Degradation of zearalenone in swine feed and feed ingredients by Bacillus subtilis ANSB01G

2014 ◽  
Vol 7 (2) ◽  
pp. 143-151 ◽  
Author(s):  
Y.P. Lei ◽  
L.H. Zhao ◽  
Q.G. Ma ◽  
J.Y. Zhang ◽  
T. Zhou ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Culture Supernatant ◽  
Antimicrobial Activities ◽  
Proteinase K ◽  
Rrna Gene ◽  
Distillers Grains With Solubles ◽  
Dried Distillers Grains ◽  
Cell Extracts ◽  
Gene Sequence Analysis ◽  
And Staphylococcus Aureus

Zearalenone (ZEA) and its derivatives are mycotoxins that can cause oestrogenic effects and impair the reproductive physiology of animals, especially in female swine. Strategies to reduce or eliminate ZEA contamination in foods and feeds are very much needed. Among 36 bacterial isolates obtained from a variety of animal intestinal chyme, mouldy foods and feeds, soils, etc., five isolates demonstrated the ability to reduce more than 50% of ZEA in a liquid medium; ANSB01G isolate taken from normal broiler intestinal chyme reduced ZEA the most, by 88.65%. Using physiological, biochemical, and 16S rRNA gene sequence analysis methods, the ANSB01G isolate was identified as Bacillus subtilis. Under simulated intestinal tract conditions, the ANSB01G B. subtilis isolate degraded 84.58, 66.34 and 83.04% of ZEA in naturally contaminated maize, dried distillers’ grains with solubles, and swine complete feed, respectively. The highest degradation of ZEA occurred when the mycotoxin was co-incubated with the whole bacterial culture, resulting in a reduction of 88.65%, followed by 75.60% using culture supernatant, 26.11% using cell extracts, and 15.06% using viable cells. Treatments consisting of both heating and addition of proteinase K significantly reduced the rate of ZEA degradation in the culture supernatant, indicating that the ZEA degradation might be enzymatic. B. subtilis ANSB01G displayed resistance to simulated gastrointestinal tract environments and antimicrobial activities against several common bacterial pathogens, including Escherichia coli, Salmonella typhimurium and Staphylococcus aureus. These properties of B. subtilis ANSB01G suggest the possibility of its potential to effectively degrade ZEA in feed and to develop functional feed products for livestock industries.

Removal of ochratoxin A by a carboxypeptidase and peptides present in liquid cultures of Bacillus subtilis CW14

2018 ◽  
Vol 11 (4) ◽  
pp. 559-570 ◽  
Author(s):  
H.N. Hu ◽  
X. Jia ◽  
Y.P. Wang ◽  
Z.H. Liang
Keyword(s):  
Bacillus Subtilis ◽  
Ochratoxin A ◽  
Culture Supernatant ◽  
Physical Adsorption ◽  
Proteinase K ◽  
Small Peptides ◽  
Liquid Cultures ◽  
Ochratoxin Α ◽  
Peptide Complexes

Ochratoxin A (OTA) is an important mycotoxin that contaminates a variety of agricultural products. The cell-free supernatant of Bacillus subtilis CW14 liquid cultures were reported previously to be capable of removing OTA efficiently. In this work, we examined several substances that are probably involved in this removal of OTA using in vitro experiments. The strain CW14 culture supernatant that was separated by ultrafiltration showed that the fractions collected at >10 kDa and <3 kDa had a significant ability to reduce OTA (84.9 and 74.8%, respectively) when incubated with 6 μg/ml OTA at 37 °C for 24 h. A putative metalloenzyme was responsible for the activity of the >10-kDa fraction, which was confirmed by the detrimental effects of heat treatments or addition of SDS, proteinase K, or EDTA. Subsequently, a carboxypeptidase (CP) gene that was likely related to the enzymatic conversion of OTA by the >10-kDa fraction was cloned from the B. subtilis CW14 genome, and over-expressed in Escherichia coli. The recombinant CP degraded 71.3% of OTA at 37 °C for 24 h, and ochratoxin α (OTα) was confirmed as a degradation product. From the <3-kDa fraction, some small peptides (1.7 kDa >Mw >0.7 kDa) were purified and decreased OTA by 45.0% under the same conditions, but no product was detected. These peptides were presumed to be capable of binding OTA due to their affinity with the OTA molecule, and the OTA-peptide complexes escaped from the extraction procedures for OTA quantification. These results indicated there was a probable synergistic effect that was involved in removal of OTA by the strain CW14 culture supernatant, which included enzymatic degradation by a CP and physical adsorption by some small peptides.

scholarly journals Genotyping and Toxigenic Potential of Bacillus subtilis and Bacillus pumilus Strains Occurring in Industrial and Artisanal Cured Sausages

2004 ◽  
Vol 70 (9) ◽  
pp. 5168-5176 ◽  
Author(s):  
Alessandra Matarante ◽  
Federico Baruzzi ◽  
Pier Sandro Cocconcelli ◽  
Maria Morea
Keyword(s):  
Bacillus Subtilis ◽  
Dna Sequences ◽  
Bacillus Pumilus ◽  
Meat Products ◽  
Rrna Gene ◽  
Emerging Pathogens ◽  
Specific Pcr ◽  
Gene Sequence Analysis ◽  
Rapd Pcr ◽  
Cured Meat

ABSTRACT Artisanal and industrial sausages were analyzed for their aerobic, heat-resistant microflora to assess whether new emerging pathogens could be present among Bacillus strains naturally contaminating cured meat products. Sixty-four isolates were characterized by randomly amplified polymorphic DNA (RAPD)-PCR and fluorescent amplified fragment length polymorphism (fAFLP). The biotypes, identified by partial 16S rRNA gene sequence analysis, belonged to Bacillus subtilis, Bacillus pumilus, and Bacillus amyloliquefaciens species. Both RAPD-PCR and fAFLP analyses demonstrated that a high genetic heterogeneity is present in the B. subtilis group even in strains harvested from the same source, making it possible to isolate 56 different biotypes. Moreover, fAFLP analysis made it possible to distinguish B. subtilis from B. pumilus strains. The strains were characterized for their toxigenic potential by molecular, physiological, and immunological techniques. Specific PCR analyses revealed the absence of DNA sequences related to HBL, BcET, NHE, and entFM Bacillus cereus enterotoxins and the enzymes sphingomyelinase Sph and phospholipase PI-PLC in all strains; also, the immunological analyses showed that Bacillus strains did not react with NHE- and HBL-specific antibodies. However, some isolates were found to be positive for hemolytic and lecithinase activity. The absence of toxigenic potential in Bacillus strains from the sausages analyzed indicates that these products can be considered safe under the processing conditions they were produced; however, great care should be taken when the ripening time is shortened, particularly in the case of traditional sausages, which could contain high amounts of Bacillus strains and possibly some B. cereus cells.

scholarly journals Diversity of Actinobacteria Isolated from Date Palms Rhizosphere and Saline Environments: Isolation, Identification and Biological Activity Evaluation

2020 ◽  
Vol 8 (12) ◽  
pp. 1853
Author(s):  
Omar Messaoudi ◽  
Joachim Wink ◽  
Mourad Bendahou
Keyword(s):  
Antimicrobial Activities ◽  
Bioactive Metabolites ◽  
Multi Drug Resistance ◽  
Rrna Gene ◽  
Date Palms ◽  
Saline Environments ◽  
Taxonomic Analysis ◽  
Algerian Sahara ◽  
Gene Sequence Analysis ◽  
Rare Actinobacteria

The diversity of cultural Actinobacteria in two types of Algerian Sahara environments, including saline environments and date palms rhizosphere, was investigated. In this study, a total of 40 strains of actinomycetes was isolated from different soil samples, using a rehydration and centrifugation method. Molecular identification, based on 16S rRNA gene sequence analysis, revealed that these isolates were affiliated to six clusters corresponding to eight genera, including Streptomyces, Nocardiopsis, Saccharopolyspora, Actinomadura, Actinocorallia, Micromonospora, Couchioplanes, and Planomonospora. A taxonomic analysis, based on the morphological, physiological, biochemical, and molecular investigation, of selected strains, which belong to the rare Actinobacteria, was undertaken. Four strains (CG3, A111, A93, and A79) were found to form distinct phyletic lines and represent new actinobacterial taxa. An assessment of antimicrobial proprieties of the 40 obtained actinomycetes strains, showed moderate to strong antimicrobial activities against fungi and bacteria. This study demonstrated the richness of Algerian Sahara with rare Actinobacteria, which can provide novel bioactive metabolites, to solving some of the most challenging problems of the day, such as multi-drug resistance.

scholarly journals Phylogenetic Framework and Biosurfactant Gene Expression Analysis of Marine Bacillus spp. of Eastern Coastal Plain of Tamil Nadu

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Sreethar Swaathy ◽  
Varadharajan Kavitha ◽  
Arockiasamy Sahaya Pravin ◽  
Ganesan Sekaran ◽  
Asit Baran Mandal ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Coastal Plain ◽  
Tamil Nadu ◽  
Bacterial Species ◽  
Individual Species ◽  
Rrna Gene ◽  
Gene Sequence Analysis ◽  
Diversity Assessment ◽  
Bacillus Spp

The present study emphasizes the diversity assessment of marine Bacillus species with special reference to biosurfactant production, respective gene expression, and discrimination among Bacillus licheniformis and Bacillus subtilis. Among the 200 individual species of eastern coastal plain of Tamil Nadu screened, five biosurfactant producing potential bacterial species with entirely different morphology were selected. Biochemical and 16S rRNA gene sequence analysis suggested that all the said five species belong to Bacillus genera but differ in species levels. Biosurfactant of all the five species fluctuates in greater levels with respect to activity as well as to constituents but showed partial similarity to the commercially available surfactin. The expression of srf gene was realized in all of the five species. However, the sfp gene expression was observed only in three species. In conclusion, both B. licheniformis and B. subtilis demonstrate srf gene; nevertheless, sfp gene was expressed only by Bacillus subtilis.

scholarly journals A Novel Protein-Deamidating Enzyme fromChryseobacterium proteolyticum sp. nov., a Newly Isolated Bacterium from Soil

2000 ◽  
Vol 66 (8) ◽  
pp. 3337-3343 ◽  
Author(s):  
Shotaro Yamaguchi ◽  
Masaaki Yokoe
Keyword(s):  
New Species ◽  
Culture Supernatant ◽  
Industrial Applications ◽  
Phenotypic Characterization ◽  
Rrna Gene ◽  
Dna Relatedness ◽  
Rice Field Soil ◽  
Physiological And Biochemical Characteristics ◽  
Gene Sequence Analysis ◽  
Novel Protein

ABSTRACT A novel protein-deamidating enzyme, which has potential for industrial applications, was purified from the culture supernatant ofChryseobacterium proteolyticum strain 9670Tisolated from rice field soil in Tsukuba, Japan. The deamidating activities on carboxybenzoxy (Cbz)-Gln-Gly and caseins and protease activity were produced synchronously by the isolate. Both deamidating activities were eluted as identical peaks separated from several proteases by phenyl-Sepharose chromatography of the culture supernatant. The enzyme catalyzed the deamidation of native caseins with no protease and transglutaminase activities. Phenotypic characterization and DNA analyses of the isolate were performed to determine its taxonomy. Physiological and biochemical characteristics, 16S rRNA gene sequence analysis, and DNA-DNA relatedness data indicated that the isolate should be placed as a new species belonging to the genus Chryseobacterium. The isolate showed no growth on MacConkey agar and produced acid from sucrose. The levels of DNA-DNA relatedness between the isolate and other related strains were less than 17%. The name Chryseobacterium proteolyticum is proposed for the new species; strain 9670 is the type strain (=FERM P-17664).

scholarly journals PSVII-8 Efficacy of corn dried distillers grains with solubles as a replacement for soybean meal in a Boer goat diet

2019 ◽  
Vol 97 (Supplement_2) ◽  
pp. 161-162
Author(s):  
Rachel J Sorensen ◽  
Savannah C Stewart ◽  
Cassandra K Jones ◽  
Alison R Crane ◽  
T G Nagaraja ◽  
...  
Keyword(s):  
Soybean Meal ◽  
Animal Feed ◽  
Distillers Grains ◽  
Rrna Gene ◽  
Soluble Fiber ◽  
Fecal Microbiome ◽  
Boer Goat ◽  
Distillers Grains With Solubles ◽  
Dried Distillers Grains ◽  
Boer Goats

Abstract Due to increased use of dried distillers grains with solubles (DDGS) in animal feed and accessibility of ethanol plants in the Midwest, this study evaluated the effect of feeding DDGS in place of soybean meal (SBM) on the fecal microbiome of Boer goats. Twenty-four Boer goat kids (apx. 70 d of age; 28.21 ± 0.96 kg) were blocked by BW and randomly assigned to 1 of 2 treatment diets for 47 d. Treatments were 0% (0DDGS) and 100% (30DDGS) DDGS in place of SBM. Goats were placed in 8 pens (4 pens/treatment; 3 goats/pen) with ad libitum access to feed and water. Fecal pellets were collected on d 47 via rectal grab and stored at -80°C until microbiome sequencing was performed. The V4 region of the 16S rRNA gene was sequenced by MR DNA (MR DNA, Shallowater, TX) on the Illumina HiSeq 2500 platform (Illumina, Inc., San Diego, CA). Data were analyzed using ANOVA with Tukey’s test for pairwise comparisons. Genera impacted by DDGS inclusion with individual relative abundances greater than 1% included increased Ruminococcus (P = 0.01) and Methanobrevibacter (P = 0.009) and decreased Lachnoclostridium (P = 0.02). Ruminococcus and Methanobrevibacter most likely increased in 30DDGS due to greater amounts of soluble fiber passing through the rumen, thus being fermented in the hindgut. The overall percentage of the phyla Bacteroidetes (P = 0.36) and Firmicutes (P = 0.12) did not differ between treatments; however, Firmicutes:Bacteroidetes increased (P = 0.05) in the 30DDGS diet. Treatment did not impact β-diversity (P = 0.47) although species richness increased (P = 0.09) in DDGS-fed goats as more soluble fiber was available for fermentation in the hindgut. In all, results of this study found replacing SBM with DDGS did not greatly alter the fecal microbiome of Boer goats.

Detoxification of aflatoxin B1 by certain bacterial species isolated from Egyptian soil

2011 ◽  
Vol 4 (2) ◽  
pp. 169-176 ◽  
Author(s):  
M. Elaasser ◽  
R. El Kassas
Keyword(s):  
Culture Supernatant ◽  
Bacterial Species ◽  
Industrial Applications ◽  
Aflatoxin Contamination ◽  
Soil Samples ◽  
Proteinase K ◽  
Strong Inhibitor ◽  
E Coli ◽  
Screening Experiments ◽  
Cell Extracts

Aflatoxin contamination of food and grain poses a serious economic and health problem worldwide. Aflatoxin B1 (AFB1) is extremely mutagenic, toxic and a potent carcinogen to both humans and livestock. A safe, effective and environmentally sound detoxification method is needed for controlling this toxin. In this study, 21 soil samples were screened from various sources with vast microbial populations using a coumarin containing medium. Eleven bacterial isolates showing AFB1 reduction activity in a liquid culture medium were selected from the screening experiments. Isolate 12-3 and 12-5, obtained from soil samples of Kafr-Zaiat Pesticide company drainage and identified to be Pseudomonas putida and Escherichia coli, reduced AFB1 by 69.3% and 58.8%, respectively, after incubation in the liquid medium at 37 °C for 72 h. The culture supernatant of these isolates was able to reduce AFB1 effectively by 76.2% and 62.5%, respectively, whereas the viable cells and cell extracts were far less effective. Factors influencing AFB1 detoxification by the culture supernatant were investigated. The highest detoxification activity for P. putida and E. coli was 83.3% and 63.8%, respectively, at pH 8 and 30 °C for 72 h. The detoxification activity was reduced at 10, 20 and 45 °C. The Mg2+, Mn2+, Se and Cu2+ ions were activators for AFB1 detoxification. However, Zn2+ ion was a strong inhibitor. Treatments with proteinase K, proteinase K plus SDS and heating significantly reduced or eradicated the detoxification activity of the culture supernatant. In conclusion, the detoxification of AFB1 by P. putida 12-3 was enzymatic and the enzymes responsible for the detoxification of AFB1 are constitutively extracellular produced. Also, the AFB1 detoxification by E. coli was conducted by enzymes as well as by cell wall binding mechanism. Both bacteria could have great potential in industrial applications.

scholarly journals Antimicrobial Characteristics of Lactic Acid Bacteria Isolated from Homemade Fermented Foods

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Dayong Ren ◽  
Jianwei Zhu ◽  
Shengjie Gong ◽  
Hongyan Liu ◽  
Hansong Yu
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Antimicrobial Effect ◽  
Antimicrobial Activities ◽  
Proteinase K ◽  
Control Group ◽  
Rrna Gene ◽  
Fermented Foods ◽  
16S Rrna Gene Analysis ◽  
Group 10

Objective. Lactic acid bacteria (LAB) were isolated from fermented foods, such as glutinous rice dough, corn noodle, chili sauce, potherb mustard pickles, and stinky tofu, in northeast China. LAB strains with antimicrobial activities were screened, and seven of these Lactobacillus strains were identified as L. plantarum, L. pentosus, and L. paracasei through 16S rRNA gene analysis. After the supernatant of LAB was treated with proteinase K, pepsin, and papain, their antibacterial effect almost disappeared. Most strains with antibacterial activities were highly resistant to heat (65°C–121°C), acidity (pH 2–6), and alcohol. The antimicrobial effect of most strains treated with the Tween-80 surfactant was significantly reduced, and the antibacterial property of T4 was even lost. Ammonium sulfate precipitation, PCR, and nanoLC-ESI-MS/MS results confirmed that T8 produced antibacterial substances belonging to a protein family, and its zone of inhibition against pathogens significantly increased (>13 mm). In bacterial growth inhibition experiments, the colony count of Staphylococcus aureus was up to 1015 CFU/mL in the 3⁎de Man, Rogosa, and Sharpe (MRS) group, and this value was more than that in the 3⁎S6 supernatant group (1012 CFU/mL) and the control group (1010 CFU/mL) at 12 h. This study provided a basis for the selection of antimicrobial peptides and the development and utilization of LAB.

scholarly journals Evaluation of the Antimicrobial Efficacy of some Fermented Traditional Turkish Beverages with Probiotic Potentials

2021 ◽  
Author(s):  
Oluwaseun Temitope Aladeboyeje ◽  
Nazmiye Ozlem Sanli ◽  
Umut Buyuk
Keyword(s):  
Fruits And Vegetables ◽  
Health Benefit ◽  
Antimicrobial Activities ◽  
Rrna Gene ◽  
Human Pathogens ◽  
Strain Specificity ◽  
Antimicrobial Effects ◽  
Gene Sequence Analysis ◽  
Good Number ◽  
Microbiota Diversity

Turkey is a home country for a good number of fermented beverages derived from milk, cereals, fruits and vegetables, and several studies have reported the probiotic potentiality of these beverages. Probiotics, otherwise known as beneficial microorganisms possess the ability to exert antimicrobial effects, which is one of the most important selection criteria for their use in commercial products. In the current study, the antimicrobial activities of potential probiotic bacteria isolated from five fermented traditional Turkish beverages (boza, kefir, ayran, shalgam juice and hardaliye) were evaluated. The bacterial isolates were morphologically characterized and genotypically identified by 16S rRNA gene sequence analysis. The antimicrobial effects of the isolates against selected human pathogens were assessed using spot-on-the-lawn and agar well diffusion assays. Eighteen of the twenty-two strains displayed varying degrees of antagonism against the tested pathogens. Amongst the isolates, the strongest antimicrobial effects were exhibited by strains from boza, kefir and shalgam which can be attributed to their greater microbiota diversity. Strain specificity in the activities of the obtained isolates and specificity with the different indicator pathogens tested was observed. The impressive antimicrobial effects exhibited by boza, kefir and shalgam isolates offer a promising health benefit to consumers of these fermented probiotic products.

scholarly journals Chemical Constituents and Antimicrobial Activities of Canthium horridum

2010 ◽  
Vol 5 (6) ◽  
pp. 1934578X1000500 ◽  
Author(s):  
Biao Yang ◽  
Guangying Chen ◽  
Xiaoping Song ◽  
Zhong Chen ◽  
Xinming Song ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Bacillus Subtilis ◽  
Aspergillus Niger ◽  
Chemical Constituents ◽  
Antimicrobial Activities ◽  
Syringic Acid ◽  
Good Activity ◽  
First Time ◽  
And Staphylococcus Aureus

Bioassay-guided isolation studies of the extract of Canthium horridum Bl. stem led to the isolation of ten compounds: (+)-syringaresinol (1), scoparone (2), scopoletin (3), 3′-methoxy-4′-hydroxy- trans-cinnamaldehyde (4), sinapic aldehyde (5), syringic acid (6), mannitol (7), vanillic acid 4- O-β-D-glucopyranoside (8), β-daucosterol (9), and β-sitosterol (10). Compounds 1-10 were reported for the first time from this species, and compounds 1, 4, 5, 6, and 8 from the genus. The antimicrobial activities of the isolated compounds were studied; 6 had the highest activity against Bacillus subtilis, but 1 showed good activity against Escherichia coli, Bacillus subtilis and Staphylococcus aureus. Compounds 2, 4 and 6 also inhibited the growth of these three bacteria. None of the compounds demonstrated inhibitory activity against Aspergillus niger.

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