹、剩余水分、毒力及免疫原性等进行鉴定,对其适宜培养基进行了筛选.试验结果表明,该冻干菌种的形态及生化特性、培养特性、血清学特性、真空度、纯粹、剩余水分、毒力及免疫原性均符合《中华人民...     

环介导等温扩增联合横向流动试纸条可视化检测海豚链球菌方法的建立

海豚链球菌(Streptococcus iniae)是一种革兰氏阳性球菌,呈β溶血,可感染多种淡水和海洋鱼类。本研究利用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)进行核酸扩增,通过横向流动试纸条方法 (lateral flow dipstick,LFD)实现检测,建立了一种可应用于海豚链球菌快速检测的LAMP-LFD技术。该技术以海豚链球菌促旋酶B亚单位(gyrase subunit B,gyr B)基因为检测靶标,设计3对引物进行由生物素标记的LAMP扩增反应,产物经异硫氰酸荧光素(fluorescein isothiocyanate,FITC)标记的探针杂交后,在LFD上完成检测。经优化的核酸扩增最适条件为65℃反应30 min,在此条件下,阳性扩增起始时间与模板浓度之间呈典型的线性相关性。从核酸扩增反应开始到LFD显色,整个检测时程只需40 min左右,比常规PCR技术缩短约2 h。LAMP-LFD能特异性地检出海豚链球菌,针对病原纯培养物的检测灵敏度为8.70×101cfu/m L,是LAMP检测的10倍、常规PCR检测的100倍。以灵敏度浓度(8.70×101cfu/m L)的海豚链球菌基因组DNA为模板进行的LAMP-LFD结果显示该方法具有良好的重复性。针对人工污染花鲈(Lateolabrax japonicus)肝组织的检测灵敏度为4.35×103cfu/m L,同样为常规PCR检测方法的100倍。利用本方法可成功从患病花鲈的组织样品中检测出海豚链球菌,检测结果与常规的细菌分离鉴定方法结果一致。因此,利用LAMP-LFD能特异、准确、高效地检测出海豚链球菌,而且操作简单、费用低、耗时短,有望成为海豚链球菌的常规检测方法。     

猪链球菌2型次黄嘌呤核苷酸脱氢酶(IMPDH)重组蛋白的酶活性和免疫原性检测

通过PCR方法从猪链球菌2型(Streptococcus suis)05ZYH33分离株基因组中扩增出次黄嘌呤核苷酸脱氢酶基因(inosine 5-monophosphate dehydrogenase,impdh),长度1 064 bp.PCR产物和pET28a载体分别经过EcoR Ⅰ和Xho Ⅰ双酶切,连接,成功地构建了重组表达质粒pET28a-impah,并转入大肠杆菌(Escherichia coli) BL21(DE3)中,经过1 mmol/L IPTG诱导获得表达,蛋白大小35kD.表达蛋白具有良好的抗原性和酶活性.蛋白经过亲和层析纯化后,在37℃,pH7.0～9.0表现出较强酶活性,NADH A_(340)介于0.662～0.816之间.