克氏原螯虾用芽孢杆菌的特性及其安全性评价

为评估克氏原螯虾用产芽孢益生菌的特性和安全,本研究从市售益生菌产品中分离到19株芽孢杆菌属成员.通过测定抗菌活性、产纤维素酶活性、温度对生长影响来评价其益生菌的特性;通过测定芽孢对高温、低pH、胆汁酸盐的耐受、自聚集性和表面疏水性等评价其稳定性;通过溶血活性、抗生素敏感性、小鼠攻毒试验、克氏原螯虾饲养试验等评价其安全性.以抗菌和产纤维素酶活性、低温(15℃)生长快和无溶血活性这四项指标为标准进行筛选,结果表明:只有3株芽孢杆菌(B8,B20,B27)可成为益生菌候选株.在15℃低温时生长迅速、抗菌和产纤维素酶活性高的2株解淀粉芽孢杆菌(B14和B40)有溶血活性,能引起克氏原螯虾的肠炎,并导致肠道菌群中气单胞菌丰度显著上升.因此,在克氏原螯虾用芽孢杆菌益生菌的选择中,不仅要关注益生功能,更要考虑其安全性.本研究结果为芽孢杆菌在动物保护产品中科学评价和合理使用提供参考.     

斜纹夜蛾幼虫肠道细菌分离鉴定及其功能初步分析

昆虫肠道微生物对其寄主的生长发育、营养代谢、免疫以及农药抗性等方面都发挥着重要作用。为研究斜纹夜蛾Spodoptera litura幼虫肠道细菌的多样性,并为其功能验证做准备,本文利用传统微生物分离纯培养方法从斜纹夜蛾4龄幼虫肠道中共分离鉴定得到10株细菌,分别为属于变形菌门(Proteobacteria)的脱氮假单胞菌(Pseudomonas denitrificans),不动细菌(Acinetobacter sp.),肺炎克雷伯氏菌(Klebsiella pneumoniae)和肠杆菌(Enterobacter sp.);属于厚壁菌门(Firmicutes)的鸡葡萄球菌(Staphylococcus gallinarum),蒙氏肠球菌(Enterococcus mundtii),蜡样芽胞杆菌(Bacillus cereus)和枯草芽胞杆菌(Bacillus subtilis)以及放线菌门(Actinobacteria)的微杆菌(Microbacteriums sp.)和乳酪棒杆菌(Corynebacterium casei)。变形菌门和厚壁菌门是斜纹夜蛾肠道可培养细菌中的优势菌群。功能验证实验表明肠杆菌具备纤维素降解能力,微杆菌具备很强的苯酚降解能力。本研究为未来深入研究斜纹夜蛾肠道微生物的功能提供了方向和菌株材料。     

对虾白斑综合征病毒新株型WSSV-CN-Pc的毒力研究

对虾白斑综合征病毒(white spot syndrome virus,WSSV)是一种能够感染虾类并且造成其大面积死亡的环状双链DNA病毒。WSSV有多种分离株,其毒力有所差异。从克氏原螯虾(Procambarus clarkii)中分离得到1株WSSV新分离株WSSV-CN-Pc,其毒力尚不清楚。本研究采用肌肉注射和经口注射的方法,以WSSVTW型作为阳性对照,分别对克氏原螯虾(P.clarkii)和罗氏沼虾(Macrobrachium rosenbergii)进行活体实验。实验结果显示:肌肉注射WSSV-CN-Pc和WSSV-TW的克氏原螯虾均在第6天出现100%的死亡;罗氏沼虾在肌肉注射WSSV-TW后未出现死亡,但在注射WSSV-CN-Pc后的第9天死亡率达100%。经口注射WSSV-CN-Pc和WSSV-TW的克氏原螯虾均在第16天出现100%的死亡;罗氏沼虾经口注射WSSV-CN-Pc后的第19天死亡率为100%,但注射WSSV-TW的实验组并未出现死亡。结果表明,对于克氏原螯虾,WSSV-CN-Pc具有和WSSV-TW相似的毒力,而对罗氏沼虾存在明显的毒力差异。提示克氏原螯虾是WSSV传播途径中的重要因素。     

氨基酸产生菌E_(65)中质粒的分离与鉴定

在棒杆菌与短杆菌中,用微量质粒分离方法筛选到一株含多种质粒的乙酰短杆菌(Brevibacterium actylicum)E_(65)菌株,该菌株具有产氨基酸特性。进一步大量制备其质粒DNA,并进行氧化绝密度梯度超离心纯化,以及电泳分析,电镜观察,并对质粒进行分离与鉴定,从而确定了每种质粒的分子量大小。     

腈水合酶产生菌的培养及其酶活性的比较

从腈污染的土样中筛选获得,株腈水合酶活性较高的细菌,分属于棒状杆菌(Cerynebactertum sp.)、节杆菌(Arthrobacter sp.)和克雷伯氏菌(Klebsiella sp.)其中具有腈水合酶活性的克氏杆菌属菌为首次分离到。对这些菌株适宜培养条件及酶形成诱导特性的研究表明,棒状杆菌和节杆菌的腈水合酶为诱导酶,而克氏杆菌的酶为组成酶。前二个属的细菌完整细胞腈水合酶活性相近,均较后一属细菌高约10倍。     

钝齿棒杆菌溶氧诱导型启动子的筛选及其功能验证

[目的]筛选钝齿棒杆菌(Corynebacterium crenatum)SYPA5-5内源溶氧诱导型启动子,验证其在不同溶氧条件下受溶氧调控的功能.[方法]以本课题组前期利用双向电泳结合质谱鉴定技术分离鉴定出的高低供氧条件下钝齿棒杆菌SYPA5-5胞内的显著差异蛋白(27个)为基础,结合棒杆菌基因组信息分析高低溶氧状态下显著差异蛋白中单位拷贝的表达量,以其中单位拷贝表达量高的蛋白为目的蛋白,克隆目的蛋白编码基因的上游启动子,分别替换棒杆菌-大肠杆菌穿梭表达质粒pDXW-8的外源tac启动子,在大肠杆菌和钝齿棒杆菌中表达报告基因gfp和cat,并检测其表达效率.[结果]筛选出高溶氧上调蛋白转酮醇酶和延胡索酸水化酶作为目的蛋白,克隆其启动子区命名为P-tkt和P-fum.在重组大肠杆菌和重组钝齿棒杆菌中,P-tkt在高溶氧条件下CAT活性是低溶氧条件下CAT活性的2.8和3.2倍.该结果在5L发酵罐中得到验证.[结论]启动子P-tkt为首次筛选到的钝齿棒杆菌内源性高溶氧诱导型启动子,适用于高溶氧条件发酵生产氨基酸的菌株中,有利于在高供氧条件下提高棒杆菌中氨基酸的合成代谢能力.     

新疆发酵乳品中乳酸菌的分离鉴定及其耐药性

目的对新疆传统发酵乳品中乳酸菌进行分离鉴定并检测其耐药性。方法利用传统形态学鉴定法和生化鉴定等方法对新疆发酵乳中乳酸菌进行鉴定,采用纸片扩散法对分离鉴定的菌进行耐药性分析。结果从新疆发酵乳品中共分离出8株乳酸菌,经鉴定分别为瑞士乳杆菌(Lactobacillus helveticus)、植物乳杆菌(Lactobacillus plantarum)、马乳酒样乳杆菌(Lactobacillus kefianofaciens)、乳酸乳球菌(Lactococcus lactis)、副干酪乳杆菌(Lactobacillus paracasei)、副干酪乳杆菌类坚韧亚种(Lactobacillus paracasei subsp.tolerans)、哈尔滨乳杆菌(Lactobacillus harbinensis)、希氏乳杆菌(Lactobacillus hilgardii),并且发现8株乳酸菌对万古霉素、庆大霉素、阿莫西林、多西环素、环丙沙星、阿奇霉素、头孢他啶、头孢孟多具有一定敏感性。结论新疆发酵乳品中以乳杆菌居多,对常见抗生素具有一定的敏感性。     

广西番茄内生细菌的多样性和数量动态

为了探明内生细菌在番茄中的分布和数量变化规律,有目的地筛选防治番茄青枯病的内生细菌,我们对广西可培养的番茄内生细菌的类群和数量动态进行了调查。从广西部分县市采集的303个番茄样本中分离到624株内生细菌菌株,初步确定有芽孢杆菌(Bacillus)、假单胞菌(Pseudomonas)、黄单胞菌(Xanthomonas)、棒杆菌(Corynebacterium)、土壤杆菌(Agrobacterium)、微杆菌(Microbacterium)、肠杆菌(Enterobacter)和欧文氏菌(Erwinia)8个属,其中以芽孢杆菌、假单胞菌和土壤杆菌为芽孢杆菌为优势类群。番茄内生细菌在植株器官中的分布以根部数量最多,其次是茎和叶。内生细菌的总量在番茄生育期的变化趋势是从苗期到花期数量上升,而从结果期到成熟期数量逐渐下降。多数内生细菌种群的数量变化动态符合细菌总量的变化趋势,只有微杆菌在番茄植株整个生育期中始终保持下降的趋势。春季种植的番茄植株的内生细菌类群数量比秋季种植的少。     

棒杆菌宿主中质粒不稳定性的研究

重组质粒pNAR4是由钝齿棒杆菌质粒pNAT65和大肠杆菌质粒pACYCl84构建的穿梭质粒。质粒pNAR4转化不同棒杆菌,在钝齿棒杆菌T6—13和答氨酸棒杆菌l0l47中为结构型不稳定,在钝齿棒杆菌B9中为分离型不稳定。而pNAT65转化谷氨酸棒杆菌10147后,转化于中的质粒分子大小及主要酶切位点与pxz10145相同。DNA杂交实验结果表明．在10147菌中有一种与z10145高度同源的超螺旋DNA组分,而这一组分与pxz10145的来源宿主中的另一小质粒具有相同的分子量。提出了质粒结构不稳定与宿主中存在的pxz10145高度同源的小质粒(超螺旋组分)有关,并提出产生质粒分子结构反复变化原因的假设。     

直链烷基苯磺酸钠(LAS)降解菌的筛选及其降解特性的初步研究

从洗衣粉生产废水中分离到一株高效降解LAS(Linear Sodium A]kylbenzenesulfonate)菌GZ6,经初步鉴定其为杰氏棒杆菌(Corynebactenunjeikeiun GZ6).该菌株最高可以降解700mg/L左右的LAS,降解LAS的最适pH值和温度分别为7.0和30℃,最适LAS浓度为400 mg/L,所需时间为24h,降解率可达98.7%.实验还表明一些重金属离子如-Hg2+、Co2+、Cd2+等对该菌株降解LAS有不同程度的抑制作用.     

Idiopathic granulomatous mastitis. Report of a case diagnosed with fine needle aspiration cytology.

BACKGROUND: Idiopathic granulomatous mastitis (IGM) is a benign, inflammatory breast disease of unknown etiology. Although it is rare, it frequently presents in a manner similar to that of breast carcinoma. CASE: A 41-year-old female developed unilateral idiopathic granulomatous mastitis, diagnosed by fine needle aspiration cytology. The clinical presentation and mammographic findings were suspicious for carcinoma. Fine needle aspiration cytology showed granulomatous inflammation. Histopathologic examination revealed a noncaseating, granulomatous lesion. Further clinical, radiologic and laboratory investigations disclosed no etiology. Therefore, we considered the case to be idiopathic granulomatous mastitis. CONCLUSION: Cytologically it may be difficult to distinguish IGM from carcinoma of the breast. Typical cytologic findings of the lesion are helpful to rule out cancer. In the differential diagnosis, all known causes of granulomatous changes have to be excluded before a diagnosis of idiopathic granulomatous mastitis is made.     

Ultrafast pyrosequencing of Corynebacterium kroppenstedtii DSM44385 revealed insights into the physiology of a lipophilic corynebacterium that lacks mycolic acids

Corynebacterium kroppenstedtii is a lipophilic corynebacterial species that lacks in the cell envelope the characteristic alpha-alkyl-beta-hydroxy long-chain fatty acids, designated mycolic acids. We report here the bioinformatic analysis of genome data obtained by pyrosequencing of the type strain C. kroppenstedtii DSM44385 that was initially isolated from human sputum. A single run with the Genome Sequencer FLX system revealed 560,248 shotgun reads with 110,018,974 detected bases that were assembled into a contiguous genomic sequence with a total size of 2,446,804bp. Automatic annotation of the complete genome sequence resulted in the prediction of 2122 coding sequences, of which 29% were considered as specific for C. kroppenstedtii when compared with predicted proteins from hitherto sequenced pathogenic corynebacteria. This comparative content analysis of the genome data revealed a large repertoire of genes involved in sugar uptake and central carbohydrate metabolism and the presence of the mevalonate route for isoprenoid biosynthesis. The lack of mycolic acids and the lipophilic lifestyle of C. kroppenstedtii are apparently caused by gene loss, including a condensase gene cluster, a mycolate reductase gene, and a microbial type I fatty acid synthase gene. A complete beta-oxidation pathway involved in the degradation of fatty acids is present in the genome. Evaluation of the genomic data indicated that lipophilism is the dominant feature involved in pathogenicity of C. kroppenstedtii.     

Fine needle aspiration cytology of granulomatous mastitis. Report of a case and review of the literature

A case of granulomatous mastitis diagnosed by fine needle aspiration (FNA) cytology alone is reported. A 24-year-old pregnant woman presented with a six-week history of a rapidly enlarging breast mass, considered clinically to be malignant. The FNA smears contained numerous aggregates of epithelioid histiocytes admixed with Langhans' and foreign-body giant cells, lymphocytes, neutrophilic leukocytes and apoptotic debris, leading to a diagnosis of granulomatous mastitis. The features of granulomatous mastitis and its differential diagnoses are discussed.     

IL-6/STAT3信号途径介导肉芽肿性乳腺炎的作用研究

目的:探讨IL-6/STAT3信号途径介导肉芽肿性乳腺炎的机制研究及临床意义。方法:收集我院2017年1月至2017年6月收治入住的30例肉芽肿性乳腺炎患者及30例普通患者的血清样本,对肉芽肿性乳腺炎患者同时采集炎性肿块组织样本及距离炎性肿块边缘大于等于3 cm且镜检为正常乳腺组织的样本。采用ELISA法检测血清样本中的IL-6的表达水平,同时采用Western blot检测组织样本中IL-6和pSTAT3蛋白的表达水平并分析二者的相关性。结果:与普通患者血清样本相比,肉芽肿性乳腺炎患者血清中IL-6的表达水平明显增高(P0.05),肉芽肿性乳腺炎组织样本中IL-6和pSTAT3蛋白的表达水平明显高于毗邻的正常乳腺组织(P0.01),IL-6表达与组织STAT3磷酸化水平具有明显的正相关性(r=0.353,P0.05)。结论:IL-6/STAT3信号途径可能介导肉芽肿性乳腺炎的发生发展。     

Granulomatous inflammation of the breast in a pregnant woman: report of a case with fine needle aspiration diagnosis

BACKGROUND: Granulomatous inflammation of the breast is an inflammatory process with multiple etiologies. It can accompany breast carcinoma or be idiopathic. It often presents clinically in a fashion mimicking carcinoma. Idiopathic granulomatous mastitis is strongly associated with lactation and is reported to occur in postpartum patients. This is the second fine needle aspiration (FNA) report of idiopathic granulomatous inflammation in the breast of a pregnant woman. CASE: A 27-year-old, 7-month-pregnant woman presented with a hard nodule in her right breast; on ultrasound examination it showed mixed echogenicity, suspicious for carcinoma. FNA showed granulomatous inflammation. The smears were highly cellular, with many clusters of and single epithelioid cells displaying moderate pleomorphism and prominent nucleoli in a background composed of neutrophils, plasma cells, lymphocytes and multinucleated cells. Core needle biopsy revealed a nonnecrotizing, granulomatous lesion. CONCLUSION: The diagnosis of granulomatous inflammation can be challenging, and the cytologic features can be difficult to separate from those of carcinoma. The relatively rare occurrence of this lesion and its cytologic features make it a potentially difficult diagnosis and diagnostic pitfall.     

Idiopathic granulomatous mastitis masquerading as carcinoma of the breast: a case report and review of the literature

Background

Idiopathic granulomatous mastitis is an uncommon, benign entity with a diagnosis of exclusion. The typical clinical presentation of idiopathic granulomatous mastitis often mimics infection or malignancy. As a result, histopathological confirmation of idiopathic granulomatous mastitis combined with exclusion of infection, malignancy and other causes of granulomatous disease is absolutely necessary.

Case Presentation

We present a case of a young woman with idiopathic granulomatous mastitis, initially mistaken for mastitis as well as breast carcinoma, and successfully treated with a course of corticosteroids.

Conclusion

There is no clear clinical consensus regarding the ideal therapeutic management of idiopathic granulomatous mastitis. Treatment options include expectant management with spontaneous remission, corticosteroid therapy, immunosuppressive agents and extensive surgery for refractory cases.

     

Cytomorphology of tuberculous mastitis. A report of nine cases with fine needle aspiration cytology

Nine cases of tuberculosis of the breast were diagnosed by fine needle aspiration (FNA) cytology over a six-month period. All cases presented with a breast lump, three of which clinically simulated carcinoma. FNA cytology showed a picture of suppurating granulomatous mastitis; the diagnosis of tuberculosis was established after the demonstration of acid-fast bacilli in the aspirated material.     

Fine needle aspiration cytology in the diagnosis of tuberculous mastitis

OBJECTIVE: To study the relationship between granulomas in the breast and tuberculous mastitis. STUDY DESIGN: Retrospective analysis of 22 breast aspirates that showed epithelioid cell granulomas. The aspirates were reviewed and the cytomorphologic findings summarized. RESULTS: Aspiration cytology revealed epithelioid cell granulomas along with giant cells, necrosis and inflammatory cell infiltrate. Overall acid-fast bacillus (AFB) positivity was 22.7%. AFB positivity was greater in the presence of necrosis when epithelioid cells were absent. CONCLUSION: In a country like India, the diagnosis of granulomatous mastitis must be made with caution, even in the absence of AFB. Only after a sufficient trial of antituberculosis treatment has been given and the patient fails to respond should an alternative diagnosis be suggested.     

Silicone Mastitis in “Topless” Waitresses and Some Other Varieties of Foreign-body Mastitis

Three cases of disfiguring prosthetogenic mastitis are reported.In two of the cases repeated injections of silicone fluid were followed by extensive bilateral granulomatous mastitis. The axillary lymph nodes were enlarged in both cases: in one of them a node was examined histologically and showed a remarkable proliferation of vacuolated histiocytes in the sinuses and medulla. Both patients had worked in “topless restaurants” and had been obliged to seek artificial means to maintain the excessively large bust that was necessary in that environment.The third patient''s mastitis followed implantation of polyethylene prostheses designed to be varied in bulk by injection or withdrawal of saline. A foreign-body reaction developed round the prostheses. In addition, a sinus developed in a needle track on one side; this was accompanied by a local fungal and bacterial infection (Rhizopus species and Pseudomonas pyocyanea respectively).The indications for introducing foreign materials into the breasts to augment their size have to be weighed against the potential dangers that such procedures bring. The biological effects of injected silicones, particularly their long-term effects, are very imperfectly known. Statements implying that these compounds are harmless when injected are not correct. Among the hazards are (a) local sclerosing granulomatous reactions with permanent scarring, (b) histiocytosis secondary to carriage of silicones from their site of inoculation to regional lymph nodes and possibly to other parts of the lymphoreticular system, and, perhaps, (c) carcinogenic activity.     

Inflammatory Lesions of the Breast: Diagnosis By Fine Needle Aspiration

Amongst 1061 breast lesions diagnosed by fine needle aspiration (FNA) over a period of 6 years (1985-1990), 128 were reported to be showing changes consistent with an inflammatory lesion. On review, the cytodiagnosis was found to be inaccurate in 31 cases. The cytological features of the 97 cases that were correctly reported are described in this report. The cytological diagnoses issued in these 97 cases were acute mastitis or breast abscess (57 cases) and tuberculous mastitis (30 cases). Non-specific chronic mastitis and miscellaneous conditions accounted for four and six cases respectively. Acid fast bacilli (AFB) were demonstrated in 28.0% of tuberculous mastitis cases and 10.0% of those diagnosed as acute mastitis or breast abscess. FNA cytology was found to be useful for the diagnosis of inflammatory lesions of breast and their classification, as only five out of 57 cases of acute mastitis/breast abscess and one out of 30 tuberculous mastitis cases were suspected on clinical grounds.