1.Isolation of a foodborne Bacillus cereus strain and its effect on intestinal mucosal immunity-associated factors and gut microbial community in mice.
Li GAO ; Shan HE ; Lili WANG ; Yuting LIU ; Tong WEN
Chinese Journal of Biotechnology 2023;39(4):1759-1772
		                        		
		                        			
		                        			Bacillus cereus is a common foodborne pathogen. Accidently eating food contaminated by B. cereus will cause vomiting or diarrhea, and even death in severe cases. In the present study, a B. cereus strain was isolated from spoiled rice by streak culture. The pathogenicity and drug resistance of the isolated strain were analyzed by drug sensitivity test and PCR amplification of virulence-associated gene respectively. Cultures of the purified strain were injected intraperitoneally into mice to examine their effects on intestinal immunity-associated factors and gut microbial communities, to provide references for the pathogenic mechanism and medication guidance of these spoilage microorganisms. The results showed that the isolated B. cereus strain was sensitive to norfloxacin, nitrofurantoin, tetracycline, minocycline, ciprofloxacin, spectinomycin, clindamycin, erythrocin, clarithromycin, chloramphenicol, levofloxacin, and vancomycin, but resistant to bactrim, oxacillin and penicillin G. The strain carries seven virulence-associated genes including hblA, hblC, hblD, nheA, nheB, nheC and entFM, which are involved in diarrhea-causing toxins production. After infecting mice, the isolated B. cereus strain was found to cause diarrhea in mice, and the expression levels of immunoglobulins and inflammatory factors in the intestinal mucosae of the challenged mice were significantly up-regulated. Gut microbiome analysis showed that the composition of gut microbial community in mice changed after infection with B. cereus. The abundance of the uncultured_bacterium_f_Muribaculaceae in Bacteroidetes, which is a marker of body health, was significantly decreased. On the other hand, the abundance of uncultured_bacterium_f_Enterobacteriaceae, which is an opportunistic pathogen in Proteobacteria and a marker of dysbacteriosis, was significantly increased and was significantly positively correlated with the concentrations of IgM and IgG. These results showed that the pathogenic B. cereus carrying diarrhea type virulence-associated gene can activate the immune system by altering the composition of gut microbiota upon infection.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Mice
		                        			;
		                        		
		                        			Bacillus cereus/metabolism*
		                        			;
		                        		
		                        			Food Microbiology
		                        			;
		                        		
		                        			Immunity, Mucosal
		                        			;
		                        		
		                        			Diarrhea
		                        			;
		                        		
		                        			Microbiota
		                        			;
		                        		
		                        			Enterotoxins/genetics*
		                        			
		                        		
		                        	
2.Prokaryotic expression and biological activities of the hemolysin BL subunit of a pathogenic Bacillus cereus of cattle origin.
Yunjiao CHEN ; Yunjiang HE ; Qinglei MENG ; Zhilin LIU ; Xin ZHANG ; Zelin JIA ; Jiayu CUI ; Xueli WANG
Chinese Journal of Biotechnology 2023;39(12):4939-4949
		                        		
		                        			
		                        			Bacillus cereus belongs to Gram-positive bacteria, which is widely distributed in nature and shows certain pathogenicity. Different B. cereus strains carry different subsets of virulence factors, which directly determine the difference in their pathogenicity. It is therefore important to study the distribution of virulence factors and the biological activity of specific toxins for precise prevention and control of B. cereus infection. In this study, the hemolysin BL triayl was expressed, purified, and characterized. The results showed that the bovine pathogenic B. cereus hemolysin BL could be expressed and purified in the prokaryotic expression system, and the bovine pathogenic B. cereus hemolysin BL showed hemolysis, cytotoxicity, good immunogenicity and certain immune protection in mice. In this study, the recombinant expression of hemolysin BL triayl was achieved, and the biological activity of hemolysin BL of bovine pathogenic ceroid spore was investigated. This study may facilitate further investigating the pathogenic mechanism of B. cereus hemolysin BL and developing a detection method for bovine pathogenic B. cereus disease.
		                        		
		                        		
		                        		
		                        			Cattle
		                        			;
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Mice
		                        			;
		                        		
		                        			Bacterial Proteins/metabolism*
		                        			;
		                        		
		                        			Bacillus cereus/metabolism*
		                        			;
		                        		
		                        			Hemolysin Proteins/metabolism*
		                        			;
		                        		
		                        			Virulence Factors/metabolism*
		                        			;
		                        		
		                        			Enterotoxins/metabolism*
		                        			
		                        		
		                        	
3.First comprehensive report of bacteria spp. associated with cloaca of Laudakia nupta (Sauria: Agamidae) in Iran using molecular studies
Farkhondeh SAYYADI ; Nasrullah RASTEGAR-POUYANI ; Mehri AZADBAKHT ; Khosrow CHEHRI
Laboratory Animal Research 2019;35(1):9-12
		                        		
		                        			
		                        			Iran bears a remarkable variety of reptiles. One of the lizard families occurring in Iran is the Family Agamidae which is widely are distributed throughout the old world. The large-scaled rock agamid, Laudakia nupta, is one of the well-known agamid. There are few reports of cloacal microbial on reptiles hence their function in cloacae remains unknown. Laudakia nupta usually live in rural and urban areas and close vicinity to man, they are likely to play an important role in the spread of disease that may be caused by these microorganisms and their transmission to man. Therefore, the aim of this study was to identify the bacterial flora colonizing the cloacal region of Laudakia nupta using molecular studies. The cloacal fluids were directly placed on nutrient agar (NA) plates and incubated at 25 ± 2 ℃ for 48 h. The resulting bacterial colonies were transferred to fresh nutrient agar (NA) plates for molecular studies. Twelve isolates were obtained from 17 specimens of Laudakia nupta. All bacteria isolates were identified as Bacillus subtillis (5), Bacillus cereus (4), Bacillus sp. (1), Pseudomonas putida (1), and Pseudomonas sp. (1) based on partial sequences of the 16 s rRNA gene. This is the first comprehensive report of bacteria spp. associated with cloaca of Laudakia nupta using molecular studies. In this research, we found that Laudakia nupta can be a carrier of bacteria which can transfer microorganisms to hosts.
		                        		
		                        		
		                        		
		                        			Agar
		                        			;
		                        		
		                        			Bacillus
		                        			;
		                        		
		                        			Bacillus cereus
		                        			;
		                        		
		                        			Bacteria
		                        			;
		                        		
		                        			Cloaca
		                        			;
		                        		
		                        			Colon
		                        			;
		                        		
		                        			Genes, rRNA
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Iran
		                        			;
		                        		
		                        			Lizards
		                        			;
		                        		
		                        			Pseudomonas
		                        			;
		                        		
		                        			Pseudomonas putida
		                        			;
		                        		
		                        			Reptiles
		                        			
		                        		
		                        	
4.Intraventricular Vancomycin Therapy for Intractable Bacillus cereus Ventriculitis
Jong Woo HAHN ; Hee young JU ; Meerim PARK ; Eun Sang YI ; Byung Kiu PARK ; Sang Hoon SHIN ; Sang Hyun LEE ; Hyeon Jin PARK ; Ji Man KANG
Pediatric Infection & Vaccine 2019;26(2):124-128
		                        		
		                        			
		                        			Bacillus cereus causes serious central nervous system infections, especially in immunocompromised patients. Successful treatment requires adequate antimicrobial concentrations in the cerebrospinal fluid; however, in some cases, achieving this with systemic treatment alone is difficult. We treated intractable B. cereus ventriculitis with intraventricular vancomycin, with no major adverse events.
		                        		
		                        		
		                        		
		                        			Bacillus cereus
		                        			;
		                        		
		                        			Bacillus
		                        			;
		                        		
		                        			Central Nervous System Infections
		                        			;
		                        		
		                        			Cerebral Ventriculitis
		                        			;
		                        		
		                        			Cerebrospinal Fluid
		                        			;
		                        		
		                        			Immunocompromised Host
		                        			;
		                        		
		                        			Injections, Intraventricular
		                        			;
		                        		
		                        			Pharmacokinetics
		                        			;
		                        		
		                        			Vancomycin
		                        			
		                        		
		                        	
5.Antimicrobial Activities of Omija Extracts Against Bacillus cereus and Escherchia coli.
Youngah YOO ; Heejin HAM ; Insil YU ; Donghyiun YOOK ; Sujin KIM
Journal of Bacteriology and Virology 2018;48(1):31-36
		                        		
		                        			
		                        			Omija berry (Schisandra fructus) is a traditional Korean fruit, which contains lots of medicinal ingredients. In order to analyze whether Omija contains antibacterial components, we extracted Omija using five different methods including water precipitation, ethanol precipitation, hot water extract, methanol-ultrasonication, and water-ultrasonication, and examined their antimicrobial activities against Escherichia coli (E. coli), and Bacillus cereus (B. cereus). Most of Omija extracts did not inhibit bacteria growth in the paper disc diffusion assay except hot water extract for 60 minutes. Hot water extract for 60 minutes made clear inhibition zone around the disc at all the concentrations (x1, x1/10, x1/100) in both E. coli and B. cereus. However, hot water extracts for 90 minutes and 120 minutes showed antimicrobial activities only at x1 stock solution, which indicates that 60 minutes extraction at hot water is the best method to obtain Omija extract with the best antimicrobial activities in disc diffusion assay. In turbidimetric assay, water extract in soak, hot water extract, and methanol extract in ultrasonicator inhibited growth of both E. coli and B. cereus. Ethanol extract in soak and water extract in ultrasonicator had no effect on bacterial growth in both E. coli and B. cereus. In this study, we found that Omija extracts showed antimicrobial activities against E. coli and B. cereus. Therefore, water soluble materials of Omija can be used as a drink supplement and developed as an antibacterial cleanser.
		                        		
		                        		
		                        		
		                        			Bacillus cereus*
		                        			;
		                        		
		                        			Bacillus*
		                        			;
		                        		
		                        			Bacteria
		                        			;
		                        		
		                        			Diffusion
		                        			;
		                        		
		                        			Escherichia coli
		                        			;
		                        		
		                        			Ethanol
		                        			;
		                        		
		                        			Fruit
		                        			;
		                        		
		                        			Methanol
		                        			;
		                        		
		                        			Methods
		                        			;
		                        		
		                        			Water
		                        			
		                        		
		                        	
6.Anti-Bacterial Effect of Lactobacillus rhamnosus Cell-Free Supernatant Possessing Lysozyme Activity Against Pathogenic Bacteria
Jiyeon LEE ; Hyeji LIM ; Misook KIM
Journal of the Korean Dietetic Association 2018;24(4):330-343
		                        		
		                        			
		                        			Recently, there has been a growing demand for natural preservatives because of increased consumer interest in health. In this study, we produced Lactobacillus rhamnosus cell-free supernatant (LCFS) and evaluated and compared its antimicrobial activity with existing natural preservatives against pathogenic microorganisms and in chicken breast meat contaminated with Escherichia coli and Staphylococcus aureus. Lactobacillus rhamnosus cell-free supernatant possessed 30 units of lysozyme activity and contained 18,835 mg/L of lactic acid, 2,051 mg/L of citric acid and 5,060 mg/L of acetic acid. Additionally, LCFS inhibited the growth of fourteen pathogenic bacteria, S. aureus, Bacillus cereus, Listeria monocytogenes, Vibrio parahaemolyticus, Listeria innocua, S. epidermidis, L. ivanovii, E. coli, Pseudomonas aeruginosa, Shigella sonnei, Shi. flexneri, Proteus vulgaris, Pseudomonas fluorescens, and Klebsiella pneumoniae. The antibacterial activity of LCFS was stronger than that of egg white lysozyme (EWL), Durafresh (DF) and grapefruit seed extract (GSE). Additionally, LCFS maintained its antimicrobial activity after heat treatment at 50℃~95℃ and at pH values of 3~9. Moreover, LCFS inhibited the growth of E. coli and S. aureus in chicken breast meat. In conclusion, it is expected that LCFS, which contains both lysozyme and three organic acids, will be useful as a good natural preservative in the food industry.
		                        		
		                        		
		                        		
		                        			Acetic Acid
		                        			;
		                        		
		                        			Bacillus cereus
		                        			;
		                        		
		                        			Bacteria
		                        			;
		                        		
		                        			Breast
		                        			;
		                        		
		                        			Chickens
		                        			;
		                        		
		                        			Citric Acid
		                        			;
		                        		
		                        			Citrus paradisi
		                        			;
		                        		
		                        			Egg White
		                        			;
		                        		
		                        			Escherichia coli
		                        			;
		                        		
		                        			Food Industry
		                        			;
		                        		
		                        			Hot Temperature
		                        			;
		                        		
		                        			Hydrogen-Ion Concentration
		                        			;
		                        		
		                        			Klebsiella pneumoniae
		                        			;
		                        		
		                        			Lactic Acid
		                        			;
		                        		
		                        			Lactobacillus rhamnosus
		                        			;
		                        		
		                        			Lactobacillus
		                        			;
		                        		
		                        			Listeria
		                        			;
		                        		
		                        			Listeria monocytogenes
		                        			;
		                        		
		                        			Meat
		                        			;
		                        		
		                        			Muramidase
		                        			;
		                        		
		                        			Proteus vulgaris
		                        			;
		                        		
		                        			Pseudomonas aeruginosa
		                        			;
		                        		
		                        			Pseudomonas fluorescens
		                        			;
		                        		
		                        			Shigella sonnei
		                        			;
		                        		
		                        			Staphylococcus aureus
		                        			;
		                        		
		                        			Vibrio parahaemolyticus
		                        			
		                        		
		                        	
7.Chemical Composition and Antimicrobial Efficacy of Helminthostachys zeylanica against Foodborne Bacillus cereus
Tong Woei YENN ; Leong Chean RING ; Khairul Azly ZAHAN ; Muhammad Sharir ABDUL RAHMAN ; Wen Nee TAN ; Bintul Jauza' SHAIK ALAUDIN
Natural Product Sciences 2018;24(1):66-70
		                        		
		                        			
		                        			Helminthostachys zeylanica is a rare plant grows in lightly shaded areas. The fern was traditionally used as antipyretic and antiphlogistic agents. This study was aimed to evaluate the antibacterial potential of H. zeylanica on foodborne Bacillus cereus. The chemical composition of its ethanolic extract was also determined. The plant samples were collected at Kampung Kebun Relong, Kedah, Malaysia. The ethanolic extract showed significant inhibitory activity on B. cereus with a sizeable clear zone detected on disc diffusion assay. On broth microdilution assay, the MIC of the extract on B. cereus was 6.25 mg/ml and the MBC was 12.5 mg/ml. The inhibitory activity of the extract on B. cereus was bactericidal. In the growth dynamic study, the antibacterial efficacy of the extract was concentration dependent, where a lower colony forming unit count was obtained with increased extract concentration. The SEM micrograph of extract treated B. cereus cells showed invaginations of cell wall. The bacterial cell structure collapsed after 24 h exposure to the extract. The GCMS analysis of the extract showed that the major constituents of the extract were phenol (36.26%) and quercetin (29.70%). This study is important as it shows the potential use of H. zeylanica as an effective agent to control B. cereus related infections.
		                        		
		                        		
		                        		
		                        			Bacillus cereus
		                        			;
		                        		
		                        			Bacillus
		                        			;
		                        		
		                        			Cell Wall
		                        			;
		                        		
		                        			Diffusion
		                        			;
		                        		
		                        			Ethanol
		                        			;
		                        		
		                        			Ferns
		                        			;
		                        		
		                        			Gas Chromatography-Mass Spectrometry
		                        			;
		                        		
		                        			Malaysia
		                        			;
		                        		
		                        			Phenol
		                        			;
		                        		
		                        			Plants
		                        			;
		                        		
		                        			Quercetin
		                        			;
		                        		
		                        			Stem Cells
		                        			
		                        		
		                        	
8.A Conformational Isomer of Soulattrolide from the Stem Bark of Calophyllum symingtonianum and Its Antibacterial Activity
Deny SUSANTI ; Nissad ATTOUMANI ; Muhammad TAHER ; Mohd Fazlin REZALI ; Md Hossain SOHRAB ; Choudhury Mahmood HASAN ; Zainul AMIRUDDIN ZAKARIA
Natural Product Sciences 2018;24(1):47-53
		                        		
		                        			
		                        			Callophylum symingtonianum (Guttiferae), an evergreen broad-leaved tree that usually grows in hill forests, can be found distributed in the Malay Peninsula. The barks, leaves, flowers and seeds is often used medicinally to treat diarrhea and rheumatism. In the present study, we isolated two inophyllum type coumarins, 12-O-ethylinophyllum D (1) and iso-soulattrolide (2) from the stembarks of C. symingtonianum together with their antibacterial activity. The compounds were isolated by chromatographic methods on a silica gel. The structures were established by spectroscopic methods including UV, IR, (1D and 2D) NMR and mass spectrometry as well as by comparison with several literature sources. The antibacterial activity of those compounds was tested using a disc-diffusion assay against Staphylococcus aureus, Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa. Both compound exhibited mild inhibition against P. aeruginosa with both 111 µg/ml MIC value. Compound 2 also inhibits S. aureus with 25 µg/ml MIC value.
		                        		
		                        		
		                        		
		                        			Bacillus cereus
		                        			;
		                        		
		                        			Calophyllum
		                        			;
		                        		
		                        			Coumarins
		                        			;
		                        		
		                        			Diarrhea
		                        			;
		                        		
		                        			Escherichia coli
		                        			;
		                        		
		                        			Flowers
		                        			;
		                        		
		                        			Forests
		                        			;
		                        		
		                        			Malaysia
		                        			;
		                        		
		                        			Mass Spectrometry
		                        			;
		                        		
		                        			Pseudomonas aeruginosa
		                        			;
		                        		
		                        			Rheumatic Diseases
		                        			;
		                        		
		                        			Silica Gel
		                        			;
		                        		
		                        			Staphylococcus aureus
		                        			;
		                        		
		                        			Trees
		                        			
		                        		
		                        	
9.New Azafluorenone Derivative and Antibacterial Activities of Alphonsea cylindrica Barks.
Munirah Abdul TALIP ; Saripah Salbiah Syed Abdul AZZIZ ; Chee Fah WONG ; Khalijah AWANG ; Humera NAZ ; Yuhanis Mhd BAKRI ; Mohamad Syahrizal AHMAD ; Marc LITAUDON
Natural Product Sciences 2017;23(3):151-156
		                        		
		                        			
		                        			A phytochemical study of Alphonsea cylindrica King (unreported) has led to the isolation of six alkaloids. The compounds were identified as kinabaline (1; azafluorenone alkaloid), muniranine (2), O-methylmoschatoline (3; oxoaporphine alkaloid), lysicamine (4), atherospermidine (5) and N-methylouregidione (6; 4, 5-dioxoaporphine alkaloid). The structures of the isolated compounds were determined based on the spectroscopic techniques and by comparison with data reported in the literature. Alkaloid 2 was isolated as a new derivative of azafluorenone while alkaloids 1, 3 - 6 were isolated for the first time from Alphonsea species. In addition, alkaloid 3 and 4 showed inhibition zone against Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus cereus in disc diffusion test. The minimum inhibition concentration (MIC) values of lysicamine (4) against S. aureus, B. cereus and P. aeruginosa were found to be smaller than O-methylmoschatoline (3). Therefore, the reported antibacterial activity showed the potential of this plant as natural antibacterial agent and supported the documented traditional use of Alphonsea sp. in the treatment of diarrhea and fever.
		                        		
		                        		
		                        		
		                        			Alkaloids
		                        			;
		                        		
		                        			Bacillus cereus
		                        			;
		                        		
		                        			Diarrhea
		                        			;
		                        		
		                        			Diffusion
		                        			;
		                        		
		                        			Fever
		                        			;
		                        		
		                        			Plants
		                        			;
		                        		
		                        			Pseudomonas aeruginosa
		                        			;
		                        		
		                        			Staphylococcus aureus
		                        			
		                        		
		                        	
10.Distributions of Bacillus cereus, Pseudomonas, Enterococcus, and coliforms Isolated from Agricultural Products.
Journal of Bacteriology and Virology 2017;47(3):139-147
		                        		
		                        			
		                        			A survey was conducted on the bacterial contamination levels from 128 agricultural products, and identified 31 coliforms, 34 Bacillus spp., 35 Pseudomonas spp., and 3 Enterococcus spp.. Antibiotic susceptibility testing of eight Escherichia coli (E. coli) isolates, thirteen Bacillus cereus (B. cereus) isolates and seven Pseudomonas aeruginosa (P. aeruginosa) reveled that none of the E. coli isolates shows multidrug resistance pattern. However, one B. cereus isolate was resistant to Amoxicillin/clavulanic acid + Ampicillin/sulbactam + Cephaloxin, and twelve B. cereus isolates were resistant to Amoxicillin/clavulanic acid + Ampicillin/sulbactam + Cephaloxin + Tetracycline. The data from the current study points out that many agricultural products contain multidrug resistant bacteria, and it can be suggested for the need of proper guidelines and special management on the use of antibiotics in agricultural products.
		                        		
		                        		
		                        		
		                        			Anti-Bacterial Agents
		                        			;
		                        		
		                        			Bacillus cereus*
		                        			;
		                        		
		                        			Bacillus*
		                        			;
		                        		
		                        			Bacteria
		                        			;
		                        		
		                        			Drug Resistance, Multiple
		                        			;
		                        		
		                        			Enterococcus*
		                        			;
		                        		
		                        			Escherichia coli
		                        			;
		                        		
		                        			Pseudomonas aeruginosa
		                        			;
		                        		
		                        			Pseudomonas*
		                        			;
		                        		
		                        			Tetracycline
		                        			
		                        		
		                        	
            
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