scholarly journals Identification of pBC218/pBC210 Genes of Bacillus cereus G9241 in Five Florida Soils Using qPCR

2014 ◽  
Vol 2014 ◽  
pp. 1-7
Author(s):  
Vicki Ann Luna ◽  
Kimmy Nguyen ◽  
Damian H. Gilling
Keyword(s):  
Public Health ◽  
Bacterial Species ◽  
Geographical Area ◽  
Soil Samples ◽  
Open Reading Frames ◽  
Public Health Importance ◽  
Potential Threat ◽  
Gram Negative ◽  
Positive Results ◽  
Reading Frames

The distribution of the virulent plasmid pBC210 of B. cereus that carries several B. anthracis genes and has been implicated in lethal anthrax-like pulmonary disease is unknown. We screened our collection of 103 B. cereus isolates and 256 soil samples using a quantitative PCR (qPCR) assay that targeted three open reading frames putatively unique to pBC210. When tested with DNA from 2 B. cereus strains carrying pBC210, and 64 Gram-positive and 55 Gram-negative bacterial species, the assay had 100% sensitivity and specificity. None of the DNA from the B. cereus isolates yielded positive amplicons but DNA extracted from five soils collected in Florida gave positive results for all three target sequences of pBC210. While screening confirms that pBC210 is uncommon in B. cereus, this study is the first to report that pBC210 is present in Florida soils. This study improves our knowledge of the distribution of pBC210 in soils and, of public health importance, the potential threat of B. cereus isolates carrying the toxin-carrying plasmid. We demonstrated that sequences of pBC210 can be found in a larger geographical area than previously thought and that finding more B. cereus carrying the virulent plasmid is a possibility in the future.

Comparison of Bacterial Species Isolated from Ground Beef, Textured Soy Protein and Ground Beef Extended with Textured Soy Protein1,2

1978 ◽  
Vol 41 (12) ◽  
pp. 961-964 ◽  
Author(s):  
JAMES F. FOSTER ◽  
LINDA S. GUTHERTZ ◽  
RICHARD C. HUNDERFUND ◽  
JAMES L. FOWLER
Keyword(s):  
Public Health ◽  
Soy Protein ◽  
Bacterial Species ◽  
Ground Beef ◽  
Public Health Importance ◽  
Gram Negative ◽  
Salmonella Enteriditis ◽  
Public Health Hazards ◽  
Positive Isolate ◽  
Gram Negative Organisms

A survey of the bacterial species of public health importance which could be isolated from ground beef (GB), textured soy protein (TSP) and ground beef extended with TSP (SGB) after 3 and 10 days of storage at 4 C was conducted. Escherichia coli was the most frequent gram-negative isolate from GB and SGB. Few gram-negative organisms were found in TSP. Clostridium perfringens was the most frequent gram-positive isolate from GB and SGB while Bacillus sp. was isolated most frequently from TSP. Salmonella enteriditis ser. worthington was isolated from GB and TSP. These products contained a wide variety of microorganisms, some of which might result in a food-associated infection or intoxication. However, if properly handled and cooked before consumption, these products should present few public health hazards.

scholarly journals Bacteriophages LIMElight and LIMEzero of Pantoea agglomerans, Belonging to the “phiKMV-Like Viruses”

2011 ◽  
Vol 77 (10) ◽  
pp. 3443-3450 ◽  
Author(s):  
Evelien M. Adriaenssens ◽  
Pieter-Jan Ceyssens ◽  
Vincent Dunon ◽  
Hans-Wolfgang Ackermann ◽  
Johan Van Vaerenbergh ◽  
...  
Keyword(s):  
Soil Samples ◽  
Pantoea Agglomerans ◽  
Open Reading Frames ◽  
Human Pathogen ◽  
Content Type ◽  
Host Diversity ◽  
Double Stranded Dna ◽  
Opportunistic Human Pathogen ◽  
Reading Frames

ABSTRACTPantoea agglomeransis a common soil bacterium used in the biocontrol of fungi and bacteria but is also an opportunistic human pathogen. It has been described extensively in this context, but knowledge of bacteriophages infecting this species is limited. Bacteriophages LIMEzero and LIMElight ofP. agglomeransare lytic phages, isolated from soil samples, belonging to thePodoviridaeand are the firstPantoeaphages of this family to be described. The double-stranded DNA (dsDNA) genomes (43,032 bp and 44,546 bp, respectively) encode 57 and 55 open reading frames (ORFs). Based on the presence of an RNA polymerase in their genomes and their overall genome architecture, these phages should be classified in the subfamily of theAutographivirinae, within the genus of the “phiKMV-like viruses.” Phylogenetic analysis of all the sequenced members of theAutographivirinaesupports the classification of phages LIMElight and LIMEzero as members of the “phiKMV-like viruses” and corroborates the subdivision into the different genera. These data expand the knowledge ofPantoeaphages and illustrate the wide host diversity of phages within the “phiKMV-like viruses.”

scholarly journals Genome Sequences of Bacteriophages KaiHaiDragon and OneinaGillian, Isolated from Microbacterium foliorum in Riverside, California

2019 ◽  
Vol 8 (15) ◽  
Author(s):  
Gillian Miller ◽  
Steven Tran ◽  
Rhiannon Abrahams ◽  
Daniel Bazan ◽  
Ethan Blaylock ◽  
...  
Keyword(s):  
Soil Samples ◽  
Open Reading Frames ◽  
Genome Sequences ◽  
Bacterial Host ◽  
Content Type ◽  
Reading Frames

KaiHaiDragon and OneinaGillian are two bacteriophages which have been recovered from soil samples using the bacterial host Microbacterium foliorum. Their genome lengths are 52,992 bp and 61,703 bp, with 91 and 104 predicted open reading frames, respectively.

scholarly journals Evaluation of the NG-Test MCR-1 Lateral Flow Assay and EDTA-Colistin Broth Disk Elution Methods To Detect Plasmid-Mediated Colistin Resistance among Gram-Negative Bacterial Isolates

2020 ◽  
Vol 58 (4) ◽  
Author(s):  
Alexander J. Fenwick ◽  
Yehudit Bergman ◽  
Shawna Lewis ◽  
Rebecca Yee ◽  
Anne-Catrin Uhlemann ◽  
...  
Keyword(s):  
Public Health ◽  
Lateral Flow ◽  
Test Method ◽  
Health Concern ◽  
Infection Prevention And Control ◽  
Colistin Resistance ◽  
Gram Negative ◽  
Content Type ◽  
Percent Agreement ◽  
Positive Results

ABSTRACT Plasmid-mediated colistin resistance (PMCR) is a global public health concern, given its ease of transmissibility. The purpose of this study was to evaluate two methods for the detection of PMCR from bacterial colonies: (i) the NG-Test MCR-1 lateral flow immunoassay (LFA; NG Biotech, Guipry, France) and (ii) the EDTA-colistin broth disk elution (EDTA-CBDE) screening test method. These methods were evaluated using a cohort of contemporary, clinical Gram-negative bacillus isolates from 3 U.S. academic medical centers (126 isolates of the Enterobacterales, 50 Pseudomonas aeruginosa isolates, and 50 Acinetobacter species isolates; 1 isolate was mcr positive) and 12 mcr-positive CDC-FDA Antibiotic Resistance (AR) Isolate Bank isolates for which reference broth microdilution colistin susceptibility results were available. Eleven (4.6%) isolates were strongly positive by the MCR-1 LFA, with an additional 8 (3.4%) isolates yielding faintly positive results. The positive percent agreement (PPA) and negative percent agreement (NPA) for MCR-1 detection were 100% and 96.1%, respectively. Upon repeat testing, only a single false-positive MCR-2 producer remained, as the isolates with initially faintly positive results were negative. The EDTA-CBDE screening method had an overall PPA and NPA of 100% and 94.3%, respectively. The NPA for the EDTA-CBDE method was slightly lower at 94.2% with Enterobacterales, whereas it was 96.0% with P. aeruginosa. The MCR-1 LFA and EDTA-CBDE methods are both accurate and user-friendly methods for the detection of PMCR. Despite the rarity of PMCR among clinical isolates in the United States, these methods are valuable tools that may be implemented in public health and clinical microbiology laboratories to further discern the mechanism of resistance among colistin-resistant Gram-negative isolates and to detect PMCR for infection prevention and control purposes.

scholarly journals A Francisella tularensis Pathogenicity Island Required for Intramacrophage Growth

2004 ◽  
Vol 186 (19) ◽  
pp. 6430-6436 ◽  
Author(s):  
Francis E. Nano ◽  
Na Zhang ◽  
Siobhán C. Cowley ◽  
Karl E. Klose ◽  
Karen K. M. Cheung ◽  
...  
Keyword(s):  
Francisella Tularensis ◽  
Pathogenicity Island ◽  
Amino Acid Sequences ◽  
Open Reading Frames ◽  
Intracellular Pathogen ◽  
Type B ◽  
Gram Negative ◽  
Virulent Type ◽  
Highly Virulent ◽  
Reading Frames

ABSTRACT Francisella tularensis is a gram-negative, facultative intracellular pathogen that causes the highly infectious zoonotic disease tularemia. We have discovered a ca. 30-kb pathogenicity island of F. tularensis (FPI) that includes four large open reading frames (ORFs) of 2.5 to 3.9 kb and 13 ORFs of 1.5 kb or smaller. Previously, two small genes located near the center of the FPI were shown to be needed for intramacrophage growth. In this work we show that two of the large ORFs, located toward the ends of the FPI, are needed for virulence. Although most genes in the FPI encode proteins with amino acid sequences that are highly conserved between high- and low-virulence strains, one of the FPI genes is present in highly virulent type A F. tularensis, absent in moderately virulent type B F. tularensis, and altered in F. tularensis subsp. novicida, which is highly virulent for mice but avirulent for humans. The G+C content of a 17.7-kb stretch of the FPI is 26.6%, which is 6.6% below the average G+C content of the F. tularensis genome. This extremely low G+C content suggests that the DNA was imported from a microbe with a very low G+C-containing chromosome.

scholarly journals Identification of Methyl Halide-Utilizing Genes in the Methyl Bromide-Utilizing Bacterial Strain IMB-1 Suggests a High Degree of Conservation of Methyl Halide-Specific Genes in Gram-Negative Bacteria

2001 ◽  
Vol 67 (4) ◽  
pp. 1959-1963 ◽  
Author(s):  
Claire A. Woodall ◽  
Karen L. Warner ◽  
Ronald S. Oremland ◽  
J. Colin Murrell ◽  
Ian R. McDonald
Keyword(s):  
Energy Source ◽  
Methyl Bromide ◽  
Open Reading Frames ◽  
Binding Motif ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
High Sequence Homology ◽  
Methyl Halide ◽  
High Degree ◽  
Reading Frames

ABSTRACT Strain IMB-1, an aerobic methylotrophic member of the alpha subgroup of the Proteobacteria, can grow with methyl bromide as a sole carbon and energy source. A single cmugene cluster was identified in IMB-1 that contained six open reading frames: cmuC, cmuA, orf146, paaE, hutI, and partialmetF. CmuA from IMB-1 has high sequence homology to the methyltransferase CmuA from Methylobacterium chloromethanicum and Hyphomicrobium chloromethanicum and contains a C-terminal corrinoid-binding motif and an N-terminal methyltransferase motif. However,cmuB, identified in M. chloromethanicumand H. chloromethanicum, was not detected in IMB-1.

scholarly journals Screening the Milieu of an abattoir for Bacteria of Public health importance in Makurdi, Benue State, Nigeria

2017 ◽  
Vol 7 (2) ◽  
pp. 1
Author(s):  
Nseabasi Maina N ◽  
E. G Vinkings ◽  
I U Bassey ◽  
A. A Unimke ◽  
L.O Abulawor
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Nalidixic Acid ◽  
Antibiotic Sensitivity ◽  
Soil Samples ◽  
Bacillus Species ◽  
Bacterial Cells ◽  
Pseudomonas Sp ◽  
Bacillus Sp ◽  
Public Health Importance

The study investigated the bacteriological content in a major abattoir located approximately 300 meters from a densely populated and cultivated area along the Benue River. Enumeration of bacterial cells from samples in the study yielded relatively high mean count of 9.4 x 105 and 7.3 x 105 from effluent and soil samples respectively. Bacteria isolated from both samples included; Escherichia coli, Streptococcus sp, Salmonella sp, Pseudomonas sp, Shigellasp, Enterobacter sp, Staphylococcus sp, Bacillus sp, Brucella sp, Proteus sp, Micrococcus sp etc. Escherichia coli recorded an occurrence of 18.53% in effluents and 16.16% in soil while Proteus species and Brucella sp had an occurrence of 9.59% and 1.39% respectively in soil samples. Antibiotic sensitivity screening using seveenteen (17) antibiotics disc (Optun Nig.) viz: Tarivid (10 µg), peflacine (10 µg), Agumentin (30 µg), Gentamycin (10 µg), Streptomycin (30 µg), Ceporex (10 µg), Nalidixic acid (30 µg), Ciprofloxacin (10 µg), Norfloxacin (10 µg), Rifampicin (µg), Erythromycin (µg), Chloramphenicol (µg), Ampiclox (30 µg), Levofloxacin (10µg). A marked level of resistance was observed among the isolates. However, Escherichia coli indicated sensitivity to peflacine, Shigellasp indicated sensitivity to augmentin, Enterococcus and Bacillus species indicated sensitivity to ciprofloxacin and streptomycin respectively.

scholarly journals Analysis of the pmsCEAB Gene Cluster Involved in Biosynthesis of Salicylic Acid and the Siderophore Pseudomonine in the Biocontrol Strain Pseudomonas fluorescensWCS374

2001 ◽  
Vol 183 (6) ◽  
pp. 1909-1920 ◽  
Author(s):  
Jesús Mercado-Blanco ◽  
Koen M. G. M. van der Drift ◽  
Per E. Olsson ◽  
Jane E. Thomas-Oates ◽  
Leendert C. van Loon ◽  
...  
Keyword(s):  
Salicylic Acid ◽  
Histidine Decarboxylase ◽  
Bacterial Species ◽  
Open Reading Frames ◽  
Putative Promoter Region ◽  
E Coli ◽  
Low Levels ◽  
Reading Frames ◽  
Siderophore Activity

ABSTRACT Mutants of Pseudomonas fluorescens WCS374 defective in biosynthesis of the fluorescent siderophore pseudobactin still display siderophore activity, indicating the production of a second siderophore. A recombinant cosmid clone (pMB374-07) of a WCS374 gene library harboring loci necessary for the biosynthesis of salicylic acid (SA) and this second siderophore pseudomonine was isolated. The salicylate biosynthesis region of WCS374 was localized in a 5-kb EcoRI fragment of pMB374-07. The SA and pseudomonine biosynthesis region was identified by transfer of cosmid pMB374-07 to a pseudobactin-deficient strain of P. putida. Sequence analysis of the 5-kb subclone revealed the presence of four open reading frames (ORFs). Products of two ORFs (pmsC andpmsB) showed homologies with chorismate-utilizing enzymes; a third ORF (pmsE) encoded a protein with strong similarity with enzymes involved in the biosynthesis of siderophores in other bacterial species. The region also contained a putative histidine decarboxylase gene (pmsA). A putative promoter region and two predicted iron boxes were localized upstream of pmsC. We determined by reverse transcriptase-mediated PCR that thepmsCEAB genes are cotranscribed and that expression is iron regulated. In vivo expression of SA genes was achieved in P. putida and Escherichia coli cells. In E. coli, deletions affecting the first ORF (pmsC) diminished SA production, whereas deletion of pmsBabolished it completely. The pmsB gene induced low levels of SA production in E. coli when expressed under control of the lacZ promoter. Several lines of evidence indicate that SA and pseudomonine biosynthesis are related. Moreover, we isolated a Tn5 mutant (374-05) that is simultaneously impaired in SA and pseudomonine production.

scholarly journals Genomic and Proteomic Characterization of Bacteriophage BH1 Spontaneously Released from Probiotic Lactobacillus rhamnosus Pen

Viruses ◽  
2019 ◽  
Vol 11 (12) ◽  
pp. 1163 ◽  
Author(s):  
Piotr Jarocki ◽  
Elwira Komoń-Janczara ◽  
Marcin Podleśny ◽  
Oleksandr Kholiavskyi ◽  
Monika Pytka ◽  
...  
Keyword(s):  
Lactobacillus Rhamnosus ◽  
Gc Content ◽  
Probiotic Strain ◽  
Open Reading Frames ◽  
Potential Threat ◽  
Genetic Switch ◽  
Important Virulence Factor ◽  
Inducing Factors ◽  
Reading Frames

Lactobacillus rhamnosus Pen is a human endogenous strain used for the production of probiotic formula, which is effective in the prevention of antibiotic-associated diarrhoea. Our study showed that this probiotic strain releases bacteriophage BH1 without the addition of any inducing agent. Our research revealed that phage BH1 has a circular genome with a length of 40721 nt and a GC content of 44.8%. The genome of phage BH1 possesses 57 open reading frames which could be divided into functional modules associated with DNA packaging, morphogenesis, lysis, integration, genetic switch, and replication. In spite of similarity in morphology and genomic organization, comparative analysis revealed substantial genetic diversity and mosaic genomic architecture among phages described for the Lactobacillus casei group. Additionally, qPCR and ddPCR analysis confirmed earlier microscopic observations indicating that L. rhamnosus Pen liberates bacteriophage particles during growth. This occurs spontaneously, and is not a result of external inducing factors. For samples collected after 4 and 24 h of L. rhamnosus Pen culture, the number of attB and attP copies increased 2.5 and 12 times, respectively. This phenomenon, by introducing resistance to other phages or enhancing the biofilm-forming capabilities, may increase the survivability of microorganisms in their natural ecological niche. Conversely, spontaneous phage induction may be an important virulence factor for bacteria, posing a potential threat for the human host.

scholarly journals Emergence of Azithromycin Resistance Mediated by Phosphotransferase-Encoding mph(A) in Diarrheagenic Vibrio fluvialis

mSphere ◽  
2019 ◽  
Vol 4 (3) ◽  
Author(s):  
Goutam Chowdhury ◽  
Thandavarayan Ramamurthy ◽  
Amit Ghosh ◽  
Shanta Dutta ◽  
Eizo Takahashi ◽  
...  
Keyword(s):  
Public Health ◽  
Antibiotic Resistance ◽  
Bacterial Species ◽  
Antibiotic Resistance Gene ◽  
Public Health Importance ◽  
Class 1 Integron ◽  
Content Type ◽  
Genetic Lineages ◽  
Vibrio Fluvialis ◽  
Azithromycin Resistance

ABSTRACT The azithromycin resistance conferred by phosphotransferase is encoded in the gene mph(A). This gene has been discovered in and reported for many bacterial species. We examined the prevalence of azithromycin resistance in Vibrio fluvialis (AR-VF) isolated during 2014 to 2015 from the hospitalized acute diarrheal patients in Kolkata, India. Most of the V. fluvialis isolates are identified as the sole pathogen (54%). The prevalence of AR-VF was higher in 2015 (19 [68%]) than in 2014 (9 [32%]). Among AR-VF isolates, the azithromycin MICs ranged from 4 to >256 mg/liter. Twenty-eight of the 48 (58%) V. fluvialis isolates harbored the gene mph(A) and phenotypically resistant to azithromycin. All the AR-VF isolates remained susceptible to doxycycline. In addition to azithromycin, other antimicrobial resistance-encoding genes of AR-VF were also characterized. All the AR-VF isolates were positive for class 1 integron, and most of them (17/28) carried the dfrA1 gene cassettes. Only one isolate was positive for the ereA gene, which encodes resistance to erythomycin. The majority of the isolates were resistant to β-lactam antibiotics (blaOXA-1 [96%], blaOXA-7 [93%], and blaTEM-9 [68%]) and aminoglycoside actetyltransferase, conferring resistance to ciprofloxacin-modifying enzyme [aac(6′)Ib-cr] (96%). Analyses by pulsed-field gel electrophoresis (PFGE) showed that the AR-VF isolates belonged to different genetic lineages. This is the first study to report azithromycin resistance and the presence of the mph(A) gene in V. fluvialis isolates. Circulation of AR-VF isolates with high azithromycin MICs is worrisome, since it may limit the treatment options for diarrheal infections. IMPORTANCE The progressive rise in antibiotic resistance among enteric pathogens in developing countries is becoming a big concern. India is one of the largest consumers of antibiotics, and their use is not well regulated. V. fluvialis is increasingly recognized as an emerging diarrheal pathogen of public health importance. Here we report the emergence of azithromycin resistance in V. fluvialis isolates from diarrheal patients in Kolkata, India. Azithromycin has been widely used in the treatment of various infections, both in children and in adults. Resistance to azithromycin is encoded in the gene mph(A). Emerging azithromycin resistance in V. fluvialis is a major public health challenge, and future studies should be focused on identifying ways to prevent the dissemination of this antibiotic resistance gene.

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