Biofilm formation of Bacillus cereus under food-processing-related conditions

Food Science and Biotechnology - This study aims to understand the biofilm formation abilities of eight Bacillus cereus strains under food-industry-related conditions. Biofilms were grown in...     

Effect of Bacillus cereus exocellular factors on human intestinal epithelial cells

To gain insight on the biological effects of the exocellular factors produced by Bacillus cereus, culture filtrate supernatants of different strains were coincubated with differentiated Caco-2 cells. Exocellular factors were able to detach enterocyte-like cells from the substratum after 1 h of incubation. In addition, microvilli effacing and dramatic changes on the cellular surface of enterocytes were found after incubation periods as short as 20 min. Since cell detachment was not inhibited by fetal calf serum, thiol activated cholesterol-binding cytolysin, cereolysin O, does not seem to be involved. Also, translocation of phosphatidylserine from the inner to the outer leaflets of the plasma membrane was demonstrated by using fluorescein isothiocyanate (FITC)-Annexin V. In contrast to the high capability of detaching Caco-2 cells shown by all the strains under study, the mitochondrial dehydrogenase activity was lowered by culture filtrate supernatants in a strain-dependent manner. For strain M2, the decrease in dehydrogenase activity was already evident after 30 min of incubation. Production of biologically active factors depends on the growth phase, and maximal activity was found in late exponential-early stationary phases. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of concentrated exocellular factors showed a very complex scenery supporting the multifactorial character of the biological activity of B. cereus.     

乳酸对蜡样芽胞杆菌的抑制研究

本文研究了乳酸在营养肉汤培养基对蜡样芽胞杆菌的抑制,以及前期乳酸浸泡大豆后对蜡样芽胞杆菌在天培中生长情况的影响.研究发观,在营养肉汤培养基中,当采用乳酸调整pH时,在pH值为4.5时可以较好地抑制蜡样芽孢杆菌的生长.在天培的制作过程中,在前期未加乳酸浸泡处理的天培中,菌数可达108cfu/g并最终导致天培的酸败;在前期加乳酸浸泡处理过的天培中,蜡样芽胞杆菌亦可以生长,但菌数有所下降,为106cfu/g,乳酸浸泡对抑制天培中蜡样芽胞杆菌的生长起到一定的作用.     

Effect of dairy product environment on the growth of Bacillus cereus

pH is one of the most important parameters to manage bacterial replication in foodstuffs. In this study, the ability of 2 Bacillus cereus strains, 1 clinical human isolate (GPe2) and 1 isolate from a dairy product (D43), were investigated for in vitro growth at different pH values (from 3.5 to 7.5) at 2 temperatures (15 and 37°C), showing their ability to grow from 5.5 to 7.5 and from 5.0 to 7.5, respectively. The ability of spores of these 2 microorganisms to germinate in different typologies of dairy products (unflavored yogurt, Taleggio cheese, mascarpone cheese, and raw and pasteurized milk) was also investigated by inoculating the spores and maintaining the products at 15°C. No growth was observed in yogurt, likely due to the combined effect of low pH (<5) and the presence of natural microflora. An inhibitory action of the natural microflora on the growth of B. cereus was also hypothesized for Taleggio cheese and raw milk, as these substrates were characterized by a high natural lactic acid bacteria population and permissive pH values (5.8/6.8 in Taleggio cheese, >7 in raw milk). In pasteurized milk and mascarpone cheese, where pH was not restrictive for B. cereus growth and where no significant natural microflora was present, growth occurred rapidly up to loads close to 7 log cfu/g.     

Influence of carvacrol on growth and toxin production by Bacillus cereus

The natural antimicrobial compound carvacrol was investigated for its effect on diarrheal toxin production by Bacillus cereus. Carvacrol (0-0.06 mg/ml) reduced the viable count and the maximal specific growth rate (mumax) of B. cereus in BHI broth. The total amount of protein was not affected by carvacrol. However, a sharp decrease (80%) in diarrheal toxin production was observed in the presence of 0.06 mg/ml carvacrol. Carvacrol also inhibited toxin production in soup, but approximately 50-fold higher concentrations were needed to achieve the same effect as in broth. From this study it can be concluded that carvacrol can be added to food products at doses below the MIC value, thereby reducing the risk of toxin production by B. cereus and increasing the safety of the products.     

The inhibition of the growth of Bacillus cereus in liver sausage

The growth of Bacillus cereus is a problem in liver sausage especially when the sausages are stored at high temperatures. Even concentrations of greater than 10(6)/g have been detected. In this study we found that when combining glucono-delta-lactone, sodium erythorbate and citric acid with sodium nitrite and salt the growth of B. cereus could be delayed or totally inhibited.     

Influence of pH and temperature on growth of Bacillus cereus in vegetable substrates

Bacillus cereus is a food-borne pathogen which most often contaminates foods of plant origin. Spores of psychrotrophic strains have the ability to germinate and grow at refrigeration temperatures in different vegetable substrates, such as carrot broth, zucchini broth, and cooked carrot purée. In some circumstances, factors such as pH, heat treatment, and storage temperature play a fundamental role in controlling the growth of these psychrotrophic strains and in extending the shelf life of refrigerated, minimally processed vegetable-based products in relation to pathogenic spore-forming bacteria. The combination of mild acidification (pH 5.0) and refrigeration (相似文献   

全谷物体外对肠道菌群影响的研究

选取了5种谷物(燕麦、大麦、全麦、糙米、黑米)和一种精致谷物(大米)为研究对象,在体外研究了其对肠道菌群的影响,并对其成分与肠道菌群发酵前后变化量进行了相关性分析。结果表明,全谷物样品发酵后双歧杆菌数量显著增加,肠杆菌科和梭状芽孢杆菌数量显著下降,除黑米外其他谷物发酵液中乳酸杆菌数量显著增加;发酵后大米中双歧杆菌和乳酸杆菌数量有所下降,肠杆菌科和梭状芽孢杆菌数量显著上升,特别是肠杆菌数量由7.231 g(cfu)/mL增长到8.041 g(cfu)/mL。     

鲜湿米粉中蜡样芽孢杆菌生长预测模型构建

测定了不同储藏温度(5、10、15、20、25、30、35℃)下鲜湿米粉中蜡样芽孢杆菌的生长数据,利用Gompertz模型、Baranyi模型、Logistic模型、MMF模型构建鲜湿米粉中芽孢杆菌生长预测初级模型。结果表明,Baranyi模型与实际测试数据符合度最好,选用Baranyi作为初级预测模型。进一步利用Baranyi模型参数,选用Ratkowsky平方根方程建立了鲜湿米粉中芽孢杆菌生长预测次级模型。鲜湿米粉中芽孢杆菌生长预测模型表明,鲜湿米粉应在15℃以下储藏,芽孢杆菌在鲜湿米粉中的最低生长温度为5.0℃左右。     

蜡样芽孢杆菌产磷脂酶D发酵条件优化

通过单因素和正交试验,对蜡样芽孢杆菌（Bacillus cereus）产磷脂酶D发酵条件进行了优化。结果表明,培养基最佳组成为卵黄10 g/L、蛋白胨15 g/L、牛肉粉2 g/L、MgSO4·7H2O 1 g/L、氯化钠3 g/L;最佳发酵条件为：培养基初始pH8.0,接种量1.0%,于36 ℃,200 r/min培养8 h。在最适发酵条件下磷脂酶D酶活达4.21 U/mL。     

产磷脂酶菌株蜡样芽胞杆菌AF-1发酵条件的研究

以蜡样芽胞杆菌(Bacillus cereus) AF-1为发酵菌株,对其发酵产磷脂酶培养基和发酵条件进行优化。结果表明,最佳培养基组成为：葡萄糖2%,蛋白胨3%,磷酸氢二钠1.5%,磷酸二氢钾0.3%,硫酸镁0.06%,氯化钙0.01%;15 L发酵罐发酵条件为发酵时间30 h,装液系数为0.67,接种量为10%,初始pH值为7.5,发酵温度为35 ℃,通气量为0.3 m3/h,在此最佳条件下得到的发酵液磷脂酶活力为41.57 U/mL。     

Effect of temperature on growth characteristics of Bacillus cereus TZ415

The effect of temperature on the maximal specific growth rate was studied in Bacillus cereus between 5 and 40 degrees C cultivated in courgette broth and rich medium (J broth). B. cereus grown from 5 to 38 degrees C in rich medium. No growth was observed in courgette broth below 10 degrees C. The Arrhenius plot was fitted from experimental data of B. cereus grown in rich medium and at regulated pH, oxygen and temperature. Two domains which are separated by a critical temperature around 13 degrees C can be distinguished with regard to temperature dependence of maximal specific growth rate. Over the cold domain from 5 to 13 degrees C, the temperature characteristic was 2.6 fold higher than over the sub-optimal domain from 13 to 38 degrees C suggesting that the growth temperature regulates several metabolic pathways.     

Influence of controlled lactic fermentation on growth and sporulation of Bacillus cereus in milk

The growth and sporulation of Bacillus cereus NVH 45 in a fermentor with controlled pH or simulated pH conditions were investigated. The study was carried out in a fermentor to measure the influence of a rapid and a slow lactic acid production on the inhibition of B. cereus in a controlled environment during the initial part of fermentation and to observe if other factors than lactic acid influenced the inhibition. In the controlled pH experiments the pH was allowed to decrease to an end pH 5.0, 5.5 or 6.0 either by Lactobacillus casei 2756 (a fast acid producer) or Lactobacillus acidophilus NCFB 1748 (a slow acid producer). In co-cultures of Lb. casei 2756 and B. cereus NVH 45, low numbers (10-70 cfu/ml) of B. cereus NVH 45 were observed at end pH 5.5 (72 h) while at pH 5.0 no viable cells (<10 cfu/ml) were detected (48-72 h). B. cereus NVH 45 did not sporulate in co-culture with Lb. casei 2756. In co-culture with Lb. acidophilus NCFB 1748, B. cereus NVH 45 sporulated and survived as spores. In these co-cultures B. cereus NVH 45 grew to higher maximum counts (>10(7) cfu/ml) than with Lb. casei 2756 (<10(7) cfu/ml). Significantly different amounts of lactic acid were observed between the two co-cultures after 7 and 12 h. A rapid decrease of pH appears to prevent B. cereus from sporulating and it seems that it is enough to just reach pH 5.0 rapidly and keep that pH to achieve the desirable inhibition of B. cereus. In the simulated pH experiments B. cereus NVH 45 was inoculated in the fermentor and the different pH developments from different LAB strains were monitored by addition of lactic acid. These experiments showed the same tendency: a fast pH reduction during the initial hours of fermentation, simulating lactococci, resulted in complete inhibition of B. cereus NVH 45 (<10 cfu/ml). However, when simulating the pH development of the two different Lactobacillus strains, complete inhibition of B. cereus NVH 45 was not seen. In co-cultures competition for nutrients with consequences for cell density appears to be important. Based on these results it seems that B. cereus must reach a certain density to induce sporulation.     

Atmospheric oxygen and other conditions affecting the production of cereulide by Bacillus cereus in food

Factors influencing the production of cereulide, the emetic toxin of Bacillus cereus in food and laboratory media were investigated, using liquid chromatography-ion trap mass spectrometry and sperm motility inhibition bioassay for detection and quantitation. Oxygen was essential for production of the emetic toxin by B. cereus. When beans, rice or tryptic soy broth were inoculated with cereulide producing strains B203, B116 (recent food isolates) or the strain F-4810/72, high amounts (2 to 7 microg ml(-1) or g(-1) wet wt) of cereulide accumulated during 4-day storage at room temperature. In parallel cultures and foods, stored under nitrogen atmosphere (> 99.5% N2), less than 0.05 microg of cereulide ml(-1) or g(-1) wet wt accumulated. The outcome of the bioassay matched that of the chemical assay, with no indication of interference by substances in the rice or beans. Boiling for 20 to 30 min did not inactivate cereulide or cereulide producing strains in rice or the beans. Adding l-leucine and l-valine (0.3 g l(-1)) stimulated cereulide production 10- to 20-fold in R2A and in rice water agar. When the B. cereus strains were grown on agar media under permissive conditions (air, room temperature), cereulide was produced overnight with little or no increase when the incubation was extended to 4 days. In broth culture, the production of cereulide started later than 16-24 h. Anoxic storage prevented cereulide production also when the amino acids had been supplied. Packaging with modified atmosphere low in oxygen may thus be used to reduce the risk of cereulide formation during storage of food.     

Survival and growth of Bacillus cereus in Egyptian bread and its effect on bread staling

Bread was made from dough previously inoculated with spores of three Bacillus cereus strains at different inoculation levels. Counts of survivors and bread staling were determined in bread during storage at 25 degrees C. The total aerobic bacteria, sporeformer and B. cereus counts in seven different wheat flours were 3.0 x 10(6) to 9.1 x 10(7), 3.2 x 10(5) to 1.5 x 10(6) and 50-100 colony forming units (CFU/g). About one in a thousand bacteria were found to survive baking. On the other hand, no multiplication took place, however, until the bread had been stored for more than 3 days. Also, survival and multiplication of B. cereus spores was shown to depend on the kind of strains and type of bread. The addition of B. cereus strains to the dough gave bread with higher freshness than the control sample. This was quite noticeable in inoculated pan bread during storage. It is possible to conclude from the results that B. cereus survive the baking process particularly if original contamination is high. Also increasing the freshness of the bread does not necessarily mean better ingredient and processing conditions.     

Effect of carbon dioxide on the growth of Bacillus cereus spores in milk during storage

The effects of the addition of 11.9 mM CO2 on the growth of Bacillus cereus spores inoculated into sterile homogenized whole milk at 101 and 106 spores/ml and stored at 6.1 degrees C, was examined weekly for 35 d. Colony-forming units from CO2 treated inoculated milks decreased over 35 d at a rate similar to that of untreated inoculated milk, as defined by linear regression. Plate counts for treated and control milks inoculated at 10(1) cfu/ml were not significantly different on sampling d 0, 14, 21, and 28. Plate counts at d 7 were significantly different and counts at d 35 were at undetectable levels for both treated and control milks. Plate counts for milk inoculated at 10(6) cfu/ml were not significantly different on d 0, 28, and 35; they were significantly different on d 7, 14, and 21. There was no consistency as to whether the control or test milks were higher in counts on days when the differences were significant. Added CO2 reduced the pH of the milk from an average value of 6.61 to an average value of 6.31; however, this drop did not correlate with changes in any other parameter measured. These data suggest that moderate levels of CO2 do not enhance the outgrowth of B. cereus spores over long-term storage and do not increase the risk of foodborne illness due to the organism.     

蜡样芽胞杆菌生长特性及大蒜精油对其抑菌活性的研究

主要研究了37℃条件下3种蜡样芽胞杆菌在营养肉汤中的生长状况,建立了37℃下蜡样芽胞杆菌在营养肉汤中的Boltzmann牛长模犁,3条牛长曲线相关系数鼯均大于0．97;检测了不同培养时间蜡样芽胞杆菌的产芽胞情况,结果表明1号菌株和14号菌株较早产芽胞,培养相同时间,产芽胞数：1号菌株,4号菌株〉标准菌株;采用牛沣杯法和平板计数法研究了大蒜精油对蜡样芽胞杆菌的抑制效果,结果表明浓度为lO。的人蒜精油对3种蜡样芽胞杆菌都有很好的抑制效果,3种菌株中l号菌株最难抑制。     

Emetic toxin-producing strains of Bacillus cereus show distinct characteristics within the Bacillus cereus group

One hundred representative strains of Bacillus cereus were selected from a total collection of 372 B. cereus strains using two typing methods (RAPD and FT-IR) to investigate if emetic toxin-producing hazardous B. cereus strains possess characteristic growth and heat resistance profiles. The strains were classified into three groups: emetic toxin (cereulide)-producing strains (n=17), strains connected to diarrheal foodborne outbreaks (n=40) and food-environment strains (n=43), these latter not producing the emetic toxin. Our study revealed a shift in growth limits towards higher temperatures for the emetic strains, regardless of their origin. None of the emetic toxin-producing strains were able to grow below 10 degrees Celsius. In contrast, 11% (9 food-environment strains) out of the 83 non-emetic toxin-producing strains were able to grow at 4 degrees Celsius and 49% at 7 degrees Celsius (28 diarrheal and 13 food-environment strains). non-emetic toxin-producing strains. All emetic toxin-producing strains were able to grow at 48 degrees Celsius, but only 39% (16 diarrheal and 16 food-environment strains) of the non-emetic toxin-producing strains grew at this temperature. Spores from the emetic toxin-producing strains showed, on average, a higher heat resistance at 90 degrees Celsius and a lower germination, particularly at 7 degrees Celsius, than spores from the other strains. No difference between the three groups in their growth kinetics at 24 degrees Celsius, 37 degrees Celsius, and pH 5.0, 7.0, and 8.0 was observed. Our survey shows that emetic toxin-producing strains of B. cereus have distinct characteristics, which could have important implication for the risk assessment of the emetic type of B. cereus caused food poisoning. For instance, emetic strains still represent a special risk in heat-processed foods or preheated foods that are kept warm (in restaurants and cafeterias), but should not pose a risk in refrigerated foods.     

保健食品中检出蜡样芽孢杆菌的情况分析

目的了解保健食品的卫生状况。方法依照国家标准GB/T 4789.14—2003和进出口商品检验行业标准SN 0176—92,对市场随机抽检的12个品种225批保健食品检出的芽孢杆菌采用全自动微生物分析系统仪进行鉴定研究。结果确认有12批保健食品被检出蜡样芽孢杆菌,其中检出率较高的有螺旋藻片和钙片,检出率分别为60.0%和21.1%。结论分析结果可为保健食品的生产、消毒灭菌、卫生学检验标准及监督管理等方面提供参考。     

蜡样芽胞杆菌生长/非生长界面模型的建立和评价

蜡样芽胞杆菌是软烤虾仁产品的主要变质菌,它是一种条件致病菌,通过产生腹泻毒素和呕吐毒素导致食物中毒。该研究旨在建立一种概率模型来预测出蜡样芽胞杆菌的生长/非生长情况或者生长概率。用lo-gistic回归模型建立不同温度、水分活度和pH环境因子作用下蜡样芽胞杆菌的生长/非生长界面模型。实验结果表明蜡样芽胞杆菌在脑心浸液肉汤培养基中生长的最低温度为9.99℃,最低水分活度为0.931,最小pH值为4.5。在此基础上建立的蜡样芽胞杆菌生长/非生长界面模型的χ2=49.73,P<0.000 1。用logistic回归模型建立的生长/非生长模型拟合效果达到极显著水平。模型的预测值同时很好地量化了环境因子对蜡样芽胞杆菌的协同作用,为软烤虾仁产品中蜡样芽胞杆菌的生长/非生长界面模型的建立提供了参考。