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1.
Enteropathogenic Escherichia coli strains may produce a cholera-like, heat-labile enterotoxin (LT) as a virulence factor. The gene that codes for LT can be purified by recombinant DNA techniques and used as a genetic probe for DNA hybridization. These probes detect enterotoxigenic strains as well as strains that may not manifest toxin production but carry the genetic information to do so. In this study, 13 laboratories tested 3 known and 25 unknown (10 positive and 15 negative) cultures of E. coli for the presence of the LT gene. The isolates had been tested and classified by the mouse Y-1 adrenal cell test and an enzyme-linked immunosorbent assay. Cultures were spotted on nitrocellulose filters on MacConkey agar and incubated. Colonies were lysed in situ and their DNA was hybridized to 32P-labeled, purified LT gene DNA (provided to the collaborators). Positive colonies were identified by autoradiography. Of 325 samples, 315 (96.9%) were identified correctly and 10 were misclassified; there were 6 false negative and 4 false positive identifications. Chi-square values indicated that the method agreed with the previous classification and was equally efficient in distinguishing positive and negative samples (95.7 and 98.1%, respectively). The method has been adopted official first action.  相似文献   

2.
Nine laboratories evaluated the ability of the MICRO-ID test system to correctly identify members of the Enterobacteriaceae. A total of 78 isolates representing 11 genera of enterics that had been previously isolated from foods were used in the collaborative study. The collaborators streaked each isolate on plate count agar and incubated the plates overnight at 35 degrees C to check purity of the isolates. Then they proceeded with the method in which an isolated colony is transferred to a plate count agar slant and is incubated 18-24 h at 35 degrees C. Growth from the slant is emulsified in 3.5 mL physiological saline to a density comparable to a McFarland No. 2 tube and is then used to inoculate the test (MICRO-ID) strip. The strip is incubated 4 h at 35 degrees C and the reactions are read and recorded. Isolates are identified by using an octal code and the test kit manual. The system correctly identified 98.8% of the Salmonella isolates, 97.7% of the E. coli isolates, and 84.6% of the other 9 enteric genera tested. The system has been approved interim official first action as an alternative to conventional biochemical tests (1) for presumptive generic identification of food-borne Salmonella and for screening and elimination of non-Salmonella isolates; (2) for identification of E. coli from foods; and (3) for presumptive generic identification of other Enterobacteriaceae isolated from foods.  相似文献   

3.
2000~2005年,从猪大肠杆菌病 (colibacillosis)中分离到一些表达K88菌毛的大肠杆菌(Escherichia coli ),这些分离株只与K88 a因子单抗反应,而不与K88 b、c和d因子单抗反应。分离的菌株 (13/16)以O149为常见血清型,且全部拥有STb毒素基因,通过K88常规血清交叉吸收试验、SDS-PAGE和Western印迹,表明这些菌株不仅与K88ac参考菌株C83907制备的c因子血清反应,而且与以分离株SEC586制备且经K88ab、K88ac和K88ad参考菌株吸收后的血清也反应,表明这些分离株仍为K88ac大肠杆菌。对新近分离的SEC464、SEC525、SEC586、SEC799和EC910株及80年代我国分离的TM128株的K88主要亚单位结构基因faeG进行克隆、测序,发现新近分离株的faeG基因由846对核苷酸组成,编码菌毛主要亚单位的261个氨基酸及21个氨基酸的信号肽,比国内外报道的K88ac FaeG亚单位 (262个氨基酸)少了一个氨基酸,比K88ab、K88ad (264个氨基酸)少了3个氨基酸。TM128株的FaeG氨基酸序列与K88ac(M29375)的同源性为100.0%,SEC464、SEC525、SEC586、SEC799和SEC910株的FaeG亚单位氨基酸序列的同源性为97.7%~99.6%,它们与K88ac的同源性为94.6%~96.6%;与K88ab的同源性为90.0%~91.6%;与K88ad的同源性为87.0%~88.9%。 结果表明新分离的K88ac大肠杆菌黏附素主要亚单位已发生了部分变异。  相似文献   

4.
2006年5月~2007年3月间,对百色市所辖12个县(区)的86个养鸭场(户)多个品种的鸭群进行鸭疫里默氏杆菌(R iem erella ana tip estif er,RA)病发生和流行情况的调查。结果从186羽具有浆膜炎病变的死鸭及其同群病鸭的肝和脑中分离到62株RA可疑菌株,通过生化试验证明它们基本符合RA的特征;应用建立的PCR技术均可从这些菌株扩增到RA的特异性基因片断;通过与阳性血清所做的玻片凝集试验证明,分离出的RA菌株全部属于血清1型;在鸭疫里默氏杆菌阳性病料中,RA单一感染的占25.81%(16/62)、RA与大肠杆菌混合感染率56.45%(35/62)、RA与沙门氏杆菌混合感染率17.74%(11/62);病例中大肠杆菌单独感染占6.99%(13/186);2~5周龄雏鸭最易感染,北京鸭最易感染;药敏试验表明,大多数RA分离株对头孢类药物表现高敏,但总体上耐药性比较严重;分离株的致病性试验结果显示,RA的致病力比较强,与大肠杆菌的混合感染的致病力更强。本研究证明鸭疫里默氏杆菌病已成为百色肉鸭养殖业最常见的、危害最大的传染病之一。  相似文献   

5.
南宁市商业肉鸭鸭疫里默氏杆菌病的调查研究   总被引:4,自引:1,他引:4  
2004年12月~2005年12月间,对南宁市及其周边地区26个商业肉鸭群鸭疫里默氏杆菌(R iem erella ana tip estif er)病的发生情况进行了调查。从167羽具有浆膜炎病变的病死鸭中,应用巧克力培养基的分离培养,以及对分离株进行的生化试验,细菌外膜蛋白基因的聚合酶链式反应(PCR)扩增的鉴定,结果分离到74株鸭疫里默氏杆菌;通过玻片凝集试验鉴定,所有分离株均属于血清2型;在鸭疫里默氏杆菌阳性病料中同时分离到2 3株大肠杆菌,混合感染率高达3 1%;脑组织的分离率明显高于肝脏组织的;在2~6周龄的发病时间段里,肉鸭对鸭疫里默氏杆菌的感染表现出明显的年龄抵抗力;常用药物的敏感性试验表明大多数分离株对头孢唑啉、头孢啦啶、头孢哌酮表现高度敏感。  相似文献   

6.
pp. 865–874
Occurrence and survival of fecal-contamination indicator bacteria (coliform bacteria, Escherichia coli and Salmonella ) in various manure and compost samples collected from 23 composting facilities mostly in Kyushu were investigated by using selective media. Coliform bacteria were detected on desoxycholate agar from 11 (38%) of 29 product samples (15 cow dung manure, 4 poultry manure, 2 biosolid compost and 8 food waste compost) at a range of 102 to 106 cfu g1 dry matter. From positive samples, 21 isolates of possible coliform bacteria were purified. Among them, species of coliform bacteria ( E. coli , E. vulneria , Pantoea sp. and Buttiauxella agrestis ) were identified whereas isolates of Serratia marcescens , not coliform bacteria, were also obtained, suggesting that careful observation was necessary to avoid false positive counting due to the presence of a red colony of S. marcescens that resembled coliform bacteria. Isolates of E. coli were tested for slide aggregation with a set of antiserum against pathogenic E. coli serotypes and negative reaction was obtained for all the isolates tested. Direct detection of E. coli on Chromocult coliform agar and Salmonella on MLCB agar resulted in none and 2 (17%) of 12 samples tested, respectively. The fate of fecal-contamination indicator bacteria as above was followed during compost production on 7 cases at 6 compost facilities and 4 patterns were observed: fecal-contamination indicator bacteria 1) decreased and finally disappeared, 2) decreased once but re-growth was occurred on products, 3) decreased to some extent but remained in products, 4) was not detected throughout production. These results suggest that some fecal-contamination indicator bacteria may survive compost production and appropriate temperature control would be significant for hygiene control of manure and compost.  相似文献   

7.
The use of Fourier transform near-infrared (FT-NIR) spectroscopy and multivariate pattern recognition techniques for the rapid detection and identification of bacterial contamination in liquids was evaluated. The complex biochemical composition of bacteria yields FT-NIR vibrational transitions (overtone and combination bands) that can be used for classification and identification. Bacterial suspensions (Escherichia coli HB101, E. coli ATCC 43888, E. coli 1224, Bacillus amyloliquifaciens, Pseudomonas aeruginosa, Bacillus cereus, and Listeria innocua) were filtered to harvest the cells and eliminate the matrix, which has a strong NIR signal. FT-NIR measurements were done using a diffuse reflection-integrating sphere. Principal component analysis showed tight clustering of the bacterial strains at the information-rich spectral region of 6000-4000 cm(-1). The method reproducibly distinguished between different E. coli isolates and conclusively identified the relationship between a new isolate and one of the test species. This methodology may allow for the rapid assessment of potential bacterial contamination in liquids with minimal sample preparation.  相似文献   

8.
Twelve laboratories evaluated the Gram-Negative Identification (GNI) Card to identify members of the Enterobacteriaceae. Eighty-four isolates, previously isolated from foods, were used in the collaborative study; the isolates represented 12 genera within the Enterobacteriaceae group. Each collaborator streaked each isolate on tryptic soy agar plates for purity. In the method, plates are incubated 18-24 h at 35 degrees C. Isolated colonies are then subcultured to tryptic soy agar slants and incubated 18-24 h at 35 degrees C. An emulsion is made from the growth on the slant in 1.8 mL 0.45% sodium chloride solution. The GNI Card is filled and placed in a reader/incubator. Isolates are identified and an identification is printed. The Vitek System correctly identified 96.7% of Salmonella sp., 97.0% of Escherichia coli, and an average of 93.8% of the other enteric genera. The method using the Vitek System and GNI Card has been approved interim official first action by AOAC as a screening method for the presumptive identification of Salmonella sp., E. coli, and other Enterobacteriaceae isolated from foods.  相似文献   

9.
An UHPLC-PDA-ESI/HRMS/MS(n) profiling method was used for a comprehensive study of the phenolic components of red mustard greens ( Brassica juncea Coss variety) and identified 67 anthocyanins, 102 flavonol glycosides, and 40 hydroxycinnamic acid derivatives. The glycosylation patterns of the flavonoids were assigned on the basis of direct comparison of the parent flavonoid glycosides with reference compounds. The putative identifications were obtained from tandem mass data analysis and confirmed by the retention time, elution order, and UV-vis and high-resolution mass spectra. Further identifications were made by comparing the UHPLC-PDA-ESI/HRMS/MS(n) data with those of reference compounds in the polyphenol database and in the literature. Twenty-seven acylated cyanidin 3-sophoroside-5-diglucosides, 24 acylated cyanidin 3-sophoroside-5-glucosides, 3 acylated cyanidin triglucoside-5-glucosides, 37 flavonol glycosides, and 10 hydroxycinnamic acid derivatives were detected for the first time in brassica vegetables. At least 50 of them are reported for the first time in any plant materials.  相似文献   

10.
基因sdiA属于与群体效应相关的LuxR家族,目前已证实存在于Escherichia coli和Salmonella typhimurium基因组中。本研究从梨火疫病菌(Erwinia amylovora)中克隆到了一个sdiA的同源基因,命名为EAsdiA(GenBank登录号:AY864839),该基因与E.coli和S.typhimurium的sdiA基因在氨基酸水平上分别有45.42%和43.33%的同源性,与其它细菌的luxR同源基因的同源性更低。根据梨火疫病菌EAsdiA和其它病原细菌的luxR基因的序列比对设计了1对特异引物F-EAluxR和R-EAluxR,能够特异地检测梨火疫病菌,检测灵敏度为10个菌体,在含有梨组织浸出液中可检测到102个菌体。本研究首次从梨火疫病菌中克隆sdiA基因并作为新靶标应用于该病菌分子检测。  相似文献   

11.
从日本九州地区23处堆肥化设施采集了以牛粪、鸡粪、污泥和餐厨垃圾为原料的堆肥试样29个,调查了大肠菌群等粪便污染指标菌的残留状况。结果表明,使用DESO培养基有11个试样被检出大肠菌群,检出率达38%,菌数在102~106 cfu.g-1的范围。使用API 20E鉴定系统对4个试样中的21个分离纯化菌株进行了菌种鉴定,发现有大肠菌群属的Escherichia coli、Escherichiavulneris、Pantoea sp.和Buttiauxella agrestis,以及非大肠菌群属的Serratia marcescens等肠内细菌科细菌。采用血清凝集试验对5株E.coli的病原性检测结果均为阴性。进一步针对6处堆肥设施的堆肥发酵过程中大肠菌群的消长追踪发现,大肠菌群数从原料到成品出现了逐渐减少直至消失、暂时消失、未完全消失、完全未检出4种走势,显示即使堆肥发酵温度在60℃以上,大肠菌群也有可能通过交叉污染等途径残留在堆肥成品中。  相似文献   

12.
The prevalence and diversity of antibiotic resistant enterococci populations in samples collected four times from urban sewage treatment plant in Tehran, Iran between June 2005 and July 2006 were studied. Filtered samples were grown on mEnterococci medium containing 4 μg/ml vancomycin after which the enterococci isolates were identified to the species level. All strains were then tested for their resistance against nine antibiotics. Of the 131 isolates, 98 (75%) isolates were identified as Enterococcus gallinarum, followed by 24 (18%) and 9 (7%) for E. faecium and E. casseliflavus, respectively. All E. gallinarum isolates carried vanC1 gene with 64 (65%) and 14 (14%) isolates concomitantly harboured either vanA or vanB gene, respectively. Some E. casseliflavus concomitantly harboured vanA and vanC2 or vanB and vanC2. Typing the total enterococci isolates with a high resolution biochemical fingerprinting method showed a high diversity (D i?=?0.91). We have shown by biochemical fingerprinting the presence of highly diverse glycopeptide resistant E. gallinarum and E. casseliflavus that have captured vanA and vanB genetic determinants under natural conditions. To our knowledge this is the first report in this geographical region showing high frequency antibiotic resistant enterococcal populations in particular E. gallinarum carrying assorted vancomycin resistance genes.  相似文献   

13.
The defined substrate technology (DST) method is a reagent system designed to enumerate specific target microbes(s) from a mixture of bacteria. The system simultaneously enumerates total coliforms and Escherichia coli directly from a water sample. The reagent contains o-nitrophenyl-beta-D-galactopyranoside (ONPG), which is hydrolyzed by total coliforms to produce a yellow chromogen, and 4-methylumbeilliferyl-beta-D-glucuronide (MUG), which is hydrolyzed and fluoresces when E. coli organisms grow. Noncoliform bacteria are suppressed and cannot metabolize the indicator nutrients. Nine laboratories participated in a field evaluation of the method, which covered a wide range of surface and subsurface water sources and water-processing modalities, including the examination of natural samples. The DST system was compared to multiple-tube fermentation (MTF) (quantitative) and presence-absence (P-A) (qualitative) Standard Methods formats. Comparison of water samples from natural sources by using the most probable number (MPN) procedure showed that the DST test was equivalent to the currently used MTF test. Results from the DST and the qualitative P-A procedure showed that these tests agreed with each other in 94% of the water samples analyzed. Specificity of the DST method was established by subculturing a species consistent with a total coliform or E. coli from each positive tube. Eight laboratories participated in a collaborative study of the method. Each laboratory received 3 concentrations of E. coli (organisms/100 mL); 10 (low); 60 (medium); and 120 (high). The DST test was inoculated from a split sample of each bacterial density in parallel with Standard Methods brilliant green lactose broth.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

14.
Low availability of phosphorus(P) is a major constraint for optimal crop production, as P is mostly present in its insoluble form in soil. Therefore,phosphate-solubilizing bacteria(PSB) from paddy field soils of the Indo-Gangetic Plain, India were isolated, and their abundance was attempted to be correlated with the physicochemical characteristics of the soils. Ninety-four PSB were isolated on Pikovskaya's agar medium, and quantitative phosphate solubilization was evaluated using NBRIP medium. The isolates solubilized P up to a concentration of 1 006 μg mL~(-1) from tricalcium phosphate with the secretion of organic acids. These isolates were identified by 16 S rRNA gene sequence comparison, and they belonged to Gammaproteobacteria(56 isolates),Firmicutes(28 isolates), Actinobacteria(8 isolates), and Alphaproteobacteria(2 isolates). Phylogenetic analysis confirmed the identification by clustering the isolates in the clade of the respective reference organisms. The correlation analysis between PSB abundance and physicochemical characteristics revealed that the PSB population increased with increasing levels of soil organic carbon, insoluble P, K~+, and Mg~(2+). The promising PSB explored in this study can be further evaluated for their biofertilizer potential in the field and for their use as potent bio-inoculants.  相似文献   

15.
基于图像的昆虫远程自动识别系统的研究   总被引:9,自引:3,他引:6  
只有对害虫进行鉴定才能在农业生产中对害虫进行有目的的防治,而对昆虫进行鉴定只有少数分类专家才能完成,鉴定需求的日益增加与专家相对较少形成了一对尖锐的矛盾.该文的研究尝试为该矛盾的解决提供一条新的思路:在标准方法下获取昆虫图像,并经由Internet网络上传给自动种类识别系统服务器,从而实现远程识别.系统首先对昆虫图像进行基于形状和颜色特征值的提取.昆虫图像的形态特征值由矩形度、延长度、球状型、叶状型、似圆度和7个Hu不变矩等12个特征值组成,颜色特征值由红、绿、蓝、灰度真方图及基于红、绿的二维色度直方图特征值分别组成,然后建立径向基神经网络分类器,每一特征向量由独立的径向基神经网络做为分类器,最终识别由每个分类器识别结果的线性组合而成.采用该系统对16种昆虫进行了测试,每种昆虫取40个样本,20个用做训练、20个用做测试,准确率达到96%以上.  相似文献   

16.
Meats need to be heated to inactivate foodborne pathogens such as Escherichia coli O157:H7. High-temperature treatment used to prepare well-done meats increases the formation of carcinogenic heterocyclic amines (HCAs). We evaluated the ability of plant extracts, spices, and essential oils to simultaneously inactivate E. coli O157:H7 and suppress HCA formation in heated hamburger patties. Ground beef with added antimicrobials was inoculated with E. coli O157:H7 (10(7) CFU/g). Patties were cooked to reach 45 °C at the geometric center, flipped, and cooked for 5 min. Samples were then taken for microbiological and mass spectrometry analysis of HCAs. Some compounds were inhibitory only against E. coli or HCA formation, while some others inhibited both. Addition of 5% olive or apple skin extracts reduced E. coli O157:H7 populations to below the detection limit and by 1.6 log CFU/g, respectively. Similarly, 1% lemongrass oil reduced E. coli O157:H7 to below detection limits, while clove bud oil reduced the pathogen by 1.6 log CFU/g. The major heterocyclic amines 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) were concurrently reduced with the addition of olive extract by 79.5% and 84.3% and with apple extract by 76.1% and 82.1%, respectively. Similar results were observed with clove bud oil: MeIQx and PhIP were reduced by 35% and 52.1%, respectively. Addition of onion powder decreased formation of PhIP by 94.3%. These results suggest that edible natural plant compounds have the potential to prevent foodborne infections as well as carcinogenesis in humans consuming heat-processed meat products.  相似文献   

17.
本研究利用SL培养基从蚯蚓粪中分离到54株具有产酸性能的菌株,并以E.coli O157:H7(EDL933株)作为指示菌株,采用点种法检测分离菌株的抑菌活性。结果表明其中6个菌株对指示菌具有拮抗作用,通过形态特征,结合16S rDNA序列分析,初步鉴定该6个菌株分别为食物魏斯特菌(Listeria welshimeri)、乳酸片球菌(Pediococcus acidilactici)、短乳杆菌(Lactobacillus brevis)和格氏乳球菌(Lactococcus garvieae)。分离到的乳酸菌对E.coli O157:H7(EDL933株)具有显著的抑制作用,发酵温度和初始pH值影响发酵液的抑菌作用,优化环境因子可以促进拮抗菌对E.coli O157:H7的抑制作用。本研究为进一步分离抗菌产物用于人畜共患病的预防和治疗提供了理论依据。  相似文献   

18.
Wild Amazonian basil Ocimum micranthum Willd. (O. campechianum Mill.) Labiatae essential oil was analyzed by GC and GC-MS: 31 compounds were identified. The main components were eugenol (46.55 +/- 5.11%), beta-caryophyllene (11.94 +/- 1.31%), and beta-elemene (9.06 +/- 0.99%), while a small amount of linalool (1.49 +/- 0.16%) was detected. The oil was tested for its in vitro food-related biological activities and compared with common basil Ocimum basilicum and Thymus vulgaris commercial essential oils. Radical scavenging activity was evaluated employing 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The oil exerted a good capacity to act as a nonspecific donor of hydrogen atoms or electrons when checked in the diphenylpicrylhydrazyl assay, quenching 76,61 +/- 0.33% of the radical, with values higher than those reported by reference oils. In the beta-carotene bleaching test, the oil provided an antioxidant efficacy comparable with that of O. basilicum and T. vulgaris essential oils. These data were confirmed by photochemiluminescence, where the oil showed a remarkable antioxidant capacity (2.39 +/- 0.1), comparable to that of Trolox and vitamin E, and higher than the other essential oils. Antibacterial activity of O. micranthum essential oil was evaluated against Gram positive and Gram negative bacterial strains. The oil showed a dose-dependent antifungal activity against pathogenic and food spoiling yeasts.  相似文献   

19.
Pseudomonas putida GM6多聚磷酸盐激酶(ppk)基因的克隆及表达   总被引:4,自引:0,他引:4  
以一株高效聚磷菌Pseudomonas putida GM6为研究材料.为获得其多聚磷酸盐激酶(polyphosphate kinase,ppk)基因,并验证该基因在磷酸盐转运系统中的作用,根据已报道的ppk基因保守区域设计引物,从其总DNA中成功扩增到ppk基因的部分片段(约528 bp).随后采用快速染色体步移方法(Self-formed adaptor PCR,SEFA-PCR)技术扩增片段的上下游基因序列,将三个序列拼接,用OMIGA软件分析其ORFs,推测ppk基因全长为2 220 bp(GenBank accession number DQ133537).构建的多聚磷酸盐激酶表达菌株E. coli BL21(DE3)/ pET29a-ppk经IPTG诱导后3 h时,明显出现分子量约为81 kDa的表达产物.且表达菌株在12 h时的磷去除率高达80%(对照菌株的磷去除率仅为18%),远高于已报道的40%的去除率.这表明ppk基因在E. coli中的过量表达,导致了E. coli菌体中poly-P的大量聚集,从而大大去除了培养基中的磷酸盐.  相似文献   

20.
Fushimi sweet pepper, "Fushimi-togarashi", is one of the "Kyo-yasai", traditional vegetables, in Kyoto, Japan. The chloroform fraction of Fushimi sweet pepper showed bioantimutagenicity on UV induced mutation in Escherichia coli B/r WP2. The bioantimutagen was purified with silica gel chromatography and identified as 2, 4-nonadienal (ID(50) = 20 microg/plate) on the basis of GC retention time and EI-MS spectrum of authentic 2,4-nonadienal. The sweet pepper also contained a known bioantimutagen, benzaldehyde (ID(50) = 2 mg/plate). Additive bioantimutagenicity was also observed by 2, 4-nonadienal with benzaldehyde. 2,4-Nonadienal did not show bioantimutagenicity in an UV excision repair deficient strain, E. coli B/r WP2s uvrA(-)(). Furthermore no delay of the first cell division after UV irradiation was observed in E. coli B/r WP2. These results indicate that the bioantimutagenic activity of 2, 4-nonadienal on UV mutagenesis might depend on the excision repair system in E. coli B/r WP2.  相似文献   

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