Microbiological safety of kinema, a fermented soya bean food

Kinema is a fermented soya bean food of Nepal and the hilly regions of North-eastern States of India. Generally, the fermentation is dominated by Bacillus spp. that often cause alkalinity and desirable stickiness in the product. The present study was undertaken in a limited number of commercial (market) kinema samples to test for the presence of foodborne pathogens and their properties. Bacillus cereus was present in numbers exceeding 10⁴ cfu/g product in five of the tested 15 market samples. Enterobacteriaceae and coliform bacteria exceeded 10⁵ cfu/g in 10 of the 15 samples. Escherichia coli exceeding 10⁵ cfu/g was found in two samples. Staphylococcus aureus was not detected in any of the tested samples. Of 31 isolated typical and atypical strains of B. cereus, 18 representative strains were tested qualitatively for the ability to produce diarrhoeal type enterotoxin (BCET) using an Oxoid BCET-RPLA test kit. Overall, BCET was formed by 12 strains in BHIG (brain heart infusion broth +1% glucose), by seven strains on sterilized cooked rice, and by five strains on sterilized cooked soya beans. Semi-quantitative tests on BCET revealed that levels exceeding 256 ng/g soya beans, produced by single pure culture inoculation with the isolated B. cereus strains, were reduced to ≤ 8 ng/g by frying kinema in oil, a common procedure when making kinema curry. It was also shown in a mixed pure culture experiment that a kinema strain B. Subtilis DK-W1, is able to suppress growth and BCET formation by a selected toxin producing strain (BC7-5) of B. cereus. It is concluded that the traditional way of making kinema and its culinary use in curries is safe. However, for novel applications of kinema, safety precautions are advisable.     

Quantification of Factors Which Influence Nisin's Inhibition of CIostridium botulinum 56A in a Model Food System

We modeled nisin's anticlostridial activity and assessed the antagonistic or potentiating influences of food ingredients. The model systems contained yeast extract, proteose peptone, and glucose; were supplemented with protein (0.075, 0.75, 7.5% w/v), phospholipid (0.075, 0.75, 7.5% w/v), or soluble starch (5, 17.5, 30% w/v); and were adjusted to pH 5.5, 6.0, or 6.5. Samples inoculated with 10⁴/mL spores were incubated at 15, 25, or 35°C. Statistical analysis developed an equation (r²= 0.76) that modeled the response and identified temperature as the most significant (α 0.001) variable. Nisin lost effectiveness with increasing temperature. Nisin concentration had significant positive and phospholipid negative, linear effects. Many interactive effects were significant (α 0.20). Nisin inhibited C. botulinum until its residual level dropped below a threshold, which decreased from 154 IU/mL at 35°C to 12 IU/mL at 15°C.     

The Effect of Ozone Gas-Liquid Contacting Conditions in a Static Mixer on Microorganism Reduction

The effect of gas-liquid contacting conditions in a static mixer on ozone transfer efficiency and reduction of Bacillus subtilis spores was studied in an experimental ozone contactor. An empirical mathematical model was developed that related the transfer efficiency in the experimental system to the superficial liquid velocity in the mixer, the gas-liquid flow rate ratio and the height of the down-stream bubble column. Spore reduction was determined primarily by the dissolved ozone concentration-time (C_avgt_m) product in the reactive flow segment and was independent of the gas-liquid contacting conditions in the static mixer. In an integrated ozone contacting system, the static mixer should be designed to maximize ozone mass transfer while the reactive flow segment should be designed for efficient microorganism reduction.     

Comparative study of the elimination of toluene vapours in twin biotrickling filters using two microorganisms Bacillus cereus S1 and S2

BACKGROUND: To investigate the microbial degradation performance of organic pollutants in the atmosphere using a biotrickling filter, two microorganism strains, Bacillus cereus S1 and Bacillus cereus S2, were selected, identified and inoculated into a twin biotrickling filter for comparison. RESULTS: Both strains showed good performance towards the degradation of model organic pollutants when gas flow rates ranged from 100 to 600 L h⁻¹. For S1, the total maximum removal efficiency (RE) of toluene was maintained nearly 100% not only at gas flow rates of 100 L h⁻¹ corresponding to empty bed residence time (EBRT) 199.44 s, but also at gas flow rates of 200 L h⁻¹ (EBRT = 99.72 s) and 300 L h⁻¹ (EBRT = 66.48 s). However, S2 had a much lower degradation capability; near 100% removal efficiency was obtained only at the gas flow rate of 100 L h⁻¹ although both bacteria belong to the same Bacillus cereus. With further increase in gas flow rate, the total REs for both S1 and S2 decreased slightly at first and then dropped sharply to 46% and 35%, respectively, at an EBRT of 33.24 s, corresponding to a gas flow rate of 600 L h⁻¹. Starvation for between 2 and 10 days resulted in the re‐acclimation times of both strains ranging between 1.0 and 15.5 h. CONCLUSION: Strain S1 would be a better choice for inoculation into a biotrickling filter than strain S2, because of its much higher toluene removal capacity and rapid recovery to full performance. Copyright © 2008 Society of Chemical Industry     

胨冻样芽孢杆菌(Bacillus mucilaginosus)对烟草灰霉病菌(Botrytis cinerea)的抑制作用

本文首次报道胨冻样芽孢杆菌XDB1和D4B1对烟草灰霉病菌(Botrytis cinerea)的抑制作用。实验表明,XDB1和D4B1的发酵原液和无细胞提取物具有相同的抑菌效果,而经121℃处理30分钟后,抑菌活性丧失。D481菌株的抑菌活性高于XDB1,钾长石作为基质时的活性高于土壤矿物。但是,XDB1和D481对立枯丝核病菌(R..solani)、西瓜尖孢镰刀菌(F.oxysporum)、腐霉(Pythium sp)、小麦全蚀病菌(G.graminis)、烟炭疽病菌(C.gloeosporides)、烟草赤星病菌(A.alternata)、烟草黑胫病菌(P.nicotianae、串珠镰刀病菌(F.moniliforme)和水稻稻瘟病菌(P.grisea)无抑制作用。     

枯草杆菌碱性磷酸酶制备的最佳工艺条件及酶促反应动力学性质的研究

以枯草杆菌为菌种液体培养制备碱性磷酸酶，研究确定了培养的最佳工艺条件，并对碱性磷酸酶的酶促反应动力学性质进行了初步探讨。结果表明，枯草杆菌制备碱性磷酸酶的最佳工艺条件为：40℃，pH值7．4振荡培养10h，酶活最高。对碱性磷酸酶的动力学性质研究表明，该酶催化底物磷酸苯二钠水解反应的最适pH值8．8，最适温度52℃，米氏常数Km值为2．94mmol／L。     

Monooxygenase system of <Emphasis Type="Italic">Bacillus megaterium</Emphasis> ALA2: Studies on palmitic acid oxidation products

We identified many novel oxygenated FA produced from linoleic acid by microbial strain ALA2: 12,13,17-trihydroxy-9(Z)-octadecenoic acid (12,13,17-THOA); 12,13,16-trihydroxy-9(Z)-octadecenoic acid (12,13,16-THOA); 12-hydroxy-13,16-epoxy-9(Z)-octadecenoic acid; and 12,17;13,17-diepoxy-16-hydroxy-9(Z)-octadecenoic acid. 12,13,17-THOA, the main product, inhibits the growth of some plant pathogenic fungi. Recently, we reclassified strain ALA2 as Bacillus megaterium ALA2 NRRL B-21660 and opened a possible link with the well-studied catalytically self-sufficient P450 monooxygenase of Bacillus megaterium ATCC 14581 (NRRL B-3712) and B. subtilis strain 168 (NRRL B-4219). Now we have found that strain ALA2 also oxidizes palmitic acid into three oxygenated products: 13-, 14-, and 15-hydroxy palmitic acids. This indicates that strain ALA2 also possesses a monooxygenase system similar to the abovementioned well-known strains. These data facilitate studies on the oxygenase system of strain ALA2.     

Seawater Ozonation of Bacillus subtilis Spores: Implications for the Use of Ozone in Ballast Water Treatment

The potential of ozone for disinfection of ships’ ballast water was investigated using Bacillus subtilis spores as an indicator. The effects of pH, presence of iron, and bacterial strain on disinfection efficacy in seawater, under simulated ballast conditions, were investigated. Ozone dosages of 9 mg/L (pH 7) and 14 mg/L (pH 8.2) and 24 h contact achieved a 4-log inactivation with the various oxidant residuals formed. Iron surface at a ratio to water of 9 m²/m³ impaired the oxidant residuals and the disinfection of spores. Different strains of B. subtilis resulted in different CT values. Ozone does not seem to be a good choice for the control of spore-forming organisms in ballast water, but may be suitable for the control of other species.     

纳豆制备过程中大豆异黄酮的转化条件

使用日本古法制备纳豆,从纳豆中分离筛选出高活性的纳豆菌为实验菌株。研究了纳豆制备过程中大豆异黄酮的转化情况,发酵液中的大豆异黄酮在大豆异黄酮糖苷酶的水解作用下水解掉侧链糖基,转化为活性更高的大豆异黄酮苷元。根据薄层层析法确定了大豆异黄酮转化为大豆异黄酮苷元的最佳培养条件:培养温度为37℃,培养时间2 d,pH 7.0,含水质量分数60%。发酵液中大部分的大豆异黄酮苷转化为大豆异黄酮苷元,从而得到更利于人体吸收的大豆异黄酮苷含量较高的纳豆制品。     

纳豆芽孢杆菌菌剂的制备工艺

采用单因素及正交试验对纳豆芽孢杆菌发酵培养基、pH、温度、发酵时间等条件进行了考察,利用喷雾干燥法制备菌剂,考察了β-环糊精、蔗糖、可溶性淀粉及脱脂奶粉作为保护剂对菌剂制备的影响,并研究了储藏时间及条件对菌剂发酵活力的影响.研究结果表明:纳豆芽孢杆菌发酵培养基为黄豆浸汁培养基:黄豆以5倍水体积浸泡8~14h,121℃~126℃,0.1~0.15MPa处理1h,滤除黄豆,取豆水,加入5%的葡萄糖和0.02%的K2HPO4,调节pH至7.0;最优发酵工艺:初始pH7.0,37℃发酵10h;喷雾干燥最佳保护剂为5%脱脂奶粉;菌剂4℃冷藏180d以内不影响发酵活力.上述工艺下所制得的纳豆芽孢杆菌菌剂活菌数为6.5×108 CFU/g,发酵生产纳豆激酶性能良好.发酵生产纳豆激酶酶活为2 000~2 200IU/g.本研究为纳豆芽孢杆菌菌剂制备提供了一条较为合理的工艺路线.