Acetic acid bacteria strain typing by PFGE (pulsed-field gel electrophoresis) and bacterial cellulose production

2012 ◽  
Vol 29 ◽  
pp. S56
Author(s):  
Esin Poyrazoglu Coban ◽  
Bulent Bozdogan ◽  
H. Halil Biyik
Keyword(s):  
Acetic Acid ◽  
Bacterial Cellulose ◽  
Gel Electrophoresis ◽  
Acetic Acid Bacteria ◽  
Pulsed Field Gel Electrophoresis ◽  
Strain Typing ◽  
Cellulose Production ◽  
Bacterial Cellulose Production ◽  
Pulsed Field

Enrichment, Amplification, and Sequence-Based Typing of Salmonella enterica and Other Foodborne Pathogens

2016 ◽  
Vol 80 (1) ◽  
pp. 15-24 ◽  
Author(s):  
TOM EDLIND ◽  
JEFFREY D. BREWSTER ◽  
GEORGE C. PAOLI
Keyword(s):  
Foodborne Pathogens ◽  
Gel Electrophoresis ◽  
Pure Culture ◽  
Salmonella Enterica ◽  
Cost Effective ◽  
Pulsed Field Gel Electrophoresis ◽  
Alternative Methods ◽  
Strain Typing ◽  
Isolation And Identification ◽  
Pulsed Field

ABSTRACT Detection of Salmonella enterica in foods typically involves microbiological enrichment, molecular-based assay, and subsequent isolation and identification of a pure culture. This is ideally followed by strain typing, which provides information critical to the investigation of outbreaks and the attribution of their sources. Pulsed-field gel electrophoresis is the “gold standard” for S. enterica strain typing, but its limitations have encouraged the search for alternative methods, including whole genome sequencing. Both methods typically require a pure culture, which adds to the cost and turnaround time. A more rapid and cost-effective method with sufficient discriminatory power would benefit food industries, regulatory agencies, and public health laboratories. To address this need, a novel enrichment, amplification, and sequence-based typing (EAST) approach was developed involving (i) overnight enrichment and total DNA preparation, (ii) amplification of polymorphic tandem repeat–containing loci with electrophoretic detection, and (iii) DNA sequencing and bioinformatic analysis to identify related strains. EAST requires 3 days or less and provides a strain resolution that exceeds serotyping and is comparable to pulsed-field gel electrophoresis. Evaluation with spiked ground turkey demonstrated its sensitivity (with a starting inoculum of ≤1 CFU/g) and specificity (with unique or nearly unique alleles relative to databases of >1,000 strains). In tests with unspiked retail chicken parts, 3 of 11 samples yielded S. enterica–specific PCR products. Sequence analysis of three distinct typing targets (SeMT1, SeCRISPR1, and SeCRISPR2) revealed consistent similarities to specific serotype Schwarzengrund, Montevideo, and Typhimurium strains. EAST provides a time-saving and cost-effective approach for detecting and typing foodborne S. enterica, and postenrichment steps can be commercially outsourced to facilitate its implementation. Initial studies with Listeria monocytogenes and Shiga toxigenic Escherichia coli suggest that EAST can be extended to these foodborne pathogens.

scholarly journals Lactobacillus helveticus : Strain Typing and Genome Size Estimation by Pulsed Field Gel Electrophoresis

1997 ◽  
Vol 34 (3) ◽  
pp. 180-185 ◽  
Author(s):  
Sylvie Lortal ◽  
Annette Rouault ◽  
Stéphane Guezenec ◽  
Michel Gautier
Keyword(s):  
Genome Size ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Lactobacillus Helveticus ◽  
Strain Typing ◽  
Size Estimation ◽  
Pulsed Field

scholarly journals Screening of Acetic Acid Bacteria from Pineapple Waste for Bacterial Cellulose Production using Sago Liquid Waste

2017 ◽  
Vol 9 (3) ◽  
pp. 387 ◽  
Author(s):  
Nur Arfa Yanti ◽  
Sitti Wirdhana Ahmad ◽  
Sri Ambardini ◽  
Nurhayani Haji Muhiddin ◽  
La Ode Iman Sulaiman
Keyword(s):  
Bacterial Cellulose ◽  
Culture Media ◽  
Liquid Waste ◽  
Acetic Acid Bacteria ◽  
Industrial Sector ◽  
Production Costs ◽  
High Yield ◽  
Cellulose Production ◽  
Bacterial Cellulose Production ◽  
Pineapple Waste

<p class="IsiAbstrakInggris"><span>Bacterial cellulose is a biopolymer </span><span lang="EN-GB">produced by fermentation process with the help of bacteria. It </span><span>has numerous applications in industrial sector with its characteristic as a biodegradable and nontoxic compound in nature. </span><span lang="EN-GB">The potential application of BC is limited by its production costs, because BC is produced from expensive culture media. The use of cheap carbon and nutrient sources such as sago liquid waste is an interesting strategy to overcome this limitation. The objective of this study was to obtain the AAB strain that capable to produce bacterial cellulose from sago liquid waste. Isolation of AAB strains was conducted using CARR media and the screening of BC production was performed on Hestrin-Schramm (HS) media with glucose as a carbon source. The strains of AAB then were evaluated for their cellulose-producing capability using sago liquid waste as a substrate. Thirteen strains of AAB producing BC were isolated from pineapple waste (pineapple core and peel) and seven of them were capable to produce BC using sago liquid waste substrate. One of the AAB strains produced a relatively high BC, i.e. isolate LKN6. The result of morphological and biochemical test was proven that the bacteria was </span><em><span>Acetobacter xylinum</span></em><span lang="EN-GB">. The result of this study showed that </span><em><span>A. xylinum </span></em><span lang="EN-GB">LKN6 can produce a high yield of BC, therefore this strain is potentially useful for its utilization as a starter in bacterial cellulose production. </span></p>

Molecular strain typing ofCampylobacter jejuniby pulsed-field gel electrophoresis in a single day

2001 ◽  
Vol 47 (7) ◽  
pp. 667-669 ◽  
Author(s):  
Sophie Michaud ◽  
Robert D Arbeit ◽  
Christiane Gaudreau
Keyword(s):  
Infection Control ◽  
Molecular Epidemiology ◽  
Campylobacter Jejuni ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Control Measures ◽  
Strain Typing ◽  
High Quality ◽  
Pulsed Field ◽  
Infection Control Measures

Rapid molecular strain typing is critical for effective outbreak investigation and implementation of infection control measures. Pulsed-field gel electrophoresis is a highly discriminatory technique for Campylobacter jejuni, but generally requires 3–5 days. We describe a simplified protocol for pulsed-field gel electrophoresis that provides high quality typing of C. jejuni isolates in a single day.Key words: pulsed-field gel electrophoresis, Campylobacter jejuni, molecular epidemiology.

scholarly journals Molecular epidemiology analysis of Salmonella enterica serotype Typhi used Pulsed-Field Gel Electrophoresis (PFGE)

2014 ◽  
Vol 8 (2) ◽  
pp. 49-59
Author(s):  
Ashna J. Faik ◽  
Farook. K. Hassan ◽  
Ali Al-Zaag
Keyword(s):  
Molecular Epidemiology ◽  
Gel Electrophoresis ◽  
Salmonella Enterica ◽  
Salmonella Typhi ◽  
Pulsed Field Gel Electrophoresis ◽  
Molecular Methods ◽  
Strain Typing ◽  
Epidemiological Typing ◽  
Pulsed Field ◽  
Plasmid Profiling

Strain typing is an integral part of molecular epidemiology used to discern the clonality of Salmonella Typhi involved in local epidemics. The purpose of this study is to identify sporadic Salmonella enterica serotype Typhi by conventional and molecular methods that include characterization by Pulsed Field Gel Electrophoresis (PFGE) and to present molecular epidemiology analysis. Thirty isolstes of Salmonella typhi from sporadic clinical cases of typhoid fever were obtained. They represent cases from Baghdad, Basra, Babylon and Diala provinces during the period between June 2005 to July 2006. Two biotypes were obtained, 26 isolates under biotype I and four under biotype II. Two antibiogram patterns were obtained: twenty-nine isolates were susceptible to all antibiotic used while the remaining isolate was of different pattern. Plasmid profiling allowed little or no differentiation amongst these isolates. Only 4 (13.3%) isolates were found to contain plasmids which were of three patterns, the majority of strains 26 (86.7%) isolates did not show any plasmid. BOX-PCR fingerprinting has revealed 9 distinct patterns.Cluster analysis by UPGMA based dendrogram revealed six clusters with 90% similarity. Pulsed-field gel electrophoresis (PFGE) of digested chromosomal DNAs from these Salmonella Typhi isolates was performed using the restriction endonucleases XbaI (5'-TCTAGA-3') and SpeI (5'-ACTAGT-3'). XbaI-based analysis was superior to SpeI restriction patterns. PFGE fingerprinting with XbaI restriction have yielded sixteen distinct patterns. Cluster analysis by UPGMA based dendrogram revealed seven clusters with 90% similarity. PFGE fingerprinting with the comparative use of the XbaI and SpeI endonucleases have proved high discriminatory value to other molecular methods and a helpful tool for the epidemiological typing of Salmonella Typhi isolates.

scholarly journals Species Identification and Strain Typing of Staphylococcus agnetis and Staphylococcus hyicus Isolates from Bovine Milk by Use of a Novel Multiplex PCR Assay and Pulsed-Field Gel Electrophoresis

2017 ◽  
Vol 55 (6) ◽  
pp. 1778-1788 ◽  
Author(s):  
P. R. F. Adkins ◽  
J. R. Middleton ◽  
M. J. Calcutt ◽  
G. C. Stewart ◽  
L. K. Fox
Keyword(s):  
Multiplex Pcr ◽  
Species Identification ◽  
Gel Electrophoresis ◽  
Bovine Milk ◽  
Gene Sequencing ◽  
Pulsed Field Gel Electrophoresis ◽  
Strain Typing ◽  
Content Type ◽  
Pulsed Field ◽  
Multiplex Pcr Assay

ABSTRACTStaphylococcus hyicusandStaphylococcus agnetisare two coagulase-variable staphylococcal species that can be isolated from bovine milk and are difficult to differentiate. The objectives of this study were to characterize isolates of bovine milk origin from a collection that had previously been characterized as coagulase-positiveS. hyicusbased on phenotypic species identification methods and to develop a PCR-based method for differentiatingS. hyicus,S. agnetis, andStaphylococcus aureus. Isolates (n= 62) were selected from a previous study in which milk samples were collected from cows on 15 dairy herds. Isolates were coagulase tested and identified to the species level using housekeeping gene sequencing. A multiplex PCR to differentiateS. hyicus,S. agnetis, andS. aureuswas developed. Pulsed-field gel electrophoresis was conducted to strain type the isolates. Based on gene sequencing, 44/62 of the isolates were determined to be eitherS. agnetis(n= 43) orS. hyicus(n= 1). Overall, 88% (37/42) of coagulase-positiveS. agnetisisolates were found to be coagulase positive at 4 h. The herd-level prevalence of coagulase-positiveS. agnetisranged from 0 to 2.17%. Strain typing identified 23 different strains. Six strains were identified more than once and from multiple cows within the herd. Three strains were isolated from cows at more than one time point, with 41 to 264 days between samplings. These data suggest thatS. agnetisis likely more prevalent on dairy farms thanS. hyicus. Also, someS. agnetisisolates in this study appeared to be contagious and associated with persistent infections.

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