Isolation and characterization of antibiotic-resistance plasmids in thermophilic bacilli

1985 ◽  
Vol 31 (4) ◽  
pp. 339-345 ◽  
Author(s):  
Takayuki Hoshino ◽  
Takayuki Ikeda ◽  
Hiroyuki Narushima ◽  
Noboru Tomizuka
Keyword(s):  
Antibiotic Resistance ◽  
Southern Hybridization ◽  
Tetracycline Resistance ◽  
Restriction Endonuclease Analysis ◽  
Kanamycin Resistance ◽  
Isolation And Characterization ◽  
Thermophilic Bacilli ◽  
Resistance Plasmids ◽  
Endonuclease Analysis

Four antibiotic-resistance plasmids isolated from thermophilic bacilli were characterized in detail. Three tetracycline-resistance (Tcr) plasmids were designated as pTHT9 (7.7 kilobases (kb)), pTHT15 (4.5 kb) and pTHT22 (8.4 kb). From the results of restriction endonuclease analysis and the subsequent Southern hybridization, these were found to possess extensive genetic homology in the regions that include the replication origin and the Tcr gene. Detailed restriction maps of the smallest Tcr plasmid pTHT15 and a kanamycin-resistance (Kmr) plasmid pTHN1 (4.8 kb) were constructed. The positions of antibiotic-resistance loci and regions essential for plasmid replication were determined by cloning plasmid fragments in Bacillus subtilis. These four plasmids were found to replicate and express the resistance genes stably in both B. subtilis and B. stearothermophilus.

scholarly journals Directed Recovery and Molecular Characterization of Antibiotic Resistance Plasmids from Cheese Bacteria

2021 ◽  
Vol 22 (15) ◽  
pp. 7801
Author(s):  
Ana Belén Flórez ◽  
Lucía Vázquez ◽  
Javier Rodríguez ◽  
Baltasar Mayo
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Tetracycline Resistance ◽  
Restriction Enzyme Digestion ◽  
Erythromycin Resistance ◽  
Leading Role ◽  
Resistance Plasmids ◽  
Digestion Profile

Resistance to antimicrobials is a growing problem of worldwide concern. Plasmids are thought to be major drivers of antibiotic resistance spread. The present work reports a simple way to recover replicative plasmids conferring antibiotic resistance from the bacteria in cheese. Purified plasmid DNA from colonies grown in the presence of tetracycline and erythromycin was introduced into plasmid-free strains of Lactococcus lactis, Lactiplantibacillus plantarum and Lacticaseibacillus casei. Following antibiotic selection, the plasmids from resistant transformants were isolated, analyzed by restriction enzyme digestion, and sequenced. Seven patterns were obtained for the tetracycline-resistant colonies, five from L. lactis, and one each from the lactobacilli strains, as well as a single digestion profile for the erythromycin-resistant transformants obtained in L. lactis. Sequence analysis respectively identified tet(S) and ermB in the tetracycline- and erythromycin-resistance plasmids from L. lactis. No dedicated resistance genes were detected in plasmids conferring tetracycline resistance to L. casei and L. plantarum. The present results highlight the usefulness of the proposed methodology for isolating functional plasmids that confer antibiotic resistance to LAB species, widen our knowledge of antibiotic resistance in the bacteria that inhabit cheese, and emphasize the leading role of plasmids in the spread of resistance genes via the food chain.

Isolation and characterization of two temperate phages from Lactococcus lactis ssp. cremoris C3

1992 ◽  
Vol 38 (5) ◽  
pp. 398-404 ◽  
Author(s):  
Audrey W. Jarvis ◽  
Vaughan R. Parker ◽  
Moana B. Bianchin
Keyword(s):  
Lactococcus Lactis ◽  
Dna Hybridization ◽  
Base Plate ◽  
Restriction Endonuclease Analysis ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Isolation And Characterization ◽  
Dna Fragment ◽  
Endonuclease Analysis

Lactococcus lactis ssp. cremoris C3 was shown to contain two prophages, C3-T1 and C3-T2, both inducible bymitomycin C. The phages were morphologically indistinguishable, having an isometric head (55 nm), a noncontractile tail (142 nm), and a distinctive base plate. The phages C3-T1 and C3-T2 were differentiated from one another by restriction endonuclease analysis of their DNA, and genome sizes of 37.8 (C3-T1) and 32.5 kb (C3-T2). Southern blot DNA hybridization suggested a maximum homology between the phage genomes of 50%. Phage C3-T1 was propagated and the phage obtained was designated C3-TIℓ. Phage C3-T2 was not propagated despite tests with a large number of possible indicator strains. Restriction enzyme analysis of phage C3-T1ℓ genome yielded a 37.8-kb circular restriction map. The packaging site (pac) and site of integration into the bacterial chromosome (att) were located and an EcoRI DNA fragment containing the att site was cloned. Only one C3-T1 att site was present in the C3 chromosome. No homology was detected between DNA from C3-T1ℓ and DNA from lytic phages commonly isolated from cheese wheys. Key words: lactococcal phages, temperate phages, lysogeny, evolution.

Characterization of staphylococci contaminating automated teller machines in Hong Kong

2011 ◽  
Vol 140 (8) ◽  
pp. 1366-1371 ◽  
Author(s):  
M. ZHANG ◽  
M. O'DONONGHUE ◽  
M. V. BOOST
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Benzalkonium Chloride ◽  
Tetracycline Resistance ◽  
Chlorhexidine Digluconate ◽  
Coagulase Negative Staphylococci ◽  
Automated Teller Machines ◽  
Resistance Determinants

SUMMARYEnvironmental staphylococcal contamination was investigated by culture of 400 automated teller machines (ATMs). Isolates were characterized for antibiotic and antiseptic susceptibility, carriage of antiseptic resistance genes (QAC genes), and spa types. MRSA, which was similar to local clinical isolates, was present on two (0·5%) of the 62 (15·5%) ATMs that yielded Staphylococcus aureus. QAC genes were more common in coagulase-negative staphylococci (qacA/B 26·0%, smr 14%) than S. aureus (11·3% qacA/B, 1·6% smr). QAC-positive isolates had significantly higher minimum inhibitory concentrations/minimum bactericidal concentrations to benzalkonium chloride and chlorhexidine digluconate. QAC gene presence was significantly associated with methicillin and tetracycline resistance. Survival of staphylococci, including MRSA, on common access sites may be facilitated by low disinfectant concentrations, which select for disinfectant-tolerant strains, while co-selecting for antibiotic-resistance determinants. Disinfection procedures should be performed correctly to help prevent spread of resistant pathogens from reservoirs in the community.

scholarly journals Isolation and characterization of bacteriophages that infect Citrobacter rodentium, a model pathogen for investigating human intestinal diseases

2018 ◽  
Author(s):  
Carolina M. Mizuno ◽  
Tiffany Luong ◽  
Robert Cedarstrom ◽  
Mart Krupovic ◽  
Laurent Debarbieux ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Phage Therapy ◽  
Limit Of Detection ◽  
Lytic Cycle ◽  
Citrobacter Rodentium ◽  
E Coli ◽  
Isolation And Characterization ◽  
A New Species

AbstractEnteropathogenic Escherichia coli (EPEC) is a major etiology for diarrheal diseases among children. Antibiotics, when used appropriately, are effective; however, their overuse and misuse has led to the rise of antibiotic resistance worldwide. Thus, there are renewed efforts into the development of phage therapy. Due to the drawbacks of EPEC in vivo models, a surrogate is the mouse-restricted gut pathgoen Citrobacter rodentium. In this study, two new phages CrRp3 and CrRp10, which infect C. rodentium, were isolated and characterized. CrRp3 was found to be a new species within the genus Vectrevirus and CrRp10 is a new strain within the genus Tequatrovirus. Neither phage carries known genes associated with bacterial virulence, antibiotic resistance, or lysogeny. CrRp3 and CrRp10 appear to have independently evolved from E. coli phages. CrRp3 appears to be the more ‘potent’ being 24x more likely to find a host cell and has a shorter lytic cycle, while CrRp10 at MOI 0.001 was able to maintain bacterial density below the limit of detection after 18 h. We found that hypoxia (5% O2 and 5% CO2) inhibited CrRp3 ability to reverse exponential bacterial growth. It is unclear whether the subtle characteristic differences between CrRp3 and CrRp10 will influence treatment efficacy in future phage therapy in vivo investigations.

scholarly journals Genomic Relationships between Enterococcus faecium Strains from Different Sources and with Different Antibiotic Resistance Profiles Evaluated by Restriction Endonuclease Analysis of Total Chromosomal DNA Using EcoRI andPvuII

1999 ◽  
Vol 65 (4) ◽  
pp. 1777-1780 ◽  
Author(s):  
M. Quednau ◽  
S. Ahrné ◽  
G. Molin
Keyword(s):  
Antibiotic Resistance ◽  
Healthy Subject ◽  
Restriction Endonuclease ◽  
Enterococcus Faecium ◽  
Restriction Endonuclease Analysis ◽  
Chromosomal Dna ◽  
Genomic Relationships ◽  
Different Sources ◽  
Endonuclease Analysis

ABSTRACT Forty-seven Enterococcus faecium strains from different sources were evaluated by restriction endonuclease analysis (REA) of total chromosomal DNA. Strains from chicken, pork, and humans were clearly divided into separate clusters, whereas strains from different countries, strains with different antibiotic resistance profiles, or clinical and healthy-subject strains were not.

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