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1.
Purification of an extracellular protein exhibiting the vascular permeability activity produced by Bacillus cereus was performed by ammonium sulfate precipitation followed by chromatography on DE-32 cellulose, Sephadex G-100, and Sephadex G-75. The purified protein was found to be electrophoretically and antigenically almost homogeneous although it contained a trace of contaminant. The molecular weight of the protein was calculated to be 45,000 by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The purified protein showed vascular permeability activity and mouse lethal toxicity, and caused fluid accumulation in ligated mouse intestinal loops, whereas it did not show any hemolytic and lecithinase activities. From these findings, the purified protein is suggested to be an enterotoxin (or a diarrheagenic toxin) responsible for diarrhea caused by B. cereus in a diarrheal-type food poisoning.  相似文献   

2.
Bovine coronavirus isolated from calf faeces diseased with gastroenteritis and passaged to colostrum-free calves agglutinated mouse and rat erythrocytes. The agglutination reaction depended on temperature and took place only at a temperature of 4 degrees C. At a temperature of 37 degrees C the agglutinate broke down within 15 minutes. The coronavirus could be detected by the haemagglutination test in the contents of the small and large intestines and in the faeces of experimentally and naturally infected calves. The agglutination capacity of mouse erythrocytes was not affected by careful fixation of these erythrocytes with formalin and subsequent lyophilization and remained unchanged for as long as 52 weeks of storage at a temperature of 4 degrees C. It was demonstrated by a comparative examination of 182 samples of the faeces of calves suffering from diarrhoea that haemagglutination test was as sensitive as electron microscopy.  相似文献   

3.
Vacuole response in HEp-2 cells was induced with culture supernatants of Bacillus cereus strains isolated from outbreaks of vomiting- and diarrheal-type food poisoning grown in rice flour and laboratory media. High vacuole response was obtained with culture supernatants of B. cereus strains isolated from vomiting-type food poisoning grown in cooked rice suspension or on a cooked rice plate, whereas no response was obtained with those of the same strains grown in brain heart infusion and trypto-soya broth media. The vacuole activity appeared only after spore formation of B. cereus. The activity was stable to proteolytic enzymes, heating, and exposing to pH 2.0 and 11.0. Of 124 strains isolated from B. cereus food poisoning that were tested, the vacuole activity was observed by 68 of 110 (61.8%) of the strains isolated from the vomiting-type food poisoning but not by all strains (14 strains) from diarrheal-type ones. Moreover, the vacuole response in the HEp-2 cells was found to be induced by 56 of 76 (73.7%) of the serotype H-1 strains isolated from vomiting-type food poisoning.  相似文献   

4.
Certain properties of 22 Bacillus cereus strains isolated from different foods and food poisoning episodes were investigated in order to evaluate possible différences between strains isolated from diarrhoeal and vomiting type food poisoning outbreaks.None of the strains isolated from vomiting type episodes produced acid from salicin and mannose, whereas 80 and 40 % of the strains from diarrhoeal type outbreaks were positive, respectively. No association between the antibiotic sensitivity pattern or the fatty acid composition and the source of a strain could be found, although some strains differed from the general pattern of B. cereus in some instances. No significant differences in the production of the skin factor between strains isolated from the two types of outbreaks were found either. The findings of this study support the observation that the food environment itself essentially affects the enterotoxin formation of B. cereus.  相似文献   

5.
Many aspects of the biology and epidemiology of Trichinella infection in the horse are poorly understood, including survival of Trichinella spp in horse muscle. In this study, we have assessed the freeze tolerance of T. spiralis in horse meat stored at 5, -5, and -18 degrees C for 1 day to 24 weeks. Results demonstrate a steady reduction in the number of live ML recovered from the cold stored meat samples. On Day 1, recovery of live larvae had been reduced by 18.6%, 50.1%, and 37.2%, and by 4 weeks, recovery of larvae had been reduced by 65.4%, 66.5%, and 96.2% in samples stored at 5, -5, and -18 degrees C, respectively. Infectivity results (measured as reproductive capacity index (RCI)) from mice inoculated with larvae recovered from non-frozen meat samples at day 0 was 23.5. Following storage at -18 degrees C for one and two days, the RCIs were 2.09 and 0.99, respectively. Small numbers of infective larvae were still present in meat samples stored at -18 degrees C for 4 weeks. The RCI of ML recovered from meat samples stored at -5 degrees C was 14.99 and 6.36 at 2 weeks and 4 weeks respectively; the RCI of samples stored at 5 degrees C was 23.1 at 8 weeks, and fell rapidly thereafter (12 week RCI 1.33; 0 at 24 weeks). These data demonstrate that infective T. spiralis, a non-freeze tolerant species, can survive for at least 4 weeks in horse tissue frozen at -5 or -18 degrees C, and that the numbers of infective larvae decrease substantially by day 2 at -18 degrees C and by week 4 at -5 degrees C.  相似文献   

6.
To investigate the mode of action of Clostridium perfringens epsilon-toxin, MDCK cells were treated with purified toxin and incubated at 37 degrees C for up to 24h. Exposure to epsilon-toxin caused a time-dependent decrease in cell-cell and cell-substrate interactions. After 30min of treatment retraction of the cell body and the emission of filopodia were detectable in a number of cells. Longer exposure resulted in cell rounding and cell blebbing which reached a maximum after 5h of toxin treatment. A parallel modification in the cytoskeleton was also detected. Actin marginalization and the entanglement of microtubules and intermediate filaments were observed by fluorescence microscopy after 30min of toxin exposure. Functional alterations of the plasma membrane of MDCK cells were assessed by flow cytometry. After 10 or 30min of intoxication an increase in cell volume was detected, indicating an alteration in plasma membrane permeability. These findings provide evidence for cytoskeletal changes and plasma membrane functional alterations in the in vitro cell response to C. perfringens epsilon-toxin.  相似文献   

7.
The post-thaw motility and the acrosome integrity of semen from 4 boars frozen with a programmable freezing machine, in mini (0.25 ml) and maxi (5 ml) plastic straws and in 10 x 5 cm Teflon FEP-plastic bags (0.12 mm thick, 5 ml), were compared. The freezing of the semen was monitored by way of thermo-couples placed in the straws and the bags. Three freezing programmes were used, namely A: from +5 degrees C, at a rate of 3 degrees C/min, to -6 degrees C, held for 1 min at -6 degrees C, and followed by a cooling rate of 20 degrees C/min to -100 degrees C; B: a similar curve except that there was no holding time at -6 degrees C and that the cooling rate was 30 degrees C/min, and C: from +5 degrees C to -100 degrees C, with a cooling rate of 35 degrees C/min, followed by storage in liquid N2. Despite the freezing curve assayed, both the mini-straws and the bags depicted much shorter freezing point plateaus as compared to the maxi-straws. Post-thaw sperm motility as well as the amount of normal apical ridges were equally significantly higher when semen was frozen in mini-straws or in bags than in maxi-straws. Significant differences in these post-thawing parameters were obtained between the freezing curves used. The stepwise freezing procedure A appeared as the best alternative for boar semen, considering this in vitro evaluation.  相似文献   

8.
蜡样芽孢杆菌是一种可引起食物中毒的常见食源性细菌,属于革兰氏阳性的条件致病菌,广泛存在于土壤、空气、水及植物源、动物源加工的食品中。近年发现也能感染包括人在内的多种动物,是一种人兽共患性细菌,特别是能够引起人畜肠道疾病,导致腹泻型或呕吐型食物中毒。快速准确检测蜡样芽孢杆菌是控制其污染和感染后治疗的关键环节。作者对蜡样芽孢杆菌的检测方法进行了全面详细的总结,主要包括传统检测方法、普通PCR、多重PCR（mPCR）、实时荧光定量PCR（Real-time PCR）、叠氮溴化丙锭-定量PCR（PMA-qPCR）、微滴数字PCR技术（ddPCR）、环介导等温扩增（LAMP）及酶联免疫吸附测定（ELISA）。从传统方法到新兴技术,涉及传统检测技术、分子生物学检测和免疫学检测技术,作者主要总结了各方法的原理、检测范围,并对各方法的优缺点进行了比较。这些检测方法的灵敏度、精确度、样本要求等有所不同,可根据检测需要和条件限制进行选择。将分子生物学方法和免疫学等方法有效地结合起来,多角度多层次地对样品进行检测,可全面而准确地呈现检测结果。蜡样芽孢杆菌能产生多种毒素,这些毒素决定了其致病性,所以除了检测菌体外,还可以对毒素进行检测,有助于确定病原及其致病力。总的来说,蜡样芽孢杆菌对人畜的健康安全均构成了威胁,快速准确地检测能有效辅助治疗和提前预防,作者将主要检测方法进行了总结,希望能有助于全面准确地评估蜡样芽孢杆菌的风险,为主动监测和预警提供科学依据。  相似文献   

9.
Characterization of a parvovirus isolated from the diarrheic feces of a pig   总被引：7，自引：0，他引：7  
A small DNA virus was isolated from the feces of a sow with diarrhea and identified as a parvovirus on the basis of its properties. The virus replicated preferentially in cell cultures of swine origin, including primary porcine thyroid gland and kidney cell cultures in which the cytopathic effect developed. The virus agglutinated erythrocytes of guinea pig, mouse and human group O but not these of chicken. The growth of the virus was inhibited by 5-iodo-2'-deoxyuridine. The virus was resistant to ether and heating at 56 degrees C for 30 min and stable at pH 3.0. The buoyant density of the infectious particles was 1.40 g/ml in CsCl density gradient, and the virions were 27 nm in diameter by electron microscopy. The viral protein seemed to be separated into four polypeptides with molecular weights of 81k, 70k, 66k and 62k daltons respectively. Cross serum neutralization test demonstrated that the virus was antigenically different from porcine parvovirus as well as bovine and canine parvoviruses. These findings and the survey on neutralizing antibody distribution indicated indirectly that another parvovirus which could be antigenically distinguished from well-known porcine parvovirus had been widespread among swine in Japan.  相似文献   

10.
Pseudorabies virus hemagglutinin was readily adsorbed on mouse erythrocytes at 4, 22, or 37 degrees C, but not on cattle erythrocytes. The adsorbed hemagglutinin could not be eluted from the cells by resuspending in phosphate-buffered saline (PBS), by incubating at 37 or 50 degrees C, or by incubating in the presence of neuraminidase. The receptor on mouse erythrocytes for the hemagglutinin was inactivated by trypsin, but not by neuraminidase, sodium deoxycholate (DOC), potassium periodate (KIO4), dithiothreitol (DTT), 2-mercaptoethanol (2-ME) and formalin. The hemagglutinin was inactivated by trypsin, alpha-amylase, pepsin, DOC, KIO4, and ethylendiamine-tetraacetic acid (EDTA), but not by papain, beta-glucosidase, phospholipase C, neuraminidase, DTT, 2-ME, Tween-80, ethylether, chloroform, trichloro-trifluoroethane, beta-propiolactone and formalin, suggesting that the hemagglutinin active component involved glycoproteins. The hemagglutinin was stable at 37 degrees C for lower temperatures but not at 60 degrees C or higher. The hemagglutinin activity was resistant to ultraviolet irradiation, while the infectivity was very susceptible. The hemagglutinin and the infectivity were readily sedimented by ultracentrifugation at 48,000 x g for 3 hr. In rate zonal centrifugation of the preparation on a sucrose density gradient, the hemagglutination (HA) activity showed a sharp peak at 1.22 g/ml coinciding with the peak of infectivity. The HA activity in the peak fraction seemed to be structually associated with virus particles. After fractionation of the virus by Nonidet P-40, the HA activity was found only in the fraction of the envelope material, indicating that the hemagglutinin is situated in the viral envelop.  相似文献   

11.
嗜水气单胞菌粘附特性的研究   总被引：7，自引：1，他引：6  
不同来源的６株嗜水气单胞菌（Ａｅｒｏｍｏｎａｓｈｙｄｒｏｐｈｉｌａ）对１０种不同红细胞的血凝试验表明，Ｊ－１株能凝集所有的１０种红细胞，而与其他菌株的血凝谱有所不同。血凝图式有２种：人、鸡、麻雀的红细胞凝集快，呈大小不等的絮状；而鲫鱼、绵羊、猪、小鼠、兔、鸭、犬的红细胞凝集慢，且呈均匀颗粒状。这两种血凝均能被Ｄ－甘露糖抑制，但不能被Ｊ－１株Ｒ菌毛抑制。组织粘附试验显示，Ｊ－１株能粘附小鼠和鲫鱼的肠组织和肠绒毛。将提纯的Ｒ菌毛预处理肠组织，或用Ｄ－甘露糖或木瓜蛋白酶消化的Ｒ菌毛抗血清Ｆａｂ片段预处理菌体，都不能抑制上述的粘附作用。Ｊ－１株对ＨＥｐ－２细胞显示强粘附，菌体随机粘附在细胞上，有少数侵入细胞浆内。其余５株菌的粘附特性与Ｊ－１株不尽相同。所有６株菌对草鱼肠组织及肠绒毛、ＥＰＣ细胞（鲤鱼乳头状上皮瘤细胞系）均无粘附性。  相似文献   

12.
1. The present study compared the ability of native, heat-treated and aged turkey breast muscle proteins to undergo proteolysis by digestive tract proteases. 2. Domestic turkey toms were slaughtered under laboratory conditions. Breast muscles were excised immediately post mortem; one was placed under conditions to develop exudative meat by maintaining the muscle at 40 degrees C for at least 30 min and the other was refrigerated under commercial conditions. 3. Meat was collected and stored for 7 d at 4 degrees C. Breast samples removed at d 0 and d 7 were frozen and stored at -80 degrees C until used for determination of solubility, protein surface hydrophobicity and protein oxidation through carbonyl content. Measurements of pepsin and trypsin/chymotrypsin activities were performed in vitro on myofibrillar proteins. 4. Storage increased carbonyl content in control samples while the oxidation increase was not significant in heat-treated myofibrillar protein. Hydrophobicity was not affected by storage time or treatment or protein solubility. 5. Storage significantly increased trypsin + chymotrypsin activity only in the control group. The activities of pepsin and trypsin + chymotrypsin were negatively correlated with protein surface hydrophobicity.  相似文献   

13.
Partially purified vascular permeability (VP) factor (VPF) of Bacillus cereus induced fluid accumulation in the ligated intestinal loops of mouse (MIL) and rabbit (RIL), suggesting that the VP activity may correlate with fluid accumulation in ligated intestinal loops of these animals. Fluid accumulation was observed at 6-8 hr in 55-67% of mouse intestinal loops inoculated with 40-50 immunodiffusion units (IDU) of partially purified VPF, whereas it was found at 2 hr in all loops with 400-600 IDU of partially purified VPF. In rabbit intestinal loops with 120-190 IDU of partially purified VPF, fluid accumulation was observed at 6 hr. From these findings, VPF produced by B. cereus can be easily detected in both MIL and RIL. The intestinal tissue of mouse intestinal loops was histopathologically damaged at different concentrations of the VPF to induce fluid accumulation. With 50 IDU of partially purified VPF, severe edema was found in the laminia proprial layer and submucosa. With 600 IDU of partially purified VPF, on the other hand, severe necrosis in the surface epithelium of villus and laminia proprial layer, and hyperemia in the submucosa were observed, suggesting that partially purified VPF may be cytotoxic and/or intestinecrotic.  相似文献   

14.
Slow-reacting complement-requiring neutralizing (NT) antibody was detected in sera from cattle 2 weeks after infection with Akabane virus. Bovine sera obtained 3 or 4 weeks after infection contained slow-reacting noncomplement-requiring NT antibody. The slow-reacting complement-requiring NT antibody was sensitive to 2-mercaptoethanol (2-ME), whereas the slow-reacting noncomplement-requiring NT antibody was resistant to 2-ME. The initial phase may represent the IgM response and the later phase a change to IgG. A NT test was developed in which virus-serum mixtures were incubated at 4 degrees C for 48 h and then with complement at 37 degrees C for 60 min; this gave an improved sensitivity over the previous incubation at 37 degrees C for 60 min.  相似文献   

15.
鳝鱼嗜水气单胞菌外毒素的分离纯化及活性研究   总被引：1，自引：0，他引：1  
为探讨嗜水气单胞菌的致病机理,经饱和硫酸铵分级盐析、Sephadex G-75葡聚糖凝胶层析和PEG6000浓缩,获得层析纯的外毒素。对纯化外毒素的溶血价、溶血谱、毒素稳定性及对小鼠的致病性等生物学性状进行研究。结果表明:该纯化外毒素对多种动物红细胞有溶血作用;对小鼠有较高的致死率;溶血活性的最适pH值为5~7;对热不稳定,37℃处理3 d可保持活性,14 d后基本失活,56℃处理2 min及100℃处理1 min即基本丧失活性。  相似文献   

16.
Akabane virus was shown to lyse as well as to agglutinate pigeon erythrocytes. The hemolytic activity of the virus was markedly enhanced by repeated freeze-thawing, but its hemagglutinating activity was not affected. Hemolysis (HL) with the virus, like its hemagglutination, was affected by the NaCl concentration as well as by the pH of the diluent. HL was markedly affected by the incubation temperature, but hemagglutination (HA) was not; HL activity was highest at 37°C, somewhat lower at 25°C, very low at 4°C, and did not occur at 0°C. While pigeon erythrocytes were positive for both HL and HA, goose erythrocytes were positive for HA but negative for HL. Erythrocytes from cattle, sheep, rabbits, guinea pigs, mice and day-old chickens were tested for HA as well as for HL activity with negative results. A linear relationship was shown, in a wide range of the virus concentrations, between the percent HL and the virus concentration, as expressed on a logarithmic scale. Based on these findings we developed an assay method for Akabane virus hemolysin. Analysis by CsCl equilibrium density gradient centrifugation indicated the hemolytic as well as the hemagglutinating activity to be structurally associated with the virion. Scanning electron microscopy of pigeon erythrocytes undergoing HL with the virus revealed the appearance of a depressed area with a hole on the cell surface. The hemolytic activity of the virus was specifically inhibited by antisera to the virus and an HL-inhibition test was developed.  相似文献   

17.
Eleven Actinobacillus strains were isolated in pure culture from 12- to 13-week-old aborted swine fetuses. Apart from minor biochemical differences, they resembled strains isolated from sow vaginas by Ross et al. (1972) in the U. S. A. Some of the strains agglutinated sheep, cattle and horse but not pig, dog and chicken erythrocytes. Haemagglutination was mannose resistant and could be inhibited by specific hyperimmune serum. Heating above 70 degrees C diminished or abolished haemagglutination of the cultures. Electron microscopy showed no fimbriae on the surface of the bacteria.  相似文献   

18.
Investigaltions to determine the effect of sample storage on the concentration of copper in liver tissue and on the activity of erythrocyte superoxide dismutase were undertaken in preparation for a study of blesbok (Damaliscus pygargus phillipsi) that were suspected to be suffering from copper deficiency. Two liver samples were collected from each of 20 culled blesbok in a manner that simulated the collection of biopsies from the live animal. These samples were stored either in 10% formalin or frozen at -20 degrees C until analysed 4 1/2 months later. The effect of different methods of sample storage on superoxide dismutase activity was determined. Erythrocytes collected from 3 Jersey cows and 5 culled blesbok were washed and divided into 0.5 ml portions, stored at room temperature (approximately 20 degrees C), in a refrigerator (4 degrees C), frozen at -20 degrees C in a freezer, and in liquid nitrogen (-200 degrees C). An analysis of superoxide dismutase activity was undertaken using a commercial assay kit at intervals of 2-4 days until the levels of activity had fallen significantly. The copper concentration in formalin-preserved liver samples was significantly lower than that measured in frozen liver tissue apparently as a result of leaching. The activity of superoxide dismutase in cattle blood was unchanged for 4 days at room temperature but fell appreciably after 2 days at 4 degrees C and -20 degrees C. Enzyme activity remained unchanged for 200 days in erythrocytes stored in liquid nitrogen. Superoxide dismutase activity levels in healthy blesbok were considerably lower than those measured in Jersey cows and remained unaffected for up to 6 days in samples stored at 4 degrees C and 20 degrees C. The level of activity fell significantly thereafter. Samples stored in liquid nitrogen were unchanged after 40 days.  相似文献   

19.
The endotoxic activity of Fusobacterium necrophorum bov 5 was investigated. The supernatant (S) fluid and cell wall (CW) preparation, obtained after differential centrifugation of the ruptured cell mass, were lethal for mice. The toxicity of the S fluid was stable during prolonged storage, treatment with formalin, and heating for 15 minutes at 80, 100, and 121 C, but was destroyed by alkaline hydrolysis with 0.25 N NaOH. The toxic factor was found in a high molecular weight (MW) fraction after gel filtration. The properties exhibited by the toxic S fluid resembled those of endotoxic lipopolysaccharide (LPS). Extracted and partially purified LPS (endotoxin) from F necrophorum bov 5 demonstrated a mouse median lethal dose (mouse LD50) of 16.8 mg/kg of body weight. The toxic LPS material, a high molecular weight moiety as estimated by gel filtration, was resistant to ribonuclease (RNase), deoxyribonuclease (DNase), and pronase treatment. A positive Shwartzman reaction (median skin lesion dose (SLD50) equal to 3.32 mug/kg of body weight) and biphasic fever response (minimal dose required to produce a fever index of 40 sq cm which falls on the linear portion of dose-response curve (FL40) equal to 0.41 mug/kg of body weight) further indicated the toxin was endotoxin in nature. The LPS from F necrophorum bov 5 was less toxic than Salmonella typhimurium LPS; but had considerable toxicity for experimental animals. The toxic activity of the partially purified F necrophorum bov 5 endotoxin was separated into 2 fraction regions by diethylaminoethyl (DEAE)-cellulose chromatography. The data provide evidence for the production of a potent endotoxin, possibly composed of more than one toxic component, which may be released upon cell disruption.  相似文献   

20.
Bovine interleukin 2: production and characterization   总被引：1，自引：0，他引：1  
The production of bovine IL 2 was studied and IL 2 was partially characterized. PMA at 5 ng/ml + Concanavalin A at 5 micrograms/ml treatment of peripheral blood mononuclear cells gave a greater yield of IL 2 activity in the supernatants than Con A, PMA or sodium periodate treatments alone. Macrophage depletion increased yields as did the addition of indomethacin, a prostaglandin E2 inhibitor. Bovine IL 2 was sensitive to trypsin, relatively stable at pH 2-9, 2-ME resistant and sensitive to increasing molar concentrations of urea. The activity of bovine IL 2 was reduced by over 45% at 70 degrees C for 30 min and 95% at 90 degrees C for 30 min. Bovine IL 2 was more stable at 4 degrees C than at room temperature and the stability at room temperature could be improved by inclusion of 1% BSA. Bovine IL 2 eluted from DEAE-Sephadex as a broad peak with 0.1-0.2 M NaCl. Peak activity corresponded to a molecular weight of approximately 16,000 daltons on Sephadex G-100.  相似文献   

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