Identification ofBacillusstrains isolated from milk and cream with classical and nucleic acid hybridization methods

1994 ◽  
Vol 61 (4) ◽  
pp. 529-535 ◽  
Author(s):  
Ralf Tatzel ◽  
Wolfgang Ludwig ◽  
Karl Heinz Schleifer ◽  
Peter R. Wallnöfer
Keyword(s):  
Nucleic Acid Hybridization ◽  
23S Rrna ◽  
Dairy Production ◽  
Production Plant ◽  
Oligonucleotide Probes ◽  
Hybridization Method ◽  
Bacterial Strains ◽  
Identification Methods ◽  
Colony Hybridization ◽  
Physiological And Biochemical

SummaryA total of 529 bacterial strains have been isolated from milk and cream sampled at different sites in a dairy production plant under conditions selective for aerobic sporeforming bacteria. Identification with classical methods based on morphological, physiological and biochemical criteria showedBacillus licheniformisto be the most frequently occurringBacillussp. The investigation also revealed 62 unidentified strains. Classical identification methods were time consuming (3–7d), lacked specificity and —because of their dependence on phenotypic gene expression—sometimes produced ambiguous results. Consequently, a colony hybridization method developed for the identification ofB. licheniformisstrains and using non-radioactive labelled 23S rRNA targeted oligonucleotide probes was also used. Identification ofB. licheniformiswith borth classical and hybridization methods revealed diverging identification results for 70 strains.

Identification of Bacillus licheniformis by Colony Hybridization with 23S rRNA-targeted Oligonucleotide Probes

1994 ◽  
Vol 17 (1) ◽  
pp. 99-103 ◽  
Author(s):  
Ralf Tatzel ◽  
Wolfgang Ludwig ◽  
Karl Heinz Schleifer ◽  
Peter R. Wallnöfer
Keyword(s):  
Bacillus Licheniformis ◽  
23S Rrna ◽  
Oligonucleotide Probes ◽  
Colony Hybridization

scholarly journals Role of Broiler Carcasses and Processing Plant Air in Contamination of Modified-Atmosphere-Packaged Broiler Products with Psychrotrophic Lactic Acid Bacteria

2006 ◽  
Vol 73 (4) ◽  
pp. 1136-1145 ◽  
Author(s):  
Elina Vihavainen ◽  
Hanna-Saara Lundstr�m ◽  
Tuija Susiluoto ◽  
Joanna Koort ◽  
Lars Paulin ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
23S Rrna ◽  
Processing Plant ◽  
Modified Atmosphere ◽  
Production Plant ◽  
Meat Spoilage

ABSTRACT Some psychrotrophic lactic acid bacteria (LAB) are specific meat spoilage organisms in modified-atmosphere-packaged (MAP), cold-stored meat products. To determine if incoming broilers or the production plant environment is a source of spoilage LAB, a total of 86, 122, and 447 LAB isolates from broiler carcasses, production plant air, and MAP broiler products, respectively, were characterized using a library of HindIII restriction fragment length polymorphism (RFLP) patterns of the 16 and 23S rRNA genes as operational taxonomic units in numerical analyses. Six hundred thirteen LAB isolates from the total of 655 clustered in 29 groups considered to be species specific. Sixty-four percent of product isolates clustered either with Carnobacterium divergens or with Carnobacterium maltaromaticum type strains. The third major product-associated cluster (17% of isolates) was formed by unknown LAB. Representative strains from these three clusters were analyzed for the phylogeny of their 16S rRNA genes. This analysis verified that the two largest RFLP clusters consisted of carnobacteria and showed that the unknown LAB group consisted of Lactococcus spp. No product-associated LAB were detected in broiler carcasses sampled at the beginning of slaughter, whereas carnobacteria and lactococci, along with some other specific meat spoilage LAB, were recovered from processing plant air at many sites. This study reveals that incoming broiler chickens are not major sources of psychrotrophic spoilage LAB, whereas the detection of these organisms from the air of the processing environment highlights the role of processing facilities as sources of LAB contamination.

scholarly journals Active Antibiotics Production by Actinomycetes Indigenous To Saudi Arabia Soils

2021 ◽  
Vol 11 (6) ◽  
pp. 20-29
Author(s):  
Fetoon M ◽  
helaiwi Alk ◽  
Ismet Ara ◽  
Nadine Moubayed
Keyword(s):  
Saudi Arabia ◽  
Biochemical Properties ◽  
Soil Samples ◽  
Fungal Strain ◽  
Pathogenic Microorganisms ◽  
Bacterial Strains ◽  
Desert Soils ◽  
Growth Environment ◽  
Physiological And Biochemical ◽  
Physiological And Biochemical Properties

Streptomyces are the most popular among the Actinomycetes groups and found in soils worldwide. They form an important part of the soil ecology within the Actinomycetales order. Streptomyces are diverse as secondary antibiotic metabolites such as Novobiocin, Amphotericin, Vancomycin, Neomycin, Gentamicin, Chloramphenicol, Tetracycline, Erythromycin and Nystatin. Thus, the current study was aimed to isolate, identify and assess the active antibiotic metabolites produced by different actinomyces sp. found in Saudi Arabian soils. Six samples were collected from desert soils of the Al Thumamah area and analyzed using GS-MS. Scanning Electron Microscopy was used to identify the bacterial strains along with their antibiotic metabolites effectiveness of secondary metabolites (antibiotics) against different Gram-positive (Bacillus subtilis, Staphylococcus aureus), negative pathogens (Pseudomonas aeruginosa, Escherichia coli, Salmonella suis, and Shigella sonnei) as well as the fungal strain Candida albicans was investigated. Thirty active bacterial (F1-30) strains were isolated from the soil samples and the strains F3, F7, F22, F30 have white, gray, pink, yellow and red colours respectively. Only ten strains (F13, F14, F15, F16, FI7, F18, F19, F20, F21, and F22) were found to have antimicrobial activity against at least one pathogen. The optimum growth environment was pH 4-10, temperature (300C), and NaCl (7% w/v) concentration. According to our findings, the extreme desert environment of Al Thumamah from Saudi Arabia is rich in its actinobacterial population with diverse colouring groups and various physiological and biochemical properties. This shows it’s capability of generating secondary metabolite elements that could inhibit pathogenic microorganisms.

scholarly journals Nature of Polymorphisms in 16S-23S rRNA Gene Intergenic Transcribed Spacer Fingerprinting of Bacillus and Related Genera

2003 ◽  
Vol 69 (9) ◽  
pp. 5128-5137 ◽  
Author(s):  
Daniele Daffonchio ◽  
Ameur Cherif ◽  
Lorenzo Brusetti ◽  
Aurora Rizzi ◽  
Diego Mora ◽  
...  
Keyword(s):  
Product Formation ◽  
23S Rrna ◽  
Trna Genes ◽  
Rrna Gene ◽  
Bacterial Strains ◽  
Additional Tool ◽  
Pcr Fingerprinting ◽  
23S Rrna Gene ◽  
Ribosomal Operons ◽  
Its Pcr

ABSTRACT The intergenic transcribed spacers (ITS) between the 16S and 23S rRNA genetic loci are frequently used in PCR fingerprinting to discriminate bacterial strains at the species and intraspecies levels. We investigated the molecular nature of polymorphisms in ITS-PCR fingerprinting of low-G+C-content spore-forming bacteria belonging to the genera Bacillus, Brevibacillus, Geobacillus, and Paenibacillus. We found that besides the polymorphisms in the homoduplex fragments amplified by PCR, heteroduplex products formed during PCR between amplicons from different ribosomal operons, with or without tRNA genes in the ITS, contribute to the interstrain variability in ITS-PCR fingerprinting patterns obtained in polyacrylamide-based gel matrices. The heteroduplex nature of the discriminating bands was demonstrated by fragment separation in denaturing polyacrylamide gels, by capillary electrophoresis, and by cloning, sequencing, and recombination of purified short and tRNA gene-containing long ITS. We also found that heteroduplex product formation is enhanced by increasing the number of PCR cycles. Homoduplex-heteroduplex polymorphisms (HHP) in a conserved region, such as the 16S and 23S rRNA gene ITS, allowed discrimination of closely related strains and species undistinguishable by other methods, indicating that ITS-HHP analysis is an easy and reproducible additional tool for strain typing.

scholarly journals Antibiotic Resistance Characteristics of Environmental Bacteria from an Oxytetracycline Production Wastewater Treatment Plant and the Receiving River

2010 ◽  
Vol 76 (11) ◽  
pp. 3444-3451 ◽  
Author(s):  
Dong Li ◽  
Tao Yu ◽  
Yu Zhang ◽  
Min Yang ◽  
Zhen Li ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
River Water ◽  
Resistance Genes ◽  
Treatment Plant ◽  
Antibiotic Resistance Genes ◽  
Tetracycline Resistance ◽  
Production Plant ◽  
Bacterial Strains ◽  
High Concentration ◽  
Receiving River

ABSTRACT We characterized the bacterial populations in surface water receiving effluent from an oxytetracycline (OTC) production plant. Additional sampling sites included the receiving river water 5 km upstream and 20 km downstream from the discharge point. High levels of OTC were found in the wastewater (WW), and the antibiotic was still detectable in river water downstream (RWD), with undetectable levels in river water upstream (RWU). A total of 341 bacterial strains were isolated using nonselective media, with the majority being identified as Gammaproteobacteria. The MICs were determined for 10 antibiotics representing seven different classes of antibiotics, and the corresponding values were significantly higher for the WW and RWD isolates than for the RWU isolates. Almost all bacteria (97%) from the WW and RWD samples demonstrated multidrug-resistant (MDR) phenotypes, while in RWU samples, these were less frequent (28%). The WW and RWD isolates were analyzed for the presence of 23 tetracycline (tet) resistance genes. The majority of isolates (94.2% and 95.4% in WW and RWD, respectively) harbored the corresponding genes, with tet(A) being the most common (67.0%), followed by tet(W), tet(C), tet(J), tet(L), tet(D), tet(Y), and tet(K) (in the range between 21.0% and 40.6%). Class I integrons were detected in the majority of WW and RWD isolates (97.4% and 86.2%, respectively) but were not associated with the tet genes. We hypothesize that the strong selective pressure imposed by a high concentration of OTC contributes to the wide dissemination of tetracycline resistance genes and other antibiotic resistance genes, possibly through mobile genetic elements.

Evaluation of an Alkaline Phosphatase–Labeled Oligonucleotide Probe for the Detection and Enumeration of the Thermostable-Related Hemolysin (trh) Gene of Vibrio parahaemolyticus

2006 ◽  
Vol 69 (11) ◽  
pp. 2770-2772 ◽  
Author(s):  
JESSICA L. NORDSTROM ◽  
RACHEL RANGDALE ◽  
MICHAEL C. L. VICKERY ◽  
ANDREA M. B. PHILLIPS ◽  
SHELLEY L. MURRAY ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Vibrio Parahaemolyticus ◽  
Oligonucleotide Probe ◽  
The United States ◽  
Bacterial Strains ◽  
Specific Detection ◽  
Colony Hybridization ◽  
The United Kingdom ◽  
Pathogenic Vibrio ◽  
Reliable Methods

Reliable methods are needed to detect total and pathogenic Vibrio parahaemolyticus. One marker of V. parahaemolyticus virulence is the thermostable-related hemolysin. We developed an alkaline phosphatase–labeled DNA probe method for the specific detection and enumeration of trh-positive V. parahaemolyticus by colony hybridization. The probe was tested against a panel of 200 bacterial strains and determined to be specific for trh-positive V. parahaemolyticus. Additionally, the trh alkaline phosphatase probe colony hybridization was successfully used to detect and enumerate trh-positive V. parahaemolyticus in seafood and water samples collected from the United States and the United Kingdom.

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