Next‐Generation Sequencing Analyses of Bacterial Community Structures in Soybean Pastes Produced in Northeast China

Fermented soybean foods contain nutritional components including easily digestible peptides, cholesterol‐free oils, minerals, and vitamins. Various fermented soybean foods have been developed and are consumed as flavoring condiments in Asian regions. While the quality of fermented soybean foods is largely affected by microorganisms that participate in the fermentation process, our knowledge about the microorganisms in soybean pastes manufactured in Northeast China is limited. The current study used a culture‐independent barcoded pyrosequencing method targeting hypervariable V1/V2 regions of the 16S rRNA gene to evaluate Korean doenjang and soybean pastes prepared by the Hun Chinese (SPHC) and Korean minority (SPKM) populations in Northeast China. In total, 63399 high‐quality sequences were derived from 16 soybean paste samples collected in Northeast China. Each bacterial species‐level taxon of SPHC, SPKM, and Korean doenjang was clustered separately. Each paste contained representative bacterial species that could be distinguished from each other: Bacillus subtilis in SPKM, Tetragenococcus halophilus in SPHC, and Enterococcus durans in Korean doenjang. This is the 1st massive sequencing‐based study analyzing microbial communities in soybean pastes manufactured in Northeast China, compared to Korean doenjang. Our results clearly showed that each soybean paste contained unique microbial communities that varied depending on the manufacturing process and location.     

Prevalence,Genetic Diversity,and Antibiotic Resistance of Bacillus cereus Isolated from Korean Fermented Soybean Products

Bacillus cereus contamination is a major food safety problem for Korean fermented soybean products, but few studies have assessed its potential to cause foodborne illness. The objectives of this study were to investigate the prevalence and characteristics of B. cereus isolated from Korean fermented soybean products. B. cereus was detected in 110 of 162 (67.9%) samples. The highest B. cereus frequency was observed in deonjang (68 of 93 samples, 73.1%) and cheonggukjang (18 of 25, 72.0%); however, nonhemolytic enterotoxin was detected only in 22 of 162 samples (13.6%). Although the tested B. cereus isolates showed diverse pulsotypes according to repetitive sequence‐PCR banding patterns, they displayed similar antibiotic sensitivity spectra. The low frequency of enterotoxin detection suggests that the potential risk of B. cereus foodborne illness associated with Korean fermented soybean products is lower than generally presumed. However, considering the prevalence of B. cereus and the high content of fermented soybean products in the Korean diet, it is necessary to constantly monitor the level of contamination with B. cereus and its toxins in such Korean food products.     

Screening of Strains with fibrinolytic activity and angiotensin-converting enzyme inhibitory activity from doenjang

As a first step to develop a doenjang possessing functional properties, microbial strains possessing proteolytic activity, fibrinolytic activity, and angiotensin-converting enzyme (ACE) inhibitory activity were screened and isolated from Korean traditional doenjang. First, total 264 isolates showing proteolytic activity were isolated from 14 different doenjang samples. Among them, 4 strains producing both of fibrinolytic activity and ACE inhibitor into the culture medium were selected. The levels of ACE inhibitory effect of G19, G26, P16, and P25 strains were about 82.5, 78.5, 74.5, and 75.6% of inhibition ratio in soybean powder broth, respectively. All isolates were identified as Bacillus amyloliquefaciens based on SDS-PAGE profiles of whole cell proteins and 16S rRNA gene sequence analysis. The result implied that the selected Bacillus strains could be used for the development of doenjang possessing functional properties as potential starter.     

The members of the Bacillus cereus group are commonly present contaminants of fresh and heat-treated milk

To determine the level of milk contamination by Bacillus cereus sensu lato, 44 samples were collected from a dairy farm and two independent dairies in northeastern Poland. A total of 680 B. cereus sensu lato isolates were recovered. Based on spore counts, their highest level in milk was found during the spring and summer months. Although significant variations in chromosomal DNA polymorphisms among B. cereus sensu lato isolates were noted based on repetitive element sequence polymorphism (rep-PCR) and pulse-field gel electrophoresis (PFGE), indistinguishable B. cereus isolates were observed in all sampling points and locations. Both B. cereus sensu stricto/Bacillus weihenstephanensis and Bacillus thuringiensis cultured from milk harbored nheA, hblA, and cytK in, respectively, 80%, 55%, and 60% of the isolates. With respect to Bacillus mycoides/Bacillus pseudomycoides, 30% and 70% of theses isolates harbored, respectively, nheA and hblA. The presence of cytK was not detected in any isolate. Our data show the occurrence of potentially toxic B. cereus s.l. in both raw and heat-treated milk, thus emphasizing the requirement for precautions that prevent spore germination and vegetative proliferation by keeping the milk at low temperatures during all steps of production and dispensation to the consumers.     

Analysis and risk assessment of ethyl carbamate in various fermented foods

Ethyl carbamate (EC) was analyzed in fermented foods, comprising various matrices; subsequently, a risk assessment for EC was carried out. In total, 128 fermented foods were purchased from department stores and traditional markets in Korea. The samples comprised 55 lactic acid–fermented vegetable (kimchi), 53 fermented soybean paste (doenjang, gochujang, cheonggukjang, and ssamjang), 20 fermented fish products (jeotgal), 18 yoghurt, 37 bread, 12 cheese, and 23 vinegar samples. The level of EC in fermented soybean pastes ranged from not detectable to 240.2 ng g^?1. EC was not detected in kimchi, jeotgals, yoghurt, and cheese. Bread and vinegar contained EC up to 3.70 and 16.79 ng g^?1, respectively. The estimated daily intake and chronic daily intake of EC were 1.98 ng kg body weight (bw)^?1 day^?1, 1.50 ng kg bw^?1 day^?1, respectively. Excess cancer risk and margin of exposure of EC were 7.5 × 10^?8 and 151,515, respectively.     

Typing and evaluating heat resistance of Bacillus cereus sensu stricto isolated from the processing environment of powdered infant formula

Bacillus cereus sensu lato is one of the most harmful bacterial groups affecting the quality and safety of powdered infant formula (PIF). In this study, samples were collected from the raw materials and processing environments of PIF. A total of 84 isolates were identified as Bacillus cereus sensu stricto (B. cereus s. s.) by 16S rRNA analysis, molecular typing technology, and physiological and biochemical tests. The 84 B. cereus s. s. strains were assigned to panC group II, group III, and group IV. Then, the 7 housekeeping genes glpF, gmk, ilvD, pta, pur, pycA, and tpi were selected for multilocus sequence typing. Results showed that the 84 isolates were clustered into 24 sequence types (ST), and 14 novel ST were detected. Among the 24 ST, ST999 (19/84, 22.62%) and ST1343 (13/84, 15.48%) predominated. The correlation between processing areas and ST showed that the processing environments of the production and packing areas were the most susceptible to contamination by B. cereus s. s. Spores of these ST showed different heat resistance phenotypes evaluated by the analysis of D_T (time in minutes of spore decimal reduction at each temperature) and Z values (temperature increase required to reduce the D_T value to one-tenth of the original). Spores from group III according to panC gene analysis were the most heat resistant. These findings will help us to better understand B. cereus s. s. contamination and control in PIF processing environments.     

Genetic diversity, antimicrobial resistance, and toxigenic profiles of Bacillus cereus strains isolated from Sunsik

Bacillus cereus can cause emetic and diarrheal types of food poisoning, but little study has been done on the toxins and toxin-encoding genes of B. cereus strains isolated from Sunsik, a Korean ready-to-eat food prepared from grains, fruits, and vegetables. In this study, 39 unique B. cereus strains were isolated and identified from Sunsik samples, with an average contamination level of 10 to 200 CFU/g. The detection rates of the hblACD, cytK, and bceT genes among all the strains were 48.7, 66.7, and 87.1%, respectively. All 39 B. cereus strains carried nheABC and entFM genes, and 36 strains also had the ces gene, which encodes an emetic toxin. Nonhemolytic enterotoxin and hemolysin BL enterotoxin were produced by 39 and 26 strains, respectively. The strains were separated into 13 profiles based on the presence or absence of toxins and their genes, as determined by antibody tests and PCR analysis. Profile 1 was the largest group, comprising 30.7% (12 of 39) of the B. cereus strains tested; these strains harbored all toxins and their genes. The B. cereus strains were susceptible to most of the antibiotics tested but were highly resistant to b -lactam antibiotics. The repetitive element sequence polymorphism PCR fingerprints of the B. cereus strains were not influenced by the presence of toxin genes or antibiotic resistance profiles. Our results suggest that B. cereus strains from Sunsik could cause either the diarrheal or emetic types of food poisoning because all strains isolated contained at least one toxin and its gene, although the level of B. cereus contamination in Sunsik was low.     

Enterotoxins and emetic toxins production by Bacillus cereus and other species of Bacillus isolated from Soumbala and Bikalga, African alkaline fermented food condiments

The ability of various species of Bacillus from fermented seeds of Parkia biglobosa known as African locust bean (Soumbala) and fermented seeds of Hibiscus sabdariffa (Bikalga) was investigated. The study included screening of the isolates by haemolysis on blood agar, detection of toxins in broth and during the fermentation of African locust bean using the Bacillus cereus Enterotoxin Reverse Passive Latex Agglutination test kit (BCET-RPLA) and the Bacillus Diarrhoeal Enterotoxin Visual Immunoassay (BDEVIA). Detection of genes encoding cytotoxin K (CytK), haemolysin BL (Hbl A, Hbl C, Hbl D), non-hemolytic enterotoxin (NheA, NheB, NheC) and EM1 specific of emetic toxin producers was also investigated using PCR with single pair and multiplex primers. Of 41 isolates, 29 Bacillus belonging to the species of B. cereus, Bacillus subtilis, Bacillus licheniformis and Bacillus pumilus showed haemolysis on blood agar. Using RPLA, enterotoxin production was detected for three isolates of B. cereus in broth and all B. cereus (9) in fermented seeds. Using BDEVIA, enterotoxin production was detected in broth as well as in fermented seeds for all B. cereus isolates. None of the isolates belonging to the other Bacillus species was able to produce enterotoxins either by RPLA or BDEVIA. Nhe genes were detected in all B. cereus while Hbl and CytK genes were detected respectively in five and six B. cereus strains. A weak presence of Hbl (A, D) and CytK genes was detected in two isolates of B. subtilis and one of B. licheniformis but results were inconsistent, especially for Hbl genes. The emetic specific gene fragment EM1 was not detected in any of the isolates studied.     

Plasmid exchanges among members of the Bacillus cereus group in foodstuffs

The Bacillus cereus sensu lato group is genetically very close and possesses a remarkable plasmid gene pool that encodes a variety of functions such as virulence and self-transfer capabilities. The potential for horizontal transfer among the various subspecies of this group, which includes the human opportunistic pathogens B. cereus sensu stricto and B. anthracis as well as the biopesticide B. thuringiensis, has led to growing concerns regarding food safety and public health. In this study, the conjugative behaviour of B. thuringiensis strains was compared in LB medium, milk and rice pudding using the pXO16 and pAW63 conjugative systems, as well as the mobilisable plasmid pC194, in bi- and triparental matings. Conjugation and mobilisation of these plasmids were shown to occur at significant levels in both food products, attaining the highest transfer frequencies in milk, with an approximately ten-fold increase in conjugative transfer in this growth medium as compared to liquid LB. Furthermore, the ability of an emetic strain of B. cereus to function as either plasmid donor or recipient partner in heterologous biparental matings with B. thuringiensis was demonstrated in these food matrices.     

Identification and safety evaluation of Bacillus species occurring in high numbers during spontaneous fermentations to produce Gergoush, a traditional Sudanese bread snack

Gergoush is a naturally fermented Sudanese Bread snack produced in three fermentation steps (primary starter, adapted starter and final dough), followed by three baking steps for a half to one hour at above 200 °C. This study examines the microbiota of two sets of fermentations performed at a traditional production site in Khartoum, Sudan in 2006 and 2009, respectively. In 2006 four different milk/legume based primary starters (faba bean, chick pea, lentil and white bean) were sampled in order to enumerate and identify the Bacillus at species or group level. In 2009 specific focus was on the enumeration and safety evaluation of the dominant Bacillus cereus group species occurring during various Gergoush productions (including the three fermentations steps and after baking). In 2006, the primary starters contained Bacillus spp. in the order of between 7.7 and 8.1 log₁₀ CFU/g. Species identifications were performed by M13-PCR typing using the Escherichia coli phage M13 derived primer PM13 combined with internally transcribed 16-23S rRNA PCR, 16S rRNA gene and gyrA or gyrB gene sequencing, and selected phenotypic tests. Depending on the legume used, 40-68% of the isolates were identified as B. cereus sensu lato, 16-27% as Bacillus licheniformis, 8-32% as Bacillus subtilis and 4-20% as Bacillus sonorensis. During the second set of fermentation trials performed in 2009, the Bacillus spp. and B. cereus occurred in numbers of between 7.7-9.9 and 6.1-7.8 log₁₀ CFU/g, respectively, while no bacteria were detected after baking. A total of 180 B. cereus sensu lato isolates from four different primary starters, adapted starters and final doughs were further identified as B. cereus sensu stricto (118 isolates) and Bacillus thuringiensis (62 isolates). The safety of Gergoush was evaluated based on the counts and toxin gene profiles of the dominant B. cereus species. Considering that no bacteria survived the baking process, and that the cereulide synthetase gene cesB involved in the production of the heat stable emetic toxin cereulide was not detected in any of the investigated B. cereus isolates, the results indicate, that Gergoush produced at the traditional production site is safe for human consumption. This study is the first to identify the Bacillus of Gergoush to species level, and it is the first to perform a safety evaluation of the product, based on the dominant B. cereus species.     

Molecular and toxigenic characterization of Bacillus cereus and Bacillus thuringiensis strains isolated from commercial ground roasted coffee

Thirty samples of roasted ground coffee beans from 10 different commercial brands were analyzed to investigate the occurrence and levels of Bacillus cereus and Bacillus thuringiensis strains. Strains were evaluated for their genetic diversity by repetitive element sequence polymorphism PCR (Rep-PCR) and for their toxigenic profiles, i.e., the presence of hblA, hblC, hblD, nheA, nheB, nheC, cytK, ces, and entFM. Survival and multiplication of B. cereus sensu lato in the ready-to-drink coffee was determined to evaluate this beverage as a possible vehicle for B. cereus infection. B. cereus was detected in 17 (56.7%) of the 30 samples, and B. thuringiensis was detected in 8 (26.7%) of the 30 samples. Five samples did not produce any characteristic growth. The most common gene, entFM, was detected in 23 strains (92%). The NHE complex (nheA, nheB, and nheC genes) was found in 19 strains (76%). The HBL complex (hblA, hblC, and hblD) was found in 16 strains (64%). All strains were negative for ces. The cytK gene was found in 16 strains (64%). The computer-assisted cluster analysis of Rep-PCR profiles using a clustering criterion of 80% similarity revealed four main clusters. Cluster 1 was the predominant and comprised three B. thuringiensis strains with 100% similarity, cluster 2 comprised two B. cereus strains (100% similarity), cluster 3 comprised two B. thuringiensis strains (90% similarity), and cluster 4 comprised one B. thuringiensis strain and one B. cereus strain (85% similarity). The cluster analysis of fingerprints generated by Rep-PCR revealed a high genetic diversity among the B. cereus strains, suggesting that the contamination could have originated from different sources. In our experiments, when sugar was added and the beverage was kept in thermic bottles there was a significant increase in B. cereus sensu lato levels, which may increase the risk of food poisoning. These results highlight the need for additional studies on this subject to better evaluate coffee as a food poisoning vehicle.     

Enterotoxigenic Profiling of Emetic Toxin‐ and Enterotoxin‐Producing Bacillus cereus,Isolated from Food,Environmental, and Clinical Samples by Multiplex PCR

Bacillus cereus comprises the largest group of endospore‐forming bacteria and can cause emetic and diarrheal food poisoning. A total of 496 B. cereus strains isolated from various sources (food, environmental, clinical) were assessed by a multiplex PCR for the presence of enterotoxin genes. The detection rate of nheA, entFM, hblC, and cytK enterotoxin genes among all B. cereus strains was 92.33%, 77.21%, 59.47%, and 47.58%, respectively. Enterotoxigenic profiles were determined in emetic toxin‐ (8 patterns) and enterotoxin‐producing strains (12 patterns). The results provide important information on toxin prevalence and toxigenic profiles of B. cereus from various sources. Our findings revealed that B. cereus must be considered a serious health hazard and Bacillus thuringiensis should be considered of a greater potential concern to food safety among all B. cereus group members. Also, there is need for intensive and continuous monitoring of products embracing both emetic toxin and enterotoxin genes.     

Effect of Novel Starter Culture on Reduction of Biogenic Amines,Quality Improvement,and Sensory Properties of Doenjang,a Traditional Korean Soybean Fermented Sauce Variety

To select appropriate microorganisms as starter cultures for the reliable and reproducible fermentation of soybean fermented products of Korean Doenjang, various ratios of fungi (Aspergillus oryzae J, Mucor racemosus 15, M. racemosus 42) combined with Bacillus subtilis TKSP 24 were selected as either single, double, or multiple Meju strains for commercial mass production of Doenjang, followed by analysis of sensory characteristics. In the sensory evaluation, Doenjang BAM15‐1 and BAM42‐1, which were fermented with multiple strains (1:1:1), showed the highest sensory scores as compared to control. Based on sensory characteristics, 6 Doenjang samples were subjected to quantitative determination of amino acids, free sugars, and organic acids (volatile and nonvolatile) contents, followed by determination of biogenic amines. Total sweet taste amino acid contents were highest in BAM15‐1 and BAM42‐1 samples (333.7 and 295.8 mg/100 g, respectively) and similar that of control (391.1 mg/100 g). Samples BAM15‐1 and BAM42‐1 showed the relatively high volatile and nonvolatile organic acid contents (154.24, 192.26, and 71.31, 82.42 mg/100 g, respectively). In addition, BAM15‐1 and BAM42‐1 showed negligible biogenic amine formation, ranging from 0.00 to 1.02 and 0.00 to 3.92 mg/100 g, respectively. These findings indicate that determination of food components along with sensory and quality attributes using multiple microbial Meju strains as a starter culture may provide substantial results on improved quality fermented Doenjang products.     

INCIDENCE AND CHARACTERIZATION OF BACILLUS CEREUS IN MEAT AND MEAT PRODUCTS CONSUMED IN TURKEY

A total of 100 retail samples of meat and meat products were examined for Bacillus cereus using mannitol egg yolk polymyxin (MYP) agar as a selective isolation medium. Only 22.4% of the samples contained detectable levels of B. cereus, with counts ranging from log₁₀ 0.69 to 4.80 cfu/g, but a large number of other organisms up to log₁₀ 9.06 cfu/g were sometimes observed on the plates and may have masked the presence of B. cereus or inhibited growth. Two samples of soudjouck contained significant levels of B. cereus, sufficient enough to create a public health hazard. Selected isolates were tested for diarrheal enterotoxin production by a reversed passive latex agglutination (RPLA) test kit. Results showed no difference in the toxin production of B. cereus between beef, ground meat, soudjouck and pastrami samples. Plasmid‐profile analysis and susceptibility to the six commonly used antimicrobial agents were done on selected B. cereus isolates. About 96.4% of the isolates contained plasmids. Plasmid sizes ranged between 2.5 and 3.5 kb. The isolates showed a high rate of resistance to oxacillin (OXA) and amoxicillin (AMC) and a low frequency of resistance to the other antimicrobial agents, with all of them being susceptible to vancomycin (VAN). Approximately 54% of the isolates showed multiple resistance. There was no apparent relationship between drug resistance and carriage of plasmids.     

Toxin genes profiles and toxin production ability of Bacillus cereus isolated from clinical and food samples

Bacillus cereus can cause diarrheal and emetic type of food poisoning but little study has been done on the main toxins of food poisoning caused by B. cereus in Korea. The objective of this study is to characterize the toxin gene profiles and toxin-producing ability of 120 B. cereus isolates from clinical and food samples in Korea. The detection rate of nheABC, hblCDA, entFM, and cytK enterotoxin gene among all B. cereus strains was 94.2, 90.0, 65.8, and 52.5%, respectively. The ces gene encoding emetic toxin was not detected in all strains. Bacillus cereus strains carried at least 1 of the 8 enterotoxin genes were classified into 12 groups according to the presence or absence of 8 virulence genes. The 3 major patterns, I (nheABC, hblCDA, entFM, and cytK gene), II (nheABC, hblCDA and entFM gene), and VI (nheABC and hblCDA gene), accounted for 79.2% of all strains (95 out of 120 B. cereus isolates). Non-hemolytic enterotoxin (NHE) and hemolysin BL (HBL) enterotoxins were produced by 107 and 100 strains, respectively. Our finding revealed that NHE and HBL enterotoxins encoded by nhe and hbl genes were the major toxins among B. cereus tested in this study and enterotoxic type of B. cereus was predominant in Korea.     

ANTIMICROBIAL ACTIVITY AND PROTEIN PROFILES OF PEDIOCOCCUS SPP. ISOLATED FROM TURKISH "SUCUK"

In this study, a total of 206 lactic acid bacteria were isolated on De Man–Rogosa–Sharpe agar from 20 fermented Turkish “sucuk” samples. Twenty‐five isolates were identified as Pediococcus spp. and were tested for their antibacterial activity against Staphylococcus aureus, Micrococcus luteus, Listeria monocytogenes, Bacillus cereus, Salmonella Typhimurium and two strains of Escherichia coli. Only eight strains (32%) of pediococci showed a significant degree of antibacterial activity against L. monocytogenes and S. aureus. According to sodium dodecyl sulfate‐polyacrylamide gel electrophoresis results, among eight isolates, two showed different protein profiles. The size of proteins that made the difference was calculated from 60 to 80 kDa. After partial purification, the highest degree of inhibition against L. monocytogenes was caused by the protein fractions collected after 40% ammonium sulfate precipitation. Therefore, these isolates, which produce antimicrobial substances, have a potential especially to control L. monocytogenes in fermented meat products.     

Diversity and toxigenicity among members of the Bacillus cereus group

Members of the Bacillus cereus group were isolated from rice products by centrifugation-plating and conventional spread-plating methods. Random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) results showed broad diversity among the strains and revealed some associations among isolates from raw and cooked rice samples, at the genotypic level. A comparatively greater diversity among strains was observed in isolates from raw rice than those from cooked rice and, generally, the RAPD profiles of isolates from raw and cooked rice were different, with only a few of them common to both types of rice. The toxigenic potential of the isolates was also determined by molecular and immunoassay analyses. The results revealed that most isolates from the B. cereus group were potentially or actually toxigenic, and some isolates could produce both diarrhoeal and emetic toxins. Generally, isolates belonging to the B. cereus group with the same RAPD pattern were shown to have a similar profile of enterotoxigenicity.     

江苏省徐州市市售发酵与非发酵豆制品微生物污染调查

目的 调查江苏省徐州市市售豆制品微生物污染情况。方法 采集发酵、非发酵豆制品86份, 按照GB 4789进行大肠菌群、菌落总数(非发酵豆制品)、蜡样芽胞杆菌、金黄色葡萄球菌的定量和单核细胞增生李斯特氏菌、沙门氏菌的定性检测。结果 大肠菌群检出率为34.88%, 蜡样芽胞杆菌为12.79%, 金黄色葡萄球菌为1.16%, 其余致病菌均未检出。非发酵豆制品中大肠菌群检出率为65.91%, 计数>103 CFU/g占52.27%, 散装较预包装样品检出率高, 差异有显著意义(P<0.05); 菌落总数>105 CFU/g占75.00%; 蜡样芽胞杆菌检出率为6.82%, 104 CFU/g(均<105 CFU/g)占4.55%; 金黄色葡萄球菌检出率为2.27%。发酵类豆制品中大肠菌群检出率为2.38%, 蜡样芽胞杆菌为19.05%, 104 CFU/g占7.14%(均<105 CFU/g); 预包装、散装样品蜡样芽胞杆菌检出率差异无显著意义(P>0.05)。发酵、非发酵豆制品中大肠菌群检出率差异均有显著意义(P<0.05); 发酵豆制品合格率为97.62%, 非发酵豆制品合格率为11.36%, 差异有显著意义(P<0.05)。结论 徐州市豆制品中非发酵类卫生指标菌污染较为严重, 应加强卫生监督。     

Bacillus subtilis HJ18‐4 from Traditional Fermented Soybean Food Inhibits Bacillus cereus Growth and Toxin‐Related Genes

Bacillus subtilis HJ18‐4 isolated from buckwheat sokseongjang, a traditional Korean fermented soybean food, exhibits broad‐spectrum antimicrobial activity against foodborne pathogens, including Bacillus cereus. In this study, we investigated the antibacterial efficacy and regulation of toxin gene expression in B. cereus by B. subtilis HJ18‐4. Expression of B. cereus toxin–related genes (groEL, nheA, nheC, and entFM) was downregulated by B. subtilis HJ18‐4, which also exhibited strong antibacterial activity against B. cereus. We also found that water extracts of soy product fermented with B. subtilis HJ18‐4 significantly inhibited the growth of B. cereus and toxin expression. These results indicate that B. subtilis HJ18‐4 could be used as an antimicrobial agent to control B. cereus in the fermented soybean food industry. Our findings also provide an opportunity to develop an efficient biological control agent against B. cereus.     

Fermentation properties of low-salted Doenjang supplemented with licorice and mustard

Licorice (Glycyrrhiza glabra) and mustard (Brassica juncea) were added to doenjang to reduce salt content in traditional Korean doenjang and solve problems related to abnormal fermentation during the storing process. After supplementing the low-salted doenjang (6.3% salt) with licorice and mustard (LM), a number of fungi were reduced and lactic acid bacteria increased. Protease activity increased in the 6.3% salted doenjang regardless of additives. The amino nitrogen content increased as much as that in the low-salted doenjang. pH decreased and acidity increased at day 10. A sensory evaluation was conducted after doenjang was stored for 40 days. The 6.3% + LM had very high taste quality. LM added to lowsalted doenjang suppressed fungal growth, and promoted the growth of lactic acid bacteria. The results indicate that the salts content of doenjang could be lowered to 6.3% by adding LM with improved quality and taste, and without abnormal fermentation.