人产肠毒素大肠杆菌ST、LT—B肠毒素基因融合的研究

将人产肠毒素大肠杆菌(ETEC),编码耐热肠毒素(ST)的基因片段与编码不耐热肠毒素B亚基(LT-B)的基因进行融合,并在此基础上进行不同数目ST基因的串联,ELISA检测融合基因表达蛋白产物观察到ST与LT-B之间存在着相互影响。ST的检测滴度随基因串联个数增加而逐渐升高,而LT的ELISA滴度则减弱。说明了ST可以通过基因串联提高表达产物抗原活性,这为产肠毒素大肠杆菌多价疫苗的研制提供了重要的研究基础。     

耐甲氧西林金黄色葡萄球菌及其肠毒素的检测和耐药性分析

目的评价乳胶结合试验检测耐甲氧西林金黄色葡萄球菌(MRSA)及其肠毒素(SE),并进行耐药性分析.方法收集130株金黄色葡萄球菌临床分离株,通过药敏试验将其分为耐甲氧西林金黄色葡萄球菌和甲氧西林敏感金黄色葡萄球菌(MSSA),用反向间接血凝试验(RPHA)检测金黄色葡萄球菌肠毒素.结果67株MR-SA产肠毒素,19株MSSA产肠毒素,MRSA产肠毒素率为100%,MSSA产肠毒素率为30%.结论实验室应重视金黄色葡萄球菌肠毒素的检测.     

人产肠毒素大肠杆菌ST、LT—B肠毒素基因融合的研究

将人产肠毒素大肠杆菌(ETEC),编码耐热肠毒素(ST)的基因片段与编码不耐热肠毒素B亚基(LT—B)的基因进行融合,并在此基础上进行不同数目sT基因的串联,ELJlSA检测融合基因表达蛋白产物观察到sT与LT－B之间存在着相互影响。ST的检测滴度随基因串联个数增加而逐渐升高,而LT的ELISA滴度则减弱。说明了ST可以通过基因串联提高表达产物抗原活性,这为产肠毒素大肠杆菌多价疫苗的研制提供了重要的研究基础。     

产毒金黄色葡萄球菌鉴定及在百叶中生长与产毒特性分析

从生鲜食品中分离鉴定得到产肠毒素SEA的金黄色葡萄球菌（Staphyloccocus aureus）菌株,研究金黄色葡萄球菌在百叶中的生长变化及产肠毒素特性。将不同浓度的金黄色葡萄球菌同时接种到减菌（样品组）和未减菌（对照组）百叶中,高、低初始接种量分别控制在4.0 lg(cfu/g)和2.0 lg(cfu/g)左右,并定时追踪不同储存温度条件下百叶中金黄色葡萄球菌的活菌数及产毒情况。金黄色葡萄球菌肠毒素的检测采用商业化的ELISA试剂盒。在37 ℃储存过程中,金黄色葡萄球菌在样品组中不但能生长良好并在18 h后菌落数可达到7.0 lg(cfu/g)并被检测出肠毒素,对照组中金黄色葡萄球菌生长良好菌落数可达到7.0 lg(cfu/g)却未检测出产肠毒素。在25、15和5 ℃储存过程中,样品组和对照组百叶都未检测出肠毒素。温度对金黄色葡萄球菌在百叶中是否产肠毒素具有决定性的作用,且百叶中本身存在的杂菌可能可以抑制金黄色葡萄球菌产肠毒素。     

葡萄球菌肠毒素A单克隆抗体的制造

<正>葡萄球菌属食物中毒是一种主要的公害,美国细菌引起的食物中毒占第二位。人们用特异抗体、通常是兔抗血清进行血清学反应来检测葡萄球菌肠毒素A、B、C、D、E(以下简称SEA、B、C、D、E)。检测这些葡萄球菌肠毒素最普通的方法包括使用微量凝胶扩散系统。自从这一检测系统建立以来,人们成功地使之用于食物中毒病例。     

嗜水气单胞菌菌落多重PCR方法的建立及应用

[背景]嗜水气单胞菌(Aeromonas hydrophila)对水产动物、畜禽和人类均有致病性.基因表达的溶血素、气溶素和肠毒素是重要毒力因子,在致病性嗜水气单胞菌早期检测及防治中尤为重要.目前采用菌落直接提取DNA用于多重PCR研究的相关报道较少.[目的]基于菌落PCR方法建立针对嗜水气单胞菌溶血性基因、肠毒素基因...     

快速检测金黄色葡萄球菌肠毒素A基因方法的建立与应用

目的建立一种快速准确定量检测金黄色葡萄球菌肠毒素A的方法。方法以femB、SEA基因分别作为金黄色葡萄球菌菌株、肠毒素A的靶序列,设计合成引物和TaqM an探针;收集腹泻患者大便标本分离的金黄色葡萄球菌68株,并定量检测其肠毒素A。结果TaqM an探针荧光聚合酶链反应检测金黄色葡萄球和肠毒素A的灵敏度均为1.0×102拷贝。68株金黄色葡萄球菌中检出产肠毒素A菌株11例(16.2%,11/68),CT值为13.5~20.6。结论TaqM an探针荧光聚合酶链反应能够准确快速检测金黄色葡萄球菌肠毒素A。     

金黄色葡萄球菌肠毒素研究进展

金黄色葡萄球菌肠毒素是由金黄色葡萄球菌分泌的一类胞外毒素,其中传统型为SEA-SEE,具有催吐活性。目前也发现了一些催吐活性未知的新类型肠毒素,命名为SEls。所有肠毒素的分子量都在22-28KDa之间,由单一肽链组成,其稳定性好,可耐大多数蛋白水解酶。金黄色葡萄球菌肠毒素具有较强的超抗原特性,可促使T淋巴细胞大量增殖,同时表现出对组织相容性复合物Ⅱ类分子(MHCⅡ)等位基因的不同偏爱性。超抗原的产生和调控依赖agr系统,同时也受其它因素的调控,如一些氨基酸。因此在免疫治疗中具有着重要作用。本文从性质、组成、结构、功能、检测方法等方面对金黄色葡萄球菌肠毒素进行简要介绍,为肠毒素的研究提供依据。     

人毒素原性大肠杆菌肠毒素ST、LT-B基因的融合

将毒素原性大肠杆菌(ETEC)编码耐热肠毒素(ST)的基因片段与编码热敏肠毒素B亚基(LT—B)的基因进行融合,并在此基础上进行不同数目ST基因的串联。ELISA检测融合基因表达蛋白产物,观察到ST与LT-B之间存在着相互影响。ST的检测滴度随基因串联个数增加而逐渐升高,而LT的ELISA滴度则减弱。本研究说明ST可以通过基因串联提高表达产物抗原活性。这为毒素原性大肠杆菌多价疫苗的研制提供了重要的研究基础。     

多重PCR方法快速检测4种主要致腹泻性大肠埃希菌

肠毒素性大肠埃希菌(ETEC)、肠致病性大肠埃希菌(EPEC)、肠出血性大肠埃希菌(EHEC)和侵袭性大肠埃希菌(EIEC)是引起腹泻的主要大肠埃希菌,威胁着食品安全和人类健康,建立同时检测4种致腹泻性大肠埃希菌的方法具有重要意义。基于ETEC LT肠毒素基因、EPEC bfpA基因、EHECO抗原基因和EIEC侵袭性质粒特异性基因,设计了4对特异性引物,通过对单一PCR反应条件的优化建立了快速检测4种主要致腹泻性大肠埃希菌的多重PCR方法,彼此之间无交叉反应。该多重PCR方法具有良好的特异性和灵敏性,对24株致病菌进行检测,所试4株致腹泻性大肠埃希菌均为PCR阳性,其他菌株则为阴性。实践证明,利用所建立的多重PCR方法对124份肉类、奶类制品及人工污染样品等进行检测,检出15份阳性,与国标(GB4789.6-1994)检测结果相同。结果表明本文建立的多重PCR方法可用于ETEC、EPEC、EHEC和EIEC的单一或混合感染的鉴别诊断及食品安全风险评估,具有良好的实用性。     

Interlaboratory proficiency tests for the detection of staphylococcal enterotoxin type A in food

Staphylococcal enterotoxins (SEs) are among the leading causes of food intoxications, affecting consumer health even in nanogram (ng) amounts. In the European Union, certain food safety criteria are specified, including the absence of SEs in cheeses, milk powder and whey powder. Until 2019, the analytical reference method used was the European Screening Method, which was replaced by EN ISO 19020. For the official laboratories involved in food control, the German Reference Laboratory for coagulase-positive staphylococci including Staphylococcus aureus organized three interlaboratory proficiency tests (ILPTs) to detect SE type A in food during the years 2013–2018. The selected food products (cream cheese and vanilla pudding) were successfully tested beforehand with regard to easy handling, homogeneity and stability of the added toxin. In 2013, ILPT participants overall were not competent in detecting SE type A in food. The following factors were identified to improve the performance: (i) concentration of sample extract using dialysis; (ii) selection of a sensitive detection kit; and (iii) proper sample handling. By taking these factors into account and instructing and training the laboratories, their competence greatly improved. In 2018, all performance criteria (specificity, sensitivity and accuracy) were >90%, even at very low concentrations of SE type A of approximately 0·01 ng g^–1 food.     

A single-stranded DNA aptamer that selectively binds to Staphylococcus aureus enterotoxin B

The bacterium Staphylococcus aureus is a common foodborne pathogen capable of secreting a cocktail of small, stable, and strain-specific, staphylococcal enterotoxins (SEs). Staphylococcal food poisoning (SFP) results when improperly handled food contaminated with SEs is consumed. Gastrointestinal symptoms of SFP include emesis, diarrhea and severe abdominal pain, which manifest within hours of ingesting contaminated food. Immuno-affinity based methods directly detect, identify, and quantify several SEs within a food or clinical sample. However, the success of these assays depends upon the availability of a monoclonal antibody, the development of which is non-trivial and costly. The current scope of the available immuno-affinity based methods is limited to the classical SEs and does not encompass all of the known or emergent SEs. In contrast to antibodies, aptamers are short nucleic acids that exhibit high affinity and specificity for their targets without the high-costs and ethical concerns of animal husbandry. Further, researchers may choose to freely distribute aptamers and develop assays without the proprietary issues that increase the per-sample cost of immuno-affinity assays. This study describes a novel aptamer, selected in vitro, with affinity to staphylococcal enterotoxin B (SEB) that may be used in lieu of antibodies in SE detection assays. The aptamer, designated APT(SEB1), successfully isolates SEB from a complex mixture of SEs with extremely high discrimination. This work sets the foundation for future aptamer and assay development towards the entire family of SEs. The rapid, robust, and low-cost identification and quantification of all of the SEs in S. aureus contaminated food is essential for food safety and epidemiological efforts. An in vitro generated library of SE aptamers could potentially allow for the comprehensive and cost-effective analysis of food samples that immuno-affinity assays currently cannot provide.     

细菌生物被膜与食品生物危害

食源性病原菌生物被膜是威胁食品安全的一个重大隐患。生物被膜一旦在食品加工过程中形成,易引起相当严重的交叉污染和加工后污染,产生极大危害,从而导致一系列严重的健康问题。本文就食源性病原菌生物被膜的分布、形成机制以及防治措施进行了综述。     

EFFECTS OF FASTING AND FEEDING ON SERUM UREA AND SERUM CREATININE LEVELS IN POLAR BEARS

Polar bears were held in captivity and fasted for an average of 38 d prior to being fed for three days. Blood samples were collected prior to feeding and then at one and three days post-feeding in 1984 and at one, four, and seven days post-feeding in 1985. The ratio of serum urea to serum creatinine varied significantly in 1984, rising from a pre-feeding mean of 11.0 (SE = 2.6) to 32.0 (SE = 3.2) on the first day post-feeding and then dropped to 22.8 (SE =3.2) in the last sample. In 1985, the ratio of serum urea to serum creatinine increased from a pre-feeding mean of 15.8 (SE = 2.3) to 61.2 (SE = 10.6) after three days of feeding and dropped to a mean of 29.2 (SE = 5.1) seven days after feeding ended. Serum urea levels varied over the study period in both years. No significant variation in serum creatinine levels was found in 1984, but in 1985, serum creatinine levels demonstrated significant variation, declining from the pre-feeding mean of 1.83 mg/dl (SE = 0.29) to 0.96 mg/dl (SE = 0.12) in the last sample.
The findings suggest that polar bears can have a low serum urea to serum creatinine ratio, similar to that found in hibernating black bears, or higher ratios after feeding. Polar bears can rapidly return to a fasting serum urea to creatinine ratio after food is withheld. Polar bears may demonstrate urea conservation, similar to that found in black bears, and may be able to move between a fasting and a feeding metabolism based on food availability throughout the year, an adaptation to life on the labile sea ice.     

Staphylococcal enterotoxin superantigens

Staphylococcal enterotoxins (SE) are a family of structurally related proteins that are produced by Staphylococcus aureus. They play a role in the pathogenesis of food poisoning and are the most potent activators of T lymphocytes known. The receptors for SE on antigen-presenting cells are major histocompatibility complex class II molecules. Recent studies have shown that a complex of SE and major histocompatibility complex class II molecules is required for binding to the variable region of the T cell antigen receptor beta-chain. SE mitogenic activity is dependent on induction of interleukin 2, which may be intimately involved in the mechanism of SE toxicity. The minor lymphocyte-stimulating "endogenous" self-superantigen has recently been shown to be a retroviral gene product, so that this too is apparently a microbial superantigen. An understanding of the mechanism of action of these microbial superantigens has implications for normal and pathological immune functions.     

Main lipid classes in some species of deep-sea corals in the Newfoundland and Labrador region (Northwest Atlantic Ocean)

Corals contain large quantities of lipids in their tissues; these lipids may be either structural or for storage. Little information is available about the lipid content of deep-sea corals, as well as ratios of main lipid classes. In this study, lipid percentages of 81 deep-sea specimens were measured and the presence of six major classes, including sterols (STEROLS), free fatty acids (FFA), triacylglycerols (TG), monoalkyldiacyl glycerol (MADAG), wax (WAX), and sterol esters (SE), was assessed. Deep-sea corals had fewer lipids than their shallow water counterparts. Decision-tree analysis revealed a link between coral groups and total lipid percentages, showing that species within the same group were characterized by similar lipid amounts. Depth did not seem to impact the total lipid percentages, suggesting that deep-sea corals adapt to the differential access to food by changing the proportion of lipid classes while maintaining equivalent lipid levels. In deep-sea species, similar to their shallow water counterparts, energy seems to be stored as neutral lipids (wax esters and triacylglycerols), with the notable difference that a high proportion of MADAG is present. These compounds are less rich in energy than TG. Depth trends were found for FFA, TG and SE with an increase in percentages after 800 m suggesting a potential need for storage due to decreased food availability. A subsequent decrease after 1,100 m was observed for FFA and TG but a more detailed investigation is warranted as the number of specimens acquired from these depths was less than 20. It is nonetheless a surprising result as increased storage is expected when food sources are sparse.     

The use of a bacteriophage cocktail as a biocontrol measure to reduce Salmonella enterica serovar Enteritidis contamination in ground meat and goat cheese

We evaluated the effectiveness of phages on meats and goat cheese contaminated with Salmonella Enteritidis (SE). In meats, reductions of SE were observed during the whole experiment, while in goat cheese a reduction was only observed at day 3. We discuss the relevance of phages as a biocontrol in food.     

Lack of Staphylococcal Enterotoxin Production among Strains of Staphylococcus aureus from Bovine Mastitis in Denmark

Staphylococcal enterotoxins (SE’s) are a group of small exoproteins produced by some strains of Staphylococcus aureus. The SE’s, designated A to E according to their antigenic specificities, are important causes of food poisoning worldwide. Milk and dairy products are frequently associated with S. aureus enter-otoxin food poisoning, and it is supposed that infected milk from mastitic animals constitute the main source of enterotoxigenic S. aureus of animal origin (Bryon 1983, Gilmour & Harvey 1990, Bergdoll 1989). Indeed, S. aureus is the most common cause of bovine mastitis worldwide, and if mastitis strains produce SE this makes up an enormous reservoir of potential enterotoxin producers. The production of SE by S. aureus isolated from bovine mastitis have been investigated in several countries (Matsunaga et al. 1993, Kenny et al. 1993, Olson et al 1970, Orden et al. 1992, Olsvik et al. 1981, Adekeye 1980, Garcia et al. 1980, Abbar 1986, Harvey & Gilmour 1985). Since no studies have been performed on the prevalence of enterotoxigenic strains of S. aureus isolated from bovine mastitis in Denmark, a well characterized collection of S. aureus (Aarestrup et al. 1995) was investigated with respect to this property.     

细菌群体感应及其在食品变质中的作用

食品相关细菌引起的生物被膜形成和食品变质是食品工业中的重大问题。研究表明细菌群体感应(Quorum sensing,QS)与被膜形成、食品腐败变质密切相关。重点对细菌产生的各种QS信号分子及其在被膜形成的作用和被膜在食品工业中的重要性做了介绍。QS信号分子与食品变质密切相关,故对QS抑制剂作为新型食品防腐剂以延长储存期限及加强食品安全的前景进行了概述。     

Breeding in the monsoon: semi-annual reproduction in the Seychelles warbler ( Acrocephalus sechellensis )

Despite the absence of pronounced changes in day length, there is considerable climatological seasonality in the tropics. Its expression can be complex like in the monsoon climate of the Indian Ocean Islands. The land mass distribution on both sides of the equator leads to seasonal changes in prevailing wind direction and seasonal patterns in food supply, which are distinct between different coasts. Here we assess that territory quality of the Seychelles warbler, endemic to the small island of Cousin peaks during the South East monsoon (April to September) in most of the island. The Southeastern wind-exposed side is exceptional by being generally poor and having slightly increased food supply in the opposite season (NW monsoon; October to March). In response, the warblers in most territories breed primarily during the SE season, while on the SE shores they tend to breed in both seasons. The result is a semi-annual periodicity in breeding. Individual pairs can breed in 6 months intervals. In the poorer territories on the SE shore most of them indeed do this, while in the richer central habitats most pairs restrict themselves to the best (SE) season. The frequency of breeding thereby increases with territory quality in the SE season, and decreases with quality in the NW season. Withholding from breeding in the NW season despite the high food supply may have long-term advantages for the birds occupying the richer habitat. Such flexible adjustment of seasonality to local circumstances requires not only an immediate response mechanism but also access to accurate information about time of year. The subtle sensitivity of tropical songbirds to photoperiod, demonstrated so convincingly by Eberhard Gwinner (Biological rhythms, Hokkaido University Press, Sapporo, pp19–40, 2005), may well have profound adaptive meaning in taking temporally optimized reproductive decisions in response to both food and time of year.