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.

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.

scholarly journals Degradation of 1,2-Dibromoethane byMycobacterium sp. Strain GP1

1999 ◽  
Vol 181 (7) ◽  
pp. 2050-2058 ◽  
Author(s):  
Gerrit J. Poelarends ◽  
Johan E. T. van Hylckama Vlieg ◽  
Julian R. Marchesi ◽  
Luisa M. Freitas Dos Santos ◽  
Dick B. Janssen
Keyword(s):  
Amino Acid ◽  
Ethylene Oxide ◽  
Dna Sequences ◽  
Gene Sequence ◽  
Gene Region ◽  
Rrna Gene ◽  
Haloalkane Dehalogenase ◽  
C Terminus ◽  
Gene Sequence Analysis ◽  
Dehalogenase Gene

ABSTRACT The newly isolated bacterial strain GP1 can utilize 1,2-dibromoethane as the sole carbon and energy source. On the basis of 16S rRNA gene sequence analysis, the organism was identified as a member of the subgroup which contains the fast-growing mycobacteria. The first step in 1,2-dibromoethane metabolism is catalyzed by a hydrolytic haloalkane dehalogenase. The resulting 2-bromoethanol is rapidly converted to ethylene oxide by a haloalcohol dehalogenase, in this way preventing the accumulation of 2-bromoethanol and 2-bromoacetaldehyde as toxic intermediates. Ethylene oxide can serve as a growth substrate for strain GP1, but the pathway(s) by which it is further metabolized is still unclear. Strain GP1 can also utilize 1-chloropropane, 1-bromopropane, 2-bromoethanol, and 2-chloroethanol as growth substrates. 2-Chloroethanol and 2-bromoethanol are metabolized via ethylene oxide, which for both haloalcohols is a novel way to remove the halide without going through the corresponding acetaldehyde intermediate. The haloalkane dehalogenase gene was cloned and sequenced. The dehalogenase (DhaAf) encoded by this gene is identical to the haloalkane dehalogenase (DhaA) of Rhodococcus rhodochrous NCIMB 13064, except for three amino acid substitutions and a 14-amino-acid extension at the C terminus. Alignments of the complete dehalogenase gene region of strain GP1 with DNA sequences in different databases showed that a large part of adhaA gene region, which is also present in R. rhodochrous NCIMB 13064, was fused to a fragment of a haloalcohol dehalogenase gene that was identical to the last 42 nucleotides of thehheB gene found in Corynebacterium sp. strain N-1074.

Chromosomal organization of rRNA operons in Bacillus subtilis.

Genetics ◽  
1988 ◽  
Vol 120 (3) ◽  
pp. 625-635
Author(s):  
E D Jarvis ◽  
R L Widom ◽  
G LaFauci ◽  
Y Setoguchi ◽  
I R Richter ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Dna Sequences ◽  
Hot Spots ◽  
Genetic Locus ◽  
Rrna Gene ◽  
Origin Of Replication ◽  
Gene Sets ◽  
Bacterium Bacillus Subtilis ◽  
Bacterium Bacillus ◽  
Genomic Map

Abstract Integrative mapping with vectors containing ribosomal DNA sequences were used to complete the mapping of the 10 rRNA gene sets in the endospore forming bacterium Bacillus subtilis. Southern hybridizations allowed the assignment of nine operons to distinct BclI restriction fragments and their genetic locus identified by transductional crosses. Nine of the ten rRNA gene sets are located between 0 and 70 degrees on the genomic map. In the region surrounding cysA14, two sets of closely spaced tandem clusters are present. The first (rrnJ and rrnW) is located between purA16 and cysA14 closely linked to the latter; the second (rrnI, rrnH and rrnG) previously mapped within this area is located between attSPO2 and glpT6. The operons at or near the origin of replication (rrnO,rrnA and rrnJ,rrnW) represent "hot spots" of plasmid insertion.

Development of a Rapid On-Site Method for the Detection of Chicken Meat in Processed Ground Meat Products by Using a Direct Ultrafast PCR System

2020 ◽  
Vol 83 (6) ◽  
pp. 984-990 ◽  
Author(s):  
SUYEON SUL ◽  
MI-JU KIM ◽  
JUNG-MIN LEE ◽  
SUNG-YEON KIM ◽  
HAE-YEONG KIM
Keyword(s):  
Limit Of Detection ◽  
Meat Products ◽  
Accurate Method ◽  
Chicken Meat ◽  
Rrna Gene ◽  
Target Sequence ◽  
Ground Meat ◽  
Specific Pcr ◽  
Pcr Method ◽  
Meat Samples

ABSTRACT In this study, we developed a rapid on-site detection method by using direct ultrafast PCR coupled with a microfluidic chip to identify the presence of chicken meat in processed ground meat products. Chicken-specific PCR primer targeting mitochondrial 16S rRNA gene was newly designed, and its specificity was confirmed against 17 other animal species and 4 different chicken meat samples from different countries of origin. The sensitivity of the chicken-specific ultrafast PCR was 0.1 pg of chicken DNA. To evaluate the limit of detection of the direct ultrafast PCR method, different percentages of chicken meat mixed with pork or beef were prepared. The limit of detection of the direct ultrafast PCR method for the chicken meat–pork and chicken meat–beef mixtures was 0.1% for both raw meat and autoclaved meat. This method was used for 15 commercialized processed ground meat products. In this method, the target sequence was successfully amplified, and the presence of chicken meat in processed ground meat products was identified within approximately 25 min, including the time for sample preparation. Thus, our study shows that this developed direct ultrafast PCR method is a rapid and accurate method for on-site detection of chicken DNA in commercial food products. HIGHLIGHTS

scholarly journals Detection of pork in processed meat products by species-specific PCR for halal verification: food fraud cases in Hat Yai, Thailand

Food Research ◽  
2020 ◽  
Vol 4 (S1) ◽  
pp. 244-249 ◽  
Author(s):  
Mohd Hafidz M.M. ◽  
Makatar W.-H. ◽  
H. Adilan ◽  
T. Nawawee
Keyword(s):  
Meat Products ◽  
Food Products ◽  
Conventional Polymerase Chain Reaction ◽  
Processed Meat ◽  
Rrna Gene ◽  
Food Fraud ◽  
Street Food ◽  
Specific Pcr ◽  
Halal Food ◽  
Species Specific

Consumer confidence in halal integrity of the unique and various food products provides Hat Yai, Thailand a great potential for a global destination of Muslim-friendly tourism. Islam prohibits the consumption of pork and its derivatives in any food products. The issue of food adulteration and contamination, particularly in the processed halal meat products with pork and its derivatives, greatly concern Muslim consumers. The aim of this study was to detect the presence of pork DNA from processed meat products collected from self-proclaimed “halal” Muslim street food stalls at Hat Yai, Thailand. Thirty-six samples of various processed meat products were randomly collected from seven Muslim street food stalls including patties, meatballs, and sausages containing processed chicken, beef, or a mixture of various meats. The detection of the presence of pork and its derivatives was performed by a conventional polymerase chain reaction (PCR) technique based on the pork-specific primers for a conserved region in the mitochondrial (mt) 12S ribosomal RNA (rRNA) gene. The results revealed that three out of the thirty-six samples (8.3%) were positively identified to contain porcine DNA by the detection of the expected single band of size 387 bp. The DNA method conveniently provides reliable results for routine food analysis for halal requirement. Overall, the study highlights the importance of halal integrity between the producers, suppliers, and street food business owners to provide halal food products particularly to Muslim consumers.

Echinococcus multilocularis in a Eurasian lynx (Lynx lynx) in Turkey

Parasitology ◽  
2018 ◽  
Vol 145 (9) ◽  
pp. 1147-1150 ◽  
Author(s):  
Hamza Avcioglu ◽  
Esin Guven ◽  
Ibrahim Balkaya ◽  
Ridvan Kirman
Keyword(s):  
Echinococcus Multilocularis ◽  
Pcr Analysis ◽  
12S Rrna ◽  
Rrna Gene ◽  
Lynx Lynx ◽  
Eurasian Lynx ◽  
The North ◽  
Specific Pcr ◽  
Gene Sequence Analysis ◽  
Species Specific

AbstractEchinococcus multilocularis is the causative agent of alveolar echinococcosis (AE), one of the most threatening zoonoses in Eurasia. Human AE is widespread in the Erzurum region of Turkey, but the situation of the disease in intermediate and definitive hosts is unknown. A Eurasian lynx (Lynx lynx) was killed in a traffic accident in the north of Erzurum, and was taken to our laboratory. Sedimentation and counting technique (SCT), DNA isolation and polymerase chain reaction (PCR) analysis were performed. The SCT results showed that the lynx was infected with E. multilocularis with a medium (745 worms) worm burden. The DNA of adult worms obtained from the lynx was analyzed with a species-specific PCR, and the worms were confirmed to be E. multilocularis by 12S rRNA gene sequence analysis. This is the first report of E. multilocularis from Eurasian lynx in Turkey.

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.

A report on extensive lateral genetic reciprocation between arsenic resistant Bacillus subtilis and Bacillus pumilus strains analyzed using RAPD-PCR

2017 ◽  
Vol 107 ◽  
pp. 443-454 ◽  
Author(s):  
Sapna Khowal ◽  
Md. Zulquarnain Siddiqui ◽  
Shadab Ali ◽  
Mohd. Taha Khan ◽  
Mather Ali Khan ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Bacillus Pumilus ◽  
Rapd Pcr

A PCR Method Based on 16S rRNA Sequence for Simultaneous Detection of the Genus Listeria and the Species Listeria monocytogenes in Food Products

2003 ◽  
Vol 66 (9) ◽  
pp. 1658-1665 ◽  
Author(s):  
LILACH SOMER ◽  
YECHEZKEL KASHI
Keyword(s):  
Listeria Monocytogenes ◽  
Dna Sequences ◽  
Simultaneous Detection ◽  
Food Products ◽  
Positive Control ◽  
Sensitive Detection ◽  
Rrna Gene ◽  
16S Rrna Sequence ◽  
Selective Enrichment ◽  
Specific Pcr

The genus Listeria comprises six closely related species, of which only Listeria monocytogenes is a human pathogen. The rapid and sensitive detection of L. monocytogenes is important in the food industry as well as in medical diagnosis. In this study, a PCR-based method for the rapid, specific, and sensitive detection of L. monocytogenes in food products was developed. The PCR is based on DNA sequences and primer pairs that are found within the 16S subunit of the rRNA gene and are specific to the Listeria genus and to L. monocytogenes within the Listeria genus. The primers for the Listeria genus and for L. monocytogenes were used in the same reaction mix for their simultaneous detection. In addition, a pair of bacterial primers universal to any bacterial DNA at the 16S subunit of the rRNA gene were developed as a positive control. For the detection of Listeria and L. monocytogenes in food products, the method includes selective enrichment for Listeria followed by DNA extraction and a specific PCR reaction. The method detects 1 to 5 CFU in a 25-g sample in ≤24 h. It can be easily incorporated into the routine screening of diverse food products and readily adapted for clinical use.

scholarly journals Isolation and Characterization of Probiotic Bacillus subtilis MKHJ 1-1 Possessing L-Asparaginase Activity

2021 ◽  
Vol 11 (10) ◽  
pp. 4466
Author(s):  
Hyeji Lim ◽  
Sujin Oh ◽  
Sungryul Yu ◽  
Misook Kim
Keyword(s):  
Bacillus Subtilis ◽  
Bile Salt ◽  
Shigella Flexneri ◽  
Acid Resistance ◽  
Rrna Gene ◽  
Solution Ph ◽  
Isolation And Characterization ◽  
Gene Sequence Analysis ◽  
Asparaginase Activity ◽  
Bacillus Strains

The purpose of this study was to isolate functional Bacillus strains from Korean fermented soybeans and to evaluate their potential as probiotics. The L-asparaginase activity of MKHJ 1-1 was the highest among 162 Bacillus strains. This strain showed nonhemolysis and did not produce β-glucuronidase. Among the nine target bacteria, MKHJ 1-1 inhibited the growth of Escherichia coli, Pseudomonas aeruginosa, Shigella sonnei, Shigella flexneri, Klebsiella pneumoniae, Staphylococcus aureus, and Bacillus cereus. 16S rRNA gene sequence analysis resulted in MKHJ 1-1 identified as Bacillus subtilis subsp. stercoris D7XPN1. As a result of measuring the survival rate in 0.1% pepsin solution (pH 2.5) and 0.3% bile salt solution for 3 h, MKHJ 1-1 exhibited high acid resistance and was able to grow in the presence of bile salt. MKHJ 1-1 showed outstanding autoaggregation ability after 24 h. In addition, its coaggregation with pathogens was strong. Therefore, MKHJ 1-1 is a potential probiotic with L-asparaginase activity and without L-glutaminase activity, suggesting that it could be a new resource for use in the food and pharmaceutical industry.

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