Sequencing analysis and characterization of the plasmid pBIF10 isolated from Bifidobacterium longum

2015 ◽  
Vol 61 (2) ◽  
pp. 124-130
Author(s):  
Yongping Ma ◽  
Ting-ting Xie ◽  
Qiongwen Hu ◽  
Zongyin Qiu ◽  
Fangzhou Song
Keyword(s):  
Antibiotic Resistance ◽  
Bifidobacterium Longum ◽  
Tetracycline Resistance ◽  
Open Reading Frames ◽  
Host Cells ◽  
Sequencing Analysis ◽  
Plasmid Replicon ◽  
Full Length Sequence ◽  
Bifidobacterium Strain

A resident plasmid, pBIF10, was isolated from Bifidobacterium longum B200304, and the full-length sequence of pBIF10 was analyzed. In this sequence, we identified at least 17 major open reading frames longer than 200 bp. A tetracycline resistance gene, tetQ, was identified and verified to confer antibiotic resistance to tetracycline. The plasmid replicon with replication protein B gene (repB) and a typical iteron was identified in pBIF10. An artificial clone vector was constructed with the replicon of pBIF10; the results showed that repB controlled plasmid replication in other bifidobacteria host cells at low transformation frequency. Taken together, the analysis and characterization of pBIF10 provided necessary information for the understanding of antibiotic resistance mediated by a plasmid in a Bifidobacterium strain. GC% and repB sequence analyses indicated that pBIF10 was a molecular hybrid of at least 2 other bacterial genera plasmids.

scholarly journals Necrotic Enteritis-Derived Clostridium perfringens Strain with Three Closely Related Independently Conjugative Toxin and Antibiotic Resistance Plasmids

mBio ◽  
2011 ◽  
Vol 2 (5) ◽  
Author(s):  
Trudi L. Bannam ◽  
Xu-Xia Yan ◽  
Paul F. Harrison ◽  
Torsten Seemann ◽  
Anthony L. Keyburn ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Clostridium Perfringens ◽  
Tetracycline Resistance ◽  
Necrotic Enteritis ◽  
Sequencing Analysis ◽  
Content Type ◽  
Conjugative Plasmids ◽  
Resistance Plasmid ◽  
Large Plasmids ◽  
Beta2 Toxin

ABSTRACTThe pathogenesis of avian necrotic enteritis involves NetB, a pore-forming toxin produced by virulent avian isolates ofClostridium perfringenstype A. To determine the location and mobility of thenetBstructural gene, we examined a derivative of the tetracycline-resistant necrotic enteritis strain EHE-NE18, in whichnetBwas insertionally inactivated by the chloramphenicol and thiamphenicol resistance genecatP. Both tetracycline and thiamphenicol resistance could be transferred either together or separately to a recipient strain in plate matings. The separate transconjugants could act as donors in subsequent matings, which demonstrated that the tetracycline resistance determinant and thenetBgene were present on different conjugative elements. Large plasmids were isolated from the transconjugants and analyzed by high-throughput sequencing. Analysis of the resultant data indicated that there were actually three large conjugative plasmids present in the original strain, each with its own toxin or antibiotic resistance locus. Each plasmid contained a highly conserved 40-kb region that included plasmid replication and transfer regions that were closely related to the 47-kb conjugative tetracycline resistance plasmid pCW3 fromC. perfringens. The plasmids were as follows: (i) a conjugative 49-kb tetracycline resistance plasmid that was very similar to pCW3, (ii) a conjugative 82-kb plasmid that contained thenetBgene and other potential virulence genes, and (iii) a 70-kb plasmid that carried thecpb2gene, which encodes a different pore-forming toxin, beta2 toxin.IMPORTANCEThe anaerobic bacteriumClostridium perfringenscan cause an avian gastrointestinal disease known as necrotic enteritis. Disease pathogenesis is not well understood, although the plasmid-encoded pore-forming toxin NetB, is an important virulence factor. In this work, we have shown that the plasmid that carries thenetBgene is conjugative and has a 40-kb region that is very similar to replication and transfer regions found within each of the sequenced conjugative plasmids fromC. perfringens. We also showed that this strain contained two additional large plasmids that were also conjugative and carried a similar 40-kb region. One of these plasmids encoded beta2 toxin, and the other encoded tetracycline resistance. To our knowledge, this is the first report of a bacterial strain that carries three closely related but different independently conjugative plasmids. These results have significant implications for our understanding of the transmission of virulence and antibiotic resistance genes in pathogenic bacteria.

scholarly journals Bifidobacterium longum subsp. infantis ATCC 15697 and Goat Milk Oligosaccharides Show Synergism In Vitro as Anti-Infectives against Campylobacter jejuni

Foods ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 348 ◽  
Author(s):  
Erinn M. Quinn ◽  
Helen Slattery ◽  
Dan Walsh ◽  
Lokesh Joshi ◽  
Rita M. Hickey
Keyword(s):  
Campylobacter Jejuni ◽  
Bifidobacterium Longum ◽  
Goat Milk ◽  
Host Cells ◽  
Intestinal Cells ◽  
Milk Oligosaccharides ◽  
Bifidobacterium Strain ◽  
Ht 29 ◽  
Human Intestinal Cells

Bifidobacteria are known to inhibit, compete with and displace the adhesion of pathogens to human intestinal cells. Previously, we demonstrated that goat milk oligosaccharides (GMO) increased the attachment of Bifidobacterium longum subsp. infantis ATCC 15697 to intestinal cells in vitro. In this study, we aimed to exploit this effect as a mechanism for inhibiting pathogen association with intestinal cells. We examined the synergistic effect of GMO-treated B. infantis on preventing the attachment of a highly invasive strain of Campylobacter jejuni to intestinal HT-29 cells. The combination decreased the adherence of C. jejuni to the HT-29 cells by an average of 42% compared to the control (non-GMO treated B. infantis). Increasing the incubation time of the GMO with the Bifidobacterium strain resulted in the strain metabolizing the GMO, correlating with a subsequent 104% increase in growth over a 24 h period when compared to the control. Metabolite analysis in the 24 h period also revealed increased production of acetate, lactate, formate and ethanol by GMO-treated B. infantis. Statistically significant changes in the GMO profile were also demonstrated over the 24 h period, indicating that the strain was digesting certain structures within the pool such as lactose, lacto-N-neotetraose, lacto-N-neohexaose 3′-sialyllactose, 6′-sialyllactose, sialyllacto-N-neotetraose c and disialyllactose. It may be that early exposure to GMO modulates the adhesion of B. infantis while carbohydrate utilisation becomes more important after the bacteria have transiently colonised the host cells in adequate numbers. This study builds a strong case for the use of synbiotics that incorporate oligosaccharides sourced from goat′s milk and probiotic bifidobacteria in functional foods, particularly considering the growing popularity of formulas based on goat milk.

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 Molecular Characterization of Pneumococci with Efflux-Mediated Erythromycin Resistance and Identification of a Novel mef Gene Subclass, mef(I)

2005 ◽  
Vol 49 (12) ◽  
pp. 4999-5006 ◽  
Author(s):  
Ileana Cochetti ◽  
Manuela Vecchi ◽  
Marina Mingoia ◽  
Emily Tili ◽  
Maria R. Catania ◽  
...  
Keyword(s):  
Tetracycline Resistance ◽  
Sequence Type ◽  
Open Reading Frames ◽  
Resistant Isolate ◽  
Pfge Type ◽  
Polymorphism Analysis ◽  
Erythromycin Resistance ◽  
M Gene ◽  
Sequence Types

ABSTRACT The molecular genetics of macrolide resistance were analyzed in 49 clinical pneumococci (including an “atypical” bile-insoluble strain currently assigned to the new species Streptococcus pseudopneumoniae) with efflux-mediated erythromycin resistance (M phenotype). All test strains had the mef gene, identified as mef(A) in 30 isolates and mef(E) in 19 isolates (including the S. pseudopneumoniae strain) on the basis of PCR-restriction fragment length polymorphism analysis. Twenty-eight of the 30 mef(A) isolates shared a pulsed-field gel electrophoresis (PFGE) type corresponding to the England14-9 clone. Of those isolates, 27 (20 belonging to serotype 14) yielded multilocus sequence type ST9, and one isolate yielded a new sequence type. The remaining two mef(A) isolates had different PFGE types and yielded an ST9 type and a new sequence type. Far greater heterogeneity was displayed by the 19 mef(E) isolates, which fell into 11 PFGE types, 12 serotypes (though not serotype 14), and 12 sequence types (including two new ones and an undetermined type for the S. pseudopneumoniae strain). In all mef(A) pneumococci, the mef element was a regular Tn1207.1 transposon, whereas of the mef(E) isolates, 17 carried the mega element and 2 exhibited a previously unreported organization, with no PCR evidence of the other open reading frames of mega. The mef gene of these two isolates, which did not match with the mef(E) gene of the mega element (93.6% homology) and which exhibited comparable homology (91.4%) to the mef(A) gene of the Tn1207.1 transposon, was identified as a novel mef gene variant and was designated mef(I). While penicillin-nonsusceptible isolates (three resistant isolates and one intermediate isolate) were all mef(E) strains, tetracycline resistance was also detected in three mef(A) isolates, due to the tet(M) gene carried by a Tn916-like transposon. A similar mechanism accounted for resistance in four of the five tetracycline-resistant isolates carrying mef(E), in three of which mega was inserted in the Tn916-like transposon, giving rise to the composite element Tn2009. In the fifth mef(E)-positive tetracycline-resistant isolate (the S. pseudopneumoniae strain), tetracycline resistance was due to the presence of the tet(O) gene, apparently unlinked to mef(E).

scholarly journals Prevalence and Molecular Characterization of Tetracycline Resistance in Enterococcus Isolates from Food

2004 ◽  
Vol 70 (3) ◽  
pp. 1555-1562 ◽  
Author(s):  
Geert Huys ◽  
Klaas D'Haene ◽  
Jean-Marc Collard ◽  
Jean Swings
Keyword(s):  
Tetracycline Resistance ◽  
Recipient Strain ◽  
Sequencing Analysis ◽  
Evolutionary Modeling ◽  
Phenotypic Resistance ◽  
Homology Groups ◽  
Filter Mating ◽  
Gene Filter ◽  
Phylogenetic Lineages

ABSTRACT In the present study, a collection of 187 Enterococcus food isolates mainly originating from European cheeses were studied for the phenotypic and genotypic assessment of tetracycline (TC) resistance. A total of 45 isolates (24%) encompassing the species Enterococcus faecalis (n = 33), E. durans (n = 7), E. faecium (n = 3), E. casseliflavus (n = 1), and E. gallinarum (n = 1) displayed phenotypic resistance to TC with MIC ranges of 16 to 256 μg/ml. Eight of these strains exhibited multiresistance to TC, erythromycin, and chloramphenicol. By PCR detection, TC resistance could be linked to the presence of the tet(M) (n = 43), tet(L) (n = 16), and tet(S) (n = 1) genes. In 15 isolates, including all of those for which the MIC was 256 μg/ml, both tet(M) and tet(L) were found. Furthermore, all tet(M)-containing enterococci also harbored a member of the Tn916-Tn1545 conjugative transposon family, of which 12 erythromycin-resistant isolates also contained the erm(B) gene. Filter mating experiments revealed that 10 E. faecalis isolates, 3 E. durans isolates, and 1 E. faecium isolate could transfer either tet(M), tet(L), or both of these genes to E. faecalis recipient strain JH2-2. In most cases in which only tet(M) was transferred, no detectable plasmids were acquired by JH2-2 but instead all transconjugants contained a member of the Tn916-Tn1545 family. Sequencing analysis of PCR amplicons and evolutionary modeling showed that a subset of the transferable tet(M) genes belonged to four sequence homology groups (SHGs) showing an internal homology of ≥99.6%. Two of these SHGs contained tet(M) mosaic structures previously found in Tn916 elements and on Lactobacillus and Neisseria plasmids, respectively, whereas the other two SHGs probably represent new phylogenetic lineages of this gene.

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.

Characterizing the citrus cultivar Carrizo genome through 454 shotgun sequencing

Genome ◽  
2011 ◽  
Vol 54 (12) ◽  
pp. 1005-1015 ◽  
Author(s):  
William R. Belknap ◽  
Yi Wang ◽  
Naxin Huo ◽  
Jiajie Wu ◽  
David R. Rockhold ◽  
...  
Keyword(s):  
Rna Binding ◽  
Genomic Sequence ◽  
Populus Trichocarpa ◽  
Shotgun Sequencing ◽  
Open Reading Frames ◽  
Sequencing Analysis ◽  
Coding Region ◽  
Genome Coverage ◽  
Citrus Cultivar

The citrus cultivar Carrizo is the single most important rootstock to the US citrus industry and has resistance or tolerance to a number of major citrus diseases, including citrus tristeza virus, foot rot, and Huanglongbing (HLB, citrus greening). A Carrizo genomic sequence database providing approximately 3.5× genome coverage (haploid genome size approximately 367 Mb) was populated through 454 GS FLX shotgun sequencing. Analysis of the repetitive DNA fraction indicated a total interspersed repeat fraction of 36.5%. Assembly and characterization of abundant citrus Ty3/gypsy elements revealed a novel type of element containing open reading frames encoding a viral RNA-silencing suppressor protein (RNA binding protein, rbp) and a plant cytokinin riboside 5′-monophosphate phosphoribohydrolase-related protein (LONELY GUY, log). Similar gypsy elements were identified in the Populus trichocarpa genome. Gene-coding region analysis indicated that 24.4% of the nonrepetitive reads contained genic regions. The depth of genome coverage was sufficient to allow accurate assembly of constituent genes, including a putative phloem-expressed gene. The development of the Carrizo database ( http://citrus.pw.usda.gov/ ) will contribute to characterization of agronomically significant loci and provide a publicly available genomic resource to the citrus research community.

Incidence of antibiotic resistance and characterization of plasmids in Campylobacter jejuni strains isolated from clinical sources in Alberta, Canada

1986 ◽  
Vol 32 (1) ◽  
pp. 28-32 ◽  
Author(s):  
Diane E. Taylor ◽  
N. Chang ◽  
R. S. Garner ◽  
R. Sherburne ◽  
L. Mueller
Keyword(s):  
Antibiotic Resistance ◽  
Campylobacter Jejuni ◽  
Tetracycline Resistance ◽  
Additional Restriction ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Antibiotic Susceptibilities ◽  
Restriction Sites ◽  
High Level

Antibiotic susceptibilities of 382 strains of Campylobacter jejuni isolated in Alberta, Canada, in 1980 and 1981 were determined. Although none were resistant to erythromycin or gentamicin, 5.4 and 22% of strains were resistant to ampicillin in 1980 and 1981, respectively. Tetracycline resistance was noted in 6.8% of strains in 1980 and in 8.6% in 1981. Moreover, resistance to high-level tetracycline (32–128 μg/mL) was always mediated by a plasmid of 45–50 kilobases. Three transmissible plasmids from the Alberta strains were compared with the prototype plasmid pMAK175 by restriction enzyme analysis and some minor differences in restriction sites were noted between pMAK175 and pUA183. The two other plasmids pUA142 and pUA143 were 4 kilobases larger than pMAK175 and contained additional restriction sites. However, in all plasmids examined, the HincII and AccI fragments where the tetracycline-resistance determinant was located were shown to be conserved.

scholarly journals Ecology of Antibiotic Resistance Genes: Characterization of Enterococci from Houseflies Collected in Food Settings

2006 ◽  
Vol 72 (6) ◽  
pp. 4028-4035 ◽  
Author(s):  
Lilia Macovei ◽  
Ludek Zurek
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Protein Gene ◽  
Antibiotic Resistance Genes ◽  
Tetracycline Resistance ◽  
Surface Protein ◽  
Antibiotic Resistant ◽  
Aggregation Substance ◽  
Surface Protein Gene

ABSTRACT In this project, enterococci from the digestive tracts of 260 houseflies (Musca domestica L.) collected from five restaurants were characterized. Houseflies frequently (97% of the flies were positive) carried enterococci (mean, 3.1 � 103 CFU/fly). Using multiplex PCR, 205 of 355 randomly selected enterococcal isolates were identified and characterized. The majority of these isolates were Enterococcus faecalis (88.2%); in addition, 6.8% were E. faecium, and 4.9% were E. casseliflavus. E. faecalis isolates were phenotypically resistant to tetracycline (66.3%), erythromycin (23.8%), streptomycin (11.6%), ciprofloxacin (9.9%), and kanamycin (8.3%). Tetracycline resistance in E. faecalis was encoded by tet(M) (65.8%), tet(O) (1.7%), and tet(W) (0.8%). The majority (78.3%) of the erythromycin-resistant E. faecalis isolates carried erm(B). The conjugative transposon Tn916 and members of the Tn916/Tn1545 family were detected in 30.2% and 34.6% of the identified isolates, respectively. E. faecalis carried virulence genes, including a gelatinase gene (gelE; 70.7%), an aggregation substance gene (asa1; 33.2%), an enterococcus surface protein gene (esp; 8.8%), and a cytolysin gene (cylA; 8.8%). Phenotypic assays showed that 91.4% of the isolates with the gelE gene were gelatinolytic and that 46.7% of the isolates with the asa1 gene aggregated. All isolates with the cylA gene were hemolytic on human blood. This study showed that houseflies in food-handling and -serving facilities carry antibiotic-resistant and potentially virulent enterococci that have the capacity for horizontal transfer of antibiotic resistance genes to other bacteria.

scholarly journals Isolation and Characterization of a Single-Stranded RNA Virus Infecting the Bloom-Forming Diatom Chaetoceros socialis

2009 ◽  
Vol 75 (8) ◽  
pp. 2375-2381 ◽  
Author(s):  
Yuji Tomaru ◽  
Yoshitake Takao ◽  
Hidekazu Suzuki ◽  
Tamotsu Nagumo ◽  
Keizo Nagasaki
Keyword(s):  
Rna Virus ◽  
Burst Size ◽  
Host Cells ◽  
Sequencing Analysis ◽  
Virus Family ◽  
Isolation And Characterization ◽  
Ssrna Virus ◽  
Rhizosolenia Setigera ◽  
Virus Clone

ABSTRACT Diatoms are very significant primary producers in the world's oceans. Various environmental factors affect the depletion of diatom populations. The importance of viruses as a potential mortality source has recently been recognized. We isolated and characterized a new diatom virus (Chaetoceros socialis f. radians RNA virus [CsfrRNAV]) causing the lysis of the bloom-forming species Chaetoceros socialis Lauder f. radians (Schütt) Proschkina-Lavrenko. The virus infectious to C. socialis f. radians was isolated from water samples collected in Hiroshima Bay. Here we show the physiology, morphology, and genome characteristics of the virus clone. Virions were 22 nm in diameter and accumulated in the cytoplasm of the host cells. The latent period and the burst size were estimated to be <48 h and 66 infectious units per host cell, respectively. CsfrRNAV harbors a single-stranded RNA (ssRNA) genome and encodes at least three polypeptides of 32.0, 28.5, and 25.0 kDa. Sequencing analysis shows the length of the genome is 9,467 bases, excluding a poly(A) tail. The monophyly of CsfrRNAV and other diatom-infecting RNA viruses, Rhizosolenia setigera RNA virus and Chaetoceros tenuissimus RNA virus, was strongly supported by phylogenetic analysis based on the amino acid sequence of the RNA-dependent RNA polymerase domains. This suggested a new ssRNA virus family, Bacillariornaviridae. This discovery of CsfrRNAV may aid in further understanding the ecological dynamics of the C. socialis f. radians population in nature and the relationships between ssRNA diatom viruses and their hosts.

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